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Christensen et al.: Acquired Natural Enemies of Oxyops vitiosa 1 ACQUIRED NATURAL ENEMIES OF THE WEED BIOLOGICAL CONTROL AGENT OXYOPS VITIOSA (COLEPOTERA: CURCULIONIDAE) R OBIN M. C HRISTENSEN P AUL D. P RATT S HERYL L. C OSTELLO M IN B. R AYAMAJHI AND T ED D. C ENTER USDA/ARS, Invasive Plant Research Laboratory, 3225 College Ave., Ft. Lauderdale, FL 33314 A BSTRACT The Australian curculionid Oxyops vitiosa Pascoe was introduced into Florida in 1997 as a biological control agent of the invasive tree Melaleuca quinquenervia (Cav.) S. T. Blake. Populations of the weevil increased rapidly and became widely distributed throughout muc h of the invasive trees adventive distribution. In this study we ask if O. vitiosa has acquired natural enemies in Florida, how these enemies circumvent the protective terpenoid laden exudates on larvae, and what inuence 1 of the most common natural enemies has on O. vitiosa population densities? Surveys of O. vitiosa populations and rearing of eld-collected individuals resulted in no instances of parasitoids or pathogens exploiting weevil eggs or larvae In contrast, 44 species of predatory arthropods were commonly associated (>5 individuals when pooled across all sites and sample dates) with O. vitiosa Eleven predatory species were observed feeding on O. vitiosa during timed surveys, including 6 pentatomid species, 2 formicids and 3 arac hnids. Species with mandibulate or chelicerate mouthparts fed on adult stages whereas pentatomids, with haustellate beaks, pierced larval exoskeletons thereby bypassing the protective larval coating. Observations of predation were rare, with only 8% of timed surveys resulting in 1 or more instances of attack. Feeding by the pentatomid Podisus mucronatus Uhler accounted for 76% of all recorded predation events. Podisus mucronatus numerically responded to fourth instars but no response was observed for other life stages. Damage to M. quinquenervia plants from feeding by O. vitiosa however, was not inuenced by P. mucronatus densities, indicating that predation does not alter plant suppression. K ey Words: biological control, biotic resistance, predation, Oxyops vitiosa, Melaleuca Quinquenervia, Podisus mucronatus R ESUMEN El curculinido australiano Oxyops vitiosa Pascoe fue introducido a la Florida en 1997 como un agente de control biolgico para el rbol invasor Melaleuca quinquenervia (Cav.) S. T. Blake Poblaciones del gorgojo aumentaron rapidamente y se distribuyeron ampliamente por mucho de la distribucin del rbol invasor adventivo. En este estudio, preguntamos si O. vitiosa han adquerido enemigos naturales en la Florida, como estos enemigos evitan las secreciones de turpenoides que protejen las larvas y que inuencia tiene uno de los enemigos naturles mas comunes sobre la densidad de la poblacin de O. vitiosa ? La inspeccin de la poblacin de O. vitiosa y la cria de individuos recolectados en el campo resulto en no caso de parasitoides y patgenos usando los huevos o larvas de los gorgojos En contraste, 44 especies de artrpodos depredadores fueron comnmente asociadas (>5 individuos cuando se agregados por todos los sitios y fechas de muestreo) con O. vitiosa Se observaron once especies de depredadores alimentndose sobre O. vitiosa durante los sondeos, incluyendo 6 especies de pentatmidos 2 formcidos y 3 arcnidos. Especies con partes bucales mandbuladas y queliceradas se alimentaron sobre los estadios adultos mientras que los pentatmidos, con su pico chupador, puncharon los exo-esqueletos de las larvas asi pasando el cubertura protectiva de las larvas. Observaciones de depredacin fueron raras, con solamente 8% de los estudios que llevaron el tiempo resultaron en 1 ms instancias de ataque. La alimentacin del pentatmido Podisus mucronatus Uhler cont con 76% de los eventos de depredacin registrados Podisus mucronatus respondi numericamente al los instares de cuarto estadio pero ninguna respuesta fue observada en los otros estadios de vida. El dao a las plantas de M. quinquenervia debido a la alimentacin por O. vitiosa sin embargo, no fue inuenciado por la densidad de P. mucronatus que indica que la depredacin no altera la supresin de la planta. Acquisition of novel natural enemies may inuence the successful establishment, spread, and impact of introduced weed biological control agents in their adventive range (Goeden & Louda 1976; Semple & Forno 1987; Simberloff 1989; Cornell & Hawkins 1993; Hill & Hulley 1995; McPartland & Nicholson 2003; Norman et al. 2009; Paynter et al. 2010). Of the arthropods introduced for control of invasive plants world wide, approximately 50% suffer sufcient mortality from higher trophic levels to signicantly limit suppression of target weeds (Goeden & Louda 1976).

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2 Florida Entomologist 94(1) March 2011 The spider mite Tetranychus lintearius (Dufour), for example was introduced into New Zealand, Australia, and the United States as a biological control agent of the invasive plant Ulex europaeus L. (Fabaceae) (Hill & Stone 1985; Hill et al. 1991). Although successfully established and widely distributed, mites in each country rarely sustained sufcient population densities to provide permanent control of the target weed (Rees & Hill 2001). Subsequent studies demonstrated that a complex of native and introduced predators suppressed T. lintearius populations and limited control of the invasive weed (P eterson 1993; Peterson et al. 1994; Pratt et al. 2003). Considering the ecological risks (Carvalheiro et al. 2008) and expense of biological control, increased attention in the scientic literature has focused on predicting susceptibility of introduced biological control agents to natural enemies in the adventive range (Kuhlmann et al. 2006). Hill & Hulley (1995), for instance, demonstrated that variation in susceptibility of introduced herbivores to parasitoids is related, in part, to evolutionary strategies that render the prey less accessible, apparent, or palatable to the attacker. Along this continuum of use by natural enemies lie those species that are highly apparent yet experience relatively less attack due to the expression of chemical deterrents that render them less palatable or even toxic to prospective natural enemies. The introduced weevil Oxyops vitiosa Pascoe sequesters terpenoids from lea ves of its host plant Melaleuca quinquenervia (Cav.) S. T. Blake and larvae excrete these compounds through their integument (Wheeler et al. 2002). The consumption and expression of these terpenoids repels the red imported re ant ( Solenopsis invicta Buren) and red wing blackbird ( Agelaius phoeniceus L.) under controlled feeding tests (Wheeler et al. 2002). It remains unclear, however, if this acquired repellency confers protection from the suite of potential novel natural enemies that exist in the herbivores adventive range. Oxyops vitiosa is native to eastern Australia and is a specialist herbivore of the invasive tree M. quinquenervia (Balciunas et al. 1994). Based on its narrow host range the weevil was permitted for release in Florida in 1997 and readily established in M. quinquenervia dominated habitats (Center et al. 2000; Pratt et al. 2003). Adult weevils feed on M. quinquenervia foliage whereas larvae consume only newly-developed lea ves that are ephemerally produced in seasonal ushes at branch apices (Purcell & Balciunas 1994). Following its introduction, O. vitiosa populations increased rapidly and became widely distributed throughout muc h of the invasive trees geographic distribution in Florida (Pratt et al. 2003; Balentine et al. 2009). When considering the large densities of these herbivores in the environment, we questioned (1) whether O. vitiosa had acquired natural enemies in Florida, (2) how these enemies mitigated the defensive strategies of the herbivore and (3) what impact the most abundant of these natural enemies has on O. vitiosa population densities? M ATERIALS AND M ETHODS Surveys for natural enemies associated with O. vitiosa were conducted at 4 locations in south Florida. Site 1 was located near Ft. Lauderdale, Broward Co., FL. The site was a 0.5-ha eld consisting of 2to 5-m tall trees occurring at a density of 21,560 trees/ha. In general, M. quinquenervia trees were growing in organically rich soils typical of rec laimed glades systems. Melaleuca quinquenervia trees at site 2 occurred under a power line right of w ay near Weston, Broward Co., FL. Prior to 1997 land managers cut M. quinquenervia trees near their bases, resulting in multi-stemmed coppices The study area was 0.5 ha and trees were 2-5 m tall, occurring at a density of 2,517 trees/ha. Site 3 was located near Estero, Collier Co., FL and consisted of an 8-ha area of drained wetland converted to pasture. To suppress M. quinquenervia growth, land managers mowed trees at 6-month intervals resulting in coppices 0.5-2 m in height. These coppicing clumps formed a dense, nearly continuous canopy of leaves with 4,406 stumps/ha. In contrast to the previous sites the soil type was primarily sand, consistent with an invaded pine atwoods. Site 4 consisted of a 1-ha area within the historically mesic atwoods of the Picayune Forest near Belle Meade, Collier Co., FL. A re burned much of the M. quinquenervia dominated areas in 1998 resulting in recruitment of 129,393 trees/ha of primarily small 1-2 m tall saplings with an occasional large, mature tree interspersed (Table 1). Surveys were conducted monthly at each site from Nov 2000 through Jun 2001 and sampling occurred between 10 AM and 2 PM on days without precipitation. Sampling consisted of sweeping M. quinquenervia foliage, and occasionally trunks, with a 90-cm diameter sweep net. One sample consisted of 100 sweeps of the net in a sweeping motion of 180 with sweeps spaced ca. 1.0 m apart along a randomly selected 100-m transect. Four samples along separate transects were collected each month. The content of the net after 100 sweeps was emptied into a 4-liter sealable bag and frozen at minus 20 () C until processed. Arthropods were then separated from plant material, sorted by morphological types, and pinned or stored in 70% ethanol. One limitation of our sweep sampling method included collecting arthropods that were not closely associated with O. vitiosa but were transients merely resting on the plant foliage or disturbed from understory vegetation while sampling. Additionally, this method was biased to-

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Christensen et al.: Acquired Natural Enemies of Oxyops vitiosa 3 wards poor iers. All study sites possessed smaller trees that facilitated sampling but may have biased collections to lower rather than higher canopy dwelling species. Therefore, caution should be used when drawing inferences from these data due to the unknown relationships between sampled arthropods and O. vitiosa. For this reason, a minimum of 2 observers also searched for direct predation or parasitism for 30 min/survey at each site monthly. All specimens, except formicids, were submitted to and deposited at the Florida State Collection of Arthropods (FSCA, Division of Plant Industries, Gainesville, FL) for identication and incorporated into their taxonomic database (Costello et al. 2003). Most formicids were identied and retained by L. Davis of the Fire Ant Unit, Agricultural Research Service, USDA, Gainesville, FL. A few formicids were identied by M. Deyrup of the Archbold Biological Station, Lake Placid, FL. Several dipteran specimens were identied at the Systematic Entomology Laboratory, Agricultural Research Service, USDA, Beltsville, MD. Population densities of O. vitiosa at sites 2 and 3 were monitored by delineating a 0.5-ha study site within the existing M. quinquenervia stands, respectively Within these plots, transects were arranged in a grid pattern with 8 transects oriented east to west at 10-m intervals and points on each transect spaced 10 m apart. Beginning in Nov 2000 through Jun 2001, M. quinquenervia trees were sampled monthly at 20 randomly selected transect points Plants at each sampling point were selected based on the quarter method of vegetation sampling (Smith 1966). The area was divided into 4 quarters at each sampling point based on the 4 cardinal directions. The nearest tree to the sample point in each quarter was examined to determine the number of O. vitiosa per plant. The ordered distance method was used to quantify weevil population densities over time at sites 1 and 4 (Krebs 1999). In total, 30 points were randomly selected at each sampling interval and the nearest tree to each point was inventoried. At all sites, O. vitiosa life stages were counted along with plant resource a vailability. Resource availability was assessed on a 5-point scale based on visual estimation of percentage of suitable foliage for feeding by O. vitiosa : 0 = no suitable foliage; 1 = less than 25%; 2 = 26 to 50%; 3 = 51 to 75%; 4 = 76 to 100%. A partial correlation analysis was used to identify those predators positively associated with O. vitiosa (PROC CORR), after controlling for the inuence of site by the P ARTIAL statement (SAS 1999). For all tests, a P -value <0.05 was considered signicant evidence for association among predators and O. vitiosa However, caution should be used when interpreting these data because association is not sufcient evidence to suggest that a trophic relationship exists among the species To determine if O. vitiosa had acquired egg or larval parasitoids in its adventive range 50 eggs were collected at random from sites 1-4 at monthly intervals. Eggs were examined under a dissecting microscope (10-50X) to detect presence of larval exit holes indicating larval eclosion; eggs with exit holes were discarded. The remaining eggs were left attached to leaf material and placed in gelatin capsules. These capsules were transferred into a Petri dish (10 1.5 cm) that w as then sealed with Paralm to retain leaf moisture. Petri dishes were placed in an environmental chamber at 25 () C, with a photoperiod of 16:8 (L:D) and a relative humidity of 65% 10%. Hatching of eggs was monitored once a week. Eggs that did not hatch after 1 month were dissected. To detect larval parasitoids, 50 third or fourth instars of O. vitiosa were reared to the pupal stage Each larva was placed individually in 1 Petri dish (10 1.5 cm) with moistened lter paper and M. quinquenervia leaves. Petri dishes were sealed with P aralm and kept in an environmental chamber under the same conditions as described earlier. Host leaves were replaced every T ABLE 1. R ESEARCH SITE DESCRIPTION AND SUMMARY OF SURVEYS CONDUCTED FOR O. VITIOSA IN SOUTH F LORIDA Site GPS Coordinates Surveys conducted a HabitatHydro-period 1 N 26.05605W -80.25168 1, 2, 3, 4 Swale Short 2 N 26.03548W -80.43495 1, 2, 3, 4 Swale Medium 3 N 26.42550W -81.81033 1, 2, 3, 4, 5Mesic atwoodsShort 4 N 26.10478W -81.63392 1, 2, 3, 4 Mesic atwoodsShort 5 N 26.46017W -81.70186 4 Mesic atwoodsShort 6 N 26.54698W -81.79820 4 Wet atwoodsShort 7 N 28.47323W -81.33632 4 Upland lake Long 8 N 25.71341W -80.47949 4 Swale Medium 9 N 25.81208W -80.41780 4 Swale Medium 10 N 26.16227W -80.36269 4 Swale Long a 1 = Arthropods associated with O. vitiosa, 2 = O. vitiosa population density, 3 = O. vitiosa egg parasitism, 4 = Entomopathogens of O. vitiosa, 5 = Impacts of P. mucronatu s on O. vitiosa populations.

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4 Florida Entomologist 94(1) March 2011other day until the prepupal stage, when leaves were removed for the remainder of the study. Surveys for entomopathogens of O. vitiosa were conducted at 10 sites between Jun 2003 and Jan 2004 (Table 1). Late instars and adults were collected, packaged in an ice-cooler, transported to the laboratory, and examined by USDA/ARS insect pathologists at the Center for Medical, Agricultural, and Veterinary Entomology, Gainesville, FL. All live individuals originating from the same location were homogenized in 3-5 mL of deionized water and a sample of the crude suspension was examined with a phase-contrast microscope to searc h for pathogens, such as microsporidia, fungal spores, or occluded viruses. During initial surveys, the pentatomid bug Podisus mucronatus Uhler was commonly associated with O. vitiosa and observed feeding on larval stages of the biological control agent at each of the 4 study sites. Therefore, we quantied the population dynamics of P. mucronatu s and O. vitiosa at site 3. Sampling was conducted as described earlier except 20 transects were oriented east to west at 20-m intervals with 9-10 points on each transect spaced 20 m apart. Melaleuca quinquenervia plants were sampled at 50 randomly selected transect points every 6 weeks (approximate generation time; Purcell & Balciunas 1994) beginning in Dec 2000 and continuing through Oct 2002. As before, the nearest plant to the sample point in each quadrant was examined to determine the number and life stage of each O. vitiosa and P. mucronatus individual. In addition to these data, we also noted the amount of damage due to herbivory, plant resource availability (as described earlier), and the number of dead larvae. Herbivory damage was assessed on a 5-point scale based on visual estimation of the percentage of suitable foliage destroyed by O. vitiosa feeding: 0 = no damage; 1 = less than 25% destroyed; 2 = 26 to 50%; 3 = 51 to 75%; 4= 76 to 100% destroyed. Linear regression was used to test for a numerical response of predators to life stages of O. vitiosa (n = 6), dead larvae, food availability, and herbivory damage (i.e., aggregation of predators to patches of high prey density; Schenk & Bacher 2002). RESULTS AND DISCUSSIONBiotic resistance describes the collective inuence of parasitoides, predators, pathogens, and competitors on the establishment and proliferation of non-indigenous species, including introduced biological control agents (Simberloff & Von Holle 1999). Historically, native predators, parasitoids, and pathogens have interfered with half of the published case histories involving insect introductions for weed control (Goeden & Louda 1976). Considering this high rate of interference, we questioned if O. vitiosa had acquired natural enemies in its adventive range. We observed no instances of parasitoids (egg and larval) or pathogens exploiting O. vitiosa. Of the 1138 O. vitiosa eggs collected from the study sites (Table 1), 782 hatched and developed normally while the remaining 356 did not hatch. Of the 1266 fourth-instars collected from study sties, 913 survived to become adults. Dissection of both unhatched eggs and dead larvae yielded no evidence that mortality was due to parasitism. Similarly, no pathogens were found in the late instars and adults collected from sampled sties (Table 1). These results indicate that despite the herbivores high population densities and large geographic distribution (Pratt et al. 2003; Balentine et al. 2009), native parasitoids and pathogens have failed to exploit these lifestages of O. vitiosa. One explanation for the lack of O. vitiosa parasitization may be that native parasitoids require more than the 4 years allotted in this study to adjust behaviorally and physiologically to exploit the new host as well as produce sufcient densities to be discovered through our sampling protocols. In contrast, Hill & Hulley (1995) determined that 16 of the 40 established weed biological control agents in South Africa had acquired native parasitoids within 3 years of release. Similarly, the biological control agent Neomusotima conspurcatalis Warren acquired a suite of parasitoids within months of its release in Lygodium microphyllum (Cav.)-dominated habitats of Florida (Kula et al. 2010). These and other examples of rapid parasitoid acquisition by biological control agents (Carvalheiro et al. 2008; Paynter et al. 2010) suggest that the timing of our study was not premature but that future parasitoid (or pathogen) surveys may yield new discoveries as the region continues to recruit exotic species (Klassen et al. 2002; Dobbs & Brodel 2004; Childers & Rodrigues 2005). Surveys of O. vitiosa populations resulted in the collection of 154 species of predatory arthropods, yet only 44 had an overall abundance greater than 5 individuals when pooled across all sites and dates (Table 2). Species positively correlated with O. vitiosa (all stages) included the salticid Eris ava (Peckham & Peckham), the crab spiders Misumenops bellulus (Banks) and Misumenops sp., and the pentatomid bug Podisus mucronatus Uhler (Table 2). Although these data indicate that predators are associated with the introduced herbivore, direct observation of predation provides conclusive evidence of these novel trophic interactions. Eleven predatory species were observed feeding on O. vitiosa during timed surveys, including 6 pentatomid species ( Euthyrhynchus oridanus (L.), P. mucronatus (Say), Podisus jole (Stal), Podisus maculiventris (Say), Podisus sagitta (F.), Stiretrus anchorago (F.)), 2 formicids (Pseudomyrmex gracilis (F.), Solenopsis invicta) and 3 arachnids ( Peucetia viridans (Hentz), Latrodectus mactans (F.), Latrodectus geometri-

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Christensen et al.: Acquired Natural Enemies of Oxyops vitiosa 5cus C.L. Koch). The formicids and arachnids were observed feeding exclusively on adult weevils whereas the pentatomids attacked larvae of O. vitiosa; E. oridanus was the only species observed exploiting all active stages of the introduced herbivore. Observing predation was rare, with only 8% of timed surveys resulting in 1 or more instances of attack. Feeding by P. mucronatus accounted for 76% of all recorded predation events and the remaining species each represented <5% of the events, respectively. Ecological theory suggests that host range expansion is inuenced in part by host phylogeny, with close relatives more readily adopted than distant ones (Paynter et al. 2010). Therefore, an alternative explanation for the lack of acquiredTABLE 2. THE ABUNDANCE OF PREDACEOUS ARTHROPODS COLLECTED FROM THE INVASIVE TREE MELALEUCA QUINQUENERVIA AND THEIR CORRELATION WITH THE INTRODUCED BIOLOGICAL CONTROL AGENT OXYOPS VITIOSA. Family Species Site AbundancePartial Correlation 1234PCC P-value Alydidae Hyalymenus sp. A 11200-0.230980.2112 Anyphaenidae Hibana sp. 0009 0.045190.8093 Hibana sp. 4012 -0.066530.7222 Araneidae Acacesia hamata (Hentz) 1514220.155110.4047 Eriophora ravilla (C.L. Koch) 5101 -0.082300.6599 Neoscona sp. 271520-0.088670.6353 Neoscona arabesca (Walckenaer) 6000 -0.136060.4655 Cixiidae Bothriocera sp. 0600 -0.116740.5317 Clubionidae Clubiona sp. 0605 0.018560.9211 Formicidae Camponotus planatus (Roger) 7546200.057570.7584 Camponotus oridanus (Buckley)14002 0.094790.6120 Dolichoderus pustulatus Mayr 6000 -0.143890.4400 Paratrechina longicornis (Latreille) *8510204-0.140780.4500 Paratrechina guatemalensis (Forel) *16206 -0.086370.6441 Pseudomyrmex pallidus (F. Smith) *151100-0.131900.4794 Pseudomyrmex gracilis (Fabricius) 2106 -0.179150.3349 Solenopsis invicta Buren 81616640.026640.8869 Lycosidae Pardosa sp. 101600.331100.0688 Pirata sp. 18227220.339160.0620 Lygaeidae Oedancala crassimana (Fabricius)13280 0.338850.0622 Paromius longulus (Dallas) 7410 0.068290.7151 Membracidae Spissistilus festinus (Say) 0070 0.298640.1027 Mimetidae Mimetus sp. 102160.071100.7039 Miridae Taylorilygus pallidulus (Blanchard) 2240 0.312760.0867 Miturgidae Cheiracanthium inclusum (Hentz)961401016-0.010030.9573 Oxyopidae Peucetia viridans (Hentz) 13791140.067870.7168 Pentatomidae Podisus mucronatus Uhler 619840.527000.0023 Pisauridae Pisaurina s p. 7500 0.131990.4791 Reduviidae Zelus longipes Linnaeus 25001 -0.050460.7875 Salticidae Eris ava (Peckham & Peckham)451690.365030.0435 Hentzia palmarum (Hentz) 724510550.005400.9770 Pelegrina galathea (Walckenaer)1251400.183430.3233 Phidippus sp. 0105 -0.148070.4267 Thiodina peurpera (Hentz) 01900-0.138630.4570 Scelionidae Trissolcus sp. 0800 -0.104830.5746 Tetragnathidae Tetragnatha sp. 33851 0.112160.5480 Theridiidae Anelosimus studiosus (Hentz) 67030 0.089560.6318 Chrysso pulcherrima (Mello-Leitao) 1014 0.265300.1492 Theridion glaucescens (Becker)12001 0.130180.4852 Theridion avonotatum (Becker)24002 -0.014000.9404 Thomisidae Misumenops sp. 2354 0.305190.0950 Misumenops bellulus (Banks) 263727150.357390.0484 Misumenops sp. 1002920.397150.0269 Tmarus sp. 20152-0.241540.1905

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6 Florida Entomologist 94(1)March 2011parasitoids and pathogens may be due to the paucity of closely related species in the biological control agents adventive range. The Australian weevil O. vitiosa belongs to the tribe Goniopterini, which has no representatives in the New World (Alonso-Zarazaga & Lyal 1999). Similarly, invasion by M. quinquenervia markedly alters community structure in ways that are likely to repel habitat specialists. Therefore, the aquisition of parasitoids will likely require evolutionary rather than ecological time scales (Hill & Hulley 1995). With the exception of E. oridanus the exclusive use of adult versus larval prey observed herein may be explained by mouthpart morphologies and the antipredatory activity of the viscous coating that covers immature stages of O. vitiosa (Purcell & Balciunas 1994). Larvae of the introduced weevil sequester terpenoids from M. quinquenervia leaves and excrete these compounds through their integument (Wheeler et al. 2003). This larval coating has been shown to repel the red imported re ant (S. invicta) and likely confers protection against other mandibulate predators (Wheeler et al. 2002). However, adults and pupae lack the coating and are susceptible to predation by a range of predator types. The larval coating does not confer protection against pentatomid species observed herein. The haustellate mouthparts of pentatomid species pierce the larval integument and largely bypass the terpenoidladen coating to access the internal contents of the larval prey. Yet, mouthpart type alone does not facilitate exploitation of the abundant novel resource as other predators with haustellate mouthparts (i.e., Zelus longipes L.) occurred at the study sites but were not common or observed directly feeding on O. vitiosa larvae. Increased densities of O. vitiosa eggs, early instars, and adults did not inuence patch colonization by P. mucronatus (Table 3). A numerical response by P. mucronatus was observed, however, on plants harboring fourth instars (Table 3), indicating a preference for larger larval stages of the introduced weevil. These ndings are consistent with Hawkins et al. (1997), who reported that insect predation is higher in late developmental stages due, in part, to resource concentration and handling time. Not surprisingly, a positive relationship between P. mucronatus and larval corpses also was observed. While it is clear that P. mucronatus attacks O. vitiosa larvae and numerically responds to the single most damaging stage of the herbivore, does this predation disrupt biological control of M. quinquenervia ? We hypothesized that increases in P. mucronatus densities results in concomitant increases in predation and ultimately decreases in plant damage caused by O. vitiosa. Damage levels observed herein, however, were not inuenced by P. mucronatus densities (Table 3), indicating that predation does not alter plant suppression within the sampled patch. Similarly, the amount of plant resource availability for consumption by O. vitiosa does not vary based on predator loads, which suggest that predation does not result in a corresponding increase in undamaged plant material (Table 3). These results are supported by independent studies that also were conducted at site 3 and reported marked reductions in M. quinqueneriva growth and survival despite the presence of these predators (Center et al. 2000; Pratt et al. 2002; Pratt et al. 2004). The limited inuence of P. mucronatus on O. vitiosa population growth and herbivory is likely related to low predation rates (mean = 9.5%, SE = 0.5). The introduction of O. vitiosa has resulted in marked reductions in growth and survivorship of the invasive tree M. quinqueneriva (Pratt et al. 2003, 2005; Rayamajhi et al. 2008; Tipping et al. 2008, 2009; Balentine et al. 2009), with no direct non-target impacts to plant species in the weevils adventive range (Pratt et al. 2009). The acquisition of higher trophic levels by O. vitiosa, however, suggests that indirect effects of apparent competition may exist as predators are subsidized by the introduced weevil and their resultant increased population densities may exert asymmetrical predation on their historical prey species (Carvalheiro et al. 2008). In the absence of pre-introduction food web analyses, it remains unclear how the exploitation of O. vitiosa by native predators affects apparent competition on shared prey densities. The limited predation by generalists suggests that the strength of apparent competition is weak but additional research is needed to quantify interactions among introduced and native prey species as mitigated by common predators. ACKNOWLEDGMENTSWe thank 2 anonymous reviewers for comments on earlier versions of the manuscript. We also thank Scott Wiggers, Willey Durden, Kirk Tonkel, Andrea Kral, Carl Belnavis, Tafana Fiore, and Stacey Grassano for assisTABLE 3. LINEAR REGRESSION OF PODSUS MUCRONATUS DENSITIES ON OXYOPS VITIOSA STAGE SPECIFIC DENSITIES, PLANT QUALITY, AND FOLIAGE AVAILABILITY. Life stagedfEstimate t-valuePr > t Egg1-0.00097-0.290.7752 1st instar1-0.00185-0.240.8075 2nd instar10.013941.660.0966 3rd instar10.011751.270.2025 4th instar10.02512.880.004 Adults10.008181.060.2874 Dead10.1375516.16<.0001 Damage1-0.00265-1.310.1893 Plant foliage1-0.00176-0.940.3462

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Christensen et al.: Acquired Natural Enemies of Oxyops vitiosa 7tance with data collection and site maintenance. Mention of trade names or commercial products in this publication is solely for the purpose of providing specic information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. This research was supported, in part, by grants from the South Florida Water Management District, the Florida Department of Environmental Protection Bureau of Invasive Plant Management, and the USDA Areawide T AME Melaleuca Program (tame.ifas.u.edu).REFERENCES CITEDALONSO-ZARAZAGA, M. A., AND LYAL, C. H. C. 1999. A World Catalogue of Families and Genera of Curculionoidea (Insecta: Coleoptera) (Excepting Scolytidae and Platypodidae). Entomopraxis, Barcelona. 315p. BALCIUNAS, J. K., BURROWS, D. W., AND PURCELL, M. F. 1994. 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V. 2005. Potential pest mite species collected on ornamental plants from Central America at port of entry to the United States. Florida Entomol. 88: 408-414. COSTELLO, S. L., PRATT, P. D., RAYAMAJHI, M. B., ANDCENTER, T. D. 2003. Arthropods associated with above-ground portions of the invasive tree Melaleuca quinquenervia in south Florida, USA. Florida Entomol. 86(3): 300-322. CORNELL, H. V., AND HAWKINS, B. A. 1993. Accumulation of native parasitoid species on introduced herbivores: A comparison of hosts as natives and hosts as invaders. American Nat. 141(6): 847-865. DOBBS, T. T., AND BRODEL, C. F. 2004. Cargo aircraft as a pathway for the entry of nonindigenous pests into south Florida. Florida Entomol. 87: 65-78. GOEDEN, R. D., AND LOUDA, S. M. 1976. Biotic interference with insects imported for weed control. Annu. Rev. Entomol. 21: 325-342. HARRIS, P. 1991. Classical biocontrol of weeds: its definition, selection of effective agents, and administrative political problems. Canadian Entomol. 123: 827-849. HAWKINS, B. A., CORNELL, H. V., AND HOCHBERG, M. E. 1997. Predators, parasitoids, and pathogens as mortality agents in phytophagous insect populations. Ecology. 78(7): 2145-2152. HILL, M. P., AND HULLEY, P. E. 1995. Host-range extension by native parasitoids to weed biocontrol agents introduced to South Africa. Biol. Cont. 5: 297-302. HILL, R. L., AND STONE, C. 1985. Spider mites as control agents for weeds, pp. 443-448 In W. Helle and M. H. Sabelis [eds.], Spider Mites: Their Biology, Natural Enemies and Control, Vol. 1B. Elsevier, Amsterdam. HILL, R. L., GRINDELL, J. M., WINKS, C. J., SHEAT, J. J.,AND HAYES, L. M. 1991. Establishment of gorse spider mite as a control agent of gorse. Proc. New Zealand Weed and Pest Control Conf. 44: 31-34. KLASSEN, W., BRODEL, C. F., AND FIESELMANN, D. A. 2002. Exotic pests of plants: current and future threats to horticultural production and trade in Florida and the Caribbean Basin. Micronesica Supp. 6: 5-27. KREBS, C. J. 1999. Estimating Abundance: Line Transects and Distance Methods, Ecological Methodology. Addison Wesley Longman, Menlo Park, California. KUHLMANN, U., MASON, P. G., HINZ, H. L., BLOSSEY, B., DE CLERCK-FLOATE, R., DOSDALL, L. M., MCCAFFREY, J., SCHWARZLAENDER, M., OLFERT, O., BRODEUR, J., GASSMANN, A., MCCLAY, A., AND WIEDENMANN, R. 2006. Avoiding conicts between insect and weed biological control: selection of non-target species to assess host specicity of cabbage seedpod weevil parasitoids. J. Appl. Entomol. 130: 129-141. KULA, R. R., BOUGHTON, A. J., AND PEMBERTON, R. W. 2010. Stantonia pallida (Ashmead) (Hymenoptera: Braconidae) reared from Neomusotima conspurcatalis Warren (Lepidoptera: Crambidae), a classical biological control agent of Lygodium microphyllum (Cav.) R. Br. (Polypodiales: Lygodiaceae). Proc. Entomol. Soc. Washington 112: 61-68. MCPARTLAND, J. M., AND NICHOLSON, J. 2003. Using parasite databases to identify potential nontarget hosts of biological control organisms. New Zealand J. Bot. 41: 699-706. NORMAN, K., CAPPUCCINO, N., AND FORBES, M. R. 2009. Parasitism of a successful weed biological control agent, Neogalerucella calmariensis Canadian Entomol. 141: 609-613. PAYNTER, Q., FOWLER, S. V., GOURLAY, A. H., GROENTEMAN, R., PETERSON, P. G., SMITH, L., AND WINKS, C. J. 2010. Predicting parasitoid accumulation on biological control agents of weeds. J. Appl. Ecol. 47: 575582. PETERSON, P. G. 1993. The Potential Ability of Stethorus bidus (Kapur) to Regulate Populations of Tetranychus lintearius (Dufour). Masters Thesis, Massey University, Auckland, New Zealand. PETERSON, P. G., MEGREGOR, P. G., AND SPRINGETT, B. P. 1994. Development of Stethorus bidus in relation to temperature: implications for regulation of gorse spider mite populations. Proc. New Zealand Plant Prot. Conf. 47: 103-106. PRATT, P. D., COOMBS, E. M., AND CROFT, B. A. 2003. Predation by phytoseiid mites on Tetranychus lintearius (Acari: Tetranychidae), an established weed biological control agent of gorse (Ulex europaeus). Biol. Cont. 26: 40-47. PRATT, P. D., RAYACHHETRY, M. B., VAN, T. K., ANDCENTER, T. D. 2002. Field-based rates of population increase for Oxyops vitiosa (Coleoptera: Cucurlionidae), a biological control agent of the invasive tree Melaleuca quinquenervia Florida Entomol. 85: 286287.

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8 Florida Entomologist 94(1)March 2011PRATT, P. D., RAYAMAJHI, M. B., CENTER, T. D., TIPPING, P. W. AND WHEELER, G. S. 2009. The ecological host range of an intentionally introduced herbivore: A comparison of predicted versus actual host use. Biol. Cont. 49: 146-153. PRATT, P. D., RAYAMAJHI, M. B., VAN, T. K., CENTER, T. D.,AND TIPPING, P. W. 2005. Herbivory alters resource allocation and compensation in the invasive tree Melaleuca quinquenervia Ecol. Entomol. 30: 316-326. PRATT, P. D., SLONE, D. H., RAYAMAJHI, M. B., VAN, T. K., AND CENTER, T. D. 2003. Geographic distribution and dispersal rate of Oxyops vitiosa (Coleoptera: Curculionidae), a biological control agent of the invasive tree Melaleuca quinquenervia in south Florida. Environ. Entomol. 32: 397-406. PURCELL, M. F., AND BALCIUNAS, J. K. 1994. Life history and distribution of the Australian weevil Oxyops vitiosa (Coleoptera: Curculionidae), a potential biological control agent for Melaleuca quinquenervia (Myrtaceae). Ann. Entomol. Soc. America 87(6): 867-873. RAYAMAJHI, M. B., PRATT, P. D., CENTER, T. D., TIPPING, P. W., AND VAN, T. K. 2008. Aboveground biomass of the invasive tree melaleuca ( Melaleuca quinquenervia) before and after herbivory by adventive and introduced natural enemies: a temporal case study in Florida. Weed Sci. 56: 451-456. REES, M., AND HILL, R. L. 2001. Large-scale disturbances, biological control and the dynamics of gorse populations. J. App. Ecol. 38: 364-377. SAS. 1999. The SAS System for Windows, Version 8. SAS Institute Inc., Cary, North Carolina. SCHENK, D., AND BACHER, S. 2002. Functional response of a generalist insect predator to one of its prey species in the eld. J. Anim. Ecol. 71: 524-531. SEMPLE, J. L., AND FORNO, I. W. 1987. Native parasitoids and pathogens attacking Samea multiplicalis Guene (Lepidoptera: Pyralidae) in Queensland. J. Australian Entomol. Soc. 26: 365-366. SIMBERLOFF, D. 1989. Which insect introductions succeed and which fail? pp. 61-75 In J. A. Drake [ed.], Biological Invasions. John Wiley and Sons, New York. SIMBERLOFF, D., AND VON HOLLE, B. 1999. Positive interactions of nonindigenous species: invasional meltdown. Biol. Invasions 1: 21-32. SMITH, R. L. 1966. Ecology and Field Biology. Harper and Row, New York, N. Y. TIPPING, P. W., MARTIN, M. R., NIMMO, K. R., PIERCE, R. M., SMART, M. D., WHITE, E., MADEIRA, P. T.,AND CENTER, T. D. 2009. Invasion of a West Everglades wetland by Melaleuca quinquenervia countered by classical biological control. Biol. Cont. 48: 73-78. TIPPING, P. W., MARTIN, M. R., PRATT, P. D., CENTER, T. D., AND RAYAMAJHI, M. B. 2008. Suppression of growth and reproduction of an exotic invasive tree by two introduced insects. Biol. Cont. 44: 235-241. WHEELER G. S., MASSEY, L. M., AND SOUTHWELL, I. A. 2002. Antipredator defense of biological control agent Oxyops vitiosa is mediated by plant volatiles sequestered from the host plant Melaleuca quinquenervia. J. Chem. Ecol. 28(2): 297-315. WHEELER, G. S., MASSEY, L. M., AND SOUTHWELL, I. A. 2003. Dietary inuences on terpenoids sequestered by the biological control agent Oxyops vitiosa: Effect of plant volatiles from different Melaleuca quinquenervia chemotypes and laboratory host species. J. Chem. Ecol. 29: 188-207.

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Yang et al.: Mating Strategy of Ectropis oblique 9 COMPARATIVE MATING STRATEGIES OF MALE AND FEMALE ECTROPIS OBLIQUE (LEPIDOPTERA: GEOMETRIDAE) Y UN -Q IU Y ANG 1 X U -H UI G AO 1 Y AN -Z HUO Z HANG 3 L ONG -W A Z HANG 2 AND X IAO -C HUN W AN 1 1 Key Laboratory of Tea Biochemistry & Biotechnology, Ministry of Education and Ministry of Agriculture, Anhui Agricultural University, Hefei, Anhui 230036, P.R. China 2 Provincial Key Laboratory of Microbial Pest Control, Anhui Agricultural University, Hefei, Anhui230036, P.R. China 3 State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, P.R. China *Corresponding author; E-mail: tealab@ahau.edu.cn A BSTRACT The mating strategies of male and female Ectropis oblique Prout were investigated with the aid of male antennae as an electroantennogram (EAG) detector and capillary-GC analysis Each male was capable of mating with several females, but females that had received a spermatophore mated only once. Antennae dissected from males 0, 1, 2, 3, and 4 d post-mating and antennae from virgin males of corresponding ages displayed similar EAG responses to sex pheromone extracts from sexually active females. Pheromone extracts of mated females elicited signicantly weaker male EAG responses than the pheromone extracts of virgin females. EAG responses of males to sex pheromone extracts taken from mated females at 0, 1, 2, 3, and 4 d post-mating were consistently weak. Pheromone production in the pheromone glands of mated females was strongly suppressed and declined during each of 4 successive nights after they had mated. Key Words: EAG, GC, mating, sex pheromone emission, spermatophore R ESUMEN Las estrategias de apareamiento de machos y hembras de Ectropis oblique Prout fueron investigadas con el uso de la antena del mac ho como un detector electro-antenogramatico (EAG) y de CG capilar anlisis. Cada macho fue capaz de aparearse con varias hembras, pero las hembras que han recibido un espermatforo se aparearon solamente una vez. Las antenas diseccionadas de los machos 0, 1, 2, 3, y 4 dias despus de aparearse y las antenas de machos virgenes de edades corespondientes mostraron respuestas de EAG similares a los extractos de feromonas sexuales de hembras sexualmente activas. Los extractos de feromonas de hembras apareadas provocaron una respuesta del EAG de los machos signicativamente mas debl que los extractos de feromonas de las hembras vrgenes. Las respuestas de EAG de machos hacia los extractos de feromonas sexuales tomados de hembras apareadas a los 0, 1, 2, 3 y 4 dias despus de aparear fueron consistentemente mas debiles. La produccin de feromonas en las glandulas de feromonas de hembras apareadas fue fuertemente suprimida y declin durante cada una de las noches consecutivas despus de que las hembras se aparearon. Male insects typically mate many times during their lifetime, while females display diverse mating strategies (Arnqvist & Nilsson 2000). In some species, females need to mate once or only a few times to produce an optimal number of viable eggs; in many other species females mate frequently to maximize reproductive potential (Radwan & Rysinska 1999). No matter what strategies females use, they tend to discontinue sex pheromone production after mating, either temporarily or permanently; and this avoids problems associated with excessive male sexual harassment (Giebultowicz et al. 1991). This phenomenon was demonstrated by bioassay or chemical analysis in many studies (Webster & Card 1984; Coffelt & Vick 1987; Ahn et al. 2002). In contrast to the numerous studies on changes in female reproductive behavior after mating, few studies have focused on male response to diminished pheromone production of mated females. Recently, the electroantennogram (EAG) has been widely used in studies on semiochemical involvement in sex pheromones (Park et al. 2001; Gke et al. 2007). An EAG response prole is thought to represent the sensitivity and relative abundance of olfactory receptor neurons on the

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10 Florida Entomologist 94(1) March 2011 antennae that are tuned to the compounds tested. The EAG response amplitudes are thought to represent the quantity of semiochemicals (Pouzat & Ibeas 1989). Thus, the EAG may be used as a tool to investigate the sensitivity of males to variable amounts of sex pheromone. Ectropis oblique Prout (Lepidoptera: Geometridae) is an important tea bush pest in Southeast China. Population outbreaks can completely defoliate leaves on the bushes (Hu et al. 1994). The sex pheromone components of the female E. oblique were identified as (Z,Z,Z)-3,6,9octadecatriene and 6,7-epoxy-(Z,Z)-3,9-octadecadiene (Yao et al. 1991). Most efforts to control E. oblique populations are focused on the development of methodologies to disrupt reproduction. Therefore, understanding the behavior of E. oblique is necessary. In this study, the mating frequency and longevity of E. oblique males and females was investigated. At the same time, the different mating strategies of the two genders were examined using male antennae as EAG detectors. To verify the results of the electrophysiological analysis, changes in sex pheromone titers produced by mated and virgin females were also investigated using capillary-GC analysis. M ATERIALS AND M ETHODS Insect Culture The E. obliqu e insects were obtained from Qianshan County (31.5N, 116.3E), Anhui Province, China, and maintained for many generations in the laboratory. Larvae were reared on tea leaves. Adults and larvae were maintained in controlled conditions at 22 3C, 60-70% relative humidity, and a photoperiod of 14L:10D, with scotophase and photophase reversed from a natural light cycle to permit scotophase observations during normal working hours. Pupae were sexed based on the morphology of the 8th-10t h abdominal segments, and maintained in moist sand for eclosion. Adults were kept individually in 240-mL plastic jars and fed a 10% honey solution soaked in cotton. Effect of Mating Frequency on Longevity of Females and Males In preliminary observations of behavior, both females and males copulated during the rst scotophase after emergence, and were sexually active at the second scotophase. Therefore, a 1-d-old female (0 d after emergence) and a 2-d-old male were paired. Copulation occurred in the scotophase lasted approximately 6 h. After mating, the female insects did not resume calling during the same scotophase (Yang et al. 2008). Thus, each female in this experiment was replaced daily with another 1-dold virgin female. When the female died, the number of times it had mated was ascertained by counting the spermatophores present in her bursa copulatrix. The experiment was repeated 40 times. The number of times a male E. oblique mated was determined by keeping a record of the number of females mated during successive scotophases The experiment was also repeated 40 times. The longevity of each male and each female was recorded in order to determine whether mating affected longevity of either gender. Extraction of Sex Pheromones Active sex pheromones were extracted from the glands of the virgin females. The terminal section of the abdomen, which included the pheromone gland, was excised from the virgin female moth 6 h after the onset of the second scotophase, when the virgin female had been calling for 1 h. Experimental procedures were performed under a red light to facilitate observation without disturbing the insects. Each excised abdominal tip was immersed in 10 L of redistilled hexane for 46 h at room temperature. Then, the tip was removed and the extract without any purication was submitted for EAG or GC analysis. The procedure for extracting sex pheromones from either mated or virgin females was the same. Electroantennographic Analysis of the Effect of Mating on Pheromone Production and on Male Responsiveness at Various Days after Mating Electroantennograms (EAG) were obtained with an EAG apparatus (Syntech Co., 79199 Kirchzarten, Germany). The antennae of either mated or virgin males were excised at the bases and a few distal segments were cut off to facilitate conductivity. The antennae were then attached to the electrodes of the EAG probe with Spectra 360 Electrode Gel (Parker Laboratories Inc., Orange, New Jersey). Antennal preparations were exposed to a stream of humidied and charcoal-ltered air emitted at 4 mL s -1 after having owed through a 35-cm long glass tube (inner diameter, 8 mm; outer diameter, 10 mm). To facilitate insertion of the Pasteur pipette used to administer the pheromone test stimulus, a 3-mm hole was bored 5 cm from the outlet of the glass tube. Ten L of the extract (test stimulus) was applied to a piece of lter paper (1 5 cm1 5 cm). The lter paper w as placed in a Pasteur pipette after the solvent (hexane) had been allowed to evaporate for 5 min. Each test stimulus was delivered within a 0.5 s pulse of 4 mL s -1 of air with a stimulus controller (type CS-55) to transport the volatiles to the antenna. The EAG signal was amplied 10 through an intelligent data acquisition controller (type IDAC-2) and viewed on an oscilloscope A period of at least 30 s was allowed between 2 successive stimuli for the recovery of antennal responsiveness. Redistilled hexane (10 L) was used as a

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Yang et al.: Mating Strategy of Ectropis oblique 11 control stimulus in every test. The absolute EAG amplitude (mV) minus the solvent response was used for data analysis. First, the inuence of the males mating status on the males EAG responses to active sex pheromones was studied. To obtain mated males and females, the insects were paired in the rst scotophase and allowed to mate. Mating pairs were then removed. After mating, the mated females were used for the next experiment. With the same procedure as described above, the antennae of the mated male were dissected at 0, 24, 48, 72, and 96 h after mating for use as EAG test detectors. Sex pheromone extracts from sexually active females were used as stimuli. The EAG responses of antennae dissected from virgin males at each of these times post-mating were compared to the EAG responses of antennae obtained from mated males at corresponding times post-mating. Each treatment was repeated 6-8 times. Secondly, the inuence of the females mating status on the EAG responses of a male w as studied with sex pheromone extract of a mated female as the stimulus to elicit an EAG response from a 2-d-old virgin male. As described above, the sex pheromone of the mated females was extracted at 0, 24, 48, 72, and 96 h after mating. The EAG responses of antennae of 2-d-old virgin males exposed to pheromone extract obtained from virgin females at each of the above times post-mating were compared to the EAG responses to pheromone extract obtained from mated females at corresponding times post mating. Each treatment was also repeated 6-8 times. Pheromone Titer Analysis To assess the effect of mating on pheromone production, the sex pheromone titers of the mated and virgin females were analyzed by the procedure described above. Thus sex pheromone extract was analyzed in a gas chromatograph (Agilent 6890) equipped with a capillary column (DB5, 60m 0.5mm i.d 0.25 m lm). The oven temperature w as programmed at 50C for 2 min, then 15C min -1 to 250C and held for 5 min. The temperatures of the injector and detector were 200C and 250C, respectively. Nitrogen with a ow velocity of 40 mL min -1 was used as the carrier gas. To quantify the pheromones in the female gland, only the amount of epo3,Z6,Z9-19:H, the major sex pheromone component of E. oblique, was determined. Each treatment was repeated 6-8 times. Statistical Analysis The data were analyzed by one-way ANOVA, followed by a LSD multiple comparison test at P < 0.05 (SPSS 11.0 for Windows, 2002; SPSS Inc., Chicago, IL). R ESULTS Mating Frequencies of Females and Males The results of mating frequency are shown in Table 1. When 40 females were paired individually with 2-d-old virgin males on successive days until they died, only 1 spermatophore was detected in the abdomens of 31 females (77.5%), while no spermatophore was detected in the abdomens of the rest of the females (9, 22.5%). Thus any female that had received a spermatophore in 1 mating did not copulate again. When males were repeatedly offered 2-d-old virgin females, the numbers of males that mated various times during their lifespan and the corresponding percentages were as follows: 0 matings (10; 25.0%); 1 mating (9;22.5%); 2 matings (8; 20.0%), 3 matings (7; 17.5%), 4 matings (5; 12.5%); 5 matings (0; 0%) and 6 matings (1; 2.5%). The Effect of Mating on the Longevity of Adults Males lived signicantly longer than females (Table 1), whether mated or unmated ( P < 0.05). T ABLE 1. M ATING FREQUENCY OF E CTROPIS OBLIQUE AND ITS EFFECT ON FEMALE AND MALE LONGEVITY Mating time No. of females observed Female mating rates (%) Female Longevity (days) No. of males observed Male mating rates (%) Male longevity (days) 0 9 22.5*11.36 2.67 a10 25.014.2 0.85 b 1st 31 77.510.07 2.87 a9 22.519.6 1.82 c 2nd 0 0 ** 8 20.016.0 1.47 b 3rd 0 0 7 17.515.5 2.50 b 4th 0 0 5 12.515.5 1.50 b 5th 0 0 0 0 6th 0 0 1 2.515 *Values are mean SE. Different letters indicate signicant difference ( P < 0.05) by LSD test. ** not tested.

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12 Florida Entomologist 94(1) March 2011 In addition, the males that mated only once lived signicantly longer than unmated males or males that had mated more than once ( P < 0.05). However males that had mated 2 times did not live signicantly longer than males that had mated either 3 or 4 times The life spans of mated and unmated females did not differ signicantly. Electroantennographic Analysis of the Effect of Mating on Pheromone Production and on Male Responsiveness at Various Days after Mating The effects of mating and lapsed time after mating of males on their EAG responses to sex pheromone extracts from sexually active females are shown in Fig. 1. No signicant ( P > 0.05) differences in EAG responses to sex pheromone extracts from sexually active females were observed between the mated and virgin males The antennae of males that had been amputated 0, 1, 2, 3, and 4 d post-mating exhibited the same magnitude of the EAG responses to sex pheromone extracts from sexually active females as those of antennae of the corresponding virgin males. The male EAG responses to female sex pheromone extracts from virgin females and mated females (Fig. 2) differed profoundly ( P < 0.05) with the former evoking muc h stronger responses than the latter. Moreover the male EAG responses to sex pheromones extracted from the females at 0, 1, 2, 3, and 4 d post mating remained at very low levels (data not shown). Pheromone Titer Analysis The virgin females began to produce sex pheromones during the rst scotophase after emergence (Fig. 3). Maximal pheromone titers were present in the glands during the second and third scotophase after emergence. Thereafter the pheromone titer decreased gradually. When the females mated on the rst night after eclosion, the sex pheromone titers decreased strongly and signicantly compared with the titers of virgin females. Pheromone production in mated females remained suppressed during each of 4 successive nights after they had mated. These results are consistent with the weak male antennographic responses coinciding with the male response to mated female sex pheromones. Fig. 1. Inuence of the mating status of males on their EAG response to female sex pheromone extracts from sexually active females. Antennae of unmated males (solid bar) and virgin males (open bar) amputated 0, 1, 2, 3, and 4 d post-mating were used as EAG detectors. Data were presented as mean values SE ( n = 68) and analyzed by one-way ANOVA, followed by an LSD multiple range test ( P < 0.05). Signicant differences among various dosages of the same stimulant are indicated with different letters. Fig. 2. Inuence of differential mating status of females on male EAG responses to their pheromones. Sex pheromone extracts of mated females (solid bar) and virgin females (open bar) obtained 0, 1, 2, 3, and 4 d post-mating were used as stimuli. Data were presented as mean values SE ( n = 6-8) and analyzed by one-way A NOVA, followed by an LSD multiple range test ( P < 0.05). Signicant differences among various dosages o f the same stimulant are indicated with different letters. Fig. 3. Inuence of mating on the production of 6,7epoxy-(Z,Z)-3,9-octadecadiene, the major female sex pheromone. The female sex pheromone was extracted with hexane from pheromone glands of mated females (solid bar) and virgin females (open bar). Pheromone glands of mated females were extracted 0, 1, 2, 3, and 4 d post-mating.

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Yang et al.: Mating Strategy of Ectropis oblique 13 D ISCUSSION Our data suggest that the females in our laboratory colony of E obliqu e are monandrous. Two different views exist concerning female mating strategies Polyandry presents a variety of benets to females, including full fertilization of their egg complement, increased genetic diversity of offspring, receipt of non-sperm nutrients, and reduced chances of fertilization by sperm that are genetically defective due to age. Conversely, polyandry may decrease female tness due to the ecological cost of mating, including energy costs, and risks of physical injury and sexually transmitted pathogens and parasites (Arnqvist & Nilsson 2000). We observed that E obliqu e females after ha ving mated fended off males and did not accept a second mating partner. Mated females began laying eggs during the rst scotophase and laid nearly all of their eggs before the fourth day (data not shown). The life spans of females ranged from about to of the lengths of the life spans of males The lifespan of the female in the wild is likely to be even shorter than in the laboratory. Thus, a single mating is sufcient to fertilize nearly all eggs and minimize the above mentioned risks associated with multiple matings. Males, on the other hand, are potentially polygynous. Male polygyny is to be expected because most evolutionary theories contend that the contributions and consequences of mating are much greater for females than for males (Thornhill & Alcock 1983). This study revealed that multiple matings reduced the longevity of males but not of females. This result is in agreement with the results of studies on several other species (Proshold et al. 1982; Svensson et al. 1998). It is thought that allocation of nutritional reserves for egg development and maturation after mating may be responsible for causing the lifespan of mated females to be shorter than that of virgin females. In most species, there is a causal relationship between male calling behavior and female pheromone emission. Only when the female pheromone gland becomes exposed to emit pheromone, may the male display calling behavior. Permanent or even temporary reductions in the emissions of sex pheromones caused loss of attraction and sexual receptivity in males (Kingan et al. 1995). The present study showed that E obliqu e male EAG responses to mated female pheromone gland extracts were signicantly diminished, which could be the result of reduced pheromone release. The results support the hypothesis that mating considerably suppressed pheromone production in females. Indeed according to our capillary-GC analysis, pheromone titers in pheromone gland extracts did not increase at all up to 4 d after mating. Because the male EAG response to pheromone gland extract of already mated females did not show any increase up to 4 d after mating, it may be deduced that females may mate only once. This deduction is in accordance with the observation that each female actually mates only 1 time. Even though a few females were observed to copulate twice, but no more than 1 spermatophore was ever detected in a bursa copulatrix, possibly because the rst copulation was an unsuccessful mating. Mating-induced termination of sex pheromone production has been investigated in several moth species (Raina et al. 1994; Ando et al. 1996). The inactivation of pheromone production after copulation, which reduces the ability of females to elicit a sexual response in males, may be due to the secretion of pheromonostatic peptide (Kingan et al. 1995), or the presence of viable sperm in the spermatheca (Giebultowicz et al. 1991). The mechanisms involved in pheromone suppression after copulation in E obliqu e are currently unknown. Thus, it is suggested that further studies must be conducted on this area. The present results show that mating status did not appear to have a signicant effect on the responses of male antennae to sex pheromone extracts of sexually active females. This suggests that a past mating does not cause the male to be unresponsive to the sex pheromone, and such a male can be expected to continue to seek females for additional matings. This deduction is also in accordance with direct observations of multiple matings by males. The same mating system described for E obliqu e has been observed in some other species (Royer & McNeil 1993; Foster& Ayers 1996; Svensson et al. 1998). A CKNOWLEDGMENT The authors express gratitude to Professor M. Z. FAN for assistance during this study. R EFERENCES CITEDAHN, S. J., CHOI, M. Y., AND BOO, K. S. 2002. Mating effect on sex pheromone production of the Oriental tobacco budworm, Helicoverpa assulta. J. Asia-Pacic Entomol. 5: 43-48. ANDO, T., KASUGA, K., YAJIMA, Y., KATAOKA, H., ANDSUZUKI, A. 1996. Termination of sex pheromone production in mated females of the silkworm moth. Arch. Insect. Biochem. Physiol. 31: 207-218. ARNQVIST, G., AND NILSSON, T. 2000. The evolution of polyandry: Multiple mating and female tness in insects. Anim. Behav. 60: 145-164. COFFELT, J. A., AND VICK, K. W. 1987. Sex pheromone of Ephestia cautella (Walker) (Lepidoptera, Pyralidae): Inuence of mating on pheromone titer and release rate. J. Stored Prod. Res. 23: 119 -123. FOSTER, S. P., AND AYERS, R. H. 1996. Multiple mating and its effects in the light brown apple moth, Epiphyas postvittana (Walker). J. Insect Physiol. 42: 657667.

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14 Florida Entomologist 94(1)March 2011GIEBULTOWICZ, J. M., RAINA, A. K., UEBEL, E. C., ANDRIDGWAY, R. L. 1991. Two-step regulation of sexpheromone decline in mated gypsy moth females. Arch. Insect Biochem. Physiol. 16: 95-105. GKE, A., STELINSKI, L. L., GUT, L. J., AND WHALON, M. E. 2007. Comparative behavioral and EAG responses of female obliquebanded and redbanded leafroller moths (Lepidoptera: Tortricidae) to their sex pheromone components. European J. Entomol 104: 187-194. HU, C., ZHU, J., YE, G., AND HONG, J. 1994. Ectropis oblique Prout, a serious geometrid pest of tea bush in East China. Scientic and Technical Publishers, Shanghai. pp. 226-228. KINGAN, T. G., BODNAR, W. M., RAINA, A. K., SHABANOWITZ, J., AND HUNT, D. F. 1995. The loss of female sex pheromone after mating in the corn earworm moth, Helicoverpa zea: Identication of a male pheromonostatic peptide. Proc. Natl. Acad. Sci 92: 50825086. PARK, K. C., ZHU, J. W., HARRIS, J., OCHIENG, S. A., ANDBAKER, T. C. 2001. Electroantennogram responses of a parasitic wasp, Microplitis croceipes to host-related volatile and anthropogenic compounds. Physiol. Entomol. 26: 69-77. POUZAT, J., AND IBEAS, D. N. 1989. Electrophysiological investigations of sex pheromone reception and release in Bruchidius atrolineatus Physiol. Entomol. 14: 319-324. PROSHOLD, F. I., KARPENKO, C. P., AND GRANAM, C. K. 1982. Egg production and oviposition in the tobacco budworm: Effect of age at mating. Ann. Entomol. Soc. America 75: 51-55. RADWAN, J., AND RYSINSKA, M. 1999. Effect of mating frequency on female tness in Caloglyphus berlesei (Astigmata: Acaridae). Exp. Appl. Acarol. 23: 399409. RAINA, A. K., KINGAN, T. G., AND GIEBULTOWICZ, J. M. 1994. Mating-induced loss of sex pheromone and sexual receptivity in insects with emphasis on Helicoverpa zea and Lymantria dispar Arch. Insect. Biochem. Physiol 25: 317-327. ROYER, L., AND MCNEIL, J. N. 1993. Male investment in the European corn borer, Ostrinia nubilalis (Lepidoptera: Pyralidae): impact on female longevity and reproductive performance. Funct. Ecol. 7: 209-215. SVENSSON, M. G. E., MARLING, E., AND LFQVIST, J. 1998. Mating behavior and reproductive potential in the turnip moth, Agrotis segetum (Lepidoptera: Noctuidae). J. Insect Behav. 11: 343-359. THORNHILL, R., AND ALCOCK, J. 1983. The Evolution of Insect Mating Systems. Harvard University Press, Cambridge. WEBSTER, R. P., AND CARD, R. T. 1984. The effects of mating, exogenous juvenile hormone and a juvenile hormone analog on pheromone titer, calling and oviposition in the omnivorous tea leafroller moth (Platynota stultana). J. Insect. Physiol. 30: 113-118. YANG, Y. Q., WAN, X. C., ZHENG, G. Y., AND GAO, X. H. 2008. Preliminary study on the habit of sex behavior of Ectropis obliqua Prout. Chinese Agric. Sci. Bull. 24: 339-342 (in Chinese). YAO, E. Y., LI, Z. M., LOU, Z. Q., AND SHANG, Z. Z. 1991. Report on structural elucidation of sex pheromone components of a tea pest (Ectropis obliqua Prout). Prog. Nat. Sci. 1: 566-569.

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Farnum & Loftin: Distribution of P. curvatus and P. tricuspis in Arkansas15 DISTRIBUTION OF PSEUDACTEON CURVATUS AND PSEUDACTEON TRICUSPIS (DIPTERA: PHORIDAE) IN ARKANSAS J AKE M. F ARNUM AND K ELLY M. L OFTIN Department of Entomology, University of Arkansas, 319 Agriculture Bldg., Fayetteville, AR 72701 A BSTRACT From 1995 to 2009, four Pseudacteon species were released in the U.S. with 3 species, P. curvatus P. tricuspis and P. obtusus released in Arkansas. To determine Pseudacteon establishment and expansion, sticky traps were used to monitor phorid y species at and near 10 release sites, in counties bordering neighboring states, and along regional transects. Pseudacteon ies were captured in 16 Arkansas counties: Ashley, Chicot, Clark, Desha, Drew, Hempstead, Howard, Little River, Montgomery, Nevada, Perry, Phillips, Pike, Polk, Sevier, and Union. Pseudacteon curvatus was found in areas far from release sites, suggesting dispersal from neighboring states The range of P. tricuspis evidently also expanded from its initial release sites in southern Arkansas. Key Words: biological control, phorid y, Solenopsis invicta parasitoid R ESUMEN De 1995 hasta el 2009, cuatro especies de Pseudacteon fueron liberadas en los Estados Unidos con 3 de ellas P. curvatus P. tricuspis y P. obtusus, liberadas en el estado de Arkansas. P ara determinar el establecimiento y expansin de Pseudacteon se usaron trampas pegajosas para monitorear las especies de fridos en y alrededor de los 10 sitios donde fueron liberadas en condados fronterizos con los estados vecinos, y a lo largo de las lineas regionales. Las moscas Pseudacteon fueron capturadas en los siguientes 16 condados de Arkansas: Ashley Chicot, Clark, Desha, Drew, Hempstead, Howard, Little River, Montgomery, Nevada, Perry, Phillips, Pike, Polk, Sevier y Union. Pseudacteon curvatus fue encontrada en reas lejos de los sitios donde fue liberada, que indica que se disperso de los estados vecinos. Evidentemente, el rango de P. tricuspis tambien se expandi de los sitios donde fue liberada inicialamente en el sur de Arkansas. Pseudacteon phorid ies parasitize and kill their re ant host ( Solenopsis invicta Buren, S. ric hteri Forel and their hybrid). Due to their high host specicity (F olgarait et al. 2002; Gilbert & Morrison 1997; Morrison & Gilbert 1999; Porter 1998a, 2000; Porter & Alonso 1999; Porter & Gilbert 2004; Vazquez et al. 2004), several Pseudacteon spp. have been introduced from South America as c lassical biological control agents against imported re ants. While internal development of the y larvae eventually decapitates and kills individual ants (Porter et al. 1995a; Consoli et al. 2001), perhaps the most important effect is the disruption of the foraging behavior of the ants (Feener & Brown 1992; Orr et al. 1995; Porter et al. 1995b; Mehdiabadi & Gilbert 2002), which leads to a decrease in food uptake and a decline in colony health (Folgarait & Gilbert 1999). Four Pseudacteon species were released in the U.S. from 1995 to 2009: P. curvatus Borgmeier P. litoralis Borgmeier, P. obtusus Borgmeier, and P. tricuspis Borgmeier. Each species lls a different nic he, in terms of diurnal activity (Pesquero et al. 1996), seasonal occurrence (Fowler et al. 1995; Folgarait et al. 2003) and preferred size of host (Campiolo et al. 1994); all complementing traits for control of S. invicta and S. richteri (Morrison et al. 1997; Porter 2000; Folgarait et al. 2002, 2005). Pseudacteon ies use semiochemicals to locate their ant hosts (Orr et al. 1997; Vander Meer & Porter 2002; Morrison & King 2004; Chen & Fadamiro 2007) and then hover over the ants before a rapid aerial attack (Morrison & Porter 2005). Within a period of an hour a female y can make up to 120 oviposition attempts (Morrison et al. 1997), before either tiring or being captured and killed by the ants (Porter 1998b). A single egg laid in the thorax develops through 3 instars before decapitating the ants head, which is then used as a pupal case (Pesquero et al. 1995; Porter et al. 1995a). The rst release of P. tricuspis occurred in T exas in 1995 (Gilbert 1996), and was unsuccessful due to unfavorable conditions (Vazquez et al. 2006). The rst successful release of P. tricuspis in northern Florida w as in 1997 (Porter et al. 1999). Pseudacteon spp. have been released in 11 southern states: Alabama, Arkansas, Florida, Georgia, Louisiana, Mississippi, North Carolina, Oklahoma, South Carolina, Tennessee, and Texas (Porter et al. 1999; Graham et al. 2003; Williams & deShazo 2004; Parkman et al. 2005; Thead et al. 2005; Henne et al. 2007; Weeks & Callcott

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16 Florida Entomologist 94(1) March 2011 2008). Pseudacteon curvatus, P. obtusus, and P. tricuspis have been released in Arkansas and in adjacent states except Missouri (Clemons et al. 2003; Weeks & Callcott 2008). Dispersal rates for Pseudacteon spp. are variable and the majority of ies disperse only a few hundred meters (Morrison et al. 1999). However, some ies in each generation are known to travel 2 to 4 km or more (Porter et al. 2004) and populations of ies expanding on average 74 km over a period of 3.5 years (Porter 2010). Monitoring of Pseudacteon spp. is achieved with a variety of methods: actively through direct collections of ies at disturbed ant colonies with either manual or electrical stimulation (Barr & Calixto 2005; Morrison & Porter 2005), or passively by trapping with a sticky trap (Puckett et al. 2007). Until the current study, monitoring of these species in Arkansas had been concentrated at and near release sites to determine Pseudacteon establishment. The objective of this study w as to determine the distribution of Pseudacteon spp. in Arkansas through wider-scale monitoring. M ATERIALS AND M ETHODS Trap Design To determine presence or absence, passive trapping was used based on a modified version of a PTS (pizza tri-stand) sticky trap (Puckett et al. 2007). The modified Puckett trap (Fig. 1) consisted of a pizza tri-stand (Polyking No. 20431) covered in Tanglefoot, glued to the flat side of an inverted portion cup (Dart No. 100PC), which was hot glued to the underside of a plastic cup lid (Dart No. 8JL). This device was placed in the center of the bottom half of a plastic Petri dish (150 by 15 mm). The surfaces of the portion cup and the inner lip of the Petri dish were coated with Fluon, to prevent ants from climbing up the trap or out of the Petri dish. One trap was placed per location by rst locating a mound of substantial size and activity. The mound was then disturbed, by kicking it over creating a at surface on which the Petri dish was placed. As ants climbed into the Petri dish, a few ants were crushed by hand to induce alarm pheromone release and the trap was placed in the center of the Petri dish. A brightly colored pin ag (91 cm long) was positioned alongside the trap. Global positioning system (GPS) coordinates were recorded for each location, and a corresponding number written on the lid of the trap. Traps were retrieved 20 to 24 h after placement. At time of retrieval, an 8 oz expanded polystyrene foam cup (Dart No. 8J8) was placed over the trap, and the lid was snapped in place. The cup prevented damage and contamination to the sticky portion of the trap. Sampling at Release Locations Fourteen releases of Pseudacteon spp. were made in Arkansas from 1998 to 2009 (Table 1). Sampling along transects in the cardinal directions from the release sites began in 2002 (Pike Co.) and 2004 (Miller Co.). In 2009, the Miller, Perry, Pike, and Sevier County release sites were revaluated for this study to confirm establishment of Pseudacteon spp. A 1.6-km interval was used between traps along each transect placed in Pike, Miller, and Sevier Counties, and at 0.8-km intervals in Perry County. Transects were determined by locating roads and highways on aerial maps that radiated out from the release site in north, south, east, and west directions. One modified Puckett trap was placed at each sampling location. Sampling Bordering Counties Sampling was intended to monitor spread of established populations of Pseudacteon spp. in bordering counties/parishes of neighboring states of Louisiana (Henne et al. 2007), Mississippi (Thead et al. 2005), and Tennessee (Graham et al. 2003; Parkman et al. 2005; Weeks & Callcott 2008), and sampling packages were sent to University of Arkansas Cooperative Extension Service County Agents in imported re ant infested counties in eastern and southern Arkansas in the early summer of 2009. Each package contained four modied Puckett traps, latex gloves, and an information sheet. Instructions were to place the traps during the summer months, as previously described, at 4 locations within the county. Deployed traps were returned from 4 bordering counties: Columbia, Lafayette, Phillips, and St. Francis. Geographic coordinates were recorded on the instruction sheet with the number of the corresponding trap. Fig. 1. A modied Puckett trap.

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Farnum & Loftin: Distribution of P. curvatus and P. tricuspis in Arkansas17 Sampling Along Regional Transects Due to possible expansion of Pseudacteon spp. from neighboring states, sampling transects were identified and mapped in 3 regions: western, southeastern, and southwestern Arkansas (Fig. 2). Transects began at the state line of the adjacent state and traveled inward, with modified Puckett traps placed at 3-mile intervals and GPS coordinates recorded for each trap. Sampling was conducted for 36 h along transects from each region between 19 Sep and 3 Oct 2009. Phorid Fly Identications Upon retrieval, modified Puckett traps were taken to the University of Arkansas laboratory (Fayetteville, AR) and GPS coordinates logged into Google Earth. Traps were examined under a dissecting microscope for presence of Pseudacteon spp. The sticky portion of the trap was sprayed with liquid degreaser (Goo Gone) if fly removal was necessary. Species were identified based on the morphology of the female ovipositor (Porter & Pesquero 2001). Voucher specimens of P. curvatus P. tricuspis and P. obtusus were obtained from laboratory specimens and the phorid fly rearing facility in Gainesville, FL for identification of male and female flies. Male Pseudacteon spp. are currently difficult to identify through use of keys (Morrison et al. 1997; Porter & Pesquero 2001). R ESULTS AND D ISCUSSION Release Locations Traps placed along transects radiating from the phorid fly release sites yielded surprising results. Of the 20 traps retrieved from Sevier County, no Pseudacteon spp. were caught. Traps from Miller, Perry, and Pike Counties captured Pseudacteon spp Along the Miller County release site transects, P. curvatus was captured at 3 locations north of Texarkana, in the southeastern part of Little River County (Fig. 3). However, this was unexpected because the phorid fly species released in Miller County in 2004 was P. tricuspis T ABLE 1. H ISTORY OF P SEUDACTEON SPP RELEASES IN A RKANSAS 1998-2009. DateCountySpeciesNumber Released 1998, Jul-Aug 1 Drew P. tricuspis 1,350 2002, May 2 Pike P. tricuspis 3,000 2002, Oct 2 Bradley P. tricuspis 1,200 2003, Sep 2 Bradley P. tricuspis 1,500 2004, May 2 Miller P. tricuspis 2,580 2005, May 2 Sevier P. tricuspis 4,300 2005, Oct 2 Clark P. curvatus 8,500 3 2006, Sep 2 Perry P. curvatus 15,816 4 2007, Sep 2 Perry P. tricuspis 1,900 2008, Jun2Jefferson P. tricuspis 120 2008, Oct2Polk P. obtusus 3,20052009, May2Grant P. curvatus 25,29662009, Jun2Jefferson P. tricuspis 120 2009, Sep2Garland P. tricuspis 3601Release made by Lynne Thompson, University of Arkansas Monticello. 2Release made by Kelly Loftin, University of Arkansas Cooperative Extension Service.3Number released based on 35.3 g S. invicta workers, 800/gram and 30% parasitism.4Number released based on 65.9 g S. invicta workers, 800/gram and 30% parasitism.5Number released based on 23.7 g S. invicta workers, 450/gram and 30% parasitism.6Number released based on 105.4 g S. invicta workers, 800/gram and 30% parasitism. Fig. 2. Regional transects in Arkansas (SE1, SE2, SE3, SW1, SW2, SW3, SW4, W1, W2, W3, W4) (ESRI Inc. 2009).

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18 Florida Entomologist 94(1)March 2011(Fig. 3, I on map). While possible, it is unlikely that these captures resulted from cross contamination of the 2 species at the rearing facility. Several factors are in place to prevent cross contamination including parasitization of the ants in separate rooms and lack of available ants of preferred host size. Furthermore, voucher sampling is conducted to insure parasitization by the correct species (Amy Croft, Florida Department of Agriculture and Consumer Services, personal communication). A plausible explanation could be the expansion of P. curvatus from released and established populations in the bordering states of Louisiana, Oklahoma, and Texas (Weeks & Callcott 2008; Anne-Marie Callcott, USDAAPHIS, personal communication). Similar results were observed along the northeast transect of the Pike County release site; capture of a Pseudacteon fly (P. curvatus ) different from the species originally released (P. tricuspis ). Due to the proximity (5 km) with the Clark County release site (Fig. 3A), it is possible that this capture was from movement of P. curvatus from Clark County, although revaluation of Clark County was not conducted in 2009. In Perry County, capture of P. curvatus was recovered at the initial release site and at locations to the south (0.5 km) and east (2.2 km).Bordering CountiesPuckett traps from Columbia, Lafayette, and St. Francis Counties were devoid of Pseudacteon flies, although all traps returned from Phillips County on the Mississippi border captured P. curvatus. Phillips County is the one of 2 counties in Arkansas (Crittenden Co. the other) currently known to have only S. richteri and no record of S. invicta (Robert Vander Meer, USDA-ARS, personal communication). Sampling locations in Phillips Co. were located on the western levee of the Mississippi River, northwest of Friars Point, MS (Fig. 3D), adjacent to counties in Mississippi and Tennessee with known S. richteri populations (Streett et al. 2006; Oliver et al. 2009). The high number of P. curvatus found on 1 trap (~172) implies sufficient colonies of S. richteri to support P. curvatus populations.Regional TransectsA total of 176 modied Puckett traps were placed along transects in western, southeastern, and southwestern Arkansas (Fig. 2). In the western region of transects, 28 traps contained P. curvatus and 2 contained P. tricuspis (Fig. 3). Of the traps that captured P. tricuspis one was located 29.5 km west of the P. tricuspis release in Pike County, and the other was 46 km northwest of the release site. All 4 transects in this region included captures of P. curvatus On the 2 northerly routes (W1 and W2), traps with P. curvatus were found at regular intervals with the most easterly capture 54 km from the Arkansas/Oklahoma state line. The remaining 2 transects (W3 and W4) also had captures. Transect (W3) picked up P. curvatus 13 km from the Arkansas/Oklahoma border. The 3 traps from the most southerly transect (W4) that contained P. curvatus were located near the northern section of the Miller County release site transect. This directional pattern of recoveries supports the hypothesis of immigration from conrmed P. curvatus populations in Le Flore County, Oklahoma (Weeks & Callcott 2008) approximately 24 km northwest of Mena, Arkansas. Prevailing winds generally do not correlate with dispersion patterns (Morrison et al. 2000), as ies move close to the ground where wind is reduced. Pseudacteon curvatus but not P. tricuspis was trapped along the southeastern region transects. P. curvatus was found at 1 location in Union County, north of El Dorado, and on 12 traps along each of the other transects (SE2 and SE3) to the east (Fig. 3). From the 2 easterly transects (SE2, SE3), traps with P. curvatus were found 91 km north of the Arkansas/Louisiana border and 65 km west of the Arkansas/Mississippi border. This distribution in Arkansas is expected based on collections of P. curvatus in bordering counties along the western side of Mississippi (Adams, Bolivar, Claiborne, Coahoma, Desoto, Jefferson, Tunica, Warren, Washington, Wilkinson), and along Louisianas northern side (Claiborne, East Carroll, Morehouse, Union, West Carroll) (Anne-Marie Callcott, USDA-APHIS, personal communication). Fig. 3. Pseudacteon curvatus release sites in (A) Clark Co., (B) Grant Co., (C) Perry Co., and 2009 capture locations including (D) Phillips Co., Arkansas. P seudacteon tricuspis release sites in (E) Bradley Co., (F) Drew Co., (G) Garland Co., (H) Jefferson Co., (I) Miller Co., (J) Perry Co., (K) Pike Co., (L) Sevier Co., Arkansas and 2009 capture locations (ESRI Inc. 2009).

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Farnum & Loftin: Distribution of P. curvatus and P. tricuspis in Arkansas19Pseudacteon tricuspis was captured at 2 locations along a transect (SW3) of the southwestern region of Arkansas (Fig. 3). One of the traps was located in the northern part of Nevada County, in the city of Prescott, and the other was 18 km to the southeast. Their proximity to the Pike County release site (Fig. 3K), 20 km northwest, may suggest their origin, although P. curvatus appeared in no samples taken from the Pike County release site transect. Weather conditions may have played a role in the lack of Pseudacteon spp. present on the remaining 55 traps in this region. Temperatures for the region on 3 Oct 2009 ranged from a low of 8C to a high of 26C. Temperatures below 20C inhibit activity of Pseudacteon ies (Morrison et al. 1999; Wuellner et al. 2002). On 3 Oct 2009 temperatures were above 20C for a 7-h period midday (11:00 AM to 6:00 PM CST) and below 20C for the entire day of 4 Oct 2009, when the traps were retrieved. However, 2 traps located on the transect southeast of Prescott, AR collected P. tricuspis Weather data suggests similar conditions for the Prescott, AR area, although slightly warmer temperatures (17C) were recorded for an additional 3.5 h on the morning of 4 Oct 2009. Of the traps that collected Pseudacteon spp., no traps contained both species. Perhaps because P. tricuspis is reliant on larger ants, P. curvatus is able to establish more readily where P. curvatus and P. tricuspis overlap, and thus were not detected if present in low densities (Gilbert et al. 2008). Another factor for the lack of both species in the trap may be due to the modication of the Puckett trap. The 2 differences in the traps design were the attractant used and the placement of the trap. Traps were placed on a disturbed mound with live ants whereas Puckett traps were placed in an open area with midden (Puckett et al. 2007). The Puckett trap as originally designed captured more P. tricuspis than P. curvatus although seasonal uctuations could be a variable (Puckett et al. 2007). CONCLUSIONPassive Pseudacteon trapping with the modied Puckett trap provided advantages over direct collection from disturbed mounds. Because it is deployed quickly, multiple traps can be placed over a large area which allows continuous and simultaneous sampling (Puckett et al. 2007). The manpower needed to achieve similar coverage using observational sampling is cost prohibitive. The addition of the protective cup to the original design allowed longer storage time between collection and examination of the trap, protection of the sticky portion, and reduced contamination. The modication based on disturbed mounds and uon-coated petri dishes rather than re ant midden was advantageous in that maintenance of a re ant colony for collection of midden is no longer necessary. With this modication, trapped re ants rather than re ant midden serve as the Pseudacteon y attractant. The results suggest establishment and expansion of P. curvatus from the release site in Perry County, and P. tricuspis from the release site in Pike County. While limited, the current range of P. tricuspis in Arkansas appeared to be along a narrow 86-km band stretching from northwest Pike County to south central Nevada County, and 25 km west of the release site. The current distribution of P. curvatus in Arkansas suggested natural movement from surrounding states. Despite extensive sampling across southern Arkansas, many areas remained unsampled. Additional trapping would provide a better understanding of the distribution of Pseudacteon spp. in Arkansas. ACKNOWLEDGMENTSWe thank Anne-Marie Callcott of the USDA APHIS lab in Gulfport, MS for approving Pseudacteon spp. for release, Amy Bass, Amy Croft, and Deborah Roberts of the Florida Department of Agriculture and Consumer Services for assistance in supplying phorid ies, Ed Brown, Jerry Clemons, Randy Forst, Rex Herring, Mike McCarter, Shawn Payne, Doug Petty, Amy Simpson, Rebecca Thomas, Shaun Rhodes, Carla Vaught, Joe Vestal, and Danny Walker of the University of Arkansas Cooperative Extension Service for assistance placing phorid y traps, Michael Hamilton and Robert Goodson for collecting imported re ants for identication, and Ricky Corder of the University of Arkansas Cooperative Extension Service for assistance.REFERENCES CITEDBARR, C. L., AND CALIXTO, A. 2005. Electrical stimulation of Solenopsis invicta (Hymenoptera: Formicidae) to enhance phorid fly, Pseudacteon tricuspis (Diptera: Phoridae), detection. Southwest. Entomol. 3: 165-168. CAMPIOLO, S., PESQUERO, M. A., AND FOWLER, H. G. 1994. Size selective oviposition by phorid (Diptera: Phoridae) parasitoids on workers of the fire ant, Solenopsis saevissima (Hymenoptera: Formicidae). Etiologia. 4: 85-86. CHEN, L., AND FADAMIRO, H. Y. 2007. Behavioral and electroantennogram responses of phorid fly Pseudacteon tricuspis (Diptera: Phoridae) to red imported fire ant Solenopsis invicta odor and trail pheromone. J. Insect Behav. 20: 267-287. CLEMONS, J., MCCARTER, M., GAVIN, J., PETTY, D., LOFTIN, K., SHANKLIN, D., AND HOPKINS, J. 2003. Phorid fly releases in Arkansas: county agents perspective, pp. 40-41 In L. Greenberg, and C. Lerner [eds.], Proc. Imported Fire Ant Conference, Palm Springs, CA. CONSOLI, F. L., WUELLNER, C. T., VINSON, S. B., ANDGILBERT, L. E. 2001. Immature development of Pseudacteon tricuspis (Diptera: Phoridae), an endoparasitoid of the red Imported fire ants (Hymenoptera: Formicidae). Ann. Entomol. Soc. America 94: 97-109. ESRI, INC. 2009. ArcGIS 9.3.1. ESRI, Inc., Redlands, CA. FEENER, D. H., AND BROWN, B. V. 1992. Reduced foraging of Solenopsis geminata (Hymenoptera: Formi-

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20 Florida Entomologist 94(1)March 2011cidae) in the presence of parasitic Pseudacteon spp. (Diptera: Phoridae). Ann. Entomol. Soc. America 85: 80-84. FOLGARAIT, P. J., AND GILBERT, L. E. 1999. Phorid parasitoids affect foraging under different availability of food in Argentina. Ecol. Entomol. 24: 1-11. FOLGARAIT, P. J., BRUZZONE, O. A., PATROCK, R. J. W.,AND GILBERT, L. E. 2002. Developmental rates and host specificity for Pseudacteon parasitoids (Diptera: Phoridae) of fire ants (Hymenoptera: Formicidae) in Argentina. J. Econ. Entomol. 95: 1151-1158. FOLGARAIT, P. J., BRUZZONE, O. A., AND GILBERT, L. E. 2003. Seasonal patterns of activity among species of black fire ant parasitoid flies (Diptera: Phoridae) in Argentina explained by analysis of climatic variables. Biol. Control. 28: 368-378. FOLGARAIT, P. J., CHIRNO, M. G., PATROCK, R. J. W.,AND GILBERT, L. E. 2005. Development of Pseudacteon obtusus (Diptera: Phoridae) on Solenopsis invicta and Solenopsis richteri fire ants (Hymenoptera: Formicidae). Environ. Entomol. 34: 308-316. FOWLER, H. G., PESQUERO, M. A., CAMPIOLO, S., ANDPORTER, S. D. 1995. Seasonal activity of species of fire ants in Brazil. Cientifica. 23: 367-371. GILBERT, L. E. 1996. Prospects of controlling fire ants with parasitoid flies the perspective from research based at Brackenridge Field Laboratory, pp. 77-92 In W. E. Cohen [ed.], Texas Quail Short Course II, Texas Agricultural Extension Service, Texas A&M University, Kingsville, TX. GILBERT, L. E., AND MORRISON, L. W. 1997. Patterns of host specificity in Pseudacteon parasitoid flies (Diptera: Phoridae) that attack Solenopsis fire ants (Hymenoptera: Formicidae). Environ. Entomol 26: 1149-1154. GILBERT, L. E., BARR, C. L., CALIXTO, A. A., COOK, J. L., DREES, B. M., LEBRUN, E. G., PATROCK, R. J. W., PLOWES, R. M., PORTER, S. D., AND PUCKETT, R. T. 2008. Introducing phorid fly parasitoids of red imported fire ant workers from South America to Texas: outcomes vary by region and by Pseudacteon species released. Southwest. Entomol. 33: 15-29. GRAHAM, L. C., PORTER, S. D., PEREIRA, R. M., DOROUGH, H. D., AND KELLEY, A. T. 2003. Field releases of the decapitating phorid fly Pseudacteon curvatus (Diptera: Phoridae) for control of imported fire ants (Hymenoptera: Formicidae) in Alabama, Florida, and Tennessee. Florida Entomol. 86: 334-339. HENNE, D. C., JOHNSON, S. J., AND PORTER, S. D. 2007. Status of the fire ant decapitating fly Pseudacteon tricuspis (Diptera: Phoridae) in Louisiana. Florida Entomol. 90: 565-569. MEHDIABADI, N. J., AND GILBERT, L. E. 2002. Colony level impacts of parasitoid flies on fire ants. Proc. Biol. Sci. 269: 1695-1699. MORRISON, L. W., AND GILBERT, L. E. 1999. Host specificity in two additional Pseudacteon spp. (Diptera: Phoridae), parasitoids of Solenopsis fire ants (Hymenoptera: Formicidae). Florida Entomol. 82: 404-409. MORRISON, L. W., AND KING, J. R. 2004. Host location behavior in a parasitoid of imported fire ants. J. Insect Behav. 17: 367-383. MORRISON, L. W., AND PORTER, S. D. 2005. Phenology and parasitism rates in introduced populations of Pseudacteon tricuspis a parasitoid of Solenopsis invicta. Biocontrol. 50: 127-141. MORRISON, L. W., DALLAGLIO-HOLVORCEM, C. G., ANDGILBERT, L. E. 1997. Oviposition behavior and development of Pseudacteon flies (Diptera: Phoridae), parasitoids of Solenopsis fire ants (Hymenoptera: Formicidae). Environ. Entomol. 26: 716-724. MORRISON, L. W., KAWAZOE, E. A., GUERRA, R., ANDGILBERT, L. E. 1999. Phenology and dispersal in Pseudacteon flies (Diptera: Phoridae), parasitoids of Solenopsis fire ants (Hymenoptera: Formicidae). Ann. Entomol. Soc. America 92: 198-207. MORRISON, L. W., KAWAZOE, E. A., GUERRA, R. AND GILBERT, L. E. 2000. Ecological interactions of Pseudacteon parasitoids and Solenopsis ant hosts: environmental correlates of activity and effects on competitive hierarchies. Ecol. Entomol. 25: 433-444. OLIVER, J. B., VANDER MEER, R. K., OCHIENG, S. A., YOUSSEF, N. N., PANTALEONI, E., MREMA, F. A., VAIL, K. M., PARKMAN, J. P., VALLES, S. M., HAUN, W. G., AND POWELL, S. 2009. Statewide survey of imported fire ant (Hymenoptera: Formicidae) populations in Tennessee. J. Entomol. Sci. 44: 149-157. ORR, M. R., SEIKE, S. H., BENSON, W. W., AND GILBERT, L. E. 1995. Flies suppress fire ants. Nature. 373: 292-293. ORR, M. R., SEIKE, S. H., AND GILBERT, L. E. 1997. Foraging ecology and patterns of diversification in dipteran parasitoids of fire ants in South Brazil, genus Pseudacteon (Phoridae). Ecol. Entomol. 22: 305-314. PARKMAN, P., VAIL, K., RASHID, T., OLIVER, J., PEREIRA, R., PORTER, S. D., SHIRES, M., HAUN, G., POWELL, S., THEAD, L., AND VOGT, J. T. 2005. Establishment and spread of Pseudacteon curvatus in Tennessee, pp. 111-112 In D. J. Meloche [ed.], Proc. Imported Fire Ant Conference, Gulfport, MS. PESQUERO, M. A., PORTER, S. D., FOWLER, H. G., ANDCAMPIOLO, S. 1995. Rearing of Pseudacteon spp. (Diptera: Phoridae), parasitoids of fire ants ( Solenopsis spp.) (Hymenoptera: Formicidae). J. Appl. Entomol. 119: 677-678. PESQUERO, M. A., CAMPIOLO, S., FOWLER, H. G., ANDPORTER, S. D. 1996. Diurnal patterns of ovipositional activity in two Pseudacteon fly parasitoids of fire ants (Hymenoptera: Formicidae). Florida Entomol. 79: 455-456. PORTER, S. D. 1998a. Host specific attraction of Pseudacteon flies (Diptera: Phoridae) to fire ant colonies in Brazil. Florida Entomol. 81: 423-429. PORTER, S. D. 1998b. Biology and behavior of Pseudacteon decapitating flies (Diptera: Phoridae) that parasitize Solenopsis fire ants (Hymenoptera: Formicidae). Florida Entomol. 81: 292-309. PORTER, S. D. 2000. Host specificity and risk assessment of releasing the decapitating phorid fly Pseudacteon curvatus as a classical biocontrol agent for imported fire ants. Biol. Control. 19: 35-47. PORTER, S. D. 2010. Distribution of the Formosa strain of the fire ant decapitating fly Pseudacteon curvatus (Diptera: Phoridae) three and a half years after releases in North Florida. Florida Entomol. 93: 107112. PORTER, S. D., AND ALONSO, L. E. 1999. Host specificity of fire ant decapitating flies (Diptera: Phoridae) in laboratory oviposition tests. J. Econ. Entomol. 92: 110-114. PORTER, S. D., AND GILBERT, L. E. 2004. Assessing host specificity and field release potential of fire ant decapitating flies (Phoridae: Pseudacteon ), pp. 152-176 In R. G. Van Driesche and R. Reardon [eds.], Assessing Host Ranges for Parasitoids and Predators Used for Classical Biological Control: A Guide to Best

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Farnum & Loftin: Distribution of P. curvatus and P. tricuspis in Arkansas21Practice. FHTET-2004-03, USDA Forest Service, Morgantown, WV. PORTER, S. D., AND PESQUERO, M. A. 2001. Illustrated key to Pseudacteon decapitating flies (Diptera: Phoridae) that attack Solenopsis saevissima complex fire ants in South America. Florida Entomol. 84: 691-699. PORTER, S. D., PESQUERO, M. A., CAMPIOLO, S., ANDFOWLER, H. G. 1995a. Growth and development of Pseudacteon phorid fly maggots (Diptera: Phoridae) in the heads of Solenopsis fire ant workers (Hymenoptera: Formicidae). Environ. Entomol. 24: 475479. PORTER, S. D., VANDER MEER, R. K., PESQUERO, M. A., CAMPIOLO, S., AND FOWLER, H. G. 1995b. Solenopsis (Hymenoptera: Formicidae) fire ant reactions to attacks of Pseudacteon flies (Diptera: Phoridae) in southeastern Brazil. Ann. Entomol. Soc. America 88: 570-575. PORTER, S. D., NOGUEIRA DE S, L. A., FLANDERS, K.,AND THOMPSON, L. 1999. Field releases of the decapitating fly, Pseudacteon tricuspis p. 102 In L. Reeves [ed.], Proc. Imported Fire Ant Conference, Charleston, SC. PORTER, S. D., NOGUEIRA DE S, L. A., AND MORRISON, L. W. 2004. Establishment and dispersal of the fire ant decapitating fly Pseudacteon tricuspis in North Florida. Biol. Control. 29: 179-188. PUCKETT, R. T., CALIXTO, A., BARR, C. L., AND HARRIS, M. 2007. Sticky traps for monitoring Pseudacteon parasitoids of Solenopsis fire ants. Environ. Entomol. 36: 584-588. STREETT, D. A., FREELAND, T. B., AND VANDER MEER, R. K. 2006. Survey of imported fire ant (Hymenoptera: Formicidae) populations in Mississippi. Florida Entomol. 89: 91-92. THEAD, L. G., VOGT, J. T., AND STREETT, D. A. 2005. Dispersal of the fire ant decapitating fly, Pseudacteon curvatus (Diptera: Phoridae) in Northeast Mississippi. Florida Entomol. 88: 214-216. VANDER MEER, R. K. AND PORTER, S. D. 2002. Fire ant, Solenopsis invicta, worker alarm pheromones attract Pseudacteon phorid flies, pp. 77-80 In Proc. Imported Fire Ant Conference, Athens, GA. VAZQUEZ, R. J., PORTER, S. D, AND BRIANO, J. A. 2004. Host specificity of a biotype of the fire ant decapitating fly Pseudacteon curvatus (Diptera: Phoridae) from northern Argentina. Environ. Entomol. 33: 1436-1441. VAZQUEZ, R. J., PORTER, S. D., AND BRIANO, J. A. 2006. Field release and establishment of the decapitating fly Pseudacteon curvatus on red imported fire ants in Florida. Biocontrol. 51: 207-216. WEEKS, R. D., AND CALLCOTT, A.-M. A. 2008. USDAAPHIS and consortium efforts to establish phorid flies (Pseudacteon spp.) in imported fire ant ( Solenopsis spp.) populations in the U.S. and Puerto Rico, pp. 87-89 In Proc. Imported Fire Ant Conference, Charleston, SC. WILLIAMS, D. F., AND DESHAZO, R. D. 2004. Biological control of fire ants: an update on new techniques. Ann. Allergy Asthma Immunol. 93: 15-22. WUELLNER, C. T., DALLAGLIO-HOLVEROCEM, C. G., BENSON, W. W., AND GILBERT, L. E. 2002. Phorid fly (Diptera: Phoridae) oviposition behavior and fire ant (Hymenoptera: Formicidae) reaction to attack differs according to phorid species. Ann. Entomol. Soc. America 93: 257-266.

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22 Florida Entomologist 94(1) March 2011 MINI-ASPIRATOR: A NEW DEVICE FOR COLLECTION AND TRANSFER OF SMALL ARTHROPODS TO PLANTS M AHMUT D OGRAMACI 1* J IANJUN C HEN 2 S TEVEN P. A RTHURS 1 C INDY L. M C K ENZIE 3 F ABIELI I RIZARRY 1 K ATHERINE H OUBEN 1 M ARY B RENNAN 1 AND L ANCE O SBORNE 1 1 University of Florida, Department of Entomology and Nematology, Mid-Florida Research and Education Center, Apopka, FL 32703, USA 2 University of Florida, Department of Environmental Horticulture, Mid-Florida Research and Education Center, Apopka, FL 32703, USA 3 U.S. Horticultural Research Laboratory, ARS-USDA, Fort Pierce, FL 34945, USA A BSTRACT The process of collecting and/or infesting plants with a designated number of small arthropods in biological experiments is tedious and laborious. We developed a modied mini-aspirator, powered with a vacuum pump and tted with a specially adapted (removable) collection vial to reduce the handling effort. The efciency of the mini-aspirator was tested with the chilli thrips, Scirtothrips dorsalis Hood (Thysanoptera: Thripidae), a predatory mite Amblyseius (= Neoseiulus) cucumeris (Oudemans) (Acari: Phytoseiidae), and the insidious ower bug Orius insidiosus (Say) (Heteroptera: Anthocoridae). Using the mini-aspirator operators collected 10 A. cucumeris mites and 10 S. dorsalis thrips and transferred them onto pepper plants in 43 s and 37 s respectively, compared with 639 and 229 s, respectively, using a camels hair brush as a conventional method. The use of the mini-aspirator for collecting A. cucumeris predatory mites and S. dorsalis thrips and infesting pepper plants with them represents a 15-fold and 6-fold time sa ving, respectively. Collection of 10 O. insidiosus ower bugs took 20 s with the mini-aspirator compared with 30 s when an unmodied aspirator w as used. Proportionally, the amount of time saved with the mini-aspirator for the handling of O. insidiosus ower bugs was minimal compared with the timesavings when handling S. dorsalis thrips and the A. cucumeris predatory mites with the mini-aspirator. Additionally the mini-aspirator can be tted with a battery-powered Mini-Vac, which makes it portable for eld applications, such as in sampling eld populations when screening for pesticide resistant individuals. Key Words: mini-aspirator, Neoseiulus cucumeris Scirtothrips dorsalis Orius insidiosus manual infestation R ESUMEN El proceso de recolectar y/o infestar plantas con un cierto nmero de artrpodos pequeos en experimentos biolgicos es tedioso y laboroso. Desarrollamos una mini-aspiradora modicada, que funciona mediante una bomba de succin especialmente montada con un frasco de recoleccin especialmente adaptado (desmontable) para reducir el esfuerzo de manejo. La eciencia de la mini-aspiradora fue probada con el trips de chile, Scirtothrips dorsalis Hood (Thysanoptera: Thripidae), un caro depredador, Amblyseius (=Neoseiulus) cucumeris (Oudemans) (Acari: Phytoseiidae), y el chinche pirata diminuto, Orius insidiosus (Say) (Heteroptera: Anthocoridae). Usando la mini-aspiradora, los operadores recolectaron y transferieron 10 caros depredadores y trips de chile a plantas de chile en 43 y 37 segundos, comparado con 638 y 229 segundos usando el metodo convencional con una brocha de pintar. El uso de la mini-aspiradora para mover e infestar plantas con caros depredadores y trips representa un ahorro de 15 y 6 veces, respectivamente. La recoleccin de 10 chinches piratas diminutos tom 20 segundos con la mini-aspiradora comparada con 30 segundos cuando se uso una aspiradora no modicada. El tiempo ahorrado proporcionalmente fue mnimo comparado con la recoleccin de trips de chile y los caros depredadores con la mini-aspiradora. Adems, la mini-aspiradora puede ser mantada con una mini-bomba de succionar de batera, que la hace portable para aplicaciones en el campo, como en la evaluacin de resistencia de plaguicidas en poblaciones de campo. Manual collection and infestation of small arthropods (<2 mm) can be labor intensive and cumbersome and result in injury to the handled arthropods. Thrips and predatory mites are examples of small arthropods used by scientists in numerous experiments (Mound & Palmer 1981;

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Dogramaci et al.: Mini-aspirator for Handling Small Arthropods 23 Chiu et al. 1991; Tatara & Furuhushi 1992; Tschuchiya et al. 1995; Bournier 1999; Seal et al. 2006; Arthurs et al. 2009). Scirtothrips dorsalis Hood, chilli thrips, is one of the smallest thrips species with adults ranging from 1.5-2.0 mm. The S. dorsalis adult moves rapidly, and may jump or y when disturbed. Immature stages of S. dorsalis especially rst instars, are very small (<1 mm) and ha ve fragile easily injured bodies. Thus, an important requirement for studying small arthropods is to have a reliable method for collecting and releasing designated numbers of individuals without harm. Three methods are used for infesting plants with thrips. One is manual infestation of arthropods with a soft-bristled camels hair brush (Cloyd & Sadof 1998). This method has been used widely but is labor-intensive and time-consuming. For instance, Cloyd et al (2001) reported that infesting 50 plants eac h with 10 adult western ower thrips (WFT), Frankliniella occidentals (Pergande), required a technician 3.5 h. In addition, the process of mechanical transfer involved a risk of injury to the specimen. The second method involves placement of plants in a location where thrips are known to occur in order to allow a natural population of the pest to build-up on the test plants. Although this method is less cumbersome than manual infestation, the number of thrips transferred onto test plants cannot be accurately regulated, which introduces variation among test plants. The third method is the use of a commercial mouth operated aspirator purchased from BioQuip, Rancho Dominquez, CA. An improvement to the mouth-operated aspirator was reported by Cloyd et al. (2001), who developed a Small Insect Aspirator for collecting WFT. The latter involved the use of an aspirator from BioQuip attached to a battery operated Mini-Vac (MV Instrument, Glendale, CA 91205; http://www.minivac.com/index02.html). Use of the small insect aspirator is attended with operational difculties similar to those encountered with the use of regular aspirators, in particular the collection of extraneous materials. In our studies, we found that the wide suction tubes of the aspirators collect too many extraneous materials along with thrips (e.g., other organisms and plant debris) and sometimes causes physical damage to thrips. Additionally, in our experience the design of the collection vial does not allow the thrips to be released easily following collection. In order to overcome some of the design limitations of previous small arthropod handling devices, we developed a mini-aspirator that can be powered either with a laboratory vacuum pump or with a small portable vacuum pump and tted with a specially adapted and removable collection vial that allows rapid transfer of the collected arthropods onto plants. We compared the efciency of the mini-aspirator with a paintbrush and commercial aspirator for collecting and releasing the chilli thrips, Scirtothrips dorsalis Hood and 2 of its natural enemies a predatory mite, Neoseiulus cucumeris (Oudemans), and the ower bug, Orius insidiosus (Say). M ATERIALS AND M ETHODS Mini-aspirator The mini-aspirator was built from clear 6.35mm diam vinyl tubing tted with a 1-mL ltered pipette tip (VWR International, West Chester, PA). The intake tubing opening was reduced by using an adaptor to attach a 200-L pipette tip, which facilitated the collection of individual small arthropods (Figs. 1A and 1B). The modied mini-aspirator was powered by an electrical laboratory vacuum pump (Rocker vacuum pump, Rocker Scientic Co., Ltd., Kaohsiung, Taiwan) (Fig. 1A). To collect S. dorsalis an infested leaf w as placed under a stereomicroscope (Fig. 1C) and the desired number of thrips was captured in the collection vial for transfer onto plants. The collection vial was removed from the miniaspirator and attached to a plant with a hair clip to allow the voluntary dispersal of the thrips onto the plant (Fig. 1D). To make the mini-aspirator portable, we integrated the mini-aspirator with a Mini-Vac (MV Instrument, Glendale, CA 91205; http://www.mini-vac.com/index02.html) (Fig. 1E). However, to compensate for the reduced suction power of the Mini-Vac, the pipette tip lter was replaced with ne woven nylon fabric. The assembly of the pipette tip, collection vial, lter and vacuum tube is shown in Fig. 2. Such integration of the mini-aspirator with the battery powered Mini-Vac made the system portable for eld use. Arthropods Scirtothrips dorsalis specimens were obtained from a colony that originated from rose plants in W inter Park, FL. The colony was maintained on cotton plants, Gossypium hirsutum Deltapine 493 Conventional. The health of the colony was maintained by periodically introgressing thrips from naturally infested plants. Commercially available thrips predators (a predatory mite, N. cucumeris (Oudemans), and the insidious ower bug Orius insidiosus (Say)) were obtained from K oppert Biological Systems, Berkel en Rodenrijs, The Netherlands. Plant Material The infestation methods were tested on sweet pepper plants, Capsicum annum L. Pepper seeds were germinated on moist lter papers inside

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24 Florida Entomologist 94(1) March 2011 Petri dishes. Germinated seeds were transferred to seedling trays. Seedlings each with 4-6 fully expanded leaves were planted into 15 cm diam pots. Pepper plants at >10 leaf stage were used for the arthropod infestation experiments. Collection and Infestation of Small Arthropods with the Conventional Camels Hair Brush Collecting chilli thrips and infesting plants with them. Either 10 or 20 chilli thrips were capFig. 1. Novel minute arthropod infestation apparatus and its use. A. The complete system in use; B. The miniaspirator small arthropod collector; C. Collecting S. dorsalis from a leaf; D. Mini-aspirator along with collected arthropods attached to a plant to allow dispersal; E. The portable mini-aspirator consisting of clear 6.35 mm diam vinyl tubing tted with a 1-mL pipette tip and nylon cloth lter connected with an adaptor to a 200-L pipette tip; with suction provided by a Mini-Vac.

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Dogramaci et al.: Mini-aspirator for Handling Small Arthropods 25 tured with a moistened camels hair brush. Each of these thrips was then placed onto a pepper plant leaf. The number of seconds needed to capture and transfer the above designated number of thrips was recorded. This was repeated 8 times each for groups of 10 or 20 chilli thrips. Sorting and releasing N. cucumeris. The entire content of the pac kage containing N. cucumeris and substrate was emptied into a Petri dish (15 cm diam) lined with a lter paper To separate N. cucumeris predatory mites from the substrate, the c losed Petri dish was agitated gently several times. Under a stereomicroscope each predatory mite was collected individually from the lter paper and placed onto a pepper plant with a camels hair brush. Groups of either 10 or 20 N. cucumeris were placed on a plant. The number of seconds required to collect and release either 10 or 20 predatory mites w as recorded. This was repeated 8 times each for groups of 10 and 20 mites. Collecting and releasing O. insidiosus ower bugs on pepper plants Orius insidiosus ower bug adults in groups of either 10 or 20 were collected from a purc hased colony with a BioQuip aspirator. The contents of the package (vermiculite substrate and bugs) were emptied onto a board and bugs that crawled on the board were captured with the BioQuip aspirator. Each collection vial containing the O. insidiosus was placed at the base of a pepper plant that had been infested with thrips to allow the bugs to exit the vial and distribute onto the plant. The number of seconds required to collect and to release either 10 or 20 bugs was recorded. This was repeated 8 times each for groups of 10 or 20 O. insidiosus Use of the Mini-aspirator to Collect Small Arthropods and Transfer Them onto Plants Collecting S. dorsalis thrips and infesting plants with them Ten or 20 S. dorsalis were collected with the mini-aspirator as described above The mini-aspirator along with the collected S. dorsalis was attac hed to a pepper plant leaf with a hair clip in a manner that allowed thrips to distribute themselves on plant leaves (Fig. 1D). The time required for collection and release (the latter being the time required to attach the mini-aspirator along with the collected S. dorsalis to a pepper plant leaf) S. dor salis was recorded. This was repeated 15 times eac h for groups of 10 and 20 S. dorsalis Collecting N. cucumeris and releasing them onto plants The entire contents of the package with N. cucumeris predatory mites were emptied into a P etri dish (15 cm diam). To separate N. cucumeris mites from the packaging material, a lter paper w as placed in the Petri dish and the closed Petri dish was agitated gently several times. Under a stereomicroscope, either 10 or 20 N. cucumeris mites on the lter paper were collected with the mini-aspirator The collection tube with the N. cucumeris mites was attached to a pepper leaf as described above (F ig. 1D). The seconds required for collection and release (the latter being the time required to attach the collector containing the collected N. cucumeris to a pepper plant leaf were recorded. This was repeated 15 times each for groups of 10 and 20 N. cucumeris Collecting adult O. insidiosus ower bugs and releasing them onto plants. The mini-aspirator w as modied by enlarging the opening of the pipette tip to accommodate the bugs. The bugs were collected from among the vermiculate particles scattered on a board. Orius insidiosus in groups of either 10 or 20 were collected with the mini-aspirator The collection tube containing the bugs was then attached to a leaf with a hair clip as described previously. The number of seconds required to collect and release the bugs was recorded. This was repeated 15 times each for groups of 10 and 20 O. insidiosus Statistical Analysis The efciency (time) of the mini-aspirator was compared with the conventional method for collecting and releasing eac h of the three arthropods. The study was repeated 8 and 15 times for the conventional and new method, respectively. Data were analyzed by ANOVA procedure (PROC GLM) and means were separated by Fishers protected LSD test for all the experiments (SAS Institute 1997). R ESULTS The collection of 10 adult thrips from cotton leaves and their release onto a pepper plant using the mini-aspirator took 37 s compared with 229 s with the camels hair brush method. This represented a 6-fold reduction in infestation Fig. 2. Assembly of the pipette tip, collection vial, lter and vacuum line of the mini-aspirator.

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26 Florida Entomologist 94(1) March 2011 time when the mini-aspirator method was used (Fig. 3). The difference between the 2 methods was highly signicant ( F 1,21 = 325.66; P < 0.0001). The time required to place 20 thrips on a plant with the mini-aspirator w as 42 s, compared with 461 s by using the camels hair brush method, which represented an 11-fold reduction in infestation time when the mini-aspirator was used ( F 1,21 = 974.68; P < 0.0001) (Fig. 4). With the new method, the amount of time to collect and release per thrips decreased as number of increased. Such a reduction was not observed with the paintbrush method. Some thrips adults were also observed to be injured by the commercial aspirator (Fig. 5-A). We also observed some damaged thrips and reduced thrips activity when they were handled with a commercial aspirator (Fig. 5-B). The collection and release of 10 and 20 predatory mites required 639 and 1154 s with a moistened camels hair brush compared with 43 and 90 s, respectively, with the mini-aspirator (Figs. 3 and 4). The use of the mini-aspirator saved 15-fold and 13-fold more time in collecting and releasing 10 and 20 predatory mites, respectively, compared with the use of the paintbrush method. The difference between the two methods was highly signicant ( F 1,21 = 284.86; P = 0.0001 and F1,21 = 379.00; P = 0.0001), respectively. No predatory mite was found to be damaged by the mini-aspirator. The time required to collect and release adult O. insidiosus with a BioQuip aspirator was less than time needed for collecting and releasing by the paintbrush method. However, collecting and releasing 10 and 20 O. insidiosus with the commercial aspirator required 31 and 58 s, but only 20 and 32 s with the mini-aspirator (Figs. 3 and 4). These time differences between the 2 methods of collection and release were also signicantly different (F1,21 = 43.19; P = 0.0001) (F1,21 = 33.52; P = 0.0001), respectively. DISCUSSIONThe mini-aspirator reduces the time required to collect and release a designated number of small arthropods to the plants. The camels hair brush method in addition to being very slow can cause injury, especially to soft-bodied small arthropods. An operator using the mini-aspirator can collect and transfer small arthropods using controlled air intake velocity, which minimizes injury to the collected arthropods. To avoid injury to thrips, the vacuum was adjusted to the minimum sufcient to collect thrips. Although injury to thrips was not investigated in detail, thrips collected with the mini-aspirator were checked under a stereomicroscope and no serious thrips injury was observed. The mini-aspirator is different from the commonly available commercial aspirators, which employs larger diameter removable glass or plastic collecting vials. Initially, we used the commercial aspirators but experienced difculties in transferring the designated numbers of arthropods. The commercial aspirator available to us has a 4-mm diam collection tube that could not be adjusted for the selective collection of individual thrips; and this is a signicant limitation when working with mixed colonies of insects. Another advantage of the mini-aspirator is that unlike the traditional collection vials, the smaller removable and disposable collection tubes can easily be attached to small leaves or plant stems without disturbing the insects inside the tube. The mini-aspirator developed in this study can be powered with a Mini-Vac, which makes the technique portable for eld applications. The technique may be used to quickly census wild populations for laboratory testing or for use in insecticide efcacy trials. The mini-aspirator could also be adapted for quick pesticide resistance or efcacy trials in the eld (Rueda and Shelton 2003). The inside of the collection tube of the mini-aspirator could be treated with pesticides of interest or a treated leaf disk could be placed in the mini-aspirator before collecting small arthroFig. 3. Comparison of seconds needed for collectin g and infesting 10 O. insidiosus S. dorsalis and N. cucume ris onto plants with conventional methods (camels hair brush or commercial aspirator) and the developed mini-aspirator. Fig. 4. Comparison of seconds needed for collectin g and infesting 20 O. insidiosus S. dorsalis and N. cucume ris onto plants with conventional methods (camels hair brush or commercial aspirator) and the developed mini-aspirator.

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Dogramaci et al.: Mini-aspirator for Handling Small Arthropods27pods, and then collected arthropods would be held for a xed exposure period to quantify pesticide efcacy. This kind of monitoring would be helpful to conrm pest susceptibility to pesticides before their wide area applications. ACKNOWLEDGMENTSWe are grateful to Kenneth E. Savage, Russell D. Caldwell, and Younes Belmourd for help during this study. The study was supported by the USDA Tropical and Subtropical Agricultural Research (T-STAR) Program, American Floral Endowment and the USDA-ARS Floriculture and Nursery Research Initiative.REFERENCES CITEDARTHURS, S., MCKENZIE C. L., CHEN, J., DOGRAMACI, M., BRENNAN, M., HOUBEN, K., AND OSBORNE, L. 2009. Evaluation of Neoseiulus cucumeris and Amblyseius cucumeris (Acari: Phytoseiidae) as Biological Control Agents of Chilli Thrips, Scirtothrips dorsalis (Thysanoptera: Thripidae) on Pepper. Biol. Cont. 49: 91-96 BOURNIER, J. P. 1999. Two Thysanoptera, new cotton pests in Cote dIvorie. Annales de la Societe Entomologique de France 34: 275-281. CHIU, H. T., SHEN, S. M., AND WU, M. Y. 1991. Occurrence and damage of thrips in citrus orchards southern Taiwan. Chinese J. Entomol. 11: 310-316. CLOYD, R. A., WARNOCK, D. F., AND HOLMES, K. 2001. Technique for collecting thrips for use in insecticide efcacy trials. Hort. Sci. 36: 925-926. CLOYD, R. A., AND SADOF, C. S. 1998. Flower quality, ower numbers, and Western ower thrips density on transversal daisy treated with granular insecticides. Hort. Tech. 8: 567-570. MOUND, L. A., AND PALMER, J. M. 1981. Identication, distribution and host plants of the pest species of Scirtothrips (Thysanoptera: Thripidae). Bull. Entomol. Res. 71: 467-479. RUEDA, A., AND SHELTON, A. M. 2003. Development and evaluation of a thrips insecticide bioassay system for monitoring resistance in Thrips tabaci. Pest. Manage. Sci. 59: 553-558. SAS INSTITUTE. (1997) SAS Users Guide. SAS Institute Cary, North Carolina. SEAL, D. R., CIOMPERLIK, M. A., RICHARDS, M. L., ANDKLASSEN, W. 2006. Distribution of chilli thrips, Scirtothrips dorsalis (Thysanoptera: Thripidae), in pepper elds and pepper plants on St. Vincent. Florida Entomol. 89: 311-320. TATARA, A., AND FURUHASHI, K. 1992. Analytical study on damage to Satsuma mandarin fruit by Scirtothrips dorsalis Hood (Thysanoptera: Thripidae), with particular reference to pest density. Japanese J. Appl. Entomol. 36(4): 217-223. TSCHUCHIYA, M., MAUI, S., AND KUBOYAMA, N. 1995. Color attraction of yellow tea thrips ( Scirtothrips dorsalis Hood). Japanese J. App. Entomol. Zool. 39: 299-303. Fig. 5. Illustration of an injured S. dorsalis when collected with a regular made aspirator. A, Injury to abdomen of S. dorsalis B, Injury to wing of S. dorsalis

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28 Florida Entomologist 94(1) March 2011 TAXONOMY OF KOREAN LESTEVA WITH A DESCRIPTION OF A NEW SPECIES (COLEOPTERA: STAPHYLINIDAE: OMALIINAE) T AE -K YU K IM AND K EE -J EONG A HN Department of Biology, Chungnam National University, Daejeon 305-764, Republic of Korea A BSTRACT A taxonomic study of the genus Lesteva Latreille in Korea is presented. Four species including a new species Lesteva coreana sp. nov ., are recognized. Three species, L. cordicollis Motschulsky, L distincta Watanabe and L miyabi Watanabe, are new to the Korean fauna, and L. plagiata Sharp previously recorded from Korea is a misidentication of L miyabi A key and a comparison of morphological features of K orean Lesteva species with illustrations of the diagnostic features are provided. K ey Words: Staphylinidae, Omaliinae, Lesteva new species, Korea R ESUMEN Se presenta un estudio taxonmico del gnero Lesteva Latreille en Corea. Se reconocen cuatro especies inc luyendo una nueva especie, Lesteva coreana sp. nov Tres especies, L. cordicollis Motschulsky, L distincta Watanabe y L miyabi Watanabe son nuevas para la fauna de Corea y se determin que L. plagiata Sharp, anteriormente registrada en Corea, fue basado sobre una identicacin equivocada de L miyabi Se proveen una clave y una comparisin de las caractersticas morfolgicas de las especies de Lesteva en Corea con ilustraciones de las caractersticas diagnsticas. The genus Lesteva Latreille (tribe Anthophagini Thomson) is composed of 104 species distributed in the Holarctic and Oriental regions (Watanabe 1990, 2004, 2005; Herman 2001; Smetana 2004; Li 2005; Sharvrin et al. 2007). In East Asia, 19 and 15 species of the genus are reported in Japan and in China, respectively (Watanabe 1990, 2004; Smetana 2004; Li 2005). Lesteva plagiata Sharp recorded by Cho et al. (2002) in K orea is a misidentication of L. miyabi Watanabe. Members of Lesteva occur in montane riparian areas and are often found in moss or wet litter sometimes in caves. Adults and larvae are predators (Steel 1970; Newton et al. 2001). We have studied 20 specimens of L cordicollis Motschulsky 28 specimens of L. coreana sp. nov. 60 specimens of L distincta Watanabe and 125 specimens of L miyabi In this paper we report 4 Lesteva species ( L cordicollis L. coreana sp. nov. L distincta and L miyabi ) from Korea. A key, habitus photographs and the illustrations of diagnostic features are provided. All specimens are deposited in the Chungnam National University Insect Collection (CNUIC), Daejeon, Korea. Genus Lesteva Latreille, 1797 Lesteva Latreille, 1797: 75. Tevales Casey, 1894: 398. Synonymized by Steel, 1952: 9. Diagnosis. Body ovoid and attened, densely pubescent, covered with punctures. Head subquadrate; eyes convex, large, with pubescence between facets; ocelli distinct; temple round; vertex with 2 longitudinal depressions; gular sutures separated, divergent posteriorly; mandibles subtriangular, curved inwardly with distinct internal teeth, mola distinct; maxillary palpomere 4 as wide and about 4.0 times as long as palpomere 3; antenna extending to near middle of elytra. Pronotum convex, widest at anterior third or fourth, more narrowed posteriorly than anteriorly; mesoventrite with longitudinal carina along midline and several foveae on each side; elytra at, broader than pronotum, expanded posteriorly; legs long and slender, protarsus thin in both sexes. Abdomen broad, at and abruptly narrowed posteriorly, tergites IV-V with a pair of wing folding patches. Lesteva cordicollis Motschulsky, 1860 (F igs. 1, 5, 9, 13, 17, 19, 21-22) Lesteva cordicollis Motschulsky, 1860: 549; Sharvrin, 2001: 191. Description. Body (Fig. 1) length 3.6-4.0 mm (head to abdominal end), covered with ne punctures and pubescence, brown to dark brown and glossy; head and pronotum black, mouthparts, antennae and legs light brown. Head about 1.4 times as wide as long; eye about 3.3 times as long as temple; antennae (Fig. 5) pubescent, reaching middle of elytra, 4th antennomere 2.1 times as long as wide, 8th anten-

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Kim & Ahn: Korean Lesteva Species 29 nomere 1.8 times as long as wide. Pronotum slightly convexed with ne punctures, widest near anterior fourth with ambiguous U-depression near middle, 1.3 times as wide as long, about 1.2 times as wide and 1.4 times as long as head; scutellum (Fig. 9) subtriangular, prescutoscutellar suture gently curved, scutellar process broad subtriangular; elytra bicolor, humeral region with large yellow patch and ne punctures, posterior margin truncated, 1.1 times as wide as long, 1.5 times as wide and 1.7 times as long as pronotum (Figs. 1, 13); apex of metaventral process round (Fig. 17); external surface of metatibia with 3-4 long golden setae (Fig. 19). Abdominal segments III-VIII with microsculpture. Median lobe of aedeagus elongate, parallel-sided, apical process triangular, apical middle area elevated, internal sac backboneshaped; parameres slender, slightly longer than median lobe, four setae present with two at apex (Figs. 21-22). Materials Examined. KOREA: Gangwon Prov .: Chuncheon-si, Nam-myeon, Mt. Bonghw asan (N37.2 E 127.0 186m) 17 IX 2008, TK Kim ex under stone near stream (1 4 CNUIC); Chungnam Prov.: Daejeon, Yuseonggu, Sutong-gol, 9 V 1998, KR You, HJ Lim, HJ Kim, ex near stream (2 CNUIC); Jeonbuk Prov .: Muju-gun, Anseong-myeon, Mt. Deokyusan, Chilyeon-fall, 27 V 2005, TK Kim, ex under stone near stream (8 1 CNUIC); Jinan-gun, J eongcheon-myeon, Mt. Unjangsan, V 19 1998, YB Cho (2 CNUIC); Distribution. Korea (South), Russia (East Siberia). Lesteva coreana Kim and Ahn sp. nov. (Figs. 2, 6, 10, 14, 23-24) Description. Body (Fig. 2) length 3.1-3.5 mm (head to abdominal end), covered with ne punctures and pubescence, brown to dark brown and glossy; mouthparts, antennae and legs light brown. Head about 1.4 times as wide as long; eye about 3.1 times as long as temple; antennae (Fig. 6) pubescent, reaching middle of elytra, 4th antennomere 2.6 times as long as wide, 8th antennomere 2.4 times as long as wide. Pronotum slightly convexed with ne punctures, widest near anterior fourth with obscure U-depression near middle, 1.3 times as wide as long, about 1.2 times as wide and 1.3 times as long as head; scutellum (Fig. 10) subtriangular, prescutoscutellar suture arcuate, scutellar process narrow triangular; elytra bicolor, humeral region with indistinct yellow patch and ne punctures, posterior margin truncated, 1.1 times as wide as long, 1.4 times as wide and 1.7 times as long as pronotum (Figs. 2 and 14); apex of metaventral process round; external surface of metatibia with 3-4 long golden setae. Abdominal segments III-VIII with microsculpture. Median lobe of aedeagus narrowed apically, lateral margin weakly arcuated; basal region of parameres broad, narrowed apically, apical third constricted, slightly longer than median lobe, four setae present with two at apex (Figs. 2324). Type Series: Holotype, : KOREA: Jeonbuk Prov .: Muju-gun, Anseong-myeon, Mt. Deokyusan, Chilyeon-fall, 27 V 2005, TK Kim, ex under stone near stream; Holotype, Lesteva coreana Kim and Ahn, Desig. T.-K. Kim and K.-J. Ahn 2010. Deposited in CNUIC, Daejeon. Paratypes, same data as holotype (10 1 CNUIC), P aratype, Lesteva coreana Kim and Ahn, Desig. T .-K. Kim and K.-J. Ahn 2010. Other materials: Mt. Deokyusan, Chilyeon-fall, 22-23 V 1998, HJ Kim, ex near stream (1 CNUIC); same data as holotype (2 CNUIC); Chungnam Prov.: Daejeon, Mt. Gyeryongsan, Keumsubong, 21 V 2000, SJ Park, ex near stream (5 5 CNUIC); Yuseong-gu, Sutong-gol, 5 IX 1998, SJ Baek (1 CNUIC); Sutong-gol, 9 V 1998, KR You, HJ Lim, HJ Kim, ex near stream (1 CNUIC). Distribution. Korea (South). Remarks. The species is similar to L cordicollis but can be distinguished by the shape and structures of antennomeres scutellum, and median lobe of aedeagus (Table 1).Lesteva distincta Watanabe, 1990 (Figs. 3, 7, 11, 15, 18, 25-26) Lesteva distincta Watanabe, 1990: 178; Herman, 2001: 315; Smetana, 2004: 247 .Description. Body (Fig. 3) length 3.5-4.1 mm (head to abdominal end), covered with coarse punctures and pubescence, reddish brown to black and glossy; mouthparts, antennae and legs brown. Head about 1.2 times as wide as long; eye Figs. 1-4. Habitus. 1. Lesteva cordicollis length 3.8 mm; 2. L. coreana sp. nov., length 3.4 mm; 3. L distincta, length 4.0 mm; 4. L. miyabi length 3.9 mm.

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30 Florida Entomologist 94(1)March 2011 Figs. 5-16. 5-8. Antenna, ventral aspect. 5. Lesteva cordicollis ; 6. L. coreana sp. nov.; 7. L. distincta; 8. L. miyabi. 9-12. Scutellum, dorsal aspect. 9. L. cordicollis ; 10. L. coreana sp. nov. ; 11. L. distincta; 12. L. miyabi. 13-16. elytron, ventral aspect. 13. L. cordicollis ; 14. L. coreana sp. nov. ; 15. L. distincta; 16. L. miyabi Scales = 0.1 mm (Figs. 9-12); 0.3 mm (Figs. 5-8, 13-16).

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Kim & Ahn: Korean Lesteva Species31about 1.7 times as long as temple; antennae (Fig. 7) pubescent, reaching middle of elytra, 4th antennomere 2.1 times as long as wide, 8th antennomere 1.9 times as long as wide. Pronotum much convexed with coarse punctures, about 1.2 times as wide as long, 1.2 times as wide and 1.3 times as wide as head, widest near anterior fourth with distinct U-depression near middle; scutellum (Fig. 11) subtriangular, prescutoscutellar suture arcuate, scutellar process broad pentagonal; elytra bicolor, humeral region with reddish brown patch and somewhat coarse punctures, posterior margin round, 1.1 times as long as wide, about 1.8 times as wide and 1.9 times as long as pronotum (Figs. 3 and 15); apex of metaventral process notched (Fig. 18); external surface of metatibia with 10-14 long dark brownish setae. Abdominal segments III and VIII with microsculpture. Median lobe of aedeagus broad, basal two third parallel-sided, apical third narrowed suddenly, apical process triangular, midline area elevated with longitudinal carina, lateral margin rolled dorsally; parameres robust, symmetrical, as long as median lobe, lateral margin rolled ventrally, apical region coiling ventrally and inwardly, four setae present (Figs. 25 and 26). Materials Examined. KOREA: Gangwon Prov .: Chuncheon-si, Sabuk-myeon, Jiam-ri, 15 IV 2001, SI Lee (1 CNUIC); Chungbuk Prov.: Danyang-gun, Danyang-eup, Mt. Sobaeksan, Cheondong-area, 8-9 V 1999, US Hwang, HJ Kim, sifting (2 CNUIC); Yeongdong-gun, Sangchonmyeon, Mulhan-ri, Mt. Minjujisan, Mulhanstream (N36 E127), 16 VI 2006, TK Kim, ex under stone near stream (1 CNUIC); Mt. Manloi, 30 V 1998, HJ Lim, sifting (1 CNUIC); Chungnam Prov.: Daejeon-si, Yuseong-gu, Gung-dong, Chungnam National University (N36.7 E127.5), 18 IV 2007, HW Kim, ex near pond (3 2 CNUIC); Chungnam National University (N36.7 E127.5), 7 V 2007, YH Kim, ex near pond (7 4 CNUIC); Chungnam National University (N36.7 E127.5), 14 V 2007, HW Kim, ex near pond (12 8 CNUIC); Yuseong-gu, Deokmyeong-dong, Sutonggol, 9 V 1998, KR You, HJ Lim, HJ Kim, ex near stream (1 CNUIC); Buyeo-gun, Naesan-myeon, Mt. Wolmyeongsan, Geumgisa, 3 V-1 VI 2000, US Hwang, HJ Kim, FIT (1 CNUIC); Geumgisa, 1 VI 2000, US Hwang, HJ Kim, sifting (1 CNUIC); Jeonbuk Prov.: Buan-gun, Byeonsan-myeon, Mt. Naebyeonsan, Jikso-fall, 30 V 2001, YB Cho, sifting (1 2 CNUIC); Jinan-gun, Jeongcheon-myeon, Mt. Unjangsan, V 19 1998, YB Cho (2 CNUIC); Jeonnam Prov.: Gurye-gun, Mt. Jirisan, Tojimyeon, Piagol, 24 V 2000, HJ Kim, ex near stream (1 1 CNUIC); Piagol, 24-27 V 2000, KJ Ahn, SJ Park, US Hwang, FIT (1 CNUIC); Jindogun, Uisin-myeon, Sacheon-ri, Mt. Cheomchilsan (N34.7 E126.6 115m), 23 II 2007TABLE 1. A COMPARISON OF MORPHOLOGICAL FEATURES OF KOREAN LESTEVA SPECIES. L. cordicollisL. coreana sp. nov. L. distincta L. miyabi Ratio of eye to temple length 3.3 3.1 1.7 1.7 Antennomere 4 (length/width ratio) 2.1 2.6 2.1 1.9 Antennomere 8 (length/width ratio) 1.8 2.4 1.9 1.8 Scutellum Fig. 9 Fig. 10 Fig. 11 Fig. 12 Scutellar process broad triangular narrow triangular pentagonal pentagonal Metaventral process round (Fig. 17) round notched (Fig. 18) notched Long setae of metatibia 3-4 golden setae (Fig. 19)3-4 golden setae 10-14 dark setae 10-14 dark setae (Fig. 20) Abdominal microsculpture present on III-VIII present on III-VIIIpresent on III and VIIIpresent on III and VIII Aedeagus with carina absent absent present present Apical region of paramere straight (Figs. 21-22) straight (Figs. 23-24) coiling ventrally (Figs. 25-26) coiling ventrally (Figs. 27-28)

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32 Florida Entomologist 94(1)March 2011TK Kim, sifting, leaf litter (1 CNUIC); Yeonggwang-gun, Hongnong-eup, Sangha-ri (N35.9 E126.9), 2 V 2007, KJ Ahn, TK Kim, YH Kim, ex near stream (2 CNUIC); Hadong-gun, Hwagye-myeon, Ssanggyesa, 25 V 2000, HJ Kim, ex near stream (1 CNUIC); Gyeongbuk Prov : Cheongsong-gun, Budong-myeon, Mt. Juwangsan, 29 VI 1987, YB Cho, ex under moss (1 CNUIC); Gyeongnam Prov : Geoje-si, Yeoncho-myeon, Mt. Aengsan (N34.3 E128.6 85m), 21 I 2009, DH Lee, JH Song, ex under stone near mount stream (2 1 CNUIC). Distribution. Korea (South), Japan.Lesteva miyabi Watanabe, 1990 (Figs. 4, 8, 12, 16, 20, 27-28) Lesteva miyabi Watanabe, 1990: 175; Herman, 2001: 324; Smetana, 2004: 247. Lesteva plagiata : Cho et al., 2002: 36. Misidentication.Description. Body (Fig. 4) length 3.8-4.5 mm (head to abdominal end), covered with coarse punctures and pubescence, dark brown to black and glossy; mouthparts, antennae and legs brown to reddish brown. Head about 1.3 times as wide as long; eye about 1.7 times as long as temple; antennae (Fig. 8) pubescent, reaching middle of elytra, 4th antennomere 1.9 times as long as wide, 8th antennomere 1.8 times as long as wide. Pronotum mostly convexed with coarse punctures, widest near anterior third with distinct U-depression near middle, about 1.2 times as wide as long, about 1.1 times as wide and 1.2 times as long as head; scutellum (Fig. 12) subtriangular, prescutoscutellar suture round, scutellar process broad pentagonal; elytra unicolor with coarse punctures, posterior margin round, 1.04 times as long as wide, 1.8 times as wide and 2.0 times as long as pronotum (Figs. 4 and 16); apex of metaventral process notched; external surface of metatibia with 10-14 long dark brownish setae (Fig. 20). Abdominal segments III and VIII with microsculpture. Median lobe of aedeagus broad, narrowed apically with longitudinal carina, lateral margin almost straight; parameres robust, symmetrical, as long as median lobe, lateral margin rolled ventrally, apical region coiling ventrally and inwardly, four setae present (Figs. 27 and 28). Materials Examined. KOREA: Jeju Prov .: Jeju-si, Arail-dong, Gwaneumsa, 26 V 2003, SJ Park, ex near stream (10 9 CNUIC); Jeju-si, Bonggae-dong, Muljang-oreum, 23 V 1998, YB Cho (3 5 CNUIC); Jeju-si, Nohyeong-dong, Cheonwangsa (N33.4 E126.7 395 m), 8 XI 2006, TK Kim, ex under stone near stream (1 CNUIC); Jeju-si, Orai-dong, Eorimok (N33.0 E126.1 1000 m), 31 V 2007, TK Kim, ex under stone near stream (2 CNUIC); Seoguipo-si, Hawon-dong, Seoguipo Natural Recreation Forest (N33.2 Figs. 17-20. 17-18. Metaventrite, ventral aspect. 17. Lesteva cordicollis ; 18. L. distincta. 19-20. Metatibia (pubescence omitted), anterior aspect. 19. L. cordicollis ; 20. L. miyabi. Scales = 0.3 mm.

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Kim & Ahn: Korean Lesteva Species33E126.0 735 m), 30 V 2007, TK Kim, sifting, ood debris (1 CNUIC); Seoguipo Natural Recreation Forest (N33 E126.2 665 m), 31 V 2007, DH Lee, YH Kim, sifting, leaf litter (1 CNUIC); Bukjeju-gun, Aewol-eup, 1100-goji, 28 v 2003, CW Shin, ex near stream (2 CNUIC); 1100-goji (N33.6 E126.6 1097 m), 12 2006, TK Kim, sifting, wet leaf litter (1 2 CNUIC); 1100-goji (N33.5 E126.8 1110 m), 31 V 2007, TK Kim, sifting, leaf litter (9 6 CNUIC); Bukjeju-gun, Jocheon-eup, Goepyeongi-oreum, 23 V 2006, SJ Park, DH Lee, SI Lee, YH Kim, leaf litter (1 CNUIC); Goepyeongi-oreum (N33.7 E126.6 530 m), 8 IX 2006, DH Lee, ex leaf litter (1 5 CNUIC); Goepyeongi-oreum (N33 1.8 E126.2 539 m), 8 IX 2006, TK Kim, ex wet grit near pond (1 CNUIC); Namjeju-gun, Namwon-eup, Dongsubridge (N33.4 E126.7 640 m), 8 XI 2006, TK Kim, ex under stone near stream (3 CNUIC); Dongsu-bridge, 1 III 2007, TK Kim, ex under stone near stream (2 1 CNUIC); DongsuFigs. 21-28. Aedeagus. 21-22. Lesteva cordicollis 21. dorsal aspect; 22. lateral aspect. 23-24. L. coreana sp. nov. 23. dorsal aspect; 24. lateral aspect. 25-26. L. distincta. 25. dorsal aspect; 26. lateral aspect. 27-28. L. miyabi. 27. dorsal aspect; 28. lateral aspect. Scales = 0.3 mm.

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34 Florida Entomologist 94(1)March 2011bridge (N33.5 E126.5 635 m), 29 V 2007, TK Kim, ex under stone near stream (18 12 CNUIC); Namjeju-gun, Namwon-eup, Goepyeongi-oreum, 28 V 2003, SJ Park, CW Shin, MJ Jeon, sifting (16 11 CNUIC); Goepyeongioreum, 28 V-27 VI 2003, YB Cho, SJ Park, CW Shin, FIT (1 CNUIC); Mt. Hallasan, 900 m alt., Jejudo Is., 17 VII 1994, G. Sh. Lafer leg (1 CNUIC). Distribution. Korea (South), Japan. Remarks. Cho et al. (2002) reported L. plagiata in Korea. However, we have determined that this was a misidentication of L. miyabi based on our examination of their voucher specimen (1 : Mt. Hallasan, 900 m alt., Jejudo Is., 17. VII 1994, G. Sh. Lafer leg). The species was collected only in Jeju-do island. KEY TO THE KOREAN SPECIES OF THE GENUS LESTEVA LATREILLE1. Pronotum slightly convexed with ne punctures; prosternal process without carina; posterior margin of elytra truncated with ne punctures; apex of metaventral process round (Fig. 17); metatibia without long dark brownish setae (3-4 long golden setae present) (Fig. 19). . . . . . . . . . . . . . . . . . . . .2 Pronotum distinctly convexed with coarse punctures; prosternal process with short, sinuous longitudinal carina; posterior margin of elytra round with coarse punctures; apex of metaventral process notched (Fig. 18); metatibia with 10-14 long dark brownish setae (Fig. 20) . . . . . . . . . . . . . . . . . . . .3 2. Fourth antennomere 2.1 times as long as wide, 8th antennomere 1.8 times as long as wide (Fig. 5); scutellar process broad (Fig. 9); median lobe of aedeagus elongate, in basal three fourth parallel-sided, and in apical fourth abruptly narrowed in dorsal view (Figs. 21 and 22) . . . . . . . . . . . . . . L cordicollis Fourth antennomere 2.6 times as long as wide, 8th antennomere 2.4 times as long as wide (Fig. 6); scutellar process narrow (Fig. 10); median lobe of aedeagus narrowed apically, lateral margin weakly arcuated in dorsal view (Figs. 23 and 24) . . . . . . . . . . . . . . . . . . . . . . . . . . .L coreana sp. nov. 3. Pronotum widest at anterior fourth; elytra bicolor with reddish patch around humeral region, moderately broad and long (Fig. 3) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . L distincta Pronotum widest at anterior third; elytra unicolor, broad and long (Fig. 4). . . . . . . . . . . L. miyabiACKNOWLEDGMENTSWe thank Dr. Watanabe (Tokyo University of Agriculture, Japan) for the loan of specimens. This research was supported by the project on survey and excavation of Korean indigenous species of the National Institute of Biological Resources (NIBR) under the Ministry of Environment, Korea.REFERENCES CITEDCASEY, T. L. 1894. Coleopterological notices. V. Ann. New York Acad. Sci. 7: 281-606. CHO, Y. B., LAFER, G. S., PAIK, J. C., AND PARK, J. K. 2002. Contribution to the staphylinid fauna (Coleoptera, Staphylinidae) of Korea. Korean J. Soil Zool. 7(1-2): 35-44. HERMAN, L. H. 2001. Catalog of the Staphylinidae (Insecta: Coleoptera). 1758 to the End of the Second Millennium. I. Introduction, History, Biographical Sketches, and Omaliine Group. Bull. American Mus. Nat. Hist. 265: 309-333. LATREILLE, P. A. 1797. Prcis des Caractres Gnriques des Insectes, Disposs dans un Ordre Naturel. xiv + 201 + 7 pp. Brive: F. Bourdeaux. LI, X.-J., LI, L.-Z., AND ZHAO, M.-J. 2005. A new species of the genus Lesteva (Coleoptera: Staphylinidae: Omaliinae) from China. Entomotaxonomia 27(2): 111-113. MOTSCHULSKY, V. 1860. numration des nouvelles espces de coloptres rapportes de ses voyages. 3e partie. Bull. Soc. Imper. Nat. Moscou 33(2): 539-588. NEWTON, A. F., THAYER, M. K., ASHE, J. S., AND CHANDLER, D. S. 2001. 22. Staphylinidae, pp. 272-342 In R. H. Arnett and M. C. Thomas [eds.], American Beetles. Vol. 1. Archostemata, Myxophaga, Adephaga, Polyphaga: Staphyliniformia. CRC Press, Boca Raton, Florida. SHARP, D. 1889. The Staphylinidae of Japan. Ann. Mag. Nat. Hist. (6)3: 463-476. SHAVRIN, A. V. 2001. New and little-known species of Omaliinae from the Baikal-Transbaikal area (Coleoptera: Staphylinidae). Zoosyst. Rossica 9: 189-193. SHAVRIN, A. V., SHILENKOV, V. G., AND ANISTSCHENKO, A. V. 2007. Two new species and additional records of Lesteva Latreille, 1797 from the mountains of South Siberia (Coleoptera: Staphylinidae: Omaliinae: Anthophagini). Zootaxa 1427: 37-47. SMETANA, A. 2004. Staphylinidae, subfamily Omaliinae, pp. 237-268 In I. Lbl and A. Smetana [eds.], Catalogue of Palaearctic Coleoptera. Volume 2, Hydrophiloidea Histeroidea Staphylinoidea. Applo Books, Steustrup. STEEL, W. O. 1952. Notes on the Omaliinae (Col., Staphylinidae). 4. On the genera Lesteva Latr., Paralesteva Casey and Tevales Casey, with a key to the British species of Lesteva. Entomol. Mon. Mag. 88: 8-9. STEEL, W. O. 1970. The larvae of the genera of the Omaliinae (Coleoptera: Staphylinidae) with particular reference to the British fauna. Trans. R. Entomol. Soc. London 122(1): 1-47. WATANABE, Y. 1990. A taxonomic study on the subfamily Omaliinae from Japan (Coleoptera, Staphylinidae). Mem. Tokyo Univ. Agric. 31: 149-179. WATANABE, Y. 2004. Two new species of the genus Lesteva (Coleoptera, Staphylinidae) from the Island of Dgo of the Oki Islands, West Japan. Elytra 32(1): 71-77. WATANABE, Y. 2005. A new species of the genus Lesteva (Coleoptera, Staphylinidae) from Taiwan. Elytra 33(1): 30-33.

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Goyal et al: Corn-infesting Ulidiidae of Florida 35 DISTRIBUTION OF PICTURE-WINGED FLIES (DIPTERA: ULIDIIDAE) INFESTING CORN IN FLORIDA G AURAV G OYAL 1 G REGG S. N UESSLY 1 D AKSHINA R. S EAL 2 J OHN L. C APINERA 3 G ARY J. S TECK 4 AND K ENNETH J. B OOTE 5 1 Everglades Research and Education Center, University of Florida (UF), Institute of Food and Agricultural Sciences (IFAS), 3200 E. Palm Beach Rd., Belle Glade, FL 33430 2 Tropical Research and Education Center, UF, IFAS, 18905 S.W. 280 St., Homestead, FL 33031 3 Department of Entomology and Nematology, UF, IFAS, P.O. Box 110620, Gainesville, FL 32611 4 Division of Plant Industry, Florida Department of Agriculture and Consumer Services, P.O. Box 147100, Gainesville, FL 32614 5 Department of Agronomy, UF, IFAS, P.O. Box 110500, Gainesville, FL 32611 A BSTRACT The picture-winged y Euxesta stigmatias Loew (Diptera: Ulidiidae) has been a serious pest of sweet corn (Z ea mays L.) in Florida since 1930. Several other species in the family are known to infest corn grown in the Caribbean, Central America, and South America. Surveys were conducted throughout Florida to evaluate species richness and distribution of corn-infesting Ulidiidae. Adults were sampled with sweep nets and reared from y larvae-infested corn ears collected from representative corn elds in 16 and 27 counties in 2007 and 2008, respectively. Four Ulidiidae species were found in corn elds using both sampling techniques. Euxesta eluta Loew and Chaetopsis massyla (Walker) were found throughout the state on eld and sweet corn. Euxesta stigmatias was only found in Martin, Miami-Dade, Okeec hobee, Palm Beach, and St. Lucie Counties on eld and sweet corn. Euxesta annonae (F.) was found in sweet corn in Miami-Dade, Okeechobee, and Palm Beach Counties, but eld corn w as not sampled in these counties. Euxesta eluta E. stigmatias, and C. massyla were collected from corn throughout the corn reproductive stage Raising adults from y larvaeinfested ears provided the best method for assessing rates of ear infestation and species richness. Sweep netting did not provide reliable information on the presence or species composition of ulidiid species infestation. We report for the rst time E. annonae and E. eluta as pests of corn in Florida and the USA. K ey Words: Euxesta annonae, Euxesta eluta, Euxesta stigmatias, Chaetopsis massyla, maze R ESUMEN La mosca de alas pintadas, Euxesta stigmatias Loew (Diptera: Ulidiidae), ha sido una plaga seria de maz dulce (Z ea mays L.) en la Florida desde 1930. Varias especies de la familia Ulidiidae son conocidas de infestar maz sembrado en el Caribe y el Centroamrica y Sudamrica. Se realizaron sondeos por todo la Florida para evaluar la diversidad de especies y distribucin de moscas de la familia Ulidiidae que infestan maiz. Se muestrearon los adultos con redes de recoleccin y criandolos de mazorcas infestadas con larvas de moscas de campos representativos de maiz en 16 y 27 condados en 2007 y 2008, respectivamente. Se encontraron Euxesta eluta Loew y Chaetopsis massyla (Walker) por todo el estado en maz de campo y maz dulce Euxesta stigmatias fue encontrada solamente en los condados de Martin, Miami-Dade Okeechobee, Palm Beach y St. Lucie sobre maz de campo y maz dulce. Euxesta annonae (F.) fue encontrada en maz dulce en los condados de Miami-Dade, Okeec hobee y Palm Beach, pero no se muestrearon maz de campo en estos condados. Se recolectaron Euxesta eluta E. stigmatias y C. massyla durante toda la etapa reproductiva del maz. Euxesta annonae fue criada de mazorcas solamente de 8 a 21 dias de edad, pero los campos con mazorcas de 8-dias no fueron muestreados en los condados donde esta especie fue encontrada. El criar los adultos de mazorcas infestadas con larvas de moscas fue el mejor metodo para evaluar la taza de infestacion de las mazorcas y la diversidad de especies. Las recolecciones con redes no dieron un estimado conable para identicar infestaciones de especies de ulidiidos. Reportamos por primera vez E. annonae y E. eluta como plagas de maz en la Florida y EEUU.

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36 Florida Entomologist 94(1) March 2011 There are 671 species of Ulidiidae worldwide, but less than 10 species in 2 genera are known to damage corn (Allen & Foote 1992; Anonymous 2008c; Goyal et al. 2010; Van Zwaluwenburg 1917). Van Zwaluwenburg (1917) rst reported the pest nature of Euxesta stigmatias Loew (Diptera: Ulidiidae) (Figs. 1 g, 1h) in Puerto Rico where it damaged up to 100% of untreated corn. It was rst discovered damaging corn in Miami, Florida in 1938 (Barber 1939) and had moved north into central Florida by 1951 (Hayslip 1951). This species has become a serious pest of Florida sweet corn ( Zea mays L.) requiring multiple insecticide applications during the ear stage to maintain a marketable crop (Mossler 2008; Nuessly & Hentz 2004; Seal 1996, 2001; Seal & Jansson 1994). Sweet corn is an important crop in Florida with 22.8% of the total USA fresh market sweet corn production (Anonymous 2009). Euxesta stigmatias also has been reported infesting sweet corn in Georgia (Daly & Buntin 2005), T exas (Walter & Wene 1951), California (Fisher 1996), Guatemala (Painter 1955) and Brazil (Franca & Vecchia 1986). The insect deposits its eggs primarily on silks (styles) in the tips of ears. The larvae feed on silks, kernels, and cobs. Bailey (1940) observed disruption of pollination due to larval feeding on silks. Larvae enter through the soft pericarp of milk stage kernels to completely consume the developing embryo and endosperm (Seal & Jansson 1989). App (1938) observed larval feeding on cobs followed by mold development resulting in signicant reduction in market value Several other ulidiid species are known maize pests in the Caribbean and in the Americas south of Texas (Arce de Hamity 1986; Barbosa et al. 1986; Chittenden 1911; Diaz 1982; Evans & Zambrano 1991; Gossard 1919; Painter 1955; Wyckhuys & ONeil 2007), but only 1 other species is currently recognized as a pest in the USA. Chaetopsis massyla (Walker) (Figs. 1 a, b) was recently determined to be a primary pest of sweet corn in Florida (Goyal et al. 2010). Evidence suggesting the possibility of additional picture-winged species attacking corn in Florida include a picture of Euxesta eluta Loew (Diptera: Ulidiidae) (Figs. 1 e, f) on the cover of Ha yslips (1951) paper entitled Corn silk y control on sweet corn misidentied as E. stigmatias Examination of the Ulidiidae collection at the Division of Plant Industry in Gainesville Florida revealed that E. eluta and E. annonae (F.) (Diptera: Ulidiidae) (Figs. 1 c, d) have been collected in several Florida counties since at least 1948, but these specimens were not labeled as being collected or reared from corn. These later 2 species are recognized pests of corn in South America (Diaz 1982; Fras-L 1978). Therefore, it is possible that additional Ulidiidae species may be feeding on corn in Florida. The objective of this study was to evaluate species richness and distribution of corninfesting ulidiids throughout Florida. M ATERIALS AND M ETHODS Corn grown throughout Florida was sampled for ulidiid species. Extension personnel and researchers from all 67 Florida counties provided information on corn types and growing season needed to select representative elds. Corn elds were visited with the assistance of extension agents. One to 2 corn elds were sampled for Ulidiidae in each of 16 counties from Jul through Oct 2007 (Table 2). One to 4 corn elds were sampled for Ulidiidae in each of 27 counties during Feb through Jun 2008 (Table 3), including 10 counties visited in 2007. Adult ulidiids can be elusive and difcult to reliably observe and collect. They frequently avoid direct sunlight and walk or y away from the direct line of sight of workers approaching them. They are more easily collected from the tassels and upper leaves of corn plants in the hour just after sunrise and just before sunset, but it was not possible to sample all elds at these times. Adults can also be killed after ovipositing on a plant host before they are sampled, particularly within crops that are frequently treated with insecticides, such as sweet corn. Therefore, elds were sampled for both adults and immatures to determine whether the plants served as developmental hosts for ulidiid species and to determine the feasibility of using adult collection records for determining ear infestation. Preference was given to sampling corn that was between the silking and dough stages because both the adult and immature stages of ies can best be collected during the rst 3 weeks of corn reproduction. Neither adults nor immatures in ears were found in elds sampled before silking in Lake County (sweet corn) in 2007 and in Jefferson (eld corn) and Walton (sweet corn) Counties in 2008, therefore; data from these 3 elds were not included in the results. Sweet corn elds were preferred over eld corn for sampling because the ies cause less damage in eld corn than in sweet corn (Scully et al. 2000). Corn type (i.e., eld, sweet, Bt-enhanced, and standard corn) and variety, number of days before or after rst silk, and locations of the eld were recorded. Visual observations were taken for the presence of ulidiid adults. Flies were collected from corn elds with a sweep net (37.5 cm diameter). The sample size was adjusted depending on the estimated eld size In elds 4 ha, 3 pairs of corn rows were selected for sampling: 1 pair from each side of the eld and 1 pair in the middle of the eld. In elds >4 ha, 9 pairs of rows were selected for sampling: 1 on each side of the eld, 1 in the middle of the eld, and 6 pairs of rows randomly selected from between the eld margins. Sweep net sampling for ies was done while walking the length of the eld swinging the net 100 times between 2 rows in each pair of selected rows. Flies were preserved

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Goyal et al: Corn-infesting Ulidiidae of Florida 37 Fig. 1. Chaetopsis massyla male (a) and female (b); Euxesta annonae male (c) and female (d); E. eluta male (e) and female (f); E. stigmatias male; and (g) and female (h).

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38 Florida Entomologist 94(1) March 2011 in 70% ethyl alcohol for later identication and counting with a dissecting microscope. Identied Ulidiidae specimens housed at the Division of Plant Industry, Gainesville, FL and keys of Euxesta (Ahlmark & Steck unpublished, Curran 1928, 1934, 1935) and Chaetopsis (G. Steyskal unpublished) were used to conrm identications Corn ears were examined for the presence of y larvae in the same elds sampled with sweep nets. Ears found to contain larvae were collected and held for adult emergence to conrm species infestation. The number of ears sampled per eld was adjusted depending on the number of planted rows in each eld Fifty-six ears were examined in elds with < 90 rows and 88 ears were examined in elds with >90 rows. In a eld with <90 rows, 10 groups of 4 plants each were randomly selected for ear inspection. In a eld with 90 rows, ears were examined in every tenth row starting from the rst row and continuing to the other side of the eld (total of 10 rows). In a eld with >90 rows, 6 rows were sampled from each side of the eld (each sampled row separated by 10 rows), and 6 additional rows were randomly selected and sampled in the middle of the eld. One ear on each of 4 plants in the middle of each selected row was examined for y larvae (40 and 72 ears per eld for < 90 and >90 rows, respectively). An additional 4 plants in each corner of the eld were examined for larvae-infested ears (16 ears per eld). The top third of each infested ear was removed with a knife and placed individually in a Ziploc bag (1.83 L, S.C. Johnson & Son, Inc., Racine, WI). Two paper towels were added to each bag to reduce moisture accumulation. Bags were stored in portable coolers in the eld and during transportation back to the laboratory. Infested ears kept in the Ziploc bags were held in an air conditioned room maintained at 26.0 1C and L14:D10 h photoperiod to collect pupae for adult identication. To reduce the accumulation of moisture and associated fungus growth, bags with corn were left partially open, paper towels were changed frequently, and the air was constantly circulated by box fans. Corn ears collected on Mar 6, 2007 were placed collectively in 3.78 L Ziploc bags and then transferred to plastic containers with mesh tops. Pupae were removed from the bags and plastic containers, and placed on moistened lter paper (Whatman 3, Whatman International Ltd., Maidstone, England) in covered Petri dishes for adult emergence The dishes were sealed with Paralm (P echiney Plastic Packaging, Chicago, IL) to reduce moisture loss. Adults that emerged were preserved in 70% ethyl alcohol for later identication and counting as above. Statistical Analysis The results were tested by analysis of variance to examine the effects of sample technique, corn type (eld and sweet), corn ear age day and month of sampling (1-7, 8-14, 15-21 d) and sample year on the mean numbers of each species collected (Proc GLM, Version 9.0; SAS Institute 2008). Year was used as a random variable in the model. The mean number of ies sweep netted per pair of rows used in the data analysis was calculated for each eld by dividing the total number of ies caught in sweep nets by the number of pairs of rows sampled in that eld. The mean number of ies per infested ear was calculated for each eld by dividing the total number of ies reared from infested corn ears by the number of infested ears in each eld. Different numbers of ears and plant rows were sampled in each eld and more elds were sampled in 2008 than in 2007; therefore the results were presented as least square means rather than arithmetic means of ies caught per row and reared per corn ear. R ESULTS The mean number of ulidiid adults caught in sweep nets was signicantly affected by y species, corn type, survey year, and the species year interaction (T able 1). Signicantly more E. eluta T ABLE 1.A NALYSIS OF VARIANCE FOR FLIES CAPTURED BY SWEEP OR REARED FROM CORN EAR ON SPECIES CORN TYPE AGE OF CORN AND YEAR Sweep net Corn ears Source d fFPFP Species of Ulidiidae 3 8.12<0.0001 3.73 0.0120 Corn type 1 6.770.0099 6.53 0.0113 Age of corn 2 1.820.1638 1.09 0.3368 Year 133.17<0.000113.82 0.0003 Species corn type 3 0.680.5642 1.52 0.2091 Species age of corn 6 0.310.9304 0.67 0.6739 Species year 3 5.500.0011 2.00 0.1145 ANOVA (Proc GLM, SAS Institute 2008); denominator df = 236.

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Goyal et al: Corn-infesting Ulidiidae of Florida 39 (least squares mean SEM; 3.80 0.63) and C. massyla (3.62 0.63) were caught in sweep nets per row than E. stigmatias (1.33 0.63) and E. annonae (0.14 0.63). More adults were caught per row with sweep nets in sweet corn (2.95 0.34) than in eld corn (1.49 0.49). Sweep net counts per row were greater in 2007 (3.89 0.51) than in 2008 (0.55 0.32). The mean number of adults emerged per ear were signicantly affected by y species, corn type, and survey year (Table 1). Signicantly more E. eluta (1.41 0.30) were reared from each corn ear than E. stigmatias (0.40 0.30) and E. annonae (0.05 0.30). The mean number of C. massyla per ear (0.82 0.30) was not signicantly different than the other species More adults were reared from each corn ear in 2007 (1.19 0.25) than in 2008 (0.15 0.15). Signicantly more adults per ear were reared from sweet corn (1.02 0.16) than from eld corn (0.33 0.24). Results for species by county and reared from elds were presented separately for 2007 (Table 2) and 2008 (Table 3) due to signicant differences in mean counts between years. The correlation between adults caught in sweep nets and those reared from ears varied by species. Correlation coefcients were as follows: 0.79 ( P < 0.0001) for E. stigmatias 0.62 (P < 0.0001) for C. massyla, 0.58 for ( P < 0.0001) E. annonae and 0.51 (P < 0.51) for E. eluta.2007 Field SurveyFour Ulidiidae species were caught in sweep nets and reared from y larvae-infested ears in Florida corn during the rst survey year (Table 2). Chaetopsis massyla was collected in more counties throughout the state than other species and was netted in 100% of the sampled elds. This was followed by E. eluta, which was netted in 88% of sampled elds in all counties except Lake and Lee Counties (Table 2). Euxesta annonae and E. stigmatias were netted from only 3 counties in central and southern Florida, i.e., Miami-Dade, Okeechobee, and Palm Beach Counties. As a result of the more limited distribution, both E. annonae and E. stigmatias were netted in only 18% of elds sampled. The species netted and reared varied by corn type. Adults of E. annonae and E. stigmatias were netted only from sweet corn elds in Miami-Dade, Okeechobee, and Palm Beach Counties, but eld corn was not sampled in these counties (Table 2). Adults of E. eluta and C. massyla were netted from both eld and sweet corn elds throughout the state. Euxesta eluta and C. massyla were netted from 50 and 100% of eld corn elds, respectively, while both species were netted from 100% of sweet corn elds. The percentage of ulidiid-infested ears ranged from 5% in Escambia to 38% in Santa Rosa County (Table 2). Euxesta eluta and C. massyla were reared from ears collected from all but Lee, Lake and St. Johns Counties. These 2 species were reared from infested ears in 82% of corn elds statewide. Euxesta annonae and E. stigmatias were reared only from corn ears collected from Miami-Dade, Okeechobee, and Palm Beach Counties, amounting to only 18% of elds sampled. Adults of E. eluta and C. massyla were reared from both eld and sweet corn ears. Euxesta annonae and E. stigmatias emerged from sweet corn ears in elds from Miami-Dade, Okeechobee, and Palm Beach Counties, but eld corn elds were not sampled in these counties. Euxesta eluta and C. massyla were each reared from 50% of eld corn and 92% of sweet corn elds. Euxesta annonae and E. stigmatias were reared from 100% of the sweet corn elds in above mentioned Counties. The age of sampled corn in 2007 ranged from 4 to 21 d after rst silk (Table 2). Chaetopsis massyla was sweep netted in elds of all ages sampled. Euxesta eluta was sweep netted in elds 721 d after rst silk. Euxesta annonae and E. stigmatias were sweep netted from elds 8 to 21 d after rst silk, but no elds <8 d after rst silk were sampled in counties infested with these 2 species. Euxesta eluta and C. massyla emerged from corn ears collected from elds 4 to 21 d after rst silk, while E. annonae and E. stigmatias from ears collected 8 to 21 d after rst silk (Table 4). Corn ears were not collected from elds <8 d after rst silk in counties with E. annonae and E. stigmatias .2008 Field SurveyThe same 4 species were again collected in sweep nets and reared from y-larvae infested ears in Florida corn during the second study year (Table 3). Ulidiid adults were netted in 23 of 27 counties sampled in 2008. No adult picturewinged ies were captured in corn in Dixie, Jackson, Sumter, Taylor or Volusia Counties. Chaetopsis massyla was collected from more counties than other species throughout the state and was netted in 66% of the elds sampled. Euxesta eluta was netted in 49% of the elds sampled. Chaetopsis massyla was the only species collected from corn in Alachua, Jefferson and Marion Counties, while E. eluta was the lone species collected from corn in Okaloosa County. Euxesta stigmatias was netted only in Martin, Okeechobee, Palm Beach, and St. Lucie Counties amounting to only 11% of the elds sampled. Euxesta annonae was not collected in sweep samples in 2008. Euxesta eluta, E. stigmatias and C. massyla were netted from eld and sweet corn elds (Table 3). Euxesta eluta were netted from 27 and 59%, and C. massyla from 40 and 78% of the eld and sweet corn elds throughout the state, respectively. Euxesta stigmatias was caught from 100 and 67% of the eld and sweet corn elds, respectively, in Martin,

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40 Florida Entomologist 94(1)March 2011TABLE 2. ULIDIIDAE SPECIES COLLECTED IN FIELDS OR REARED FROM INFESTED EARS IN FLORIDA, 2007. County-eld no. Corn type1Sample date Ear age (d)2No. rows sampled with sweep net Mean no. adults captured per 100 sweeps3No. ears sampled (no. infested) Mean no. adults emerged per ear (per infested ear) E. annonaeE. elutaE. stigmatiasC. massyla E. annonaeE. elutaE. stigmatiasC. massyla AlachuaSwt16 Aug15-2130.01.80.0 4.256 (6)0.0 (0.0)0.9 (8.3)0.0 (0.0)1.4 (13.5) BradfordSwt17 Oct15-2190.00.90.0 2.988 (14)0.0 (0.0)0.7 (4.4)0.0 (0.0)1.6 (9.9) Miami-DadeSwt6 Mar15-21 32.826.011.211.756 (16)0.4 (1.3)17.5 (61.3)5.6 (19.4)5.9 (20.7) Escambia 1Fld2 Aug8-14 30.011.50.0 3.856 (5)0.0 (0.0)0.2 (2.2)0.0 (0.0)0.3 (3.2) Escambia 2Bt swt2 Aug7 90.05.80.0 6.956 (3)0.0 (0.0)0.3 (6.3)0.0 (0.0)0.6 (10.3) GadsdenSwt17 Sep7 30.06.30.0 4.856 (6)0.0 (0.0)1.1 (10.3)0.0 (0.0)1.0 (9.3) HolmesSwt16 Oct15-21 30.03.70.0 2.356 (11)0.0 (0.0)0.6 (3.2)0.0 (0.0)0.6 (2.8) Lake Fld14 Sep4-5 90.00.00.0 3.256 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Lee Fld17 Oct15-21 30.00.00.0 1.556 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) LibertyFld13 Sep15-2130.02.30.0 1.856 (4)0.0 (0.0)0.3 (4.3)0.0 (0.0)0.5 (7.3) Marion Swt4 Sep8-1430.016.50.926.056 (11)0.0 (0.0)4.2 (21.2)0.0 (0.0)7.1 (36.3) OkeechobeeSwt18 Sep8-1431.10.91.0 2.988 (6)0.1 (1.3)0.6 (8.3)0.3 (4.3)0.8 (12.3) Palm BeachSwt14 Nov8-14 31.721.333.518.356 (17)1.7 (5.5)5.8 (19.1)8.8 (29.1)1.8 (5.9) Santa Rosa Swt3 Aug8-14 30.03.70.0 8.356 (21)0.0 (0.0)4.5 (12.1)0.0 (0.0)1.1 (3.0) St. JohnsSwt13 Sep15-21 30.010.70.011.356 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Suwannee Swt13 Sep15-21 30.011.50.0 2.756 (12)0.0 (0.0)2.8 (13.2)0.0 (0.0)4.6 (21.3) WashingtonSwt2 Aug15-21 30.04.30.0 3.388 (16)0.0 (0.0)11.3 (62.3)0.0 (0.0)3 (16.3)1Corn type: Fld = field corn; Swt = sweet corn; Bt swt = Bacillus thuringiensisenhanced sweet corn.2Estimated days after first silk at time of sampling.30 = no flies were detected in sweep nets.

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Goyal et al: Corn-infesting Ulidiidae of Florida 41TABLE 3. ULIDIIDAE SPECIES COLLECTED IN FIELDS OR REARED FROM INFESTED EARS IN FLORIDA, 2008. County-eld no. Corn type1Sample date Ear age (d)2No. rows sampled with sweep net Mean no. adults captured per 100 sweeps3No. ears sampled (no. infested) Mean no. adults emerged per ear (per infested ear) E. annonaeE. elutaE. stigmatiasC. massyla E. annonaeE. elutaE. stigmatiasC. massyla AlachuaSwt4 Jun730.00.00.0 1.356 (8)0.0 (0.0)0.2 (1.4)0.0 (0.0)0.0 (0.0) Bradford 1Swt23 Jun18-21 30.02.30.0 8.756 (15)0.0 (0.0)3.3 (12.3)0.0 (0.0)1.1 (4.3) Bradford 2Swt23 Jun10-1430.03.30.0 5.756 (17)0.0 (0.0)10.2 (33.5)0.0 (0.0)0.4 (1.5) ColumbiaSwt24 Jun8-1490.01.10.0 2.288 (2)0.0 (0.0)0.1 (2.5)0.0 (0.0)0.05 (2.0) Dixie Fld4 Jun2-390.00.00.0 0.088 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Gilchrest 1Swt23 Jun15-21 90.00.70.0 1.388 (6)0.0 (0.0)0.6 (9.3)0.0 (0.0)0.8 (11.3) Gilchrest 2Bt d23 Jun790.00.00.0 2.188 (4)0.0 (0.0)0.1 (2.3)0.0 (0.0)0.2 (5.3) Gilchrest 3Bt d23 Jun790.01.80.0 0.888 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Hamilton 1Swt24 Jun730.01.30.0 1.356 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Hamilton 1Fld24 Jun14 30.00.00.0 0.056 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) HendrySwt26 Feb15-2190.04.60.0 1.488 (18)0.0 (0.0)2.7 (13.3)0.0 (0.0)4.4 (21.4) Holmes 1Swt26 Jun2130.08.30.0 0.756 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Holmes 2Fld26 Jun2190.02.10.0 2.988 (9)0.0 (0.0)0.4 (3.4)0.0 (0.0)0.1 (1.4) Jackson 1Bt d5 Jun2-390.00.00.0 0.056 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Jackson 2Bt d5 Jun2-390.00.00.0 0.056 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Jefferson -1Fld25 Jun8-1490.00.00.0 0.088 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Jefferson 1Swt25 Jun1430.00.00.0 0.756 (10)0.0 (0.0)0.4 (2.4)0.0 (0.0)0.0 (0.0) Lafayette 1Swt24 Jun1430.01.00.0 1.356 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Lafayette 2Swt24 Jun1430.00.00.0 1.756 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Lafayette 3Fld24 Jun1490.00.00.0 0.088 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Lake 1Swt6 Jun2190.00.00.0 5.256 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Lake 2Swt6 Jun2190.02.00.0 1.856 (22)0.0 (0.0)1.7 (4.4)0.0 (0.0)4.1 (10.4) MarionBt d3 Jun1430.00.00.0 *56 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Martin 1Swt11Mar730.03.01.0 1.356 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Martin 2Swt11 Mar1-230.00.00.0 0.056 (9)0.0 (0.0)0.4 (2.3)3.1 (19.0)0.5 (3.3) Martin 3Swt11 Mar1-290.00.00.0 0.988 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Martin 4Swt11 Mar1430.00.014.3 4.756 (8)0.0 (0.0)0.0 (0.0)0.5 (3.4)0.0 (0.0) Nassau 1Swt23 Jun15-2130.0*0.0 *56 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Nassau 2Fld23 Jun15-2190.00.00.0 0.088 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Okaloosa 1Swt5 Jun530.02.00.0 0.056 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Okaloosa 2Swt5 Jun1030.00.00.0 0.056 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0)1Corn type: Fld = field corn; Swt = sweet corn; Bt swt = Bacillus thuringiensisenhanced sweet corn.2Estimated days after first silk at time of sampling.30 = no flies were detected in sweep nets; = fly species was observed only, not collected.

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42 Florida Entomologist 94(1)March 2011OkeechobeeSwt19 Apr1490.03.67.2 1.988 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Palm Beach Swt10 Apr15-2130.011.320.712.756 (15)0.9 (3.2)3.3 (12.3)1.1 (4.3)2.5 (9.2) Santa Rosa 1Bt swt5 Jun1030.00.00.0 1.756 (16)0.0 (0.0)16.1 (56.4)0.0 (0.0)0.4 (1.4) Santa Rosa 2Swt5 Jun1430.0*0.0 *56 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) Santa Rosa 3Bt swt5 Jun15-2130.02.30.0 *56 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) St. Johns 1Bt swt6 Jun8-1430.06.30.0 *56 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) St. Johns 2Bt swt6 Jun1430.00.00.0 0.056 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) St. LucieBt d29 May1490.00.04.1 5.756 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) SumterSwt6 Jun15-2130.00.00.0 0.056 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) SuwanneeFld4 Jun15-2130.01.00.0 1.056 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) TaylorFld25 Jun15-2130.00.00.0 0.056 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) VolusiaSwt6 Jun2130.00.00.0 0.056 (1)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.02 (1.0) WalkullaSwt25 Jun1430.02.00.0 *56 (6)0.0 (0.0)0.3 (3.2)0.0 (0.0)0.3 (2.3) WaltonFld26 Jun1490.01.80.0 0.088 (4)0.0 (0.0)0.1 (2.3)0.0 (0.0)0.0 (0.0) WaltonSwt26 Jun15-2130.0*0.0 2.056 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) WaltonSwt26 Jun15-2190.00.00.0 0.088 (0)0.0 (0.0)0.0 (0.0)0.0 (0.0)0.0 (0.0) TABLE 3. (CONTINUED) ULIDIIDAE SPECIES COLLECTED IN FIELDS OR REARED FROM INFESTED EARS IN FLORIDA, 2008. County-eld no. Corn type1Sample date Ear age (d)2No. rows sampled with sweep net Mean no. adults captured per 100 sweeps3No. ears sampled (no. infested) Mean no. adults emerged per ear (per infested ear) E. annonaeE. elutaE. stigmatiasC. massyla E. annonaeE. elutaE. stigmatiasC. massyla1Corn type: Fld = field corn; Swt = sweet corn; Bt swt = Bacillus thuringiensisenhanced sweet corn.2Estimated days after first silk at time of sampling.30 = no flies were detected in sweep nets; = fly species was observed only, not collected.

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Goyal et al: Corn-infesting Ulidiidae of Florida 43Okeechobee, Palm Beach, and St. Lucie Counties. No E. annonae adults were netted in eld or sweet corn elds. Ulidiid-infested ears were found in 13 of 27 counties sampled (Table 3). The percentage of ulidiid infested ears ranged from 2% in Volusia to 39% in Lake County. Only E. eluta were reared from corn ears collected from Alachua, Jefferson, and Walton Counties. Chaetopsis massyla was the only species reared from corn ears collected from Volusia County. Euxesta eluta and C. massyla were reared from 32% and 28% of the corn elds sampled throughout the state. Euxesta eluta were reared from 20 and 38% and C. massyla from 13 and 34% of the eld and sweet corn elds, respectively throughout the state. Euxesta stigmatias was only reared from infested sweet corn ears in Martin and Palm Beach Counties amounting to 6% of the total elds sampled. Adults of E. annonae were only reared from infested sweet corn ears collected from Palm Beach County amounting to approximately 2% of the total elds sampled. Field corn elds were not sampled in the counties where E. stigmatias and E. annonae were reared from ears. The age of corn ears in surveyed elds ranged from 1 to 21 d after rst silk (Table 3). More E. eluta and C. massyla were caught in sweep nets and reared from corn ears 15 to 21 d post-silking compared to 0 to 14 d post-silking. More E. stigmatias were caught in sweep nets and reared from corn ears in elds with 15 to 21 d post-silking ears than in younger elds. In counties where E. annonae was found, it was only reared from elds sampled 15 to 21 d after rst silk. DISCUSSIONThe results of this 2-year study conrmed that several species of Ulidiidae ies were infesting corn in Florida. Ulidiidae ies were found infesting both sweet and eld corn elds across the Florida panhandle from Escambia to Nassau Counties and south through the peninsula to Miami-Dade County. Flies were collected in sweep nets or reared from corn ears from 29 out of 33 sampled counties during the 2 survey years (Fig. 2). Flies were more common in the 2007 compared to 2008 surveys probably due to differences in sampling times. Corn elds in 2007 were largely sampled from Aug to Oct, except for Miami-Dade County that was sampled in Mar. In contrast, surveys were conducted from Feb to Jun in 2008. The ies may be more common in midsummer through fall months in northern Florida. While more research has been conducted on E stigmatias than the other species, it was found to be much less common than C. massyla and E. eluta in this survey. Euxesta eluta and C. massyla were distributed in most elds sampled throughout the state in both years, while E. stigmatias and E. annonae were found in only several counties of southern Florida (Martin, Miami-Dade, Okeechobee, Palm Beach, and St. Lucie). The distribution of alternate host plants and differences in acceptable temperature ranges for each species may explain some of the variation present in the distribution of ulidiids infesting corn in Florida. Euxesta eluta, E. stigmatias and C. massyla were collected from both eld and sweet corn, while E. annonae was collected only from sweet corn elds. Sweet corn is mostly grown in southern Florida in comparison to northern Florida where eld corn predominates (Anonymous 2008a). However, E. stigmatias was not collected or reared from sweet corn elds in northern Florida. Fras-L (1978) in Chile found that higher temperature and lower relative humidity led to greater numbers of E. annonae while the reverse led to greater numbers of E. eluta. Sampling with both sweep nets and collecting infested corn ears gave a more complete picture ofTABLE 4. PERCENTAGE OF FIELDS WITH ULIDIIDAE SPECIES SWEEP NETTED OR REARED FROM CORN EARS BY EAR AGESpecies Sweep netted Ear age (d) Reared from corn ears Ear age (d) 0 to 7 d 8 to 14 d 15 to 21 d 0 to 7 d 8 to 14 d 15 to 21 d 2007 E. annonae 0 100 100 0 100 100 E. eluta 67 100 89 67 100 78 E. stigmatias 0 100 100 0 100 100 C. massyla 100 100 100 67 100 78 2008 E. annonae 000001 0 0 E. eluta 36 42 64 27 32 35 E. stigmatias 33 100 100 33 33 100 C. massyla 54 68 71 18 21 41

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44 Florida Entomologist 94(1)March 2011y distribution in Florida corn elds than either sampling technique alone Low correlation values indicate that sweep netting is not an efcient method to estimate ulidiid species infesting corn ears. The relationship between sweep nets and y species that emerged from infested ears accounted for >60% of the variation for E. stigmatias and C. massyla, but <60% for E. eluta and E. annonae There were also a few locations where ies were observed but not collected with sweep nets. These were the places where ies were uncommon (1 or 2 per site) and netting was not the best sampling technique for insects at low densities. Seal et al. (1996) found that E. stigmatias congregated on the top of plants late in the evening. Fly species in our study may have been more active or more accessible with nets at times of the day other than when sampling was conducted. Therefore, sweep netting can be used to indicate the potential for ear infestation, but the identication of adults reared from infested ears is currently the only method available for differentiating the species developing within ears. The external physical characteristics of the immature stages of Ulidiidae infesting Florida corn are currently being examined by the authors to determine the possibility of using eggs, larvae or pupae for the identication of species of ies infesting corn. Euxesta eluta, E. stigmatias and C. massyla were collected from corn throughout the reproductive stage of corn. Adult E. annonae may be present in fields during the first week of silking, but only fields 8 d after first silk were sampled in counties where this species was found. In general, there was a tendency for greater infestation by all 4 species as sweet corn ears neared harvest and as field corn ears approached the dough stage. The authors also have frequently reared E. eluta, E. stigmatias, and C. massyla from tassels and stems of corn plants. Therefore, the potential host period on this crop is longer than just the reproductive stage. Fig. 2. Distribution of Ulidiidae species infesting corn in Florida by county during the 2007-2008 surveys. Symbols in gure represent species collected using sweep nets and reared from corn ears in each of the sampled (shaded) counties.

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Goyal et al: Corn-infesting Ulidiidae of Florida 45This is the rst report of E. annonae infesting corn in Florida and the USA. This species was not common in any location but was always netted from elds and reared from ears along with other Ulidiidae species. Euxesta annonae was the least collected species in sweep nets and it was reared from corn ears collected only from the southern end of the Florida peninsula (Fig. 2). Euxesta annonae is also reported as a pest of corn in Chile (Fras-L 1978). The authors have frequently observed E. annonae on Annona spp. (Magnoliales: Annonaceae) and Chinese long bean, Vigna unguiculata ssp sesquipedalis (L.) Verdc. (Fabales: Fabaceae) in southern Florida and reared E. annonae adults from eld collected Annona spp. fruit (Magnoliales: Annonaceae). Plants of Annona spp. are recorded in several southern and central Florida counties (Brevard, Broward, Collier, De Soto, Glades, Hendry, Highlands, Indian River, Lee, Manatee, Martin, Miami-Dade, Monroe, Palm Beach, and St. Lucie) (Wunderlin & Hansen 2008) where they may provide alternative food resources for this species. The authors have reared this species from decaying corn stalks and from spiny amaranth, Amaranthus spinosus L. (Caryophyllales: Amaranthaceae) roots collected from the eld at Belle Glade, Florida. Euxesta eluta was widely collected in this study from fields sampled throughout Florida (Fig. 2). These flies were commonly observed in fields and as many as 62 were reared from an individual ear. While this is the first known record of E. eluta being a pest of corn in Florida and the USA, its image in Hayslip (1951) suggests that it was present in Florida corn fields >50 yr ago, but incorrectly identified as E. stigmatias. The wide distribution of E. eluta in Florida and its discovery on both sweet and field corn indicates this fly is a much greater threat to corn than E. stigmatias, which is found in a much smaller portion of Florida. Euxesta eluta was recognized as infesting corn in Puerto Rico >60 yr ago (Wolcott 1948) and has been recorded as an ear pest in Ecuador (Evans & Zambrano 1991), Chile (Fras-L 1978; Olalquiaga 1980), Peru (Diaz 1982), Argentina (Arce de Hamity 1986), and Brazil (Franca & Vecchia 1986). Euxesta eluta is a pest of loquat, Eriobotrya japonica (Thumb.) Lindl. (Rosales: Rosaceae) in Alachua County, Florida (Anonymous 2008b). Loquat is grown as a dooryard plant and is distributed in several counties throughout the state (Wunderlin & Hansen 2008). Euxesta stigmatias was found in sweep net collections and reared from corn ears from southern and central Florida counties only (Fig. 2). Weather differences in southern and northern Florida may explain part of the variation in distribution of the species. Adult E. stigmatias have been reared from damaged or decayed inflorescences of sorghum, Sorghum bicolor (L.) Moench (Cyperales: Poaceae), tomato fruit, Lycopersicon esculentum L. (Solanales: Solanaceae) (Seal & Jansson 1989), and decaying carrot roots, Daucus carota L. (Apiales: Apiaceae) (Franca & Vecchia 1986). Chaetopsis massyla was caught in sweep nets and reared from corn ears in the majority of surveyed counties (Fig. 2). This fly was common in field and sweet corn fields throughout the year in southern Florida counties. The relative abundance and development range across corn types indicates this species is a much greater threat to Florida corn than previously recognized. Its habit of feeding on a range of monocots may help explain its widespread distribution throughout Florida. Allen & Foote (1992) reported it to be a secondary invader of wetland monocots. Chaetopsis massyla has been reared from cattail, Typha spp. (Typhales: Typhaceae) in California (Keiper et al. 2000). Typha spp. are found in most Florida counties except Flagler, Gadsden, Glades, Hernando, and Suwannee (Wunderlin & Hansen 2008). The authors made several personal observations of C. massyla plant associations during the course of this statewide survey. Chaetopsis massyla was frequently observed by the authors on cattail plants on ditch banks and feeding on sugary exudates from sugarcane plants (a complex hybrid of Saccharum spp.) in Belle Glade (Palm Beach County). Chaetopsis massyla adults were reared from sugarcane stems that were actively infested with the sugarcane borer, Diatraea saccharalis (F). (Lepidoptera: Crambidae) collected by the authors in November 2009 from sugarcane fields at Clewiston (Hendry County) and Sebring (Highlands County), Florida. Chaetopsis massyla was also successfully reared by the authors from otherwise healthy sugarcane stems exposed to colonies in the laboratory in which 0.5 cm diam holes were drilled in billets to simulate emergence and frass evacuation holes produced by D. saccharalis Other plants from which C. massyla has been reared include hairy sedge, Carex lacustris Willd. (Cyperales: Cyperaceae) (Allen & Foote 1992), Narcissus spp. (Liliales: Liliaceae) (Blanton 1938) and onions, Allium cepa L. (Asparagales: Alliaceae) (Merrill 1951). Carex spp. are found throughout the state while the distribution of Narcissus spp. is considered to be limited to Alachua, Calhoun, Escambia and Leon Counties (Wunderlin & Hansen 2008). Two additional Chaetopsis spp have been reported feeding on corn, but neither was found in this 2-year survey of Florida corn elds. Large populations of y larvae that were discovered in corn stalks within tunnels likely produced by European corn borer, Ostrinia nubilalis Hbner (Lepidoptera: Pyralidae) in Ohio were reared to adults and identied as Chaetopsis aenea (Wiedemann) by Gossard (1919). Larvae of C. fulvifrons (Macquart) were reared from within the tunnels

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46 Florida Entomologist 94(1)March 2011of southwestern corn borer, Diatraea grandiosella Dyar (Lepidoptera: Crambidae), in the Texas high plains (Knutson 1987). Langille (1975) reported that Chaetopsis spp. larvae were commonly associated with diapausing D. grandiosella within corn stalks in Missouri and hypothesized that ulidiid larvae feed on the decaying stalks or microbial growth within the bored stalks. In conclusion, 4 species of picture-winged ies were found infesting corn in Florida. Evidence presented herein is the rst known documentation for E. annonae and E. eluta as pests of corn in Florida and the USA. The 4 species were not uniformly distributed throughout Florida corn growing regions. Euxesta eluta and C. massyla were found infesting eld and sweet corn throughout Florida. Euxesta stigmatias was only found infesting corn in Martin, Miami-Dade, Okeechobee, Palm Beach, and St. Lucie Counties. Euxesta annonae (F.) was found in sweet corn in MiamiDade, Okeechobee, Palm Beach Counties, but eld corn was not sampled in these counties. Euxesta eluta, E. stigmatias and C. massyla were collected from corn throughout the corn reproductive stage. Euxesta annonae was reared from 8-21 d old ears only, but elds with ears <8 d old were not sampled in the counties where this species was found. The relative abundance of E. eluta and C. massyla in Florida eld and sweet corn indicates the need for more research into their biology and ecology. The discovery of E. eluta and C. massyla attacking corn ears in many of the northernmost Florida counties suggests that further surveys of corn growing areas across the borders into neighboring states is warranted to determine the extent of corn infesting picture-winged y infestations in the southern U.S. The statewide distribution of E. eluta and C. massyla in reproducing corn also suggests that studies should be conducted to evaluate additional food sources that support these species in the absence of corn. ACKNOWLEDGEMENTSWe thank Harsimran K. Gill and Bijayita Thapa for assistance in rearing Ulidiidae from collected ears, and Nicholas A. Larsen for help contacting extension agents and eld navigation. We acknowledge Jaya Das for help with the Florida county maps. The photographic assistance of Lyle Buss was instrumental in producing the y images used in this report. We are also thankful to University of Florida Cooperative Extension Agents and researchers for their help in selecting corn elds and arranging the visits, and corn growers of Florida for allowing us to survey their elds. This research was made possible by a Hand Fellowship awarded by the Dolly and Homer Hand Group.REFERENCES CITEDALLEN, E. J., AND FOOTE, B. A. 1992. Biology and immature stages of Chaetopsis massyla (Diptera: Otitidae), a secondary invader of herbaceous stems of wetland monocots. Proc. Entomol. Soc. Washington 94: 320-328. ANONYMOUS. 2008a. Florida Statistical Abstract. University of Florida, Gainesville, FL. ANONYMOUS. 2008b. Florida Cooperative Agricultural Pest Survey Program. Quarterly Report No. 2-2008. Division of Plant Industry, Florida Department of Agriculture and Consumer Services, Gainesville, FL. ANONYMOUS. 2008c. Biosystematic Database of World Diptera. Available at http://www.sel.barc.usda.gov/ diptera/names/Status/bdwdstat.htm (veried 10 Nov 2009). ANONYMOUS. 2009. USDA vegetables 2008 summary, January 2009. Available at http://usda.mannlib.cornell.edu/usda/current/VegeSumm/VegeSumm-0128-2009.pdf (veried 10 Nov 2009). APP, B. A. 1938. Euxesta stigmatias Loew, an otitid y infesting ear corn in Puerto Rico. J. Agric. Univ. Puerto Rico 22: 181-187. ARCE DE HAMITY, M. G. 1986. Biology of Euxesta eluta (Diptera: Ulidiidae): Behavior in the attack and putrefaction of corn ears. Acta Zool. Lilloana 38: 119128. BAILEY, W. K. 1940. Experiments in controlling corn ear pests in Puerto Rico. Puerto Rico Agric. Exp. Stn. Circular no. 23. BARBER, G. W. 1939. Injury to sweet corn by Euxesta stigmatias Loew in southern Florida. J. Econ. Entomol. 32: 879-880. BARBOSA, P., SEGARRA-CARMONA, A. E., AND COLONGUASP, W. 1986. Eumecosomyia nubila (Wiedemann), a new otitid in Puerto Rico, with notes on the habits of the dipteran species complex of corn. J. Agric. Univ. Puerto Rico 70: 155-156. BLANTON, F. S. 1938. Some dipterous insects reared from narcissus bulbs. J. Econ. Entomol. 31: 113-116. CHITTENDEN, F. H. 1911. Some insects injurious to truck crops Notes on various truck-crop insects. USDA Bur. Entomol. 82: 90-90. CURRAN, C. H. 1928. Insects of Porto Rico and the Virgin Islands: Diptera or two-winged ies. New York Acad. Sci. Scientic Survey of Porto Rico and the Virgin Islands 11: 1-118. CURRAN, C. H. 1934. The families and genera of North American Diptera, American Museum of Natural History, New York, NY 512 p. CURRAN, C. H. 1935. New American Diptera. American Mus. Novit. 812: 1-24. DALY, T., AND BUNTIN, G. D. 2005. Effect of Bacillus thuringiensis corn for lepidopteran control of nontarget arthropods. Environ. Entomol. 34: 1292-1301. DIAZ, W. 1982. Danos de Euxesta eluta y E. mazorca (Dipt: Otitidae) sobre maices amilaceos en la Costa Central del Peru. Rev. Peruvian Entomol. 1: 51-53. EVANS, D. C., AND ZAMBRANO, E. 1991. Insect damage in maize of highland Ecuador and its signicance in small farm pest management. Trop. Pest Manage 37: 409-414. FISHER, E. 1996. Two new insect pests of corn in California. New Pest/Disease Advisory, 31 December 1996. State of California, Dept. Food & Agric., Div. Plant Ind. FRANCA, F. H., AND VECCHIA, P. T. D. 1986. Damages caused by Euxesta stigmatias on carrot roots in commercial seed eld. Pesq. agropec. bras., Braslia 21: 789-791.

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Goyal et al: Corn-infesting Ulidiidae of Florida 47FRAS-L, D. 1978. Ecological studies in Euxesta eluta and Euxesta annonae (Diptera: Otitidae). Agric. Tcnica (Chile) 38: 109-115. GOSSARD, H. A. 1919. Insects resembling the European corn borer. Mon. Bull. Ohio Agric. Exp. Stn. 4: 372-379. GOYAL, G., NUESSLY, G. S., STECK, G. J., SEAL, D. R., CAPINERA, J. L., AND BOOTE, K. J. 2010. New report of Chaetopsis massyla (Diptera: Ulidiidae) as a primary pest of corn in Florida. Florida Entomol. 93: 198-202. HAYSLIP, N. C. 1951. Corn silk y control on sweet corn, pp. 1-6. Univ. Florida Agric. Exp. Stn. Circ S-41, Gainesville, FL. KEIPER, J. B., SANFORD, M., JIANNINO, J., AND WALTON, W. E. 2000. Invertebrates inhabiting wetland monocots damaged by Lepidoptera. Entomol. News 111: 348-354. KNUTSON, A. 1987. Dynamics and natural enemies of the southwestern corn borer in the Texas High Plains. PhD Dissertation, Texas A & M University, College Station, TX. LANGILLE, R. N. 1975. Observations on the overwintering survival and spring development of the southwestern corn borer, Diatraea grandiosella Dyar. PhD Dissertation, University of Missouri, Columbia, MO. MERRILL, JR., L. S. 1951. Diptera reared from Michigan onions growing from seeds. J. Econ. Entomol. 14: 1015-1015. MOSSLER, M. A. 2008. Crop prole for sweet corn in Florida. CIR 1233, University of Florida. Available at http://edis.ifas.u.edu/pi034 (veried 10 Nov 2009). NUESSLY, G. S., AND HENTZ, M. G. 2004. Contact and leaf residue activity of insecticides against the sweet corn pest Euxesta stigmatias Loew (Diptera: Otitidae). J. Econ. Entomol. 97: 496-502. OLALQUIAGA, G. F. 1980. Aspectos tosanitarios de la isla de pascua. Rev. Chil. Entomol. 10: 101-102. PAINTER, R. H. 1955. Insects on corn and teosinte in Guatemala. J. Econ. Entomol. 48: 36-42. SAS INSTITUTE. 2008. Proc users manual, version 9th ed. SAS Institute, Cary, NC. SCULLY, B. T., NUESSLY, G. S., AND BEIRIGER, R. L. 2000. Resistance in maize to Euxesta stigmatias Loew (Diptera: Otitidae). J. Entomol. Sci. 35: 432443. SEAL, D. R. 1996. Insect control in sweet corn, 1995. Arthropod Manage Tests 21: 111-111. SEAL, D. R. 2001. Control of the corn silk y using various insecticides, 2000. Arthropod Manage. Tests 26: E31. Entomol. Soc. Am. Available at http:// www.entsoc.org/Protected/ AMT/Amt26/INDEX.ASP (veried 10 Nov 2009). SEAL, D. R., AND JANSSON, R. K. 1989. Biology and management of corn silk y, Euxesta stigmatias Loew (Diptera: Otitidae), on sweet corn in southern Florida. Proc. Florida State Hort. Soc. 102: 370-373. SEAL, D. R., AND JANSSON, R. K. 1994. Insect control in sweet corn, 1991. Arthropod Manage Tests 19: 9696. SEAL, D. R., JANSSON, R. K., AND K. BONDARI. 1996. Abundance and reproduction of Euxesta stigmatias (Diptera: Otitidae) on sweet corn in different environmental conditions. Florida Entomol. 79: 413-422. VAN ZWALUWENBURG, R. H. 1917. Report of the Entomologist: A new corn pest. Puerto Rico Agric. Exp. Stn. 31-34. WALTER, E. V., AND WENE, G. P. 1951. Tests of insecticides to control larvae of Euxesta stigmatias and Megaselia scalaris J. Econ. Entomol. 44: 998-999. WOLCOTT, G. N. 1948. The insects of Puerto Rico. J. Agric. Univ. Puerto Rico 32: 417-748. WUNDERLIN, R. P., AND HANSEN, B. F. 2008. Atlas of Florida Vascular Plants. Institute for Systematic Botany, University of South Florida, Tampa, FL. WYCKHUYS, K. A. G., AND ONEIL, R. J. 2007. Local agroecological knowledge and its relationship to farmers pest management decision making in rural Honduras. Agric. Hum. Values 24: 307-321.

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48 Florida Entomologist 94(1) March 2011 A CHECKLIST AND KEY TO SPECIES OF THE GENUS BETACIXIUS MATSUMURA (HEMIPTERA: FULGOROMORPHA: CIXIIDAE) WITH DESCRIPTIONS OF TWO NEW SPECIES FROM GUIZHOU PROVINCE, CHINA P EI Z HANG 1, 2, 3 AND X IANG -S HENG C HEN 1, 2, 1 The Provincial Key Laboratory for Agricultural Pest Management of Mountainous Region, Guizhou University, Guiyang, Guizhou 550025, P.R. China 2 Institute of Entomology, Guizhou University, Guiyang, Guizhou 550025, P.R. China 3 Xingyi Normal University for Nationalities, Xingyi, Guizhou 562400, P.R. China *Corresponding author; E-mail: chenxs3218@163.com A BSTRACT Two new species of Betacixius Matsumura, 1914 (Hemiptera: Fulgoromorpha: Cixiidae: Cixiini), B. bispinus Zhang and Chen sp. nov. (China: Guizhou) and B. agellihamus Zhang and Chen sp. nov. (China: Guizhou), from southwest China, are described and illustrated. A key for identifying 23 known species of Betacixius is provided. K ey Words: Hemiptera, Fulgoroidea, Cixiidae, Oriental region, Betacixius, new species, China R ESUMEN Se describen e ilustran dos nuevas especies de Betacixius Matsumura, 1914 (Hemiptera: Fulgoromorpha: Cixiidae: Cixiini), B. bispinus Zhang y Chen sp. nov. (China: Guizhou) y B. agellihamus Zhang y Chen sp. nov. (China: Guizhou) del suroeste de China. Se provee una clave para identicar las 23 especies conocidas de Betacixius. The cixiid planthopper genus Betacixius (Cixiinae: Cixiini) was established by Matsumura (1914) for the type species, B. ocellatus Matsumura, 1914, from Japan. To date, 21 species with 2 subspecies have been recorded worldwide and all species occur in the southern region (Matsumura 1914; Schumacher 1915; Metcalf 1936; Jacobi 1944; Fennah 1956; Hori 1982; Chou et al. 1985, 1988; Tsaur et al. 1991; Hua 2002). During the course of studying species biodiversity of the suborder Auchenorrhyncha in southwest China, 2 specimens belonging to undescribed species of the genus Betacixius were found. The purpose of this paper is to describe these 2 new species and to provide an identication key to all species of Betacixius M ATERIALS AND M ETHODS Morphological terminology follows Lcker et al. (2006). Dry specimens were used for the description and illustration. External morphology was observed under a stereoscopic microscope and characters were measured with an ocular micrometer. The genital segments of the examined specimens were macerated in 10% KOH and drawn from preparations in glycerin jelly with the aid of a Leica MZ 12.5 stereomicroscope. Illustrations were scanned with Canon CanoScan LiDE 200 and imported into Adobe Photoshop 8.0 for labeling and plate composition. Specimens examined are deposited in the Institute of Entomology, Guizhou University, Guiyang, Guizhou Province, China (IEGU). D ESCRIPTIVE T AXONOMY Betacixius Matsumura, 1914 (Figs. 1-25) Betacixius Matsumura 1914: 412; Chou et al. 1985: 23; Tsaur et al. 1991: 27. Type species: Betacixius ocellatus Matsumura 1914, by original designation. Description. This is a redescription incorporating the descriptions previously published by Chou et al. (1985) and Tsaur et al. (1991) as follows. Body Size and Coloration. Small to mediumsize cixiids (4.3-7.3 mm). Body coloration varying from green, brown to fulvous, mostly bearing special markings on anteclypeus; lateroapical parts of frons, otherwise unicolorous throughout. Head and Thorax. Head including eyes slightly narrower than pronotum. Vertex much wider than long in midline, widest basally or

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Zhang & Chen: New Species of Betacixius from China 49 Figs. 1-13. Betacixius bispinus Zhang and Chen sp. nov. 1. Head and thorax, dorsal view; 2. Frons; 3. Forewing; 4. Male genitalia, lateral view; 5. Pygofer and genital styles, ventral view; 6. Anal segment, dorsal view; 7. Anal segment, caudal view; 8. Connective, caudal view; 9. Right genital style, ventral view; 10. Aedeagus, left side; 11. Aedeagus, right side; 12. Aedeagus, dorsal view; 13. Aedeagus, ventral view. Scale bars = 0.25 mm (Figs. 6, 7), 0.5 mm (Figs. 1, 2, 4, 5, 8-13), 1 mm (Fig. 3).

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50 Florida Entomologist 94(1) March 2011 apically, lateral carinae moderately elevated, disc shallowly hollowed on each side of median carina. Frons rounded at base, usually with incomplete median carina not reaching anterior margin of vertex, lateral carinae slightly elevated below level of eyes, with small median ocellus and semicircular frontoclypeal suture. Clypeus tricarinate, convex to midline. Pronotum small, with distinct median carina, intermedian carinae curving laterad, angularly rounded posteriorly. Mesonotum tricarinate, convex between lateral carinae, attened posteromedially. Forewings broadest at apical third, rounded at apex, with 4-5 subapical cells and 8-9 apical cells, hyaline, sometimes with an oblique band or ocellated stripe. Hind tibia with 2-4 lateral spines and 6 apical spines. Chaetotaxy of hind tarsus 7/7. Male Genitalia. Pygofer symmetrical, Ushaped, with thumb-shaped dorsolateral angles in ventral view. Medioventral process triangular or subtriangular in ventral view, generally wider at base than long in midline. Anal segment tubular. Genital styles symmetrical in ventral view. Aedeagus slender in lateral view. Distribution. Oriental and Palaearctic regions. Remarks. This genus may be easily distinguished from other genera of Cixiini by the presence of 4-5 subapical cells and 8-9 apical cells on the forewing, vertex much wider than long in midline, frons with incomplete median carina distinct near frontoclypeal suture, and chaetotaxy of hind tarsus 7/7. The 2 new species, B. bispinus Zhang and Chen sp. nov. (China: Guizhou) and B. agellihamus Zhang and Chen sp. nov. (China: Guizhou), t into the genus by the presence of features as above W ORLD C HECKLIST OF S PECIES OF B ETACIXIUS M ATSUMURA B. bispinus Zhang and Chen sp. nov. ; southwestern China (Guizhou). B. brunneus Matsumura (1914); China (Taiwan), Japan. B. clypealis Matsumura (1914); China (Taiwan). B. clypealis vittifrons Matsumura (1914); China (Taiwan). B. delicatus Tsaur & Hsu (1991); China (Taiwan). B. euterpe Fennah (1956); China (Guangdong). B. agellihamus Zhang and Chen sp. nov. ; southwestern China (Guizhou). B. avovittatus Hori (1982); China (Taiwan). B. fuscus Tsaur & Hsu (1991); China (Taiwan). B. herbaceus Tsaur & Hsu (1991); China (Taiwan). B. kumejimae Matsumura (1914); South China, Japan (Okinawa). B. maculosus Tsaur & Hsu (1991); China (Taiwan). B. michioi Hori (1982); China (Taiwan). B. nelides Fennah (1956); China (Zhejiang, Guangdong). B. nigromarginalis Fennah (1956); China (Hubei). B. obliquus Matsumura (1914); South China, Japan (Gifu). B. obliquus pallens Matsumura (1914); Japan (Tokyo, Harima, Kumanoto) B. ocellatus Matsumura (1914); China (Taiwan), Japan. B. pallidior Jacobi (1944); South China (Fujian). B. rinkihonis Matsumura (1914); China (Taiwan), Japan. B. robustus Jacobi (1944); South China (Fujian). B. shirozui Hori (1982); China (Taiwan). B. sparsus Tsaur & Hsu (1991); China (Taiwan). B. tonkinensis Matsumura (1914); South China, Vietnam. B. transversus Jacobi (1944); South China (Fujian). K EY TO S PECIES OF THE G ENUS B ETACIXIUS M ATSUMURA 1. Forewings with markings (Figs. 3, 16, 24, 25). . . . . . . . . . . . . . . . . . . . . . . . . . .2 Forewings without any markings. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .9 2. Forewings with a large ocellate marking in apical half (Figs. 16 and 25). . . . . . . . . . . . . . . .3

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Zhang & Chen: New Species of Betacixius from China 51 Figs. 14-23. Betacixius agellihamus Zhang and Chen sp. nov. 14. Head and thorax, dorsal view; 15. Frons; 16. Forewing; 17. Male genitalia, lateral view; 18. Pygofer and genital styles, ventral view; 19. Anal segment, dorsal view; 20. Anal segment, caudal view; 21. Genital styles, ventral view; 22. Aedeagus, dorsal view; 23. Aedeagus, right side. Scale bars = 0.25 mm (Figs. 18-21), 0.5 mm (Figs. 14, 15, 17, 22, 23), 1 mm (Fig. 16).

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52 Florida Entomologist 94(1) March 2011 Forewings without such a marking in apical half (Figs. 3 and 24). . . . . . . . . . . . . . . . . . .6 3. Forewings with an oblique brown band extending from clavus across middle of corium . . . . B. tonkinensis Forewings without such a band (Figs. 16 and 25) . . . . . . . . . . . . . . . . . . . . . . . . . 4 4. Flagellum of aedeagus with 1 spine, hook-shaped (Figs. 22 and 23) . . . . . . . B. agellihamus sp. nov. Flagellum of aedeagus with 2 spines, not hook-shaped. . . . . . . . . . . . . . . . . . . . . . . 5 5. Aedeagus with 2 L-shaped processes apically . . . . . . . . . . . . . . . . . . . . . .B. maculosus Aedeagus with 1 nearly straight and 1 arched process apically . . . . . . . . . . . . . . . B. ocellatus 6. Forewings with an oblique band extending from stigma passing through its middle part . . . . . . . . .7 Forewings without such a band (Figs. 3 and 24) . . . . . . . . . . . . . . . . . . . . . . . . .12 7. Frons with median carina distinct on apical third; hind basitarsus much longer than the 2nd and 3rd segment put together. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B. michioi Frons and hind basitarsus without features as above. . . . . . . . . . . . . . . . . . . . . . . . 8 8. Forewings with apical cells of M and Cu strongly infuscate . . . . . . . . . . . . . . . B. transversus Forewings with apical cells not infuscate. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 9. Forewings with apical margin black or distinctly darkened. . . . . . . . . . . . . . . . . . . . .10 Forewings with apical margin fuscous or not distinctly darkened. . . . . . . . . . . . . . . . . . .11 10. Frons with a pallid spot at centre of lateral margins, clypeus dark, mesonotum testaceous . . .B. kumejimae Frons without such spots; mesonotum, except scutellum, castaneous-piceous . . . . . . . . . .B. euterpe 11. Forewings with an oblique dark band extending from clavus into centre of corium, slightly distad of level of union of claval veins. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .B. obliquus Forewings with a spot near sutural margin of clavus near union of claval veins, no oblique dark band at this level extending into corium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B. pallidior 12. Forewings with a long black stripe from base, along clavus extending to Rs . . . . . . . . . . .B. fuscus Forewings without such a stripe (Figs. 3 and 24) . . . . . . . . . . . . . . . . . . . . . . . . .13 13. Fore tibiae with black, longitudinal stripes. . . . . . . . . . . . . . . . . . . . . . . . . . . .14 Fore tibiae without such stripes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .15 14. Midand hindtibiae with black, longitudinal stripes . . . . . . . . . . . . . . . . . . B. delicatus Midand hindtibiae without such stripes . . . . . . . . . . . . . . . . . . . . . . . B. sparsus 15. Forewings infuscated at base, extending along clavus to end of Cu1; mesonotum with a large, very distinct brown marking between lateral carinae . . . . . . . . . . . . . . . . . . . . . . . . . .B. shirozui Forewings and mesonotum without spots as above (Figs. 3 and 24) . . . . . . . . . . . . . . . . .16 16. Forewings with apical margin black or very dark (Figs. 3 and 24) . . . . . . . . . . . . . . . . . .17 Forewings with apical margin not particularly dark. . . . . . . . . . . . . . . . . . . . . . . .18 17. Medioventral process of pygofer in ventral view right-angled triangular, pointed at apex (Fig. 5); on ventral margin, periandrium with lobate processes, 2 broad processes basally, bending forward, directed ventrocephalad, agellum semisclerotised, with several serrated processes near apex (Figs. 10 and 12) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B. bispinus sp. nov. Medioventral process in ventral view subtriangular, rounded at apex; periandrium without lobate processes, but with serrated processes basally, agellum, without any processes . . . . . . . . . . . B. rinkihonis 18. First apical cell of forewing piceous . . . . . . . . . . . . . . . . . . . . . . . . . . B. robustus A dark suffusion over all apical cells and across base of forewing . . . . . . . . . . . . . . .B. nelides

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Zhang & Chen: New Species of Betacixius from China 5319. Aedeagus with a curved spine on left near apex and a short ledge in a similar position on right; agellum arising above left margin, sides parallel for most of length, distally a short curved spine directed cephalad, and a subquadrate plate with a stout spine directed ventrad . . . . . . . . . . . . . B. nigromarginalis Aedeagus and agellum without features as above . . . . . . . . . . . . . . . . . . . . . . . .20 20. Frons without median carina . . . . . . . . . . . . . . . . . . . . . . . . . . . . .B. clypealis Frons with median carina. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .21 21. Frons with white or yellowish lateroapical parts; anteclypeus entirely black . . . . . . . B. avovittatus Frons and clypeus unicolorous . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .22 22. Body pale brown; agellum of aedeagus with 2 processes on right side . . . . . . . . . . . .B. brunneus Body green; agellum of aedeagus with one process on each side . . . . . . . . . . . . . B. herbaceus Betacixius bispinus Zhang and Chen sp. nov. (Figs. 1-13, 24)Description. Body length (from apex of vertex to tip of forewings): male 5.0-6.1mm ( n = 2), female 5.5-6.8mm ( n = 7); forewing length: male 4.0-4.9mm (n = 2), female 4.5-5.7mm (n = 7). Coloration. General color brown. Body slightly covered with powdery wax. Eyes yellowish brown to blackish brown; ocelli reddish yellow. Vertex yellowish brown. Pronotum blackish brown except median carina yellowish brown. Mesonotum blackish brown, with yellowish brown area posteromedially, carinae concolorous. Frons yellowish brown except lateral carinae blackish brown. Postclypeus yellowish brown, anteclypeus blackish brown. Rostrum generally yellowish brown, blackish brown near tip. Forewings pale brown, hyaline; veins brown, tubercles dark brown; stigma black; clavus with a short transversal brown band, just distad of fork PCu+A1. Hind-tibiae yellowish brown, lateraland apicalspines yellowish brown basally, black apically; platellae of tarsi dark brown. Abdomen black ventrally. Head and Thorax. Vertex narrowing to apex as shown in Figs. 1 and 24, wider than distance beFigs. 24-25. Body of adult in dorsal view. 24. Betacixius bispinus Zhang and Chen sp. nov.; 25. Betacixius agellihamus Zhang and Chen sp. nov.

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54 Florida Entomologist 94(1)March 2011tween eyes, 2.5 times wider than long in midline; anterior margin arched convex, with small emargination at midpoint, posterior margin arched concave; median carina indistinct, lateral carinae without branches, subapical carina absent. Median ocellus very small, located the centre of the frontoclypeal suture. Pedicel of antenna 1.6 times longer than wide. Frons broad, with small spots on middle area, widest at level of lateral ocelli, narrowing to both ends, 1.25 times longer than wide in midline; median carina indistinct, extending from slightly above level of lateral ocelli to median ocellus, lateral carinae distinct and ridged, arched convex; anterior margin arched concave. Clypeus with median carina distinct and elevated throughout, widest at level of endpoints of frontoclypeal suture; lateral carinae distinct and elevated. Rostrum relatively short, reaching hind coxae, apical and subapical segments equally long. Pronotum short and narrow, collarlike, twice as long as vertex in midline; median carina distinct and complete; intermedian carinae corrugated and curving into posterior margin which is concave in obtuse angle. Mesonotum 1.89 times longer than pronotum and vertex combined; 3 longitudinal carinae all reaching anterior and posterior margins, median carina indistinct on posteromedian area, which bears transverse striations. Forewings 2.22 times longer than wide, with sparse setae on surface, tubercles along veins, claval veins without tubercles; 2 indistinct subapical lines of cross veins; fork Sc+RP distad of fork CuA1+CuA2; r-m crossvein slightly distad of fork MA+MP; RP apically bid, MA apically bid, MP apically bid; fork PCu+A1 slightly basad of centre of clavus; Sc+R and M fused at superior-outside angle of basal cell; fork MA1+MA2 distad of fork MP1+MP 2. 2nd hindtarsus with 5 platellae; hind-tibia with 4 lateral spines, 6 apical spines: 2 large, 1 medium, 3 small, divided into 2 groups. Male Genitalia. In ventral view, pygofer stout, slightly concave medially, widening laterally; dorsal margin caudad obliquely raised in lateral view, inferior part with bristles; lateral lobes symmetrical, medium-inferior part arched convexly in lateral view. Medioventral process right-angled triangular in ventral view, relatively broad, 1.5 times wider than long, distance between tips of 2 lateral lobes 3.06 times as long as width of medioventral process; narrow triangular in lateral view, covered basally. Anal segment short and stout as shown in Figs 4, 6 and 7; 2.13 times longer than wide in dorsal view; incompactly connected with pygofer, freely movable; anal style, nger-like apically, not beyond anal segment; anal opening nearly subelliptical in dorsal view. Genital styles as shown in Figs. 4, 5 and 9, in ventral view, widening to apex, apical margin truncated, with sharp angle outside, internal processes broad, touching each other; in lateral view, ventral margin curving upward, outside slightly corrugated, dorsal margin smooth and bent forming a right angle approximately; incompactly connected with connective, freely movable; apical margin with bristles in ventral view. Aedeagus broad, short, connected with anal segment by 2 points; each side with broad spine arising at apex of aedeagal shaft, curving upward; periandrium ventrally with lobate processes, basally 2 broad processes, bending forward, directed ventrocephalad. Connective anchor-like, relatively long, the width of aedeagal shaft 1.65 times as wide as width of connective plus ventral arm. Flagellum semi-sclerotised, structure simple, generally curving left, with several serrated processes near apex. Type Material. Holotype: Mayanghe National Natural Reserve (600m), Yanhe County, Guizhou Province, China, 5-12 June 2007, X.-S. Chen. Paratypes: 1 7 same data as holotype. Etymology. The name is derived from the Latin words bi(double) and spinus (spine), which refers to the 2 spines on ventral margin of periandrium. Distribution: Southwest China (Guizhou Province). Remarks. This new species is similar in appearance to B. rinkihonis but differs from the latter in the shape of the medioventral process and the anal segment and by having 2 spines on the ventral margin of the periandrium and several serrated processes near apex of agellum.Betacixius agellihamus Zhang and Chen sp. nov. (Figs. 14-23, 25)Description Body length (from apex of vertex to tip of forewings): male 5.0-5.8mm ( n = 13), female 5.4-6.2mm (n = 15); forewing length: male 4.5-5.0mm (n = 13), female 4.6-5.1mm (n = 15). Coloration. General color brown. Body covered with powdery wax. Median area of eyes black, ventral margin yellow, other part reddish brown to blackish brown. Median ocellus pale yellow, semihyaline; lateral ocelli red. Vertex pale yellow. Pronotum pale yellow. Mesonotum brown. Frons generally brown, bright yellow above frontoclypeal suture; postclypeus yellow to brown, with oblique streaks; anteclypeus black. Apical segment of rostrum brown, subapical segment yellow. Forewings pale brown, semihyaline, with ocellated marking as shown in Figs. 16 and 25; veins and tubercles brown; stigma brown to dark brown; clavus with dark brown stain on apical third, sometimes extending to end of clavus; clavus suture brown. Hind-tibia brown, lateraland apical-spines brown at base, black apically; membranous tooth of tarsi dark yellow. Abdomen black ventrally.

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Zhang & Chen: New Species of Betacixius from China 55Head and Thorax. Vertex narrowing towards apex as shown in Figs. 14 and 25, wider than distance between eyes, 2.3 times wider than long in midline, separated into 2 hollow areolets by median carina; anterior margin generally arched convex slightly, slightly concave at midpoint, posterior margin concave in obtuse angle; median carina distinct and complete, subapical carina absent. Median ocellus indistinct, in the centre of frontoclypeal suture. Frons with well-distributed small spots, widest at level of antennae, narrowing to both ends, length equal to width; median carina distinct and elevated near frontoclypeal suture; lateral carinae slightly S-shaped, elevated; apex of frons elevated and lobate; anterior margin semicircle concave. Median carina of clypeus distinct and elevated; lateral carinae of postclypeus elevated. Rostrum reaching hindfemora, apical and subapical segments equally long. Pronotum short and narrow, collar-shaped, 2.25 times longer than vertex in midline; median carina distinct and elevated; posterior margin concave in obtuse angle. Mesonotum 1.44 times longer than pronotum and vertex combined; three longitudinal carinae elevated except for median carina weakly elevated at base, all reaching anterior and posterior margins. Forewings 2.2 times longer than wide; surface of Forewings with setae, basal part less, apical part more; veins with distinct tubercles, C vein with 34 tubercles, claval apically vein with tubercles; 2 indistinct subapical lines of cross veins; fork Sc+RP distad of fork CuA1+CuA2; r-m crossvein distad of fork MA+MP; RP apically bid, MA apically bid, MP apically bid; fork PCu+A1 slightly basad of centre of clavus; Sc+R and M fused at superior-outside angle of basal cell; fork MA1+MA2 distad of fork MP1+MP2. 2nd hindtarsus with 3 platellae; hind-tibia with 3 lateral spines, 6 apical spines, being divided into 2 groups by a relatively wide gap, one group with 3 equal spines, arranged closely, the other group with 1 large and 2 small, arranged sparsely. Male Genitalia. In ventral view, pygofer stout, shallowly U-shaped, slightly widening from base to end, ventral margin slightly concave; dorsal margin caudad obliquely upward in lateral view; lateral lobes symmetrical, inferior half slightly arched concave in lateral view. Medioventral process mastoid in ventral view, with bristles, 1.25 times wider than long, distance between tips of 2 lateral lobes 2.4 times as long as width of medioventral process; tongue-shaped in lateral view. Anal segment short and stout as shown in Figs 17, 19 and 20; in dorsal view 2 times longer than wide; compactly connected with pygofer, immovable; anal styles, not beyond anal segment; anal opening pear-like in dorsal view. Genital styles as shown in Figs 17, 18 and 21, in ventral view, widening to apex, internal processes broad, not touching each other; in lateral view, ventral margin curving upward, dorsal margin strongly bending upward; compactly connected with connective, immovable. Aedeagus stout, structure simple; each side with a broad spine at apex of aedeagal shaft, right one curving dorsad, left one relatively straight, directed up-cephalad. Connective relatively slender, the width of aedeagal shaft 1.33 times as wide as width of connective plus ventral arm. Flagellum strongly sclerotised, freely movable, structure simple, with a barbshaped spine at apex. Type Material. Holotype: Leigongshan National Natural Reserve, Leishan County, Guizhou Province, China, 13 May 1985, Z.-Z. Li. Paratypes: 7 9 same data as holotype; 2 3 Guiyang, Guizhou Province, China, June 1983, Students of Grade 79, Major Plantprotecting; 2 1 Huaxi (1000m), Guiyang, Guizhou Province, 20 May 2007, Q.-Z. Song; 1 2 Forest Park (1000m), Guiyang, Guizhou Province, China, 20 May 2007, X.-S. Chen. Etymology. The name is derived from the Latin words agell (agellate) and hamus (hook), which refers to the hook-like spine of agellum. Distribution: Southwest China (Guizhou Province). Remarks. This new species is similar in appearance to B. ocellatus but differs from the latter in the shape of the anal segment and the number of spines on the agellum. ACKNOWLEDGMENTSWe are grateful to Prof. Zi-Zhong Li, Ms. QiongZhang Song (Institute of Entomology, Guizhou University, China) for providing valuable specimens. We thank Prof. Dr. Shun-Chern Tsaur (Research Center for Biodiversity, Academia Sinica, Taibei, Taiwan), Kun-Wei Huang (National Museum of Natural Science, Taichung, Taiwan) and Ms. Gail Charabin (Saskatoon Research Centre, Agriculture and Agri-Food Canada) for obtaining literature. This work was supported by the National Natural Science Foundation of China (No.30560020, 31060290), the China Postdoctoral Science Foundation (No. 2005037111), the Program for New Century Excellent Talents in University (NCET07-0220), the Specialized Research Fund for the Doctoral Program of Higher Education (No. 20060657001), and the International Science and Technology Cooperation Program of Guizhou (20107005).REFERENCES CITEDCHOU, I., LU, J.-S., HUANG, J., AND WANG, S.-Z. 1985. Homoptera, Fulgoroidea. Economic Insect Fauna of China. Fasc. 36. Science Press, Beijing pp. 1-152. CHOU, I. WANG, Y.-L, HUANG, B.-K., AND YUAN, X.-Q. 1998. Homoptera: Fulgoroidea: Cixiidae, pp. 379-382 In B.-K Huang [ed.], Insect Fauna of Fujian Province, Volume II. Fuzhou, China, Science and Technology Press.

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56 Florida Entomologist 94(1)March 2011FENNAH, R. G. 1956. Fulgoroidea from Southern China. Proc. California Acad. Sci. 28: 441-527. HORI, Y. 1982. The Genus Betacixius Matsumura, 1914 (Homoptera: Cixiidae) of Formosa, pp. 175-182 In M. Sat, Y. Hori, Y. Arita and T. Okadome [eds.], Special issue to the memory of retirement of Emeritus Professor Michio Chj. Association of the Memorial Issue of Emeritus Professor M. Chj C/O Biological Laboratory, Nagoya Womens University, Nagoya, Japan. HUA, L.-Z. 2002. Catalogue of Insects of China I. Guangzhou, China: Sun Yat-Sen University Press. JACOBI, A. 1944. Die Zikadenfauna der Provinz Fukien in Sdchina und ihre tiergeographischen Beziehungen. Mitteilungen der Mnchner Entomologischen Gesellschaft 34: 5-66. LCKER, B., FLETCHER, M. J., LARIVIRE, M.-C., ANDGURR, G. M. 2006. The Australian Pentastirini (Hemiptera: Fulgoromorpha: Cixiidae). Zootaxa 1290: 1-138. MATSUMURA, S. 1914. Die Cixiinen Japans. Annotationes Zoologicae Japanenses, Tokyo 8: 393-434 METCALF, Z. P. 1936. General Catalogue of the Homoptera Fascicle IV Fulgoroidea, Part 2, Cixiidae. pp 269. SCHUMACHER, F. 1915. Der gegenwrtige Stand unserer Kenntnis von der Homopteren-Fauna der Insel Formosa unter besonderer Bercksichtigung von Sauterschem Material. Mitteilungen aus dem Zoologischen Museum in Berlin. Berlin 8: 73-134 TSAUR, S.-C., HSU, T.-C., AND VAN STALLE, J. 1991. Cixiidae of Taiwan, Part V. Cixiini except Cixius. J. Taiwan Museum 44: 1-78.

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Scheffrahn & Crowe: Termites on Boats 57 SHIP-BORNE TERMITE (ISOPTERA) BORDER INTERCEPTIONS IN AUSTRALIA AND ONBOARD INFESTATIONS IN FLORIDA, 1986-2009 R UDOLF H. S CHEFFRAHN 1 AND W ILLIAM C ROWE 2 1 Fort Lauderdale Research and Education Center, University of Florida, Institute of Food and Agricultural Sciences, 3205 College Avenue, Davie, Florida, 33314, U.S.A. rhsc@u.edu 2 Australian Quarantine and Inspection Service, P.O. Box 222, Hamilton Central, Queensland, 4007, Australia bill.crowe@aqis.gov.au A BSTRACT Alate termite ights from mature colonies infesting marine vessels is a primary mechanism for anthropogenic transoceanic establishment of invasive termite species. A taxonomic review is given of 133 recorded termite infestations onboard vessels in Australia and Florida between 1986 and 2009. The differing governmental approaches to regulating entry by foreign boats appears to reect the relative frequency of exotic termite establishments in Australia and Florida. Key Words: invasive species, biosecurity, overwater dispersal, Kalotermitidae, Rhinotermitidae, Termitidae R ESUMEN El vuelo de termitas aladas de colonias maduras que infestan barcos es el mecanismo principal para el movimiento transocenico y establecimiento de especies de termitas invasoras. Se provee una revisin taxonmica de 133 infestaciones de termitas encontradas en barcos en Australia y la Florida entre 1986 y 2009. Los diferentes enfoques gubernamentales en regular la entrada de barcos extranjeros tienden a reejar la frecuencia relativa de establecimiento de termitas exticas en Australia y la Florida. Natural overwater dispersal of infested otsam and anthropogenic dispersal by maritime vessels are the 2 primary means by which termites are transported across distant sea barriers (Scheffrahn et al. 2009). The cessation of rapid late Pleistocene/Holocene sea level rise at about 7K years before present (ybp, Fleming et al. 1998) predates the rst known long-distance human maritime voyages by some 3.5K ybp (Anderson et al. 2006). Therefore, contemporary nonathropogenic termite distributions were established between these 2 periods. Distant termite dispersal by otsam can be presumed to be a very rare event with a success rate inversely proportional to distance. Establishment of the depauparate native terrestrial faunas on distant oceanic volcanic islands such as Hawaii was the result of transoceanic dispersal (Cowie & Holland 2006). Nearshore islands like the Krakatau can be colonized much more frequently by both otsam transport and cross-water termite dispersal ights (Gathorne-Hardy & Jones 2000). Shipboard transport of termite colonies, where success is not affected by travel distance, has been suspected in recent (Gay 1967; Scheffrahn & Su 2005) and early (Scheffrahn et al. 2009) transoceanic termite establishments. Vessels can be colonized during construction (usually only Kalotermitidae) or by alates (all taxa) ying onboard during dockage, either on water or in dry dock. Hochmair & Scheffrahn (2010) showed a strong correlation between land-borne infestations of Coptotermes spp. in Florida and their distance to maritime boat doc kage suggesting that marine vessels are predominant vehicles for dissemination of this pest genus. Within the last century, 6 exotic termite species have become established in Florida (Scheffrahn et al. 1988; Scheffrahn et al. 1992; Scheffrahn et al. 2002; Scheffrahn & Su 1995; Su et al. 1997), more than any other state or territory in America, followed by Hawaii with 5 species (or 4 species if Zootermopsis angusticollis Hagen is not established, Woodrow et al. 1999; Yeap et al. 2007). Australia has a slightly greater human population than Florida, a much larger tidal coastline, and both share similar economies, climates, pleasure boating industries, and proximities to tropical nations to the north and south, respectively. Yet, in the last century, a single exotic termite, Cr. brevis (Walker), has become established in Australia (Peters 1990). As for other suspected exotic Cryptotermes Gay & Watson (1982) determined that Cr. cynocephalus Light and Cr. domesticus (Haviland) are endemic to northeast Australia. Gay (1967) reported that Cr. dudleyi

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58 Florida Entomologist 94(1) March 2011 Banks, an exotic drywood termite from Southeast Asia, was already established in Darwin by 1913. In a further attempt to understand the dynamics and taxa involved in exotic termite establishments, we provide a summary of onboard termite infestations in Florida and border interceptions in Australia and we contrast the regulatory procedures used for boats arriving from foreign ports. M ATERIALS AND M ETHODS Termite specimens from Australia were found during interceptive inspections by WC and other Australian Quarantine and Inspection Service (AQIS) personnel. Florida samples were collected by or submitted to RHS by pest control professionals and boat owners or operators. In both cases, onboard specimens were collected and stored in ethanol. Identications were made by the authors using voucher specimens from their respective collections. Other information sought included date of collection, given in Table 1 only in years, location collected (city), vessel origin (if known), and vessel and/or infestation type. For species identication, samples were required to contain morphologically robust winged imagos and/or soldiers. Workers were identied morphologically only to genus. R ESULTS AND D ISCUSSION During 1986-2009, 74 and 59 termite incidents onboard boats were recorded in Australia and Florida, respectively (Table 1). The Australian records are comprehensive and represent all known AQIS interceptions. The Florida incidents represent an informal and very incomplete sampling of the actual number of boat infestations occurring around the State. Three vessels were infested simultaneously by 2 species and each is recorded as a separate incident in Table 1. Unlike Australia where only Cr. brevis is established, most boats in Florida are infested in their home w aters where exotic species abound. This phenomenon enhances the spread of termites in Florida from boat-to-land or land-to-boat by dockside dispersal ights and also elevates the likelihood that boats voyaging from Florida could spread termites to foreign ports. Although long, open ocean voyages are not the norm for Florida boaters, some will island-hop throughout the West Indies. One yacht, suspected of acquiring Co. gestroi Wasmann (= havilandi Kirton & Brown 2003) during a winter doc kage in the Turks and Caicos Islands (Scheffrahn & Su 2005) was simultaneously infested with Incisitermes minor (Hagen). It was presumed that the latter species infested this boat while under construction in San Diego California. The subterranean genus Coptotermes (Family Rhinotermitidae) w as observed in 53% of all boat infestations (Fig. 1) with Co. formosanus Shiraki being the most common onboard pest in both Florida (27 records) and Australia (13 records) followed by Co. gestroi with 15 and 4 records, respectively Three other infestations by subterranean termites were recorded including 2 by Reticulitermes virginicus (Banks) in Florida and 1by Heterotermes sp. in a boat that sailed from Florida to Grand Ca yman Island. The second most prevalent genus, at 30% of boat infestations, was Cryptotermes (Family Kalotermidae). Australian interceptions yielding 8 infestations eac h of Cr. brevis (Walker) and Cr. domesticus (Haviland), 3 infestations of Cr. dudleyi Banks, 2 of Cr. cynocephalus Light, and 15 Cr species undetermined. Only 3 infestations of Cr. brevis were recorded from Florida; however, fumigations for this species are so routine in Florida that samples are seldom collected for identication. One pest control company in Fort Lauderdale estimates that it is contracted to fumigate about 15 boats a year for drywood termites (read Cr. brevis, Edwards, J. K., personal communication). On the other hand, 6 infestations of I. minor were recorded in both Australia and Florida. Alates of I. minor are muc h more robust and dark (reddish pronotum and head) than Cr. brevis and have a different ight season and diel periodicity Therefore, I. minor ights prompt elevated identication requests by the Florida pest control industry Australia recorded a single shipboard infestation each of I. immigrans (Snyder) and I sp., while in Florida, a single infestation of I. snyderi (Light) w as observed on a houseboat in Key West. The most unexpected nd of this study was a mature infestation in 2009 of Rugitermes panamae (Snyder) from an itinerant yac ht intercepted while visiting Bundaberg Australia. The yacht was apparently infested by this dampwood species during a voyage in 2003 to Central America. Colonies of the predominantly arboreal genus Nasutitermes (Termitidae) were found on boats 3 times during the last 25 years Nasutitermes acajutlae (Holmgren) was found twice and N. nigriceps (Haldeman) once in Florida. Although not recorded in Table 1, N. corniger (Motschulsky) was found infesting 2 boats in dry doc k in Dania Beach, Florida, as part of a land-borne infestation of this pest (Scheffrahn et al. 2002). We suggest herein that the difference in the number of exotic termite species established in Florida versus Australia is attributable, at least in part, to differing laws and regulations intended to exclude exotic pests. The U.S. Customs and Border Patrol (CBP) requires that pleasure vessels arriving in the U.S. from a foreign port must report their arrival by telephone and be directed, with passengers and crew, to the nearest port of entry or nearest designated reporting location for a CBP face-to-face interview and/or vessel inspection (Anonymous 2009). Inspections focus on im-

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Scheffrahn & Crowe: Termites on Boats 59 T ABLE 1. T ERMITE INFESTATIONS BY GENUS AND SPECIES ONBOARD VESSELS DURING 1986-2009 ( VOUCHER SPECIMENS IN THE UNIVERSITY OF F LORIDA TERMITE COLLECTION OR AQIS RECORDS ). Vessel location where termites found Vessel origin 1 GenusSpeciesYearVessel comments Islamorada Key, FL Coptotermesformosanus 1986boat North P alm Beach, FLTennessee Coptotermesformosanus 1995boat F ort Lauderdale, FL Coptotermesformosanus 199511 m boat J acksonville, FL Coptotermesformosanus 1997cable ship Lighthouse P oint, FL Coptotermesformosanus 1998boat Hypoluxo FL Coptotermesformosanus 199826 m boat P alm Beach, FL Coptotermesformosanus 19999 m boat Brunswic k, Georgia Coptotermesformosanus 199910 m boat P alm Beach Gardens, FL Coptotermesformosanus 20009 m boat Hillsborough Beac h, FL Coptotermesformosanus 2000large boat Hallandale FL Hong Kong Coptotermesformosanus 200023 m boat T ampa, FL Coptotermesformosanus 200110 m boat P ompano Beach, FL Coptotermesformosanus 200116 m boat T ampa, FL Coptotermesformosanus 200210 m boat F ort Lauderdale, FL Coptotermesformosanus 200226 m speed boat F ort Lauderdale, FL Coptotermesformosanus 200215 m cabin cruiser Holmes Beac h, FL Coptotermesformosanus 2002boat Dania Beac h, FL Coptotermesformosanus 200311 m boat Hollywood, FL Coptotermesformosanus 2004boat F ort Lauderdale, FL Coptotermesformosanus 2004boat F ort Lauderdale, FL Coptotermesformosanus 200418 m shing yac ht Fort Lauderdale, FL Coptotermesformosanus 200515 m sailboat J acksonville Beach, FL Coptotermesformosanus 200613 m boat Marathon K ey, FL Coptotermesformosanus 2006small boat F ort Lauderdale, FL Coptotermes formosanus 200815 m boat V olusia County, FL Hong Kong Coptotermes formosanus 200818 m cabin cruiser P anama City, FL Coptotermes formosanus 20089 m boat Lake P ark, FL Coptotermes formosanus 20088 m boat F ort Pierce, FL Jamaica Coptotermesgestroi 1991Boat Hollywood, FL Virgin Gordo, B.V.I. Coptotermesgestroi 1995boat in dry dock F ort Lauderdale, FLTurks, Caicos Coptotermesgestroi 200127 m yacht K ey West, FL Coptotermesgestroi 200315 m sailboat K ey West, FL Coptotermesgestroi 2005houseboat, nest with queen K ey West, FL Coptotermesgestroi 20058 m motor boat Tequesta, FL Coptotermesgestroi 20059 m shing boat Key Largo, FL Cuba Coptotermesgestroi 2006Sailboat Key West, FL Coptotermesgestroi 2007Sailboat Key West, FL Coptotermesgestroi 2007Boat Miami Beach, FL Coptotermesgestroi 200712 m shing boat transom Stock Island Key, FLKey West Coptotermesgestroi 200715 m cabin cruiser St. Petersburg, FL Coptotermesgestroi 200711 m Yacht Boca Chica Key, FL Key West Coptotermesgestroi 2007sailboat Stock Island (Key West), FL Coptotermesgestroi 2007Boat Franklin, LouisianaFlorida Cryptotermesbrevis 200017 m boat Marathon Key, FL Cryptotermesbrevis 2005Sailboat Cudjoe Key, FL Cryptotermesbrevis 2006Boat Grand Cayman, Cayman Is.Florida Heterotermes sp. 1995Boat1Unknown if blank.2dead imagos only, no live infestation.

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60 Florida Entomologist 94(1)March 2011Fort Lauderdale, FL Incisitermesminor 2000Boat Miami, FL Los Angeles, CA Incisitermesminor 200026 m boat Fort Lauderdale, FLSan Diego, CA Incisitermesminor 200127 m yacht Marathon Key, FL Taiwan Incisitermesminor 2006Boat Dania Beach, FL western Mexico Incisitermesminor 200720 m boat St. Augustine, FL FL Keys Incisitermesminor 2008Boat Key West, FL Incisitermessnyderi 2000Houseboat Fort Lauderdale, FLSt. Thomas U.S.V.I. Nasutitermesacajutlae 200215 m boat Jacksonville, FL Puerto Rico Nasutitermesacajutlae 2002container on ship Fort Lauderdale, FL Nasutitermesnigriceps 1996Sailboat Key West, FL Reticulitermesvirginicus 2000Houseboat Jacksonville Beach, FL Reticulitermesvirginicus 2003Boat Darwin, NT, AUS China Coptotermesformosanus 1994boat (refugee) Perth, WA, AUS Hong Kong Coptotermesformosanus 2000Boat Brisbane, Qld, AUS China Coptotermesformosanus 2002boat, breglass Sydney, NSW, AUS Coptotermesformosanus 2003Yacht Brisbane, Qld, AUS Hong Kong Coptotermesformosanus 2003boat Brisbane, Qld, AUS China Coptotermesformosanus 2003Yacht Brisbane, Qld, AUS USA / Japan Coptotermesformosanus 20059 m boat Brisbane, Qld, AUS USA Coptotermesformosanus 2005boat (with I. minor) Townsville, Qld, AUSHong Kong/Asia Coptotermesformosanus 2005Boat Bundaberg, Qld, AUSHawaii Coptotermesformosanus 2006itinerant yacht Brisbane, Qld, AUS Japan Coptotermesformosanus 2007Boat Newcastle, NSW, AUSChina Coptotermesformosanus 2008Boat Brisbane, Qld, AUS USA Coptotermesformosanus 2009itinerant yacht Darwin, NT, AUS Thailand Coptotermesgestroi 1986Yacht Darwin, NT, AUS Thailand Coptotermesgestroi 1994Boat Bundaberg, Qld, AUSMarshall Islands Coptotermesgestroi 1996Yacht Brisbane, Qld, AUS China Coptotermesgestroi 2003Boat Brisbane, Qld, AUS USA Coptotermes sp. 2002Yacht Cairns, Qld, AUS AUS Coptotermes sp. 2005dinghy from TI to Cairns Sydney, NSW, AUS Unknown Coptotermes sp. 2006navy boat Brisbane Qld, AUS New Caledonia Coptotermes sp. 2008boat (returning AUS yacht) Brisbane, Qld, AUS Taiwan Coptotermes sp. 2008Boat Brisbane, Qld, AUS Hong Kong Coptotermes sp. 2005yacht (ybridge in lockers) Mackay, Qld, AUS Taiwan Coptotermes sp. 2005Boat Brisbane, Qld, AUS France Coptotermes sp. 2008Yacht Brisbane, Qld, AUS USA Coptotermes sp. 2008Boat Perth, WA, AUS Singapore Coptotermestravians? 2002boat, breglass & wood Townsville, Qld, AUS Cryptotermesbrevis 1989Yacht Brisbane, Qld, AUS USA Cryptotermesbrevis 2003wooden yacht (with I. Minor) Brisbane, Qld, AUS USA Cryptotermesbrevis 2005Superyacht Cardwell, Qld, AUS Cryptotermesbrevis 2006Trimaran Brisbane, Qld, AUS South Africa Cryptotermesbrevis 2007Boat Brisbane, Qld, AUS USA Cryptotermesbrevis 2008Boat Airlie Beach, Qld, AUSUSA Cryptotermesbrevis 2009 Catalina 400 MK II TABLE 1. (CONTINUED) TERMITE INFESTATIONS BY GENUS AND SPECIES ONBOARD VESSELS DURING 1986-2009 (VOUCHER SPECIMENS IN THE UNIVERSITY OF FLORIDA TERMITE COLLECTION OR AQIS RECORDS). Vessel location where termites foundVessel origin1GenusSpeciesYearVessel comments1Unknown if blank.2dead imagos only, no live infestation.

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Scheffrahn & Crowe: Termites on Boats 61Mackay, Qld, AUS USA Cryptotermesbrevis? 200928 m super yacht Bundaberg, Qld, AUSUSA Cryptotermescavifrons12008Yacht Darwin, NT, AUS Indonesia Cryptotermescynocephalus 2005foreign shing vessel Darwin, NT, AUS Indonesia Cryptotermescynocephalus 2009foreign shing vessel Darwin, NT, AUS Indonesia Cryptotermesdomesticus 1986Yacht Darwin, NT, AUS Indonesia Cryptotermesdomesticus 1987yacht Brisbane, Qld, AUS Vanuatu Cryptotermesdomesticus 1999boat Sydney, NSW, AUS Cryptotermesdomesticus 2003yacht Broome, WA, AUS P apela, Roti, Indonesia Cryptotermesdomesticus 2005foreign shing vessel Gove, NT, AUS Karja Sama, Indonesia Cryptotermesdomesticus 2006foreign shing vessel Gove, NT, AUS Indonesia Cryptotermesdomesticus 2006foreign shing vessel Broome, WA, AUS Indonesia Cryptotermesdomesticus 2007foreign shing vessel Darwin, NT, AUS Indonesia Cryptotermesdudleyi 1994boat Darwin, NT, AUS Philippines Cryptotermesdudleyi 2006boat Darwin, NT, AUS Indonesia Cryptotermesdudleyi 2008foreign shing vessel Darwin, NT, AUS Indonesia Cryptotermes sp. 1993boat Gove, NT, AUS Indonesia Cryptotermes sp. 1993foreign shing vessel Darwin, NT, AUS Vietnam Cryptotermes sp. 2001boat Gove, NT, AUS Indonesia Cryptotermes sp. 2004foreign shing vessel Thursday Island, AUSIndonesia Cryptotermes sp. 2004foreign shing vessel Darwin, NT, AUS Indonesia Cryptotermes sp. 2004boat Darwin, NT, AUS Indonesia Cryptotermes sp. 2005foreign shing vessel Broome, WA, AUS P apela Roti, Indonesia Cryptotermes sp. 2006foreign shing vessel Broome, WA, AUS Indonesia Cryptotermes sp. 2007foreign shing vessel Bundaberg, Qld, AUSUSA Cryptotermes sp. 2007boat Broome, WA, AUS Sulawesi, Indonesia Cryptotermes sp. 2005foreign shing vessel Broome, WA, AUS Indonesia Cryptotermes sp. 2005foreign shing vessel Broome, WA, AUS Indonesia Cryptotermes sp. 2005foreign shing vessel Darwin, NT, AUS Indonesia Cryptotermes sp. 2005foreign shing vessel Broome WA, AUS Indonesia Cryptotermes sp. 2009foreign shing vessel Brisbane, Qld, AUS Thailand Drepanotermes2sp. 2008boat Bundaberg, Qld, AUSHawaii Incisitermesimmigrans 2007boat Brisbane, Qld, AUS Incisitermesminor 2001yacht Brisbane, Qld, AUS USA Incisitermesminor 2003wooden yacht (with Cr. brevis) Brisbane, Qld, AUS USA Incisitermesminor 2005boat (with Co formosanus) TABLE 1. (CONTINUED) TERMITE INFESTATIONS BY GENUS AND SPECIES ONBOARD VESSELS DURING 1986-2009 (VOUCHER SPECIMENS IN THE UNIVERSITY OF FLORIDA TERMITE COLLECTION OR AQIS RECORDS). Vessel location where termites foundVessel origin1GenusSpeciesYearVessel comments1Unknown if blank.2dead imagos only, no live infestation.

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62 Florida Entomologist 94(1)March 2011migration compliance by the passengers and crew, possible illegal contraband, and agricultural pests in cargo. Structural and household pests, which are usually disassociated with cargo and dwell within the vessels own structure, are not mandated for inspection. In contrast to Florida practices, passengers and crew aboard vessels arriving to Australia from a foreign port must obtain clearance by the Australian Customs and Border Protection Service and the Australian Quarantine and Inspection Service (AQIS). Vessels with timber in their cargo or construction must also be inspected by AQIS. The level of AQIS inspection required will depend on the amount of timber present and the construction/re-t and sailing history of the vessel. The inspection can be conducted by an AQIS quarantine ofcer or AQIS entomologist with or without a licensed pest control professional and approved termite detection method. If termites are found upon inspection, the vessel must be fumigated with methyl bromide (AQIS method T9047) or sulfuryl uoride (AQIS method T9090) at the owners expense (Anonymous 2010).Brisbane, Qld, AUS Fiji (made in USA) Incisitermesminor 2006super yacht Bundaberg, Qld, AUSUSA Incisitermesminor 2006trimaran Cairns, Qld, AUS USA Incisitermesminor 2009trimaran Broome, WA, AUS P apela Roti, Indonesia Incisitermes sp. 2006foreign shing vessel Bundaberg, Qld, AUSCentral Amer. Rugitermespanamae 2009Yacht TABLE 1. (CONTINUED) TERMITE INFESTATIONS BY GENUS AND SPECIES ONBOARD VESSELS DURING 1986-2009 (VOUCHER SPECIMENS IN THE UNIVERSITY OF FLORIDA TERMITE COLLECTION OR AQIS RECORDS). Vessel location where termites foundVessel origin1GenusSpeciesYearVessel comments1Unknown if blank.2dead imagos only, no live infestation. Fig. 1. Frequency of termite genera collected on vessels in Australia and Florida. Other includes Drepanotermes and Rugitermes from Australia and Heterotermes from Florida.

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Scheffrahn & Crowe: Termites on Boats 63ACKNOWLEDGMENTSWe thank boat owners and pest control companies in Florida for submitting termite samples to RHS.REFERENCES CITEDANDERSON, A., CHAPPELL, J., GAGAN, M., AND GROVE, R. 2006. Prehistoric maritime migration in the Pacific islands: An hypothesis of ENSO forcing. The Holocene 16: 1-6. ANONYMOUS. 2009. Code of Federal Regulations (United States) 19CFR4.2. ANONYMOUS. 2010. ICON Condition C9645. Australian Quarantine and Inspection Service. COWIE, R. H., AND HOLLAND, B. S. 2006. Dispersal is fundamental to biogeography and the evolution of biodiversity on oceanic islands. J. Biogeography 33: 193-198. FLEMING, K., JOHNSTON, P., ZWARTZ, D., YOKOYAMA, Y., LAMBECK, K., AND CHAPPEL, J. l998. Rening the eustatic sea-level curve since the last glacial maximum using farand intermediate-eld sites. Earth and Planetary Science Letters 163: 327-342. GATHORNE-HARDY, F. J., AND JONES, D. T. 2000. The recolonization of the Krakatau islands by termites (Isoptera), and their biogeographical origins. Biol. J. Linnean Soc. 71: 251-267. GAY, F. J. 1967. A world review of introduced species of termites. CSIRO Bulletin Melbourne, Australia 286: 1-88. GAY, F. J., AND WATSON, J. A. L. 1982. The genus Cryptotermes in Australia (Isoptera: Kalotermitidae). Australian J. Zool. Supplementary Series 30, 88: 164. HOCHMAIR, H. H., AND SCHEFFRAHN, R. H. 2010. Spatial association of marine dockage with land-borne infestations of invasive termites in urban South Florida. J. Econ. Entomol. 103: 1338-1346. KIRTON, L. G., AND BROWN, V. K. 2003. The taxonomic status of pest species of Coptotermes in Southeast Asia: Resolving the paradox in the pest status of the termites, Coptotermes gestroi C. havilandi and C. travians (Isoptera: Rhinotermitidae). Sociobiol. 42: 4363. PETERS, B. C. 1990. Infestations of Cryptotermes brevis (Walker) (Isoptera: Kalotermitidae) in Queensland, Australia. 1. History, detection and identication. Australian Forester 53: 79-88. SCHEFFRAHN, R. H., AND SU, N.-Y. 1995. A new subterranean termite introduced to Florida: Heterotermes Froggatt (Rhinotermitidae: Heterotermitinae) established in Miami. Florida Entomol. 78: 623-627. SCHEFFRAHN, R. H., AND SU, N.-Y. 2005. Distribution of the termite genus Coptotermes (Isoptera: Rhinotermitidae) in Florida. Florida Entomol. 88: 201-203. SCHEFFRAHN, R. H., CABRERA, B. J., KERN JR., W. H.,AND SU, N.-Y. 2002. Nasutitermes costalis (Isoptera: Termitidae) in Florida: First record of a non-endemic establishment by a higher termite. Florida Entomol. 85: 273-275. SCHEFFRAHN, R. H., KRECK, J. RIPA, R., AND LUPPICHINI, P. 2009. Endemic origin and vast anthropogenic dispersal of the West Indian drywood termite. Biol. Invasions 11: 787-799. SCHEFFRAHN, R. H., MANGOLD, J. R., AND SU, N.-Y. 1988. A survey of structure-infesting termites of peninsular Florida. Florida Entomol. 71: 615-630. SU, N.-Y., SCHEFFRAHN, R. H., AND WEISSLING, T. 1997. A new introduction of a subterranean termite, Coptotermes havilandi Holmgren (Isoptera: Rhinotermitidae) in Miami, Florida. Florida Entomol. 80: 408-411. WOODROW, R. J., GRACE, J. K., AND YATES III, J. R. 1999. Hawaiis Termites: An Identication Guide. Honolulu (HI): University of Hawaii. 6 p. (Household and Structural Pests; HSP-1). YEAP, B.-K., OTHMAN, A. S., LEE, V. S., AND LEE, C.-Y. 2007. Genetic relationship between Coptotermes gestroi and Coptotermes vastator (Isoptera: Rhinotermitidae). J. Econ. Entomol. 100: 467-474.

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64 Florida Entomologist 94(1) March 2011 FIRST RECORD OF THE GENUS ADOXOMYIA (DIPTERA: STRATIOMYIDAE) WITH FOUR SPECIES FROM TURKEY T URGAY STNER 1 AND A BDULLAH H ASBENLI 2 1 Seluk University, Faculty of Science, Department of Biology, Alaaddin Keykubat Kamps, 42100 Seluklu, Konya, Turkey E-mail: turgayustuner@gmail.com 2 Gazi University, Faculty of Arts and Sciences, Department of Biology, 06500 Teknikokullar, Ankara, Turkey. A BSTRACT Adoxomyia aureovittata (Bigot, 1879), A. cinerascens (Loew, 1873), A. obscuripennis (Loew, 1873) and A. sarudnyi (Pleske, 1903) are recorded from Turkey for the rst time. Both sexes of the rst 3 species and the male of A sarudnyi are redescribed and photographs of all species are provided. The distributions of all species are briey discussed. The male genitalia and some other diagnostic characters of all the examined species are illustrated. An identication key to all East-Mediterranean species was constructed and is included in this report. Key Words: Adoxomyia aureovittata, A. cinerascens, A. obscuripennis, A. sarudnyi, new records distribution, Turkey R ESUMEN Se registran por primera vez Adoxomyia aureovittata (Bigot, 1879), A. cinerascens (Loew, 1873), A. obscuripennis (Loew, 1873) y A. sarudnyi (Pleske, 1903) para Turquia. Se proveen redescripciones y fotos de ambos sexos de las primeras 3 especies y del mac ho de A sarudnyi Se discutan la distribucin de todas las especies Se ilustran las genitalias de los machos y algunas de las caracteristicas diagnsticas de las especies examinadas. Una clave para la identicacin de todas las especies de la regin Este del Mediterrneo es incluida. The family Stratiomyidae belongs to the suborder Brachycera in Diptera (Rozkosny 1982). This large family includes more than 2650 species in 375 genera composed of 12 subfamilies worldwide of which 426 species in 55 genera in 7 subfamilies occur in the Palaearctic region (Woodley 2001). So far 48 species in 14 genera and 7 subfamilies ( Beridinae Pachygastrinae Clitellariinae Hermetina e, Sarginae Stratiomyinae Nemotelinae ) have been recorded in Turkey (Rozkosny & Nartshuk 1988; Woodley 2001; stner et al. 2002, 2003; stner & Hasbenli 2003, 2004). The subfamily Clitellariinae contains 50 genera worldwide, 10 genera in the Palaearctic region and 1 genus ( Pycnomalla ) in Turkey (Woodley 2001; stner et al. 2002). The genus Adoxomyia (Kertsz, 1907) belongs to the subfamily Clitellariinae and inc ludes 36 described species. They are distributed in the Palaearctic region (16 species), the Nearctic region (13 species), the Neotropical region (4 species), the Oriental region (3 species) and the Afrotropical region (2 species) (Woodley 2001; Hauser 2002; Nartshuk 2004). Palearctic species of Adoxomyia are found mainly in south-eastern Europe Transcaucasus, Near East, Central Asia and China (Rozkosny 1983; Rozkosny & Nartshuk 1988; Nartshuk 2004). Adoxomyia had not been recorded in Turkey before this report. The larvae of Adoxomyia are known only for some Nearctic and one Oriental species; they were found in decaying cacti and nests of pack rats ( Neotoma sp.) (McFadden 1967; James & McF adden 1969). In addition to the 4 species recorded in Turkey at least 5 additional species may occur here. A. dahlii (Meigen 1830) is known from southern Europe (incl. Ukraine), Armenia and Israel. A. rucornis (Loew 1873) occurs in Azerbaijan, Iran and Kyrgyzstan. A. hermonensis Lidner, 1975 and A. paleastinensis Lindner, 1937 were described from Israel and A. transcaucasica Nartshuk, 2004 is based on types from Armenia and Azerbaijan. According to a recent paper by Nartshuk (2004), A. portschinskii (Pleske) is a mere synonym of A. dahlii (Meigen). M ATERIALS A total of 16 specimens (12 males and 4 females) were collected by hand net at Antalya, Isparta, and Konya in Turkey between 1999 and 2001. Some specimens of Adoxomyia were caught sunbathing on stones or on the ground in dry creek beds The specimens are deposited in the collection of the Seluk University Department of Biology in Konya, Turkey. The fact that previously the genus Adoxomyia had not been recorded from Turkey reects the poor knowledge of the Turkish Stratiomyidae fauna.

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stner & Hasbenli: First Records of Adoxomyia Species in Turkey65 THE GENUS ADOXOMYIA Kertsz 1907 The generic name Adoxomyia was proposed by K ertsz (1907) and according to the catalog publishad a year later (Kertsz 1908) this genus embraced 23 species. In 1923, Kertsz tried to establish a new separate genus, Euclitellaria Kertsz, but it was not accepted by Pleske (1925) and subsequent authors The last comprehensive key to the Palaearctic species is that by Lindner (1937), who followed Pleskes concept of Adoxomyia in a broad sense. Besides the work by Rozkosny (1983), which treated the European species some other relevant papers were chiey devoted to descriptions of separate additional species (uchi 1938; Dusek & Rozkosny 1963; Lindner 1975; Nartshuk 2004). In Turkey, the species belonging to Adoxomyia are mid-sized (6-11 mm) with a predominantly blac k body. The eyes, which are contiguous in males and widely separated in females, are covered with dense hairs. The antennae are relatively long, predominantly black, but can be redorange to dark brown in some species. Scape and pedicel are of equal length. The agellum consists of 8 agellomeres. No thoracic spines but two scutellar spines are present. All of 4 M-veins reach the wing margin. K EY TO THE E AST M EDITERRANEAN S PECIES OF A DOXOMYIA The following key is based partly on Lindner (1937). The male of A. hermonensis and the female of A. palaestinensis are unknown. 1.Legs completely black . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2 Legs bicoloured or mainly . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 2.Antenna entirely black. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 At least basal half of antennal agellum red . . . . . . . . . . . . . . . . . A. rucornis (Loew 1873) 3. Scutellar spines short, slender and bare, basal 3-4 agellomeres in female unusually broad . . . . . . . .4 Femora Scutellar spines longer, thickened and haired, basal 3-4 agellomeres not as broad (Figs. 19 and 20) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .A obscuripennis (Loew 1873) 4. Female eyes black haired, postocular band wider than scape is long; male unknown . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . hermonensis Lindner 1975 Female eyes white haired, postocular band as wide as scape is long. . . . A. transcaucasica Nartshuk 2004 5. Legs entirely yellow . . . . . . . . . . . . . . . . . . . . . . . . . . .A. sarudnyi (Pleske 1903) At least femora black. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .6 6.Antenna black. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7 Abdomen with silverish white hair patches . . . . . . . . . . . . . . A. palaestinensis Lindner 193 8. Abdomen with golden yellow hair patches . . . . . . . . . . . . . . . .A. aureovittata (Bigot 1879) Abdomen with silverish white (rarely coppery) hair patches. . . . . . . . . . . . . . . . . . . . . 9 9.Male agellum almost cylindrical, female apical agellomere at base half as broad as scape at distal end . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A. dahlii (Meigen 1830) Male agellum distinctly swollen in middle, female apical agellomere broader, at most slightly narrower than scape at distal margin (Figs. 17 and 18) . . . . . . . . . . . . . . . .A. cinerascens (Loew 1873) Redescription of the 4 Adoxomyia species recorded in Turkey 1. Adoxomyia aureovittata (Bigot 1879); see F igs. 1-4, 15-16 and 22-23. Male : Head transversely oval, in dorsal view. Eyes touc hing on frons. Hairs on eyes as long as pedicel, dense and black. Black postocular area swollen in lower half and narrowed in upper part, covered with appressed yellowish hairs. Face, cheeks and posteroventral part of head covered with hairs as long as scape, erect, black and partly brown. Antenna entirely black and more slender in male than in female. Scape and pedicel black with black and erect hairs being as long as scape. Flagellum about four times as long as scape and pedicel combined, the rst 5 agellomeres with dense yellowish pubescence. Scutum black, with semi-erect and black hairs. Scutellum

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66 Florida Entomologist 94(1) March 2011 and scutellar spines mainly black but tip of scutellar spines brown. Scutellum covered with semi-appressed sparse black hairs. Legs black and yellow. Coxa, trochanter, and femur black except for yellow bases of femora. Tibia mainly black, its both ends narrowly yellowish brown. Tarsi yellow, front tarsus darkened dorsally as well as tarsomeres 3-5 of mid and hind legs. Coxa, trochanter, and femur with semi-erect black hairs. Tibia covered with appressed yellowish hairs. Tarsi with appressed golden yellowish hairs. Abdomen entirely black, abdominal pile black except for golden yellow lateral markings on tergite 4 and a transverse, golden yellow band on tergite 5. Female : Eyes densely black haired, hairs only one-fourth as long as pedicel. Black postocular area approximately as wide as fore tibia and covered with appressed yellowish hairs. Frons about 1/3 of head width, with ne longitudinal groove in middle, black, densely punctate, with yellowish hairs. Face black, with yellowish hairs on sides below antennae. Remainder of face, cheeks and posteroventral part of head covered with erect, black hairs being as long as scape. Antenna inFigs. 1-14. Adoxomyia aureovittata: 1male in dorsal view, 2female in dorsal view, 3male in lateral view, 4female in lateral view; 5-8 Adoxomyia cineracens 5male in dorsal view, 6female in dorsal view, 7male in lateral view, 8female in lateral view; 9-12 Adoxomyia obscuripennis 9male in dorsal view, 10female in dorsal view, 11male in lateral view, 12female in lateral view; 13-14 Adoxomyia sarudnyi 13male in dorsal view, 14male in lateral view (Scale 1 mm).

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stner & Hasbenli: First Records of Adoxomyia Species in Turkey67 serted at middle of head prole, partly black. Scape black but reddish brown at tip. Pedicel and rst 5 agellomeres dark red, last 3 agellomeres black. Scape and pedicel with erect, black hairs as long as antenal scape. Flagellum about 4.5 times as long as both basal antennal segments combined, rst 5 agellomeres densely golden yellow dusted. Thorax black, scutum with 2 golden yellow dusted longitudinal stripes which can be reduced to absent in some specimens (especially smaller ones). Scutellum black with appressed golden yellow hairs. Tips of scutellar spines brown. Legs black and yellow, femora black except for yellow base, tibiae mainly darkened, with both ends broadly yellowish brown on fore and mid legs. Hind tibia mainly black, with both ends broadly yellowish brown. Tarsi yellow, with dorsally darkened tarsomeres 3-5. All legs with yellowish hairs that are semierect on femora and appressed on tibiae and tarsi. Abdomen black but lateral markings on tergites 2-4 and a triangular apical spot on tergite 5 with golden yellow hair patches. Adoxomyia aureovittata (Bigot 1879) was described from an unknown locality Apparently, this species is distributed in the eastern part of the Mediterranean area. In addition to Turkey it was also found in Greece (M. Hauser, personal communication). Material Examined : Turkey: Konya, Hadim, between Tosmur and Gevne Village, Gevne Valley, 1450-2020 m, 10 June 1999, 1 male and 1 female; Konya, Taskent, Begreli Village, Gevne Valley, 1570 m, 10 August 2001, 1 male; all T. stner leg Distribution : Greece, Turkey. 2. Adoxomyia cinerascens (Loew 1873); see Figs 5-8, 17-18 and 24-25. Male : Hairs above compound eyes as long as pedicel, densely black. Postocular area black, covered with pale yellowish, appressed hairs. Frontal triangle black with dense, erect, pale yellowish hairs being about 1.5 times as long as the scape. Hairs on black face erect, as long as pedicel, pale yellowish. Antenna as long as head in lateral view. Scape, pedicel and rst 3 basal agellomeres brownish orange, rest of agellum black. Hairs on scape and pedicel erect, as long as scape, pale yellowish. Thorax including scutellum black, with dense yellowish hairs. Scutellar spines yellow. Wing veins brown. Legs bicoloured, coxae black, trochanters brownish, femora and tibiae mainly black, partly yellow at tips. Fore and hind tarsi yellowish on inner surface and darkened on outer surface, mid tarsi yellow but basal 2 tarsomeres darkened on outer surface. Femora with sparse semi-erect pale yellow hairs. Tibiae and tarsomeres with dense appressed yellow hairs. Abdomen mainly black, with transverse, pale yellow hair band on posterior margin of tergite 4. Female : Hairs on eyes black, about 0.3 times as long as pedicel. Postocular area black, covered with pale yellow and semi-appressed hairs. Frons about 0.3 of head-width, shining black and densely punctate, with ne longitudinal groove in middle and with sparse pale yellow hairs. Face black, with whitish, dense, erect hairs being as long as pedicel. Antenna long, about 1.5 times as long as head in lateral view, bicolored and in male more slender than in female. Scape brownish orange on lower surface but darkened on upper surface. Pedicel and rst 3 agellomeres brownish orange, rest of agellum black. Flagellomeres 2-3 darkened on outer surface. Thorax black, with appressed dense pale yellow hairs, but tip of postpronotal callus brownish. Scutellum black with pale yellow hairs, scutellar spines yellow. Wing veins brown. Legs bicoloured. Coxae black, trochanters yellow. Femora and tibiae mainly black and partly yellow on tips. Fore and hind tarsi yellowish on inner surface, darkened on outer surface, basal 2 tarsomeres of mid tarsi yellow, other tarsomeres darkened on outer surface. Femora with semi-erect pale yellow hairs. Tibiae with appressed yellowish hairs. Tarsomeres with appressed yellow hairs. Abdomen mainly black but with pale yellow hair patches at posterior margin of tergite 4. Material Examined : Turkey: Antalya, Gndogmus district, Gneycik Village, Topraktepe place, elev. 200 m, 23 June 1999, 7 males, 12 July 2000, 1 male; Konya, Taskent, Begreli Village, Gevne Valley, elev. 1585 m, 10 July 2000, 1 female; all T. stner leg. Distribution : Palaearctic: Iran, Israel, Kazakhstan, Kyrgyzstan, Tajikistan (Kertsz 1908; Lindner 1937, 1974, 1975; Rozkosny & Nartshuk 1988; Woodley 2001). This is the rst record for the fauna of Turkey. 3. Adoxomyia obscuripennis (Loew 1873); Figs 912, 19-20 and 28-29. Male: Head transverse, hemispherical, eyes touching on frons. Hairs on eyes dense, black, as long as scape. Frontal triangle black, with silverish white pubescence. Face slightly produced in lateral view, with dense, black hairs as long as scape Cheeks and posteroventral part of head with erect black hairs. Black postocular area prominent but narrower than pedicel in upper half and somewhat swollen in lower half, about as wide as antennal scape is long, covered with appressed silverish white hairs. Antenna entirely black, agellomeres 1-3 thickened, following agellomeres small and slender, last agellomere longer than 4 preceding combined. Thorax completely black. Scutum covered with long erect

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68 Florida Entomologist 94(1)March 2011 Figs. 15-21. Adoxomyia Antennae: 15A. aureovittata male, 16A. aureovittata female, 17A. cineracens male, 18A. cineracens female, 19A. obscuripennis male, 20A. obscuripennis female, 21A. sarudnyi male (Scala 1 mm.): 22-23 Adoxomyia aureovittata male terminalia: 22dorsal part of male genitalia, 23ventral part of male genitalia; 24-25 Adoxomyia cineracens male terminalia: 24dorsal part of male genitalia, 25ventral part of male genitalia; 26-27 Adoxomyia sarudnyi male terminalia: 26dorsal part of male genitalia, 27ventral part of male genitalia; 2829 Adoxomyia obscuripennis male terminalia: 28dorsal part of male genitalia, 29ventral part of male genitalia (Scale 0.1 mm.).

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stner & Hasbenli: First Records of Adoxomyia Species in Turkey69black hairs and short appressed silverish white hairs. Scutellum with very strong and thick, black scutellar spines. Legs black, but knees brown. Femora with semi-erect silverish white hairs. Tibiae with semi-appressed, dense, silverish white hairs. Tarsomeres black on outer surface, brown on inner surface, with semi-appressed, dense, silverish white hairs. Halteres pale yellow. Abdomen entirely black, tergites 1-2 with erect, moderately long, white hairs. White hairs also distinct on distal half of tergite 4 and on entire tergite 5. Female: Hairs above compound eyes dense and black, as long as antennal scape. Frons black, as wide as agellum is long, with ne median groove. Frontal hairs as long as antennal scape, pale. Face black, covered below antennae with long black hairs. Cheeks and posteroventral part of head with erect, black and yellowish hairs. Postocular area as wide as agellum, black and with appressed, dense, silverish white hairs. Antenna slender, about 1.1 times as long as head. Scape, pedicel and agellomeres 4-7 shining black, three basal agellomeres black, whitish grey dusted. Last agellomere about 1.5 times as long as 4 preceding. Thorax black, covered with appressed, silverish white hairs. Scutellum, including strong and short scutellar spines, black. Top of postpronotal callus and postalar callus brownish. Legs black but knees brown, femora with semierect white hairs, tibiae with semi-appressed, dense, silverish white hairs. Tarsomeres black on outer surface, brown on inner surface, with semiappressed, dense, silverish white hairs. Halteres pale yellow. Wings transparent and partly brownish, with brown veins. Abdomen entirely black, with erect, moderately long, silverish white hairs on sides of tergites 2-3. White hairs also developed on distal half of tergite 4 and on entire tergite 5. Material Examined: Turkey: Isparta, Yalva, The Sultan Mountains, elev. 1660 m, 29 May 2001, 1 male, 2 females, T. stner leg. Distribution : Palaearctic: Azerbaijan, Kazakhstan, Russia, Tajikistan, Uzbekistan (Kertsz 1908, 1923; Lindner 1937; Nartshuk 2004; Pleske 1925; Rozkosny 1983; Rozkosny & Nartshuk 1988; Woodley 2001). Adoxomyia obscuripennis (Loew 1873) is recorded from Turkey for the rst time. 4. Adoxomyia sarudnyi (Pleske 1903); Figs 13-14, 21 and 26-27. Male: Head transverse, hemispherical. Eyes touching on frons, dense, black eye hairs about 0.3 times as long as scape. Frontal triangle shining black, with ne median groove and white pile in upper part. Face, cheeks and posteroventral part of head black with whitish hairs. Postocular area black, about 0.4 times as wide as length of antennal scape, somewhat swollen in lower half, about 0.75 times as long as scape, covered with dense, silverish white hairs. Antenna relatively long, about twice as long as head in lateral view. Scape and pedicel orange but darkened on outer surface, with strong, erect, black hairs. First 3 agellomeres orange on inner surface but darkened on outer surface, rest of agellum black. Thorax black, with short appressed yellowish golden hairs and long semi-appressed black hairs intermixed. Tops of postpronotal callus reddish brown. Scutellum black and scutellar spines yellow. Wings transparent brownish, basal wing veins bright orange and distal veins brown, wing tip much darker and contrasting to clear wing base. Legs entirely bright yellow to orange except for black coxae. Fore tarsi yellowish on inner surface, darkened on outer surface. Mid and hind tarsi yellow, but last three tarsomeres darkened on outer surface. Femora with semi-erect, sparse, yellowish hairs. Tibia with appressed, dense, yellowish hairs. Tarsi with thick, adpressed, dense, yellowish hairs. Abdomen black with posterolateral, silverish white lateral markings on tergirtes 3-4. Material Examined: Turkey: Konya, Taskent, Begreli Village, Gevne Valley, elev. 1570 m, 1 July 2001, 1 male, T. stner leg. Distribution : Palaearctic: Afghanistan, Iran (Kertsz 1908, 1923; Lindner 1937; Pleske 1925; Rozkosny & Nartshuk 1988; Woodley 2001). DISCUSSIONThe genus Adoxomyia was previously unknown from Turkey. This fact is fairly surprising because many species were recorded from adjacent countries and are known from southern, often arid parts of the Palaearctic region. Therefore it was to be expected that at least some species of this genus would be found in Turkey as well. That is why in this report we constructed an actual identication key to the all East-Mediterranean species. Due to intense collecting efforts in many different localities in Turkey, 4 species of this genus were collected. The most remarkable record is Adoxomyia aureovittata which was described as Euparyphus aureovittatus by Bigot in 1879 from an unknown locality. The record from Turkey represents the rst evidence that it is a Palaearctic species and an unpublished record from Greece (Hauser, personal communication) conrms that this species probably has an EastMediterranean distribution. Adoxomyia cineracens is distributed in Transcaspia, in the Near East (Iran, Israel) and Central Asia. The type locality is Kizilkum (Kazakhstan). Our record thus closes the distribution gap between the known records from Israel and the type locality.

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70 Florida Entomologist 94(1)March 2011Our record of Adoxomyia obscuripennis represents the most southern and western point of its range and the second evidence of its occurrence in the western part of Asia (cf. a record from Azerbaijan in Nartshuk 2004). Adoxomyia sarudnyi was only known from Afghanistan and Iran. Our record in Turkey represents the most western locality of this very rare species. ACKNOWLEDGMENTSOur thanks to Dr. Martin Hauser and Prof. Dr. R. Rozkosny for critically reviewing the manuscript.REFERENCES CITEDDUSEK, J., AND ROZKOSNY, R. 1963. Revision mitteleuropischer Arten der Familie Stratiomyidae (Diptera) mit besonderer Bercksichtigung der Fauna der CSSR I. Acta Societatis Entomologicae Cechosloveniae 60(3): 202-221. HAUSER, M. 2002. A new species of Adoxomyia Kertsz, 1907 (Diptera: Stratiomyidae) from Socotra, Yemen. Fauna of Arabia 19: 463-466. JAMES, M. T., AND MCFADDEN, M. W. 1969. The genus Adoxomyia in America North of Mexico (Diptera: Stratiomyidae). Journal of the Kansas Entomological Society 42: 260-276. KERTSZ, K. 1907. Eine neuer Dipteren-Gattungsname. Annalen Historico-Naturales Musei Nationalis Hungarici 5(2): 499. KERTSZ, K. 1908. Catalogus dipterorum hucusque descriptorum. Volumen III. Stratiomyiidae, Erinnidae, Coenomyidae, Tabanidae, Pantophthalmidae, Rhagionidae. Museum Nationale Hungaricum, Budapestini, 366 pp. KERTSZ, K. 1923. Vorarbeiten zu einer Monographie der Notacanthen. XLV-L. Annales Historico Naturales Musei Nationalis Hungarici, 20: 85-129. LINDNER, E. 1937. 18. Stratiomyiidae [part]. Lieferung 114. Pp. 145-176. In Lindner, E. [ed.], Die Fliegen der palaearktischen Region. E. Schweizertsche Verlagsbuchhandlung, Stuttgart, 218 pp. LINDNER, E. 1974. On the Stratiomyidae (Diptera) of the Near East. Israel J. Entomol. 9: 93-108. LINDNER, E. 1975. On some Stratiomyidae (Diptera) from the Near East. Israel J. Entomol. 10: 41-49. MCFADDEN, M. W. 1967. Soldier y larvae in America north of Mexico. Proc. U.S. Natl. Mus. 121: 1-72. NARTSHUK, E. P. 2004. New data on Adoxomyia Bezi from the Caucasus and Eastern Europe (Diptera: Stratiomyidae). Zoosystematica Rossica 12: 263-266, St. Petersburg. UCHI, Y. 1938. On some stratiomyiid ies from eastern China. J. Shanghai Sci. Inst., Section III 4: 37-61. PLESKE, T. 1925. tudes sur les Stratiomyiin de la rgion palarctiqueIII.Revue des espces palarctiques de la sous-famille des Clitellariinae. Encyclopdie Entomologique, Srie B (II), Diptera 1(3-4): 105-119, 165-188. ROZKOSNY, R. 1983. A Biosystematic Study of the European Stratiomyidae (Diptera), Volume 2: Clitellariinae, Hermetiinae, Pachygasterinae and Bibliography. Dr. W. Junk, The Hague, Boston, London, 431 pp. ROZKOSNY, R., AND NARTSHUK, E. P. 1988. Family Stratiomyidae, pp. 42-96 In A. Sos and L. Papp [eds.], Catalogue of Palaearctic Diptera. Volume 5. Athericidae-Asilidae. Akadmiai Kiad Budapest, 446 pp. STNER, T., AND HASBENLI, A. 2003. First record of the subfamily Beridinae (Diptera: Stratiomyidae) from Turkey. Studia Dipterologica 10(1): 186-188, Halle (Saale). STNER, T., HASBENLI, A., AND AKTMSEK, A. 2002. Contribution to subfamily 360 Clitellariinae (Diptera, Stratiomyidae) in Fauna of Turkey. J. Entomol. Res. Soc. 4(1): 19-24. STNER, T., HASBENLI, A., AND ROZKOSNY, R. 2003. First record of Hermetia illucens (Linnaeus, 1758) (Diptera, Stratiomyidae) from the Near East. Studia Dipterologica 10(1): 181-185, Halle (Saale). STNER, T., AND HASBENLI, A. 2004. A new species of Oxycera Meigen (Diptera: Stratiomyidae) from Turkey. Entomol. News 115(3): 163-167. WOODLEY, N. E. 2001. A World Catalog of the Stratiomyidae (Insecta: Diptera). Backhuys Publishers, Leiden, 474 pp.

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Wiesenborn: Nitrogen Contents in Riparian Arthropods 71 NITROGEN CONTENT IN RIPARIAN ARTHROPODS IS MOST DEPENDENT ON ALLOMETRY AND ORDER W ILLIAM D. W IESENBORN U.S. Bureau of Reclamation, Lower Colorado Regional Ofce, P.O. Box 61470, Boulder City, NV 89006 A BSTRACT I investigated the contributions of body mass, order, family, and trophic level to nitrogen (N) content in riparian spiders and insects collected near the Colorado River in western Arizona. Most variation (97.2%) in N mass among arthropods was associated with the allometric effects of body mass. Nitrogen mass increased exponentially as body dry-mass increased. Signicant variation (20.7%) in N mass adjusted for body mass was explained by arthropod order. Adjusted N mass was highest in Orthoptera, Hymenoptera, Araneae, and Odonata and lowest in Coleoptera. Classifying arthropods by family compared with order did not explain signicantly more variation (22.1%) in N content. Herbivore, predator, and detritivore trophic-levels across orders explained little variation (4.3%) in N mass adjusted for body mass. Within orders, N content differed only among trophic levels of Diptera. Adjusted N mass was highest in predaceous ies, intermediate in detritivorous ies, and lowest in phytophagous ies. Nitrogen content in riparian spiders and insects is most dependent on allometry and order and least dependent on trophic level. I suggest the effects of allometry and order are due to exoskeleton thickness and composition. Foraging by vertebrate predators, such as insectivorous birds, may be affected by variation in N content among riparian arthropods. Key Words: nutrients, spiders, insects, trophic level, exoskeleton, cuticle R ESUMEN Se investiguo las contribuciones de la masa de cuerpo, orden, familia y el nivel trco al contenido de nitgeno (N) en araas e insectos riparianos (que viven en la orilla del rio u otro cuerpo de agua) recolectadaos cerca del Rio Colorado en el oeste del estado de Arizona. La mayora de la variacin (97.2%) en la masa (N) entre los artrpodos fue asociado con los efectos alomtricos de la masa de cuerpo. La masa de nitrgeno aument exponencialmente con el aumento de masa-seca del cuerpo. La variacin signicativa (20.7%) en la masa N ajustada por la masa del cuerpo se explica segn el ordn del artrpodo. La masa ajustada N fue mas alta en Orthptera, Hymenptera, Araneae, Odonata y mas baja en Coleoptera. Al clasicar los artrpodos por familia comparado con el ordn no explica la variacion mayor signicativa (22.1%) en el contenido de N. Los niveles trcos de los herbvoros, depredadores y detritvoros en todos los ordenes explica la pequea variacin (4.3%) en la masa N ajustada por la masa del cuerpo. Entre los ordenes, el contenido N vara solamente entre los niveles trcos de Diptera. El valor ajustado de la masa de N fue mayor para las moscas depredadores, intermedio para las moscas detritvoras y menor para las moscas tfagas. El contenido de nitrgeno en araas e insectos riparianos es mas dependiente sobre la alometra y ordn y menos dependiente sobre el nivel trco. Sugiero que los efectos de alometra y ordn son debidos al grosor y la composicin del exo-esqueleto. El forraje por los depredadores vertebrados, como aves insectivoras, puede ser afectado por la variacin del contenido N entre los artrpodos riparianos. Nitrogen concentrations in organisms are dependent on trophic level. This is most apparent between plants and herbivores, because N comprises 0.03-7% of dry mass in plants compared with 8-14% in animals (Mattson 1980). Variation in N concentration among and within plants, and its effects on abundances of herbivores including arthropods, especially agricultural pests, has been frequently examined (reviewed in Mattson 1980; Scriber 1984). Fewer studies have considered variation in N concentration among spiders and insects. Bell (1990) and Studier & Sevick (1992) tabulated measurements of %N in various insects from different studies. Fagan et al. (2002) compared %N between arthropod herbivores and predators by analyzing data compiled from various sources. Concentrations of N in spiders and insects were dependent on trophic level after controlling for body length, representing allometry, and taxonomic group, representing phylogeny (Fagan et al. 2002). Predators generally contained higher %N than herbivores. Predaceous arthropods may concentrate N from food similar to phytophagous arthropods. Variation in N concentration among spiders and insects may affect foraging by arthropodfeeding vertebrates and the qualities of food they obtain. Diet protein has been implicated as affecting egg production (Ramsay & Houston 1997) and nestling growth (Johnston 1993) in insectivorous

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72 Florida Entomologist 94(1) March 2011 birds. Identifying sources of variation in arthropod N content may improve our understanding of the prey composition required to support species of insectivorous wildlife. I examined variation in N content among spiders and insects collected from trees and shrubs established to restore riparian habitat for insectivorous vertebrates, especially birds. Variation in N mass was partitioned into various sources. I rst determined the allometric relationship between N mass and body dry-mass. After adjusting N mass for this relationship, N contents of arthropods were compared among orders and families and among trophic levels across and within orders. I interpreted N contents in relation to exoskeleton scaling and chemical composition and concluded by applying the results to diets of insectivorous birds. M ATERIALS AND M ETHODS Arthropod Collections Spiders and insects were collected next to the Colorado River within Havasu National Wildlife Refuge in Mohave County, Arizona. Most arthropods were collected at an irrigated 43-ha riparian restoration area (34N, 114W; elevation 143 m) of planted or volunteer trees and shrubs 12 km southeast and across the river from Needles, California. Plots were planted during 20032005 with cuttings that were taken from nearby areas along the river and rooted in containers. The area is straddled by Topock Marsh (16 km 2 ) and Beal Lake (0.9 km 2 ), 2 impoundments containing mostly emergent cattails ( Typhus sp., Typhaceae) and open w ater. Undeveloped areas of the surrounding oodplain support mostly naturalized tamarisk ( Tamarix ramosissima Ledeb., T amaricaceae) shrubs. The oodplain is anked by Sonoran desertscrub dominated by creosote bush ( Larrea tridentata (DC.) Cov., Zygophyllaceaae). Maximum temperatures average 42.7C during Jul, and minimum temperatures average 5.6C during Jan at Needles (DRI 2010). I collected arthropods from plants and trapped insects in ight. Arthropods were swept with a 38-cm diameter muslin net from planted cottonwood ( Populus fremontii S. Watson, Salicaceae) and Goodding s black willow ( Salix gooddingii C. Ball, Salicaceae) trees, planted narrow-leaved willow shrubs ( Salix exigua Nutt.), volunteer honey mesquite ( Prosopis glandulosa Torrey, Fabaceae) and screwbean mesquite ( Prosopis pubescens Benth.) trees, and volunteer arrowweed shrubs ( Pluchea sericea (Nutt.) Cov., Asteraceae). I also swept arthropods from T. ramosissima bordering the plots Additional arthropods on S. exigua were swept from plants growing along a dirt irrigation canal 2 km northwest of the restoration area. Plant species were swept separately except for Prosopis spp., which grew together. Each species w as swept 10-15 min on 9 dates: 30 Apr, 14 May, 27 May, 08 Jun, 22 Jun, 30 Jun, 21 Jul, 4 Aug, and 18 Aug 2009. All plant species were in ower or fruit except for P. fremontii Arthropods swept from plants were placed into plastic bags kept in a refrigerator, and killed in a freezer. Flying insects were trapped with a Malaise trap (Santee Traps, Lexington, KY) that was placed in the center of a plot supporting S. gooddingii and P. sericea and elevated 1 m aboveground with fence posts Trapped insects were collected into a dry plastic bottle containing a nitrogen-free, diclorvos insecticide strip. Insects were trapped for 6.1-7.3 h during 0855-1640 PDT on each of the above dates except 30 Apr, 14 May, and 18 Aug 2009. Spiders and insects collected on each date were sorted under a microscope into morphotypes (similar-looking specimens). Representatives of each morphotype were placed into 70% ethanol for identication. I counted and split the remaining specimens of each morphotype into samples each with an estimated maximum dry mass of 10 mg. Individual specimens with dry masses 10 mg were placed into separate samples Arthropod samples for N analyses were cleaned by vortexing in water, transferred to lter paper with a Bchner funnel, dried 2 h at 80C, and stored in stoppered vials. Arthropod Identications and Trophic Levels Spiders and insects were identied to the lowest taxon possible, at least to family and typically to genus. Vouchers of adult insects were deposited at the Bohart Museum of Entomology, University of California, Davis, and vouchers of spiders were deposited at the California Academy of Sciences, San Francisco. Arthropod taxa were classied into the trophic levels of herbivore, predator, and detritivore based on published descriptions (Table 1). Holometabolous insects were classied by larval diet. Herbivores included consumers of pollen, nectar, or honeydew (homopteran egesta). Predators included parasites and consumers of already-dead animals. Arthropod Nitrogen Estimates The mass of N in each arthropod sample was estimated with the Kjeldahl method adapted from Isaac & Johnson (1976). Samples of dried arthropods were weighed (.01 mg) with a microbalance (model C-30, Cahn Instruments, Cerritos, CA) and ground into water with a 5-mL glass tissue homogenizer. Homogenized samples were poured and rinsed with water, to a total volume of 20 mL, into 100-ml digestion tubes. I added 6 mL of concentrated sulfuric acid, containing 4.2% selenous acid, and 3 mL of 30% hydrogen

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Wiesenborn: Nitrogen Contents in Riparian Arthropods 73 T ABLE 1. A DULT ARTHROPODS COLLECTED FROM RIPARIAN HABITAT NEAR THE C OLORADO RIVER IN A RIZONA AND ANALYZED FOR NITROGEN CONTENT Order or suborderFamily Genus 1 Source 2 No. Samples No. specimens per sampleTrophic level 3 Mean body dry mass (mg) Mean SD % N Araneae Philodromidae Philodromus E,S2 3-4 P 1.93 10.6 0.9 Salticidae Habronattus S 2 1 P 6.29 9.3 1.2 Metaphidippus 6 S 1 9 P 0.07 13.0 Thomisidae Misumenops E 2 1-2 P 2.03 12.1 1.8 2 families 4,6 S 1 6 P 2.46 14.3 3 families 5,6 S 2 6-7 P 2.35 13.8 0.1 OdonataLibellulidae Pachydiplax P 4 1 P 39.7 12.3 1.0 OrthopteraAcrididae Acridinae 7 S 6 1-3 H 13.0 13.9 2.7 Tettigoniidae Scudderia S 1 1 H 115.0 14.6 HeteropteraLargidae Largus S 1 1 H 49.2 9.2 Lygaeidae Nysius S 1 67 H 0.46 9.0 Pentatomidae Brochymena F,G,P4 1 H 55.2 11.0 1.5 Thyanta E 1 1 H 17.1 11.6 Reduviidae Pselliopus P 1 1 P 14.1 13.3 Zelus F,P,S6 1-3 P 7.20 10.5 2.0 HomopteraCicadellidae Cicadellinae E,F,G5 1-3 H 6.62 10.1 2.2 Gyponinae G 1 2 H 3.36 8.6 Opsius 6 T 4 28-41 H 0.68 11.2 1.5 Typhlocybinae F 2 19-22 H 0.35 11.4 0.0 S 1 5 H 4.37 14.6 Cixiidae Oecleus S 1 4 H 1.24 10.1 Flatidae Ormenis G,T2 2 H 5.72 8.9 1.2 Membracidae G 1 2 H 5.22 10.6 NeuropteraChrysopidae Chrysoperla F,G,S9 2-14 P 1.51 9.1 1.5 G 1 11 P 1.37 11.8 Myrmeliontidae Myrmelion F 1 1 P 8.99 12.5 1 Subfamily in Acrididae and subfamily or genus in Cicadellidae. 2 E, Salix exigua ; F, Populus fremontii ; G, Salix gooddingii ; M, Malaise trap; P, Prosopis glandulosa or P. pubescens ; S, Pluchea sericea ; T, Tamarix ramosissima 3 D, Detritivore; H, Herbivore; P, Predator. Reference for all (Borror et al. 1981) except Apioceridae (Cole 1969) and Andrenidae, Formicidae, and Tettigoniidae (Essig 1926). 4 Salticidae, Habronattus sp.; Thomisidae, Misumenops sp. 5 Araneidae, Hypsosinga sp.; Salticidae, Metaphidippus sp. & Habronattus sp.; Thomisidae, Misumenops sp. 6 Adults and immatures. 7 Immatures.

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74 Florida Entomologist 94(1) March 2011 ColeopteraBruchidae Algarobius P 1 6 H 3.01 8.3 Coccinellidae Chilocorus F,P 3 2-4 P 4.75 9.8 1.2 Hippodamia F,S 3 2-8 P 6.26 6.6 2.8 DipteraApioceridae Apiocera M 1 1 P 52.87 11.4 Asilidae Proctacanthus M 1 1 P 42.3 11.7 Dolichopodidae Asyndetus M 13 17-113 D 0.39 9.9 2.0 Lauxaniidae Homoneura F,G2 4-5 D 1.31 7.8 1.0 Minettia F,G2 2-6 D 2.37 8.1 4.6 Sarcophagidae Eumacronychia F,G1 2 P 1.68 11.5 Tabanidae Apatolestes M 1 1 P 15.0 11.6 Tabanus M 13 2-3 P 13.8 10.9 2.2 Tachinidae Zaira M 2 1-2 P 7.66 9.2 2.3 Tephritidae Acinia F 2 7-9 H 1.01 5.1 1.5 HymenopteraAndrenidae Perdita S 1 2 H 1.74 9.8 Formicidae Formica E,S4 6-16 H 0.76 10.9 1.8 Halictidae Agapostemon E 1 1 H 7.42 11.7 Dieunomia S 1 3 H 5.57 14.1 Lasioglossum E 1 9 H 2.71 16.7 Sphecidae Bembix M 1 1 P 33.5 13.4 Cerceris M 1 1 P 10.6 8.8 Tachysphex M 1 1 P 7.23 8.5 Tiphiidae Myzinum E 1 6 P 4.54 21.2 Vespidae Polistes G 1 1 P 28.8 14.0 TABLE 1. (CONTINUED) ADULT ARTHROPODS COLLECTED FROM RIPARIAN HABITAT NEAR THE COLORADO RIVER IN ARIZONA AND ANALYZED FOR NITROGEN CONTENT. Order or suborderFamily Genus1Source2No. Samples No. specimens per sampleTrophic level3Mean body dry mass (mg) Mean SD % N1Subfamily in Acrididae and subfamily or genus in Cicadellidae.2E, Salix exigua; F, Populus fremontii ; G, Salix gooddingii; M, Malaise trap; P, Prosopis glandulosa or P. pubescens ; S, Pluchea sericea ; T, Tamarix ramosissima.3D, Detritivore; H, Herbivore; P, Predator. Reference for all (Borror et al. 1981) except Apioceridae (Cole 1969) and Andrenidae, Formicidae, and Tettigoniidae (Essig 1926).4Salticidae, Habronattus sp.; Thomisidae, Misumenops sp.5Araneidae, Hypsosinga sp.; Salticidae, Metaphidippus sp. & Habronattus sp.; Thomisidae, Misumenops sp.6Adults and immatures.7Immatures.

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Wiesenborn: Nitrogen Contents in Riparian Arthropods 75peroxide and heated samples 1 h at 400C with a block digestor (model 2040, Tecator, Herndon, VA). After cooling, water was added to 60 mL. The ammonia concentration formed in the clear, digested samples was measured by colorimetry, against standards prepared from dried ammonium-sulfate, with a segmented flow analyzer (model FS-4, OI Analytical, College Station, TX). Salicylate, hypochlorite, and sodium nitroprusside were used as the indicator. I converted ammonia concentration to mg N. I adjusted estimates of mg N in arthropod samples with chitin samples containing known N masses. Chitin is a nitrogenous polysaccharide (C8H13NO5)n abundant in arthropod exoskeleton, or cuticle (Neville 1975), that typically comprises 25-40% of exoskeleton dry-mass in insects (Richards 1978). Various masses (2, 4, 8, 16, 32, 64 mg) of powdered chitin (Tokyo Chemical Industry) containing 6.89% N were weighed, placed in 20 mL water, digested, and measured for ammonia within each batch ( n = 4) of arthropod samples. I increased estimates of mg N in arthropod samples in each batch to correct for the batchs mean underestimate of %N (5.76, 6.23, 6.44, 6.08%) in chitin samples. I calculated %N in arthropod samples as 100(mg N/mg dry mass). Two arthropod samples of Acinia and Chrysoperla with unusually low N concentrations (<0.9%) were excluded as outliers. Dry mass and mg N of each arthropod sample were divided by the number of specimens in the sample to estimate dry mass and N mass per specimen.Statistical AnalysisBody masses of arthropods, transformed log(mg) to normalize residuals, were compared among trophic levels with analysis of variance (SYSTAT version 12, San Jose, CA). Nitrogen masses in spiders and insects were analyzed sequentially. I first determined the relationship between N mass and body dry mass by regressing log(mg N) against log(mg body mass) for each arthropod sample. I verified that the relationship was allometric (exponential) by testing with an approximate t test the null hypothesis that the regression coefficient b1 = 1 (Neter et al. 1996). Transformed N masses were adjusted for their allometric relationship with transformed body mass by adding the residuals from the regression to the overall mean of transformed N mass (Sokal & Rohlf 1981). Adjusted, transformed N masses were compared among arthropod orders with analysis of variance. Hemiptera were split into suborders Heteroptera and Homoptera, because the digestive systems of most homopterans have lter chambers that concentrate nitrogenous compounds (Borror et al. 1981). I tested if classifying arthropods by family instead of order or suborder explained more variation in adjusted log(mg N) with the general linear test approach (Neter et al. 1996). This approach tests if mean square error in an analysis of variance decreases signicantly when the model becomes more complex. Samples containing more than 1 family (3 samples of Araneae, or spiders) were classied only to order. Arthropod N-contents adjusted for body mass were compared among trophic levels across and within orders or suborders. I compared N masses among trophic levels across orders or suborders with analysis of variance. Separate analyses were performed within Heteroptera, Diptera, and Hymenoptera, the 3 orders or suborders with 2 or more trophic levels each containing more than 1 sample. Analyses within orders or suborders weighted adjusted values of log(mg N) by 1/s2 in each trophic level to correct for uneven variances among trophic levels (Neter et al. 1996). RESULTSCollected ArthropodsI collected 121 samples of spiders and insects containing 1,490 specimens in 9 orders or suborders, 33 families, and 43 subfamilies or genera (Table 1). All of the arthropods collected were adults except for 8 samples in 3 taxa (families, subfamilies, or genera) with adults and immatures and 6 samples in 1 taxon with only immatures. Body dry-masses of adult arthropods ranged from 0.35 mg in Typhlocybinae leafhoppers (Cicadellidae) to 115 mg in the fork-tailed bush katydid Scudderia furcata Brunner (Tettigoniidae). Two orders or suborders (Orthoptera and Homoptera) of collected spiders and insects were only herbivorous, 3 orders (Araneae, Odonata, and Neuroptera) were only predaceous, and 4 orders or suborders (Heteroptera, Coleoptera, Diptera, and Hymenoptera) included both trophic levels. All Coleoptera were predaceous except for 1 sample. The only detritivores collected were ies (Diptera). Across orders or suborders, herbivores included 42 samples in 22 taxa, predators included 62 samples in 24 taxa, and 17 samples in 3 taxa were detritivores (Table 1). Trophic levels contained arthropods with different body drymasses (F = 25.5; df = 2, 118; P < 0.001). Predators were largest (back-transformed mean = 6.37 mg) followed by herbivores (4.03 mg) and detritivores (0.55 mg).Allometric Nitrogen ContentsNitrogen mass in riparian spiders and insects was allometrically related to body dry mass (Fig. 1). Transformed N mass per specimen in arthropod samples was positively related ( F = 4, 066; df = 1, 119; P < 0.001) to transformed body drymass per specimen by:

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76 Florida Entomologist 94(1)March 2011log mg N = -1.006 + 1.039(log mg dry mass) Back-transforming this equation produced: mg N = 0.0986(mg dry mass)1.039The exponent (1.039 0.016 SD) differs from unity (t* = 2.43; df = 119; P = 0.008), verifying that the relationship is exponential rather than linear. This allometric relationship explained 97.2% of variation in N mass. Percentage of N in riparian arthropods (Table 1) increased as body mass increased.Nitrogen Content in Arthropod OrdersNitrogen mass adjusted for body mass in riparian arthropods (Fig. 2) differed (F = 3.64; df = 8, 112; P < 0.001) among orders or suborders. These taxonomic levels explained 20.7% of variation in adjusted N mass. Orthoptera (mean 14.0% N), Hymenoptera (12.4% N), Araneae (11.9% N), and Odonata (12.3% N) contained the highest adjusted N contents, and Coleoptera (8.2% N) contained the lowest adjusted N content. Orthoptera were mostly immature slant-faced grasshoppers Fig. 1. Mean N mass vs mean body dry-mass in riparian arthropods from the lower Colorado River classied by family. Abbreviations are orders or suborders (in Hemiptera): A, Araneae; C, Coleoptera, D, Diptera; He, Heteroptera; Ho, Homoptera; Hy, Hymenoptera; N, Neuroptera; Od, Odonata; Or, Orthoptera. Single point labeled Araneae represents mixed samples of Araneidae, Salticidae, and Thomisidae. Axes are log scales. Line t to transformed data by linear regression weighted by sample size.

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Wiesenborn: Nitrogen Contents in Riparian Arthropods 77 (Acridinae) along with the sole katydid S. furcata. Hymenoptera included ants (Formicidae), 2 families of bees (Andrenidae and Halictidae), and 3 families of wasps (Sphecidae, Tiphiidae, and Vespidae). Spider samples contained several families (Table 1). The only odonate collected was the dragonfly Pachydiplax longipennis Burmeister. Coleoptera included 1 sample of the herbivorous seed beetle (Bruchidae) Algarobius prosopis LeConte, collected from Prosopis spp., and 6 samples containing 2 species of predaceous ladybird beetles (Coccinellidae), Chilocorus cacti L. and the widespread Hippodamia convergens GuerinMeneville. Insects in other orders, including the 2 Hemiptera suborders, contained intermediate N concentrations (Fig. 2). Classifying arthropods by family instead of or der or suborder did not explain more variation in N mass adjusted for body mass. Error variance of adjusted N mass did not decrease ( F = 1.45; df = 26, 86; P = 0.10) when arthropods were classied by family compared with order or suborder Classifying arthropods by family instead of order or suborder explained 22.1%, a 1.4% improvement, of variation in adjusted N mass. Nitrogen Content in Trophic Levels Differences in N content among the trophic levels of herbivore, predator, and detritivore depended on classification (Fig. 2). Across orders or suborders, N mass did not vary ( F = 0.62; df = 2, 118; P = 0.54) among trophic levels. Trophic levels explained 1.0% of variation in N mass after accounting for body mass. Back-transformed means of adjusted N mass (and mean % N) were 0.413 mg (11.1% N) in herbivores, 0.397 mg (10.9% N) in predators, and 0.380 mg (9.44% N) in detritivores, the smallest arthropods collected. Within orders or suborders, N mass varied among trophic levels in Diptera ( F = 4.60; df = 2, 35; P = 0.017) but not in Heteroptera ( F = 0.62; df = 1, 12; P = 0.45) or Hymenoptera ( F = 0.13; df = 1, 11; P = 0.91). Adjusted N contents in flies (Fig. 2) were lower in herbivores (mean 5.1% N) compared with predators (10.9% N) or detritivores (9.4% N). All phytophagous flies collected were 2 samples of the fruit fly (Tephritidae) Acinia picturata (Snow), swept from P. fremontii Adjusted N concentrations in predaceous or parasitic flies (Apioceridae, Asilidae, Sarcophagidae, Tabanidae, and Tachinidae) and detritivorous flies (Dolichopodidae and Lauxaniidae) were similar. D ISCUSSION Allometric Nitrogen Contents The allometric relationship between N mass and body mass in riparian arthropods resembles a similar relationship between exoskeleton mass and body mass in terrestrial arthropods. Anderson et al. (1979) dissected the exoskeletons from 3 species of immature and adult spiders, weighing between 25 mg and 1.2 g, and determined exoskeleton dry-mass and body wet-mass were positively related by: g exoskeleton = 0.078(g body mass) 1.135 Body mass in spiders explained 94.1% (their r value squared) of variation in exoskeleton mass Anderson et al. attributed this allometric relationship to scaling. The exoskeleton of terrestrial arthropods must increase in thickness as body weight increases to support the organism and withstand the stresses of bending and twisting (Prange 1977; Anderson et al. 1979). Allometric relationships between N mass and body mass, and between exoskeleton mass and body mass, may be primarily due to exoskeleton N. Trim (1941) estimated N concentrations of 11.8% in abdominal cuticles of 2 Orthoptera species, approximating the mean concentration (10.7%) in riparian arthropods. A large proportion of N in terrestrial arthropods likely resides within the exoskeleton due to its greater density compared with internal tissues and hemolymph. The allometric relationship between exoskeleton mass and body mass may have produced the similar relationship between N mass and body mass. Fig. 2. Nitrogen mass allometrically adjusted for body mass in riparian arthropods from the lower Colorado River classied by order or suborder (in Hemiptera). Letters are means ( SE) and trophic levels: D, detritivores; H, herbivores; P, predators. Adjacent numbers are sample sizes. Y -axis is log scale.

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78 Florida Entomologist 94(1)March 2011A linear increase in N mass in internal tissues as body mass increases would dampen the exponential increase in cuticular N mass. The lower exponent relating N mass to body mass (1.039) compared with the exponent relating cuticle mass to body mass (1.135) may reect this dampening.Nitrogen Contents in Orders or SubordersExoskeleton composition may have contributed to different N concentrations among orders of spiders and insects (Fagan et al. 2002). Arthropod cuticle is composed primarily of protein and chitin (Neville 1975), and concentrations of N are higher in the former. For example, I estimated %N in arthropod cuticular protein from percentages of amino acids in pronotal and abdominal cuticles of adult Tenebrio beetles (Andersen et al. 1973; reported in Table 3.4 in Neville 1975) by assuming the amino acids were bonded into polypeptides. The estimated N concentration of cuticular protein (17.4%) exceeded that of chitin (6.89%). Based on the maximum range of chitin concentration (10-60% of dry mass) in insect cuticle (Richards 1978; see also Table 1 in Hackman 1974), and assuming cuticle is entirely chitin and protein, N concentrations in insect exoskeleton may vary from 11.1% to 16.4%. Greater concentrations of protein in arthropod cuticle, producing higher N contents, have been associated with concentrations of resilin (Andersen 1979). Resilin is a exible, elastic protein that occurs in cuticle in near-pure concentrations or combined with other proteins and chitin (Richards 1978). I estimated as above that resilin contains 19.0% N from percentages of amino acids in resilin from Schistocerca grasshoppers (Andersen 1966; reported in Table 3.4 in Neville 1975). Various mechanical structures in arthropods are elastic due to resilin (Table 2.1 in Neville 1975). Resilin is especially prevalent in the wing tendons and hinges of Odonata and Orthoptera (Andersen & WeisFogh 1964), primitive orders with synchronous ight muscles. Andersen and Weis-Fogh also detected resilin in the abdominal sclerites of Schistocerca grasshoppers, presumably allowing the abdomen to stretch. Abundances of resilin in riparian Odonata and Orthoptera may have contributed to their high N contents. Although resilin has not been found in spiders (Andersen & Weis-Fogh 1964), the high degree of abdominal stretching by spiders (Browning 1942) suggests their cuticles contain a similar elastic protein. Cuticles of Coleoptera are likely less elastic. A dominant feature of beetles is the elytra, hardened front-wings that act only to cover the folded hind-wings and abdomen. The likely absence of resilin and resultant high concentrations of chitin, in elytra may have lowered %N in Coleoptera.Nitrogen Contents in Trophic LevelsI did not detect an overall difference in N concentration among herbivorous, predaceous, and detritivorous arthropods after accounting for the allometric effects of body mass. Trophic level did not appear to generally affect arthropod %N. This contradicts the overall difference in N concentration between herbivorous and predaceous arthropods detected by Fagan et al. (2002). Different results may have been due to statistical methodology. Fagan et al. controlled for body length and taxonomic group, to account for phylogeny, whereas I controlled only for body mass. Controlling for phylogeny is difficult, because different frequencies of herbivores compared with predators among taxonomic groups cause trophic level and phylogeny to be confounded. Phylogeny and trophic level cannot be statistically separated. Similar N contents between trophic levels agree with the concept that most insects satisfy nutrient requirements by adjusting food intake (Waldbauer 1968; reviewed in Simpson et al. 1995). An example in riparian arthropods may be found in the 2 suborders of Hemiptera, insects with piercing-sucking mouthparts. Phytophagous Heteroptera, such as Lygus leaf bugs (Backus et al. 2007), typically rupture, dissolve with saliva, and ingest mesophyll from a variety of plant structures. All Homoptera are herbivorous, and many homopterans feed on phloem which is high in water and carbohydrates but low in other nutrients including N. The Opsius stactogalus Fieber leafhoppers collected here increase food intake, concentrate nutrients within their lterchamber digestive tracts (Wiesenborn 2004), and void excess water and sugars. Concentrations of N in Homoptera, phytophagous Heteroptera, and predaceous Heteroptera were similar despite different diets and physiologies. An exception was Diptera. Herbivorous ies, all Tephritidae, contained lower N concentrations than predaceous or detritivorous ies after considering body mass. Fagan et al. (2002) compared phylogenetic categories of herbivorous insects and found lower N concentrations in Diptera and Lepidoptera, combined as the recent lineage Panorpida, after accounting for body length. The database analyzed by Fagan et al. included the herbivorous ies Bibionidae, Chloropidae, and Drosophilidae, each in a different superfamily separate from Tephritidae. The diversity of phytophagous Diptera found to contain low N concentrations suggests N contents in ies generally vary by trophic level. Fagan et al. (2002) suggested several explanations for lower N contents in herbivores than in predators. These included the direct effects of diet N, indirect effects of trophic niche unrelated to diet, and selection for low body N in response to low diet N. The A. picturata tephritids that I collected de-

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Wiesenborn: Nitrogen Contents in Riparian Arthropods 79velop as larvae in the ower heads of Pluchea spp. (Foote et al. 1993), corresponding with the owering P. sericea at the study site. Infestations by A. picturata reduce seed production (Alyokhin et al. 2001), suggesting larvae eat ovaries or seeds. The species does not appear to concentrate N from food, because its N concentration (5.1%) is within the range (1-7% of dry mass) reported for seeds (Mattson 1980). The structural or biochemical features correlated with low N concentration in A. picturata and other plant-feeding ies are unknown. Low exoskeleton mass in tropical, herbivorous beetles has been attributed to low diet N, short larval-development time, and high fecundity (Rees 1986). Equivalent N concentrations in predaceous or parasitic ies and detritivorous ies suggest their diets contain similar amounts of N.Arthropod Nitrogen as a Nutrient for BirdsNot all N in arthropods is digested by insectivorous birds. Bird diets are frequently determined by identifying undigested fragments of exoskeleton in fecal samples (e.g., Wiesenborn & Heydon 2007). Digestion of arthropod cuticle by vertebrates likely depends on its sclerotization (Karasov 1990). Sclerotized proteins are bonded together, frequently with chitin, forming an irreversibly-hardened cuticle that cannot be hydrolyzed into amino acids (Richards 1978). Unsclerotized proteins, like resilin, can be hydrolyzed (Richards 1978). Relative proportions of sclerotized and unsclerotized proteins vary greatly among species (Richards 1978) producing cuticles with different digestibilities. Arthropod orders with high amounts of elastic protein, such as Odonata and Orthoptera and probably Araneae, may provide insectivorous birds with high concentrations of digestible protein. Riparian arthropods presented insectivorous birds with prey containing a range (5.1-14.0%) of N concentrations. Foraging by insectivorous birds in relation to prey N concentration can be difcult to discern, because birds frequently forage in response to prey availability which is transitory and hard to estimate. Selective foraging may be inferred by comparing arthropods eaten by adults with those concurrently captured by adults but fed to nestlings. Insectivorous nestlings depend on diet nutrients in addition to calories (Johnston 1993). Adult great tits ( Parus major L.) and blue tits (Parus caeruleus L.) in woodlands ate mostly Lepidoptera larvae but provided 3-9 day-old nestlings with more spiders, earwigs (Dermaptera), and ies (Cowie and Hinsley 1988). Including other arthropods, especially spiders, as prey may have augmented the low N content of Lepidoptera (Fagan et al. 2002). Spiders also provide different amino-acid compositions (Ramsay & Houston 2003). The importance of prey N-concentration to insectivorous birds that feed on more-diverse prey is less clear. An example is the southwestern willow ycatcher ( Empidonax traillii (Audubon) ssp. extimus Phillips), a migrant that winters in Central America and breeds in southwestern U.S. riparian habitats. Adult ycatchers ate mostly heteropterans, ies, and beetles but fed more odonates and beetles to nestlings (Drost et al. 2003). Diet N may be increased by including odonates, especially dragonies due to their large biomass. Diets of nestling ycatchers in other localities contained more Diptera than those of adults (Durst et al. 2008) or prey compositions similar to adults (Wiesenborn & Heydon 2007). The high-N orders of Araneae, Odonata, and Hymenoptera, taken together, were eaten with similar frequency by ycatchers at different localities and habitats. These orders comprised 21% of prey in California (Drost et al. 2003), 31% of prey in Arizona (Durst et al. 2008), and 21% of prey at 3 localities in Arizona and Nevada (Wiesenborn & Heydon 2007). In summary, N concentrations in riparian arthropods are primarily dependent on body mass and order and less dependent on trophic level. Variation in prey N concentration may affect foraging by insectivorous birds and the qualities of food they obtain. ACKNOWLEDGMENTSI am grateful to A. Stephenson, USBR Lower Colorado Regional Laboratory, for measuring ammonia concentrations. I appreciate the help identifying insects provided by S. L. Heydon, L. S. Kimsey, and T. J. Zavortink at the Bohart Museum of Entomology, and C. A. Tauber and P. S. Ward at the Entomology Department, UC Davis. I am grateful to J. E. OHara at Agriculture and Agrifood Canada for identifying tachinids and to D. Ubick for identifying spiders. I thank J. Allen of the U.S. Fish and Wildlife Service for the collection permit. This work was funded by the Lower Colorado River MultiSpecies Conservation Program.REFERENCES CITEDALYOKHIN, A. V., MESSING, R. H., AND DUAN, J. J. 2001. Utilization of the exotic weed Pluchea odorata (Asteraceae) and related plants by the introduced biological control agent Acinia picturata (Diptera: Tephritidae) in Hawaii. Biocontrol Sci. Technol. 11: 703-710. ANDERSEN, S. O. 1966. Covalent cross-links in a structural protein, resilin. Acta Physiol. Scand. 66 (Suppl. 263): 1-81. ANDERSEN, S. O. 1979. Biochemistry of insect cuticle. Annu. Rev. Entomol. 24: 29-61. ANDERSEN, S. O., AND WEIS-FOGH, T. 1964. Resilin. A rubberlike protein in arthropod cuticle, pp. 1-65 In J. W. L. Beament, J. E. Treherne, and V. B. Wigglesworth [eds.], Advances in Insect Physiology, vol. 2. Academic Press, London. ANDERSEN, S. O., CHASE, A. M., AND WILLIS, J. H. 1973. The amino-acid composition of cuticles from Tenebrio

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80 Florida Entomologist 94(1)March 2011molitor with special reference to the action of juvenile hormone. Insect Biochem. 3: 171-180. ANDERSON, J. F., RAHN, H., AND PRANGE, H. D. 1979. Scaling of supportive tissue mass. Q. Rev. Biol. 54: 139-148. BACKUS, E. A., CLINE, A. R., ELLERSEICK, M. R., ANDSERRANO, M. S. 2007. Lygus hesperus (Hemiptera: Miridae) feeding on cotton: new methods and parameters for analysis of nonsequential electrical penetration graph data. Ann. Entomol. Soc. America 100: 296-310. BELL, G. P. 1990. Birds and mammals on an insect diet: a primer on diet composition analysis in relation to ecological energetics, pp. 416-422 In M. L. Morrison, C. J. Ralph, J. Verner, and J. R. Jehl [eds.], Avian Foraging: Theory, Methodology, and Applications. Studies in Avian Biology, no. 13. Cooper Ornithological Society, Los Angeles, CA. BORROR, D. J., DE LONG, D. M., AND TRIPLEHORN, C. A. 1981. An Introduction to the Study of Insects, 5th ed. Saunders Philadelphia, PA. 827 pp. BROWNING, H. C. 1942. The integument and moult cycle of Tegenaria atrica (Araneae). Proc. R. Soc. London, B, Biol. Sci. 131: 65-86. COLE, F. R. 1969. The Flies of Western North America. University of California Press, Berkeley, CA. 693 pp. COWIE, R. J., AND HINSLEY, S. A. 1988. Feeding ecology of great tits (Parus major) and blue tits (Parus caeruleus ), breeding in suburban gardens. J. Anim. Ecol. 57: 611-626. DESERT RESEARCH INSTITUTE (DRI). 2010. Western U.S. Climate Historical Summaries. Western Regional Climate Center, Reno, NV [http://www.wrcc.dri.edu/ Climsum.html]. DROST, C. A., PAXTON, E. H., SOGGE, M. K., AND WHITFIELD, M. J. 2003. Food habits of the southwestern willow ycatcher during the nesting season, pp. 96103 In M. K. Sogge, B. E. Kus, S. J. Sferra, and M. J. Whiteld [eds.], Ecology and Conservation of the Willow Flycatcher. Studies in Avian Biology, no. 26. Cooper Ornithological Society, Los Angeles, CA. DURST, S. L., THEIMER, T. C., PAXTON, E. H., ANDSOGGE, M. K. 2008. Age, habitat, and yearly variation in the diet of a generalist insectivore, the southwestern willow ycatcher. Condor 110: 514-525. ESSIG, E. O. 1926. Insects of Western North America. MacMillan, New York, NY. 1035 pp. FAGAN, W. F., SIEMANN, E., MITTER, C., DENNO, R. F., HUBERTY, A. F., WOODS, H. A., AND ELSER, J. J. 2002. Nitrogen in insects: implications for trophic complexity and species diversication. American Nat. 160: 784-802. FOOTE, R. H., BLANC, F. L., AND NORRBOM, A. L. 1993. Handbook of the Fruit Flies (Diptera: Tephritidae) of America North of Mexico. Comstock, Ithaca, NY. 571 pp. HACKMAN, R. H. 1974. Chemistry of the arthropod cuticle, pp. 215-270 In M. Rockstein [ed.], The Physiology of Insecta, 2nd ed. Academic Press, New York, NY. ISAAC, R. A., AND JOHNSON, W. C. 1976. Determination of total nitrogen in plant tissue, using a block digestor. J. Assoc. Off. Anal. Chem. 59: 98-100. JOHNSTON, R. D. 1993. Effects of diet quality on the nestling growth of a wild insectivorous passerine, the house martin Delichon urbica Funct. Ecol. 7: 255-266. KARASOV, W. H. 1990. Digestion in birds: chemical and physiological determinants and ecological implications, pp. 391-415 In M. L. Morrison, C. J. Ralph, J. Verner, and J. R. Jehl [eds.], Avian Foraging: Theory, Methodology, and Applications. Studies in Avian Biology, no. 13. Cooper Ornithological Society, Los Angeles, CA. MATTSON, W. J. 1980. Herbivory in relation to plant nitrogen content. Ann. Rev. Ecol. Syst. 11: 119-161. NETER, J., KUTNER, M. H., NACHTSHEIM, C. J., ANDWASSERMAN, W. 1996. Applied Linear Statistical Models, 4th ed. McGraw-Hill, Boston, MA. 1408 pp. NEVILLE, A. C. 1975. Biology of the Arthropod Cuticle. Vol. 4 of D.S. Farner [ed.], Zoophysiology and Ecology. Springer-Verlag, New York, NY. 448 pp. PRANGE, H. D. 1977. The scaling and mechanics of arthropod exoskeletons, pp. 169-181 In T. J. Pedley [ed.], Scale Effects in Animal Locomotion. Academic Press, New York, NY. RAMSAY, S. L., AND HOUSTON, D. C. 1997. Nutritional constraints on egg production in the blue tit: a supplementary feeding study. J. Anim. Ecol. 66: 649657. RAMSAY, S. L., AND HOUSTON, D. C. 2003. Amino acid composition of some woodland arthropods and its implications for breeding tits and other passerines. Ibis 145: 227-232. REES, C. J. C. 1986. Skeletal economy in certain herbivorous beetles as an adaptation to a poor dietary supply of nitrogen. Ecol. Entomol. 11: 221-228. RICHARDS, A. G. 1978. The chemistry of insect cuticle, pp. 205-232 In M. Rockstein [ed.], Biochemistry of Insects. Academic Press, New York, NY. SCRIBER, J. M. 1984. Host-plant suitability, pp. 159-202 In W. J. Bell and R. T. Card [eds.], Chemical Ecology of Insects. Sinauer, Sunderland, MA. SIMPSON, S. J., RAUBENHEIMER, D., AND CHAMBERS, P. G. 1995. The mechanisms of nutritional homeostasis, pp. 251-278 In R. F. Chapman and G. de Boer [eds.], Regulatory Mechanisms in Insect Feeding. Chapman & Hall, New York, NY. SOKAL, R. R., AND ROHLF, F. J. 1981. Biometry, 2nd ed. W. H. Freeman, New York, NY. 859 pp. STUDIER, E. H., AND SEVICK, S. H. 1992. Live mass, water content, nitrogen and mineral levels in some insects from south-central lower Michigan. Comp. Biochem. Physiol., A, Comp. Physiol. 103: 579-595. TRIM, A. R. 1941. Studies in the chemistry of the insect cuticle: some general observations on certain arthropod cuticles with special reference to the characterization of the proteins. Biochem. J. 35: 1088-1098. WALDBAUER, G. P. 1968. The consumption and utilization of food by insects, pp. 229-288 In J. W. L. Beament, J. E. Treherne, and V. B. Wigglesworth [eds.], Advances in Insect Physiology, vol. 5. Academic Press, London. WIESENBORN, W. D. 2004. Mouth parts and alimentary canal of Opsius stactogalus Fieber (Homoptera: Cicadellidae). J. Kans. Entomol. Soc. 77: 152-155. WIESENBORN, W. D., AND HEYDON, S. L. 2007. Diets of breeding southwestern willow ycatchers in different habitats. Wilson J. Ornithol. 119: 547-557.

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Paraiso et al.: Egg Parasitoids of the Cactus Moth 81 EGG PARASITOIDS ATTACKING CACTOBLASTIS CACTORUM (LEPIDOPTERA: PYRALIDAE) IN NORTH FLORIDA O ULIMATHE P ARAISO 1 S TEPHEN D. H IGHT 2 M OSES T. K. K AIRO 1 AND S TEPHANIE B LOEM 3 1 Center for Biological Control, College of Engineering Sciences, Technology and Agriculture, Florida Agricultural & Mechanical University, Tallahassee, FL 32307 2 USDA, ARS, CMAVE, Tallahassee, FL 32308 3 USDA, APHIS, PPQ, CPHST, Plant Epidemiology and Risk Analysis Laboratory, Raleigh, NC 27606 A BSTRACT Interest in the natural enemies of Cactoblastis cactorum (Berg) has increased since the moth w as found in Florida in 1989. Previous surveys for natural enemies in Argentina identied egg parasitoids in the family Trichogrammatidae as potentially important control agents of C. cactorum A study was conducted in north Florida to identify and to assess occurrence of egg parasitoids attac king this invasive moth in its new homeland. Surveys undertaken at 6 locations in north Florida from Jul 2008 to Dec 2009 revealed that eggsticks of C. cactorum were attacked by egg parasitoids from the Trichogramma genus: T. pretiosum Riley, T. fuentesi Torre, and an additional unidentied Trichogramma species belonging to the T. pretiosum group. In order to assess the importance of these egg parasitoids, the fate of individual C. cactorum eggsticks was determined during weekly visits to each site. This assessment showed that the combined level of parasitism of C. cactorum eggsticks was very low with less than 0.2% of host eggs attac ked at any one of the 6 sites. While parasitoids attacked smaller eggsticks, there was no correlation between the numbers of eggs in an eggstick attacked with increasing number of eggs/eggstick. Comparing the mean number of eggs/eggstick, there was no difference between the 3 ight periods of C. cactorum but there was a difference between the 6 sites Based on these results, the use of Trichogramma wasps as an inundative biological control agent, complementary to the Sterile Insect Technique application is discussed. K ey Words: Cactoblastis cactorum cactus moth, Trichogramma egg parasitoids, North Florida R ESUMEN El inters en los enemigos naturales de Cactoblastis cactorum (Berg) ha aumentado desde que esta especie fue encontrada en el estado de Florida en 1989. Busquedas de enemigos naturales de C. cactorum hechas en aos pasados en Argentina identicaron a parasitoides de huevos de la familia Trichogrammatidae como enemigos naturales de posible importancia para esta especie. Llevamos a cabo un estudio en seis localidades en el norte del estado de Florida con el objetivo de identicar y evaluar la ocurrencia de parasitoides de huevos atacando a esta especie en su nueva rea de distribucin. La busqueda de parasitoides llevada a cabo entre julio del 2008 y diciembre del 2009 indico que los bastoncitos de huevos de C. cactorum son atacados por parasitoides del genero Trichogramma : T. pretiosum Riley, T. fuentesi Torre, y una especie adicional no identicada perteneciente al grupo taxonmico de Trichogramma pretiosum Para evaluar la importancia de estos parasitoides en el control de C. cactorum seguimos el destino de bastoncitos individuales a travs de visitas semanales a cada una de las localidades Estas observaciones demostraron que el grado de parasitismo en esos basoncitos es muy bajo, con menos de 0.2% de los huevos parasitados en cualquiera de las seis reas. Mientras que observamos que los parasitoides atacaron bastoncitos de huevos de tamao pequeo, no hubo correlacin entre el nmero de huevos parasitados por bastoncito y el tamao del mismo. Comparando el numero promedio de huevos por bastoncito, no detectamos diferencia en los bastoncitos ovipositados en las tres generaciones anuales de C. cactorum pero detectamos diferencias dependiendo del rea. Basado en estos resultados, discutiremos el uso de parasitoides del genero Trichogramma como agentes inundativos de control biolgico complementando la aplicacin de la Tcnica del Insecto Estril contra C. cactorum Translation provided by the authors. The cactus moth, Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae), is often cited as the perfect example of a successful weed biological control agent (Moran & Zimmermann 1984). In 1925,

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82 Florida Entomologist 94(1) March 2011 the cactus moth was introduced from its native Argentina into Australia to control prickly pear cactus, Opuntia spp., which had originally been brought into Australia for commercial purposes (Dodd 1940; Mann 1970). The cactus had become invasive and made large tracts of rangeland unt for grazing cattle. Within a few years after the introduction of C. cactorum into Australia, US $6 million worth of rangeland w as restored, equivalent to more than US $60 million in todays dollars (Dodd 1940; Williamson 2009). Based on these promising results, C. cactorum was imported from Australia to South Africa, Mauritius, and Hawaii to manage other non-native and invasive Opuntia spp. (Moran & Zimmermann 1984). In 1957, C. cactorum was introduced into several Caribbean islands (Nevis Montserrat, and Antigua) to control non-native as well as native Opuntia spp. (Simmonds & Bennett 1966). Unfortunately the implementing agencies did not fully consider the potentially injurious environmental impacts of C. cactorum if this insect were to move to neighboring countries where some species of Opuntia are important native species and some are commercially important (Stiling et al. 2004). The rst record of C. cactorum in the U.S. was from Bahia Honda K ey, Florida, in Oct 1989 (Dickel 1991). It is uncertain how the moth arrived in Florida, but several interceptions of Caribbean ornamental Opuntia spp. infested with C. cactorum were found at ports of entry in south Florida during the 1980s and 1990s (P emberton 1995; Zimmermann et al. 2001; Stiling 2002; Simonsen et al. 2008). Since its appearance in Florida, C. cactorum has become a threat to native Opuntia spp. in North America. Current management options inc lude the use of Pherocon 1-C Wing traps (Trc Incorporated, Salinas, CA) baited with a 3-component synthetic sex lure (Suterra, LLC, Bend, OR) to identify the presence of the moth, coupled with removal of infested plants to reduce C. cactorum populations (Bloem et al. 2005; Hight & Carpenter 2009). Complementary to the detection, monitoring, and removal efforts, implementation of the Sterile Insect Technique (SIT) is being used to slow the geographic expansion of C. cactorum in the U.S. (Hight et al. 2002; Bloem et al. 2005; Bloem et al. 2007). In Mexico, localized invasions of C. cactorum on 2 islands were eradicated in 2008 with a program of pheromone traps host removal, and SIT (NAPPO 2006; NAPPO 2008; NAPPO 2009). Bennett & Habeck (1995) suggested biological control as an additional control option that should be considered for C. cactorum Pemberton & Cordo (2001) reported that several larval and pupal parasitoids attac ked the cactus moth in South America, including species of Hymenoptera (Braconidae, Chalcidae, and Ichneumonidae), and 1 Diptera (Tachinidae). They also reported on 2 chalcid species ( Brachymeria ovata (Say) and B. pedalis Cresson) and 1 unidentied egg parasitoid from the family Trichogrammatidae attacking C. cactorum in Florida. Logarzo et al. (2008) found the larval parasitoid Apanteles alexanderi Brethes (Hymenoptera: Braconidae) and the egg parasitoid Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) attacking C. cactorum in Argentina. Tric hogrammatid egg parasitoids have been used successfully for inundative biological control against major lepidopteran pests such as corn borers, sugarcane borers, and cotton bollworm (Li-Ying 1994; van Lenteren 2000). Egg parasitoids are easy to rear in mass quantity in laboratory conditions and to release over wide areas. Biological control can be used to complement and synergize the application of SIT (Gurr & Kvedaras 2010). Recent studies showed that the combination of both techniques was more efcient in controlling pest population of the codling moth, Cydia pomonella (L.) (Bloem et al. 1998). Synergistic interactions between SIT and fruit y biological control with parasitoids increased the suppression of pest fruit ies even leading to eradication (Sivinski 1996; Rendon et al. 2006). SIT and biological control have been successfully combined to combat several lepidopteran pests, including C. pomonella (Bloem et al. 1998) and painted apple moth, Orgyia anartoides (Walker) (Suc kling et al. 2007). Radiation doses for sterilizing C. cactoblastis adults have been determined to produce partially sterile but more t males whic h, when mated with wild females, generate sterile offspring (Carpenter et al. 2005). The combination of egg parasitoid releases and SIT has the advantage that parasitoids manage high pest densities, while SIT works best at low pest densities. In addition, release of sterile insects provides an egg resource for egg parasitoids increasing the ratio of natural enemies to adult hosts. Egg parasitoids and sterile insects have the characteristic of being self dispersing and consequently are able to cover wide areas (Sivinski 1996). We conducted eld surveys in order to identify egg parasitoids already established in North Florida that attack C. cactorum Cactoblastis cactorum adults have 3 annual ight periods in north Florida (Apr -May, Jul-Aug, and Oct-Nov) (Hight et al. 2005; Hight & Carpenter 2009). We report on the distribution, seasonality, and parasitism parameters of the Trichogramma species attac king C. cactorum in northern Florida. The number of eggs/eggstic k was compared between different ight periods and sites to assess host egg resource for egg parasitoids. The effect of C. cactorum eggstick size on level of parasitism was evaluated by comparing number of eggs from par asitized versus un-parasitized eggsticks. These data will be benecial in promoting discussions on possible implementation of biological control for the cactus moth and, in particular, assessing

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Paraiso et al.: Egg Parasitoids of the Cactus Moth 83 the potential of an inundative biological control program against C. cactorum in North America. M ATERIALS AND M ETHODS Field surveys were carried out at 6 locations (Fig. 1) in north Florida from Jul 2008 to Dec 2009. The selection of study sites was based on existing records of infestations from the literature, personal observations from preliminary surveys, and information provided by experts. Female C. cactorum place their eggs end to end to form a c hain that looks like a short stick, and the egg mass is referred to as an eggstick. Although no extensive eld surveys were conducted from May to Jul 2008 at St. Marks and St. George Island, eggsticks with eggs that appeared parasitized were collected and held in laboratory conditions until parasitoids emerged. At survey locations, 20 to 30 healthy Opuntia spp. plants were chosen with no to minor feeding damage by cactus moth larvae and an a verage of at least 50 pads per plant. During weekly visits throughout all 3 ight periods, any new eggstick was identied by plant, pad, and its general location on the plant so the eggstick could be found during subsequent checks. A mark was made on the plant at the base of the eggstick with a felt tip pen and a red tape ag afxed to an insect pin placed near the eggstick to aid in nding the eggstick. The ag was labeled with a unique number to identify each eggstick. The oviposition preferences of C. cactorum females on host plants were recorded by c lassifying the attachment of the eggstick to either a glochid at an areole, to a spine, or on the fruit. Observations on plant habitat and host eggstick distribution within the surveyed site and within the selected plant were collected to provide additional information on the host nding behavior of egg parasitoids. The number of eggs per eggstick was determined either by a direct count or by a correlated estimate of eggstick length to egg number (2.62 0.013 eggs/mm). The ratio of eggstick length to egg number was calculated in this study by counting the number of eggs in a segment of eggstick, replicated on 20 eggsticks. Eggstick length was estimated in situ by placing a plastic string next to the eggstic k and cutting a piece of equivalent length. The length of the piece of string was then measured to the nearest 0.01 mm with a metric micrometer. Measurements of eggsticks were obtained so that the number of eggs per eggstick could be estimated if the eggstick was lost before it could be collected and directly counted. The fate of each eggstick was determined by making weekly visits to each site to evaluate the status of previously tagged eggsticks. The fate of each eggstick was categorized as follows: eggstick lost; predated (visible chewing damage) eggs in the eggstick versus non predated eggs; or parasitized eggs in the eggstick (black eggs formed before C. cactorum larvae successfully developed). Eggsticks were collected if they were damaged during evaluation or measurement, eggs of the eggstick had hatched, or eggs appeared predated or parasitized. Eggsticks with viable eggs were collected and held in small plastic cups (30 mL) under laboratory conditions (25 1 C, 16:8 L:D and 40-60% RH) to record hatch rate. Eggsticks with parasitized eggs were collected and monitored in the laboratory to determine the emergence rate, number of eggs/eggstick attacked by parasitoids, number of parasitoids emerging per parasitized egg, and to ascertain the identity of the parasitoids. Parasitoid specimens were submitted to R. Stouthamer, Department of Entomology, University of California, Riverside, for molecular identication. The sequencing of ribosomal DNA Internal Transcribed Spacer 2 (ITS 2) was used to identify the different species of egg parasitoids. Data Analysis The numbers of eggs/eggstick at different ight periods for each surveyed location and the average number of eggs/eggstick at each site were log transformed before analyses to satisfy the assumptions of the analysis of variance. One way analysis of variance (PROC GLM) was applied to the log transformed data and the separation of means was made with the least signicant difference (LSD) test. Comparison of number of eggs/ eggstick that was parasitized versus number of eggs/eggstick not parasitized was also evaluated with a one-way analysis of variance (PROC GLM). Since only a few eggsticks with parasitized eggs were collected in this study (see text below), comparisons between eggsticks with parasitized eggs were made against the same number of ranFig. 1. Locations and their coordinates surveyed for eg g parasitoids of Cactoblastis cactorum in North Florida.

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84 Florida Entomologist 94(1) March 2011 domly selected eggsticks with un-parasitized eggs. Variation between the number of eggs for parasitized eggsticks and the number of eggs for the randomly selected un-parasitized eggsticks was analyzed by a folded F test (Davis 2007). Because the variances in numbers of eggs for eggstic ks with parasitism and number of eggs in eggsticks without parasitism were not signicantly different, means of these 2 groups were compared with a two-sample t -test. A Pearsons Correlation Coefcient (r) w as calculated to determine whether the numbers of eggs parasitized were dependent on the number of eggs/eggstick. The SAS Statistical Software Version 9.2 (SAS Institute, Cary, North Carolina) was used to perform the statistical analyses. Estimates of central tendencies were reported as mean standard error of mean. R ESULTS AND D ISCUSSION Although host plant species of Opuntia stricta (Haworth) Haworth O. humifusa (Ranesque) Ranesque and O. cus-indica (L.) P. Miller varied among the different geographic regions sur veyed, the oviposition preferences of C. cactorum females was similar on the various species (Table 1). In this study, parasitized eggsticks of C. cactorum appeared mostly on the areole/glochid structure of the pads (T able 1). Altogether, 1,527 eggsticks with 91,013 C. cactorum eggs, not including 344 eggsticks missing from the eld or lost during collection, were tagged on plants of Opuntia spp. (Table 2). Of all the eggstic ks checked, 62% were collected on Okaloosa Island and had a mean of 59 (+/1.83) eggs/ eggstick. The proportion of eggsticks examined in the laboratory as percentage of all eggsticks surveyed at the 6 eld sites ranged from 53 to 100%, except for summer 2008 at St. George Island and St. Marks National Wildlife Refuge (NWR) in which only 30% and 24%, respectively, of the monitored eggsticks were examined (Table 2). The majority of the eggsticks from these 2 locations for this ight period were recorded as lost (Table 2). The cause for this high number of lost eggsticks is not clear. Several biotic and abiotic factors could have contributed to the high number of lost eggsticks. During summer 2008, 23% of eggsticks examined from St. Marks had eggs that were preyed upon compared with less than 3% in other locations. Although not directly observed at St. George Island or St. Marks, substantial predation of C. cactorum eggs by ants has been recorded in South Africa (Robertson 1984). Because the plants surveyed at St. Marks were located within 100 m of the waters of the Gulf of Mexico, strong winds characteristic of coastal regions could have knocked eggsticks off the plants. All other study sites were along the Gulf Coast; in none of them were the plants as close to the water as at St. Marks. In addition, heavy rainfall may have separated the eggsticks from plants, but we do not have any data on the severity of the rain storms at different study sites. Cactoblastis cactorum life table studies in Argentina (Logarzo et al. 2009) and South Africa (Robertson & Hoffmann 1989) identied rain and wind as major factors contributing to mortality of eggs. Surveyed sites and oviposition periods were analyzed to evaluate their inuence on number of eggs/eggstick. Eggsticks were collected for multiple oviposition periods at 3 sites (St. George Island, St. Marks, and Okaloosa Island) (Table 2). The numbers of eggs/eggsticks for the different oviposition periods were not signicantly different for St. George Island ( F = 1.84, df = 1, P = 0.18), St. Marks ( F = 93.86, df = 3, P = 0.07), or Okaloosa Island (F = 0.22, df = 3, P = 0.88). Because the numbers of eggs/eggstick for multiple oviposition periods were not different, eggsticks from all ight periods were pooled to calculate the means for those sites (St. George Island (62 2.8), St. Marks (53 2.8), and Okaloosa Island (59 1.8)). The pooled eggsticks were used to compare the number of eggs/eggstick between all 6 sites and signicant differences were found ( F = 11.44, df = 5, P < 0.0001) (Table 2). Female C. cactorum laid similar numbers of eggs/eggstick for each of the 3 oviposition periods but not at all 6 survey sites along the Florida panhandle. The longest eggsticks were observed at St. George Island, Pensacola Beach, and Okaloosa Island (Table 2). Signicantly smaller eggsticks were recorded at St. Marks and Mexico Beach (Table 2). Panacea had signicantly smaller number of eggs/eggstick than all other sites (Table 2). The cause of differences between eggsticks at the various sites was unclear. Studies in South Africa identied differences in total fecundity of C. cactorum due to host plant species, the ight period when eggs were laid, and the temperature during oviposition (Robertson 1989). We did not distinguish eggsticks collected from different host plants (Table 1). While South African female C. cactorum had signicantly higher fecundity during the summer ight (Robertson 1989), our study did not show any difference in number eggs/eggstick between ight periods in north Florida. Cactoblastis cactorum has a tendency to oviposit on plants with high nitrogen (Myers et al. 1981; Robertson 1987), but we have no direct measurements of plant quality at our sites. Comparing the number of eggs/eggstick for eggsticks that were parasitized (38 13.7) (Table 3) against un-parasitized eggsticks (61 13.1) revealed a signicant difference (pooled t test = 3.14, df = 12, P = 0.0085). Although the number of eggs/eggstick was highly variable, the variation of the number of eggs/eggstick for parasitized versus the randomly selected un-parasitized group

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Paraiso et al.: Egg Parasitoids of the Cactus Moth 85TABLE 1.SITES SURVEYED IN NORTH FLORIDA FOR TRICHOGRAMMA EGG PARASITOIDS OF CACTOBLASTIS CACTORUM EGGSTICKS ON OPUNTIA SP. AND ADDITIONAL INFORMATION ON MOTH OVIPOSITION PREFERENCE. Site GPS Coordinate Dates Eggsticks Surveyed Total Number Surveys Species of Opuntia Host Plant Number Host Plant Examined Total Number Eggsticks Evaluated Percent Eggsticks at Attachment Location1Areole/ GlochidSpineFruitMissing2Pensacola BeachN30.33525Summer 2008 10 O. stricta 20 1205034160 W87.48928(Jul 10-Sep 10, 08) O. humifusa O. cus-indica St. George IslandN29.65068Summer 2008 10 O. stricta 23 105633070 W84.9120(Jul17-Sep19, 08) Fall 2008 20 27 28891100 (Sep 25, 08-Feb 25, 09) St. Marks (NWR) N30.07772 Summer 2008 18 O. stricta 30 45801370 W84.18242(Jul 15,-Sep 12, 08) O. humifusa Fall 2008 20 30 9780220 (Oct 01, 08-Feb 25, 09) Spring 2009 13 35 47882010 (Apr 17-Jul 15, 09) Fall 2009 35 151n/an/an/an/a (Oct 07, 09-Jan 12, 10) Mexico BeachN29.94133Fall 2009 3 O. cus-indica n/a 29n/an/an/an/a W85.40636(Oct 21-Nov12, 09) Panacea N30.03127 Fall 2009 3 O. stricta n/a 65n/an/an/an/a W84.39353W84.39353 O. cus-indica Okaloosa Island N30.08674 Fall 2008 21 O. cus-indica 10 1868118.50.50 W86.37807(Oct 08, 08-Feb 27, 09) Spring 2009 14 18 308791514 (Apr 08-Jul 08, 09) Summer 2009 18 18 280771922 (Jul 01-Sep 25, 09) Fall 2009 25 151n/an/an/an/a (Sep 18, 09-Jan 12, 10)1Attachment locations of eggsticks that were not determined is indicated by n/a.2Information about eggstick attachment failed to be recorded.

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86 Florida Entomologist 94(1)March 2011TABLE 2.NUMBER OF CACTOBLASTIS CACTORUM EGGSTICKS COLLECTED, LOST IN THE FIELD, EXAMINED IN THE LABORATORY, AND MEAN NUMBER OF EGGS PER EGGSTICK SE AT DIFFERENT SITES IN NORTH FLORIDA FOR DIFFERENT OVIPOSITION PERIODS. SiteFlight Period Total Number Eggsticks Tagged Total Number (Percent) Eggsticks Lost Percent Eggsticks Examined Total Number Moth Eggs Examined Mean Number Eggs/Eggstick SE Overall Mean Eggs/ Eggstick SE at Each Site1Pensacola BeachSummer 2008 120 69 (58) 42 7,402 62+/-1.5 62 1.5 a St. George IslandSummer 2008 105 84 (70) 30 6,685 64+/-2.0 62 2.8 a Fall 2008 28 13 (46) 53 1,614 58+/-3.6 St. Marks (NWR)Summer 2008 45 35 (77) 24 3,088 68+/-2.9 53 2.8 b Fall 2008 9 4 (44) 55 513 57+/-3.3 Spring 2009 47 23 (46) 54 2,561 54+/-3.1 Fall 2009 151 0 (0) 100 6,917 46+/-1.4 Mexico BeachFall 2009 29 0 (0) 100 1,522 52+/-3.0 52 3.0 b Panacea Fall 2009 65 0 (0) 100 2,892 45+/-1.9 45 1.9 c Okaloosa IslandFall 2008 186 61 (29) 71 11,118 60+/-1.4 59 1.3 a Spring 2009 308 3 (1) 99 20,527 61+/-1.2 Summer 2009 280 21 (8) 92 17,126 60+/-1.1 Fall 2009 151 1 (0.6) 99 8,638 57+/-1.21Means with different letter are significantly different ( P < 0.05).

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Paraiso et al.: Egg Parasitoids of the Cactus Moth 87TABLE 3.LOCATION AND DATE PARASITIZED CACTOBLASTIS CACTORUM EGGSTICK WAS COLLECTED, IDENTITY OF PARASITOID SPECIES, NUMBER OF EGGS PER EGGSTICK,NUMBER OF PARASITIZED EGGS, NUMBER OF PARASITOIDS EMERGED, FEMALE RATIO, AND PARASITISM LEVEL OF EGG PARASITOIDS ATTACKING C. CACTORUM IN NORTH FLORIDA. Site Collection Date Flight Period Trichogramma sp. Number Eggs/ Eggstick Number Parasitized Eggs/Eggstick # (%) Number Parasitoids Emerged Percent Females Level of Parasitism (%) St. Marks 05/15/08Spring 08 T. pretiosum 735 (7)875n/a105/15/08 T. pretiosum 7817 (22)3485 Pensacola Beach04/22/08Spring 08unknown 88 19 (22) 18 77 n/a 08/06/08Summer 08 T. pretiosum 44 8 (18) 5 40 0.2 08/13/08 T. pretiosum 18 6 (33) 10 70 Okaloosa Island 10/16/08Fall 08 T. fuentesi 20 2 (10) 11 73 0.1 10/16/08 T. pretiosum 42 10 (24) 7 71 10/16/08 T. fuentesi 56 6 (11) 16 62 11/03/08 unknown 52 3 (6) 9 56 10/23/09Fall 09 T. fuentesi 25 13 (52) 63 89 0.11Indicates that the level of parasitism was not determined.

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88 Florida Entomologist 94(1)March 2011was similar (folded F test = 1.10, df = 6, P = 0.91), suggesting that the difference found between the two groups was not driven by unequal or extreme variation. However, there was not a signicant correlation between the number of eggs/eggstick and number of eggs parasitized by Trichogramma spp. (n = 7, r = -0.16, P = 0.74). Therefore, while female Trichogramma spp. parasitized eggsticks with fewer eggs, they did not parasitize more eggs as the number of eggs in an eggstick increased. The average number of eggs parasitized in an eggstick was 9 (.8). Ten eggsticks were found parasitized at 3 of the 6 sites surveyed (Pensacola Beach, St. Marks, and Okaloosa Island). Five of the parasitized eggsticks were found at Okaloosa Island. Parasitized eggsticks were found during all 3 oviposition periods of C. cactorum females: the spring ight (St. Marks and Pensacola Beach), summer ight (Pensacola Beach), and fall ight (Okaloosa Island). Of the 496 eggs in the 10 parasitized eggsticks, a total of 89 eggs (or 18%) were parasitized, resulting in the emergence of 181 adult parasitoids with a sex ratio of 70% () females (Table 3). The level of parasitism by Trichogramma spp., relative to the total number of eggs examined during the different ight periods for each site, was less than 0.2% of total C. cactorum eggs collected (Table 3). We did not observe any parasitized eggsticks at St. George Island, Mexico Beach, or Panacea. Two species of Trichogramma were reared from C. cactorum eggsticks in north Florida (Table 3) and identied by differences in IST2 sequences. Trichogramma pretiosum was collected at St. Marks, Pensacola Beach, and Okaloosa Island, while T. fuentesi Torre was recovered only from Okaloosa Island. It was not possible to identify 2 collections of Trichogramma spp. from Okaloosa Island; one because a good molecular sequence could not be obtained and for the other the sequence was not in the database and possibly represents a new species in the T. pretiosum group (R. Stouthamer, UCRiverside, personnel communication). More than 15 million ha of agriculture and forestry worldwide are treated annually with Trichogramma egg parasitoids (van Lenteren 2000). Trichogrammatid wasps have been used successfully in inundative release programs against lepidopteran pests in greenhouses and crop production worldwide (Smith 1996). Inundative releases of Trichogramma spp. have been implemented in Florida to control major lepidopteran pests of collards, cabbages, soybeans, bell peppers, tomatoes, corn, and tobacco production (Martin et al. 1976). Trichogramma pretiosum is commonly found in the Western hemisphere. This Trichogramma species has been released commercially against major lepidopteran pests such as cotton leafworm ( Alabama argillacea) (Hbner), corn earworm ( Helicoverpa zea) (Boddie), tomato pinworm (Keiferia lycopersicella) (Walshingham), sugarcane borers ( Diatraea spp.), and cabbage looper (Trichoplusia ni) (Hbner) (Pinto et al. 1986; Hassan 1993; LiYing 1994; Monje et al. 1999). Trichogramma fuentesi have been recorded in countries in South America (Argentina, Columbia, Mexico, Peru, and Venezuela) and in the U.S. (Alabama, California, Florida, Louisiana, New Jersey, South Carolina and Texas) (Fry 1989, Pinto 1999). Its primary hosts are species from the Noctuidae family such as H. zea and Heliothis virescens (F.) and from the Pyralidae family such as Diatrea saccharalis (F.), Ephestia kuehniella Zeller, and Ostrinia nubilalis (Hbner) (Fry 1989; Wilson & Durant 1991; Pintureau et al. 1999; Querino & Zucchi 2003). Trichogramma parasitoids also are widely used for pest control in orchards (Olkowski & Zang 1990). The observed low incidence of the wasps in natural areas might be explained by unfavorable environmental factors or natural plant chemicals (Smith 1996; Romeis et al. 1997, 1999). However, contrary to other natural enemies, Trichogramma can be easily and cheaply mass-reared for the implementation of an inundative biological control program. The potential for inundative releases of T richogramma spp. as a strategy against C. cactorum is currently being investigated with sustainable laboratory colonies of T. fuentesi originating from eld collected insects reared from parasitized C. cactorum eggsticks. Biological characteristics (sex ratio, egg load, and longevity) and different behavioral mechanisms (inuence of parasitoid age, density, and host age on parasitism) involved in host nding of T. fuentesi reared on C. cactorum eggs are being evaluated. The inundative releases of Trichogramma wasps could be integrated in the current pest management system based on SIT applications during the 3 ight periods by building Trichogramma populations. This eld survey was useful in identifying a potential inundative biological control agent that could be integrated within a pest management strategy against C. cactorum ACKNOWLEDGMENTSWe thank Shalom Benton (FAMU) for eld collection and laboratory assistance and Chris Albanese, Michael Getman, and John Mass (USDA-ARS-CMAVE, Tallahassee) for eld assistance. We thank Stuart Reitz (USDA-ARS-CMAVE, Tallahassee, FL) and Jim Nation (University of Florida) for comments on earlier drafts of this manuscript. This work is funded under the FAMUUSDA APHIS Cooperative Agreement, 07-10-81000755-CA. Mention of trade names or commercial products in this publication is solely for the purpose of providing specic information and does not imply recommendation or endorsement by the U.S. Department of Agriculture.

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D., CARPENTER, J. E., BLOEM, S., AND BLOEM, K. A. 2005. Developing a sterile insect release program for Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae): Effective overooding ratios and releaserecapture eld studies. Environ. Entomol. 34: 850856. LI-YING, L. 1994. Worldwide use of Trichogramma for biological control on different crops: A survey, pp. 3753 In E. Wajnberg, S. A. Hassan [eds.], Biological Control with Egg Parasitoids, CAB International, Wallingford. LOGARZO, G., VARONE, L., AND BRIANO, J. 2008. Cactus Moth, In Annual Report 2008 South American Biological Control Laboratory, United States Department of Agriculture, Agricultural Research Service, USDA-ARS, Hurlingham, Argentina. [Online] http:/ /www.usda-sabcl.org/projects/ AnnualReport2008PartLCACTUSMOTH.pdf. LOGARZO, G., VARONE, L., AND BRIANO, J. 2009. Cactus Moth, In Annual Report 2009 South American Biological Control Laboratory, United States Department of Agriculture, Agricultural Research Service, USDA-ARS, Hurlingham, Argentina. [Online] http:/ /www.ars.usda.gov/SP2UserFiles/Place/02110000/ CompleteAnnualReport2009.pdf. MANN, J. 1970. Cacti Naturalized in Australia and Their Control. Department of Lands, Queensland, Australia. MARTIN, P. B., LINGREN, P. D., GREEN, G. L., AND RIDGWAY, R. L. 1976. Parasitization of two species of Plusiinae and Heliotis spp. after releases of Trichogramma pretiosum in seven crops. Environ. Entomol. 5: 991-995. MONJE, J. C., ZEBITZ, C. P. W., AND OHNESORGE, B. 1999. Host and host age preference of Trichogramma galloi and T. pretiosum (Hymenoptera: Trichogrammatidae) reared on different hosts. J. Econ. Entomol. 92: 97-103. MORAN, V. C., AND ZIMMERMANN, H. G. 1984. The biological control of cactus weeds: Achievements and prospects. Biocontrol News and Infor 5: 297-320. MYERS, J. H., MONRO, J., AND MURRAY, N. 1981. Egg clumping, host plant selection, and population regulation in Cactoblastis cactorum (Lepidoptera). Oecolgia 51: 7-13. NAPPO (NORTH AMERICAN PLANT PROTECTION ORGANIZATION). 2006. Ofcial Pest Reports: Detection of an outbreak of cactus moth ( Cactoblastis cactorum) in Isla Mujeres, Quintana Roo, Mexico. [Online] www.pestalert.org NAPPO (NORTH AMERICAN PLANT PROTECTION ORGANIZATION). 2008. Ofcial Pest Reports: Eradication of cactus moth (Cactoblastis cactorum Berg) outbreak in Isla Mujeres, Quintana Roo, Mexico. [Online] www.pestalert.org NAPPO (NORTH AMERICAN PLANT PROTECTION ORGANIZATION). 2009. Ofcial Pest Reports: Detection and eradication of a cactus moth ( Cactoblastis cactorum Berg) outbreak in Isla Contoy, municipality of Isla Mujeres, Quintana Roo, Mexico. [Online] www.pestalert.org OLKOWSKI, W., AND ZHANG, A. 1990. Trichogramma modern day frontier in biological control. IPM Practitioner 12: 1-15. PEMBERTON, R. W. 1995. Cactoblastis cactorum (Lepidoptera: Pyralidae) in the United States, an immigrant biological control agent or an introduction of the nursery industry? American Entomol. 41: 230-232. PEMBERTON, R. W., AND CORDO, H. 2001. Potential and risk of biological control of Cactoblastis cactorum

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90 Florida Entomologist 94(1)March 2011(Lepidoptera: Pyralidae) in North America. Florida Entomol. 84: 513-526. PINTO, J. D., OATMAN, E. R., AND PLATNER, G. R. 1986. Trichogramma pretiosum and a new cryptic species occurring sympatrically in Southwestern North America (Hymenoptera: Trichogrammatidae). Ann. Entomol. Soc. America 79: 1019-1028. PINTO, J. D. 1999, Systematics of the North American species of Trichogramma Westwood (Hymenoptera: Trichogrammatidae). Mem. Entomol. Soc. Washington 22: 140. PINTUREAU, B., PETINON, S., AND NARDON, C. 1999. Possible function of substances excreted by Trichogramma and darkening of their hosts. Bull. Soc. Zool. France 124: 261-269. QUERINO, R. B., AND ZUCCHI, R. A. 2003. New species of Trichogramma Westwood (Hymenoptera: Trichogrammatidae) associated with lepidopterous eggs in Brazil. Zootaxa 163: 1-10. RENDON, P., SIVINSKI, J., HOLLER, T., BLOEM, K., LOPEZ, M., MARTINEZ, A., AND ALUJA, M. 2006. The effects of sterile males and two braconid parasitoids, Fopius arisanus (Sonan) and Diachasmimorpha krausii (Fullaway) (Hymenoptera), on caged populations of Mediterranean fruit ies, Ceratitis capitata (Wied.) (Diptera: Tephritidae) at various sites in Guatemala. Biol. Control 36: 224-231. ROBERTSON, H. G. 1984. Egg predation by ants as a partial explanation of the difference in performance of Cactoblastis cactorum on cactus weeds in South Africa and Australia, pp. 83-88 In E. S. Delfosse [ed.], Proc. VI Symp. Biol. Contr. Weeds. 19-25 August 1984, Vancouver, Canada. ROBERTSON, H. G. 1987. Oviposition site selection in Cactoblastis cactorum (Lepidoptera): constraints and compromises. Oecologia 73: 601-608. ROBERTSON, H. G. 1989. Seasonal temperature effects on fecundity of Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae): differences between South Africa and Australia. J. Entomol. Soc. South Africa 52: 71-80. ROBERTSON, H. G., AND HOFFMANN, J. H. 1989. Mortality and life-tables of Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae) compared on two hostplant species. Bull. Entomol. Res. 79: 7-17. ROMEIS, J. T., SHANOWER, G., AND ZEBITZ, P. W. 1997. Volatile plant infochemicals mediate plant preference of Trichogramma chilonis J. Chem. Ecol. 23: 2455-2465. ROMEIS, J. T., SHANOWER, G., AND ZEBITZ, P. W. 1999. Trichogramma egg parasitism of Helicoverpa armigera on pigeonpea and sorghum in southern India. Entomol. Exp. Appl. 90: 69-81. SIMMONDS, F. J., AND BENNETT, F. D. 1966. Biological control of Opuntia spp. by Cactoblastis cactorum in the Leeward Islands (West Indies). Entomophaga 11: 183-189. SIMONSEN, T. J., BROWN, R. L., AND SPERLING, F. A. 2008. Tracing an invasion: Phylogeographical of Cactoblatis cactorum (Lepidoptera: Pyralidae) in the United States based on Mitochondrial DNA. Entomol. Soc. America 101: 899-905. SIVINSKI, J. M. 1996. The past and potential of biological control of fruit ies, pp. 369-377 In J. B. A. McPheron and G. J. Steck [eds.], Fruit Fly Pests, A World Assessment of their Biology and Management. St. Lucie Press, Delray Beach, FL. SMITH, S. M. 1996. Biological control with Trichogramma advances, successes, and potential of their use. Annu. Rev. Entomol. 41: 375-406. STILING, P. D. 2002. Potential non-target effects of a biological control agent, prickly pear moth, Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae), in North America, and possible management actions. Biol. Invasions 4: 273-281. STILING, P. D., MOON, D., AND GORDON, D. 2004. Endangered cactus restoration: Mitigating the non-target effects of biological control agent ( Cactoblastis cactorum) in Florida. Rest. Ecol 12: 605-610. SUCKLING, D. M., BARRINGTON, A. M., CHHAGAN, A., STEPHENS, A. E. A., BURNIP, G. M., CHARLES, J. G.,AND WEE, S. L. 2007. Eradication of the Australian painted apple moth Teia anartoides in New Zealand: trapping, inherited sterility, and male competitiveness, pp. 603-615 In M. J. B. Vreysen, A. S. Robinson and J. Hendrichs [eds.], Area-Wide Control of Insect Pests. Springer, Dordrecht, The Netherlands. VAN LENTEREN, J. C. 2000. Success in biological control of arthropods by augmentation of natural enemies, pp. 77-103 In G. Gurr and S. Wratten [eds.], Biological Control: Measures of Success. Kluwer Academic Publishers, Hingham, USA. WILLIAMSON, S. H. 2009. Six ways to compute the relative value of a U.S. dollar amount, 1970 to present. [Online] http://www.measuringworth.com/uscompare/ WILSON, JR., J. A., AND DURANT, J. A. 1991, Parasites of the European corn borer (Lepidoptera; Pyralidae) in South Carolina, USA. J. Agr. Entomol. 8: 109-116. ZIMMERMANN, H. G., MORAN, V. C., AND HOFFMANN, J. H. 2001. The renowned cactus moth, Cactoblastis cactorum (Lepidoptera: Pyralidae): Its natural history and threat to native Opuntia oras in Mexico and the United States of America. Florida Entomol. 84: 543-551.

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Rengifo et al.: Host Status of Purple Passionfruit for Ceratitis capitata 91 HOST STATUS OF PURPLE PASSIONFRUIT FOR THE MEDITERRANEAN FRUIT FLY (DIPTERA: TEPHRITIDAE) J OSE A. R ENGIFO 1 J AVIER G. G ARCIA 1 J OHN F. R ODRIGUEZ 1 AND K RIS A. G. W YCKHUYS 2 1 Colombian Institute for Agriculture and Livestock, ICA, Bogota, Colombia 2 International Center for Tropical Agriculture CIAT, Cali, Colombia A BSTRACT The Mediterranean fruit y Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) is a key pest of a wide range of fruit crops and the focus of rigid quarantine restrictions and eradication measures in several countries In Colombia, the susceptibility of purple passionfruit ( Pasiora edulis f edulis Sims; Violales: Passioraceae) to C. capitata is uncertain. Field collections of fruit were made to evaluate natural infestation. Forced infestation studies were conducted in the laboratory with punctured and intact fruit to determine the acceptability of fruit at different stages of maturity and physiological suitability of fruit to development. No C. capitata larvae were found and no adults emerged from a total of 976 hand-picked fruit and 623 fallen fruit. In the meantime, trap data indicated that C. capitata is not present in the principal passionfruit production regions For intact fruit, C. capitata females oviposited exc lusively in fruit of maturity level zero, with 41.67% of fruit accepted for oviposition and an average of 183.1 33.8 eggs per fruit. No oviposition was recorded in fruit of maturity levels 2 and 4. For punctured fruit, C. capitata oviposited a total of 84,410 and 84,250 eggs into fruit of maturity levels 0 and 2, respectively, but no C. capitata adults emerged from fruit at either maturity level. Laboratory tests suggest that purple passionfruit is a non-host for C. capitata K ey Words: quarantine pest, Ceratitis capitata host status, risk analysis, fruit y R ESUMEN La mosca del Mediterrneo Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) es una plaga c lave de una amplia gama de frutales y es el foco de estrictas restricciones cuarentenarias y medidas de erradicacin en varios pases En Colombia, la susceptibilidad del maracuy morado ( Pasiora edulis f edulis Sims; Violales: Passioraceae) a C. capitata es incierta. Se hicieron colectas de frutos en campo para evaluar el nivel de infestacin. En el laboratorio se desarrollaron estudios de infestacin forzada con frutos perforados e intactos para determinar la aceptabilidad del fruto en los diferentes estados de maduracin e idoneidad siolgica del desarrollo de los frutos. No se encontraron larvas de C. capitata ni adultos emergidos en un total de 976 frutos recogidos manualmente y 623 frutos cados Mientras tanto, los datos de captura indicaron que C. capitata no est presente en las principales regiones de produccin del maracuy. Para frutos intactos, las hembras de C. capitata ovipositaron exc lusivamente frutos de nivel de maduracin cero, con 41.67% de aceptacin de frutos para oviposicin y en un rango de 183.1 33.8 huevos por fruto. No se registr oviposicin en frutos con niveles de maduracin 2 y 4. Para frutos perforados, C. capitata oviposit un total de 84,410 y 84,250 huevos dentro de frutos con nivel de maduracin 0 y 2 respectivamente pero no emergieron adultos de C. capitata de los frutos en ningn nivel de maduracin. Las pruebas de laboratorio sugieren que el maracuy morado no es hospedero para C. capitata Translation provided by the authors. Tephritid fruit ies are key pests of a wide variety of fruit species, affecting crop yield, quality of harvested produce, and (international) market access (e.g., Robinson & Hooper 1989; Aluja & Mangan 2008). Given the polyphagous nature of many fruit y species, quarantine restrictions are in place to avoid their introduction in certain countries or geographical regions. A key quarantine pest for the continental United States is the Mediterranean fruit y, Ceratitis capitata (Wiedeman), a destructive pest of multiple fruit crops worldwide (Liquido et al. 1991). In assessing risk of C. capitata arrival in the U.S. and developing associated quarantine protocols supreme precaution is taken to avoid entry of potential host fruits of this pest. Listings of the status of particular fruits as hosts of C. capitata are the cornerstone of quarantine restrictions (Liquido et al. 1991). However, current restrictions include fruit species for which there is poor information

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92 Florida Entomologist 94(1) March 2011 regarding C. capitata host status. Hence, researc h is needed to revise and update C. capitata host information and thereby improve quarantine decision making (Aluja et al. 2004; Pea et al. 2006; Jenkins & Goenaga 2008; Staub et al. 2008; De Graaf 2009; Follett et al. 2009). Purple passionfruit ( Passiora edulis f. edulis Sims) is one of several tropical fruits that is wellpositioned in local markets and gradually becoming popular internationally (Ocampo 2007; Wyckhuys et al. in press). In Colombia, purple passionfruit is mainly grown by small-scale, resourcepoor farmers on a total area of 100-400 ha. It is a protable crop and fresh fruit is increasingly being exported to northern Europe and Canada (Wyckhuys, unpublished data). Entry of fresh fruit into the continental U.S. is not permitted currently, based upon its presumed suitability as a host for Anastrepha spp. and C. capitata Liquido et al. (1991) list C. capitata as a potential pest of P. edulis but provide no evidence of adult y emergence from eld-collected fruit. Other reports indicate C. capitata is an occasional pest of Passiora sp., without specifying the exact crop species botanical form or variety (Thomas et al. 2001). Yellow passionfruit ( P. edulis f. avicarpa Degener) is reported as a possible host of C. capitata in Hawaii (Akamine et al. 1954), while many tephritids attac k certain Passiora species in Brazil (Aguiar -Menezes et al. 2002). In Colombia, national pest survey records for C. capitata maintained since 1986 have not detected this pest in the principal production regions of purple passionfruit (IC A, 2009). As a note of caution, it is important to indicate that climate change could cause altitudinal range shifts of pest tephritids and may eventually bring C. capitata into those production regions in the future (Hill et al. 2011). Considering a lack of scientic information regarding purple passionfruit host status for C. capitata and the importance of its production as source of income for rural smallholders we attempted to determine the host suitability of Colombia-grown purple passionfruit for C. capitata using standard methods (Cowley et al. 1992). This information can be used to re-evaluate the quar antine status of this fruit for market access to the United States. M ATERIALS AND M ETHODS All methodologies for host status screening were adopted from Cowley et al. (1992), taking into account parameters set by RSPM No. 30 (NAPPO 2008) and APPPC RSPM No. 4 (FAO 2005; Follett & Hennessey 2007). Field Collections Between Sep 2008 and May 2010, sampling was done during 4 distinct events in the principal purple passionfruit production regions, located in the departments of Boyac, Cundinamarca, Tolima, and Huila (Colombia). During each sampling event, 9-16 different purple passionfruit orchards were visited and fruit was collected from each orchard. Fruit samples consisted of hand-picked fruit of different maturity levels (i.e., fruit harvested from vines) and fallen fruit, collected from the ground. Fruit was sampled in a random fashion, and the number of fruit collected from each orchard depended upon phenological stage of the crop. We collected a total of 405, 285, 183, and 113 hand-picked fruit from Boyaca, Cundinamarca, Huila, and Tolima, respectively. Respective numbers of fallen fruit collected from each department were 345, 124, 96, and 58. Fruit samples were counted, weighed and taken to the Horticulture Research Center CIAA (Chia, Colombia) in ventilated plastic containers (70 50 50 cm) for further laboratory processing In the laboratory, fruit samples were kept at 22.0 2.0C, 65% RH and 12:12 L:D. Within 1 week following the collection, containers were screened for presence of fruit y puparia, and fruit were dissected to assess presence of tephritid larvae. Larvae were subsequently transferred to ventilated plastic Petri dishes with moistened vermiculite. Petri dishes were checked daily for adult emergence. We recorded the number of tephritid larvae and C. capitata adults for eac h sampling event and production region. Simultaneous with eld collections, McPhail traps (baited with protein hydrolysate; Cebofrut, AgroBiologicos SAFER, Medellin, Colombia) were deployed in orchards in each production region and visited bi-weekly to record the number of C. capitata adults. A total of 6 traps were deployed per orc hard, of which 5 were placed within the orchard itself and a sixth trap was placed outside the orchard in the dominant surrounding habitat type. To check trap attractiveness, we recorded captures of other tephritids. Laboratory Experiments Insect material was collected from coffee fruit ( Coffea arabica L.) in commercial orchards in Fredonia (Antioquia, Colombia), at 1,400 m altitude, and Medelln (Antioquia), at an altitude of 1,493 m. Upon eld collection, fruits were transferred to the ICA Entomology Laboratory in Bello (Antioquia). Each fruit was dissected and any tephritid larvae were allowed to pupariate in vermiculite. Puparia of C. capitata were subsequently taken to the Quarantine Treatment Laboratory of the Colombian Institute for Agriculture and Lifestock ICA in Mosquera (Cundinamarca) for further experimenting. Adults from eld collected puparia were exposed to mango ( Mangifera indica L.), a preferred host of C. capitata (NAPPO 2008).

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Rengifo et al.: Host Status of Purple Passionfruit for Ceratitis capitata 93 Adult ies were maintained within mesh cages (25x25x25 cm), allowed ample access to water and fed ad libitum with torula yeast and sugar. All insect developmental stages were maintained within c limate-controlled rearing chambers at 25 1C, 65 5% RH and 12:12 L:D. Second generation C. capitata adults were then used for host status trials Laboratory experiments were carried out between Oct 2008 and May 2009. Voucher specimens of study insects were kept at the ICA laboratory. All fruits used in the experiment were selected and harvested in several purple passionfruit orchards in Venecia (Cundinamarca) or mango orchards in La Mesa (Cundinamarca). Fruit of different maturity levels were selected based on commonly-used color tables for either mango or purple passionfruit (ICONTEC 1999; Pinzn et al. 2007). Prior to use in experiments, fruit was disinfected by immersion in a 0.05% sodium hypochlorite solution for 10 min. Subsequently, each fruit was dried and stored in plastic containers to use in host status trials. Fruit was used for experimenting within 72 h of harvest. Oviposition Preference Assay A total of 120 C. capitata pairs, aged 14 d, were placed within a mesh cage (70 50 50 cm) (Vidal et al. 2005) and allowed access to water and ad libitum torula yeast and sugar. Within each cage, we placed 8 purple passionfruit of eac h of 3 maturity levels (i.e., maturity 0, 2, and 4; see Pinzon et al. 2007). Purple passionfruit are approximately 5 cm in diameter. After 24 h, fruit was removed from the cages and dissected to determine the total number of C. capitata eggs. Over the course of 3 d, fruit were placed within each cage and subject to the same ovipositing C. capitata females. The experiment w as carried out with 3 replicates, thus screening 72 fruit per maturity level. The number of eggs within fruit of differing maturity level was compared by one-way analysis of variance (ANOVA). For all analyses, the statistical package SAS was used. Host Status Trials Based upon results of the previous assay, further trials were conducted to determine purple passionfruit host status to C. capitata To stimulate y oviposition, fruit was punctured with standard dissection pins (10 pinholes 1-2 mm into the fruit) before placing them within experiment cages (FAO 2005; NAPPO 2008). Purple passionfruit of maturity levels 0 and 2 were included in trials, while mango fruit (maturity degree 2 or 3) was used as a positive control. We placed 11 fruit per cage (70 50 50 cm) with 120 C. capitata pairs, aged 14-19 d and provided with water and ad libitum torula yeast and sugar. There were 3 replicates of eac h fruit type and maturity level, and simultaneous trials were conducted. In total, 990 purple passionfruit and 495 mango fruits were subjected to an infestation pressure of 10.9 C. capitata females per fruit. Over the course of 15 d, fruits within each cage were replaced on a daily basis, and subsequently kept within ventilated plastic containers. In a random fashion, a subsample of 45 fruits of either species or maturity degree was dissected upon removal from experimental cages to assess the number of C. capitata eggs. Remaining fruits were kept at 25 1C 65 5% RH and 12:12 L:D and were checked daily for larval emergence, puparia formation, or adult eclosion. After 15 d, all fruits were dissected and C. capitata larvae (per fruit) were counted and placed within vermiculite to allow pupariation. R ESULTS Field Collections From 2008 up to 2010, a total of 976 purple passionfruit were hand-picked and 623 fallen fruit were collected. No C. capitata adults emerged from any fruit. Diptera larvae were found within (immature) fruit; all of which successfully developed into lonchaeid adults. No C. capitata adults were caught in McPhail traps deployed in or near orc hards in any of the production regions. Trap effectiveness was conrmed through capture of Lonchaeidae (Diptera: Tephritoidea) at all locations. Laboratory Experiments: Oviposition Preference Assay The number of C. capitata eggs signicantly differed between fruit of distinct maturity degrees ( F = 18.84, df = 2, P < 0.0001). The highest number of eggs per fruit (183.7 33.8; mean SE) was oviposited in purple passionfruit of maturity level 0, while no eggs were laid in maturity levels 2 and 4. Host Status Trials Ceratitis capitata successfully completed its development on the preferred host mango, but no adults emerged from punctured fruit of maturity levels 0 and 2 (Table 1). Few C. capitata larvae developed in passionfruit, with larval weights ranging from 2.5 to 3.2 mg. In mango, the weight of third instars ranged from 9.7 to 10.3 mg. Of the 194 C. capitata that were obtained from passionfruit (maturity 0), <10% successfully pupariated. Puparial weights of individuals developing on passionfruit ranged from 2.3 to 3.1 mg, compared to C. capitata puparia from mango that weighed between 9.3 and 10.2 mg. Also, most C. capitata puparia that developed from passionfruit were

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94 Florida Entomologist 94(1)March 2011malformed. No adults eclosed from purple passionfruit puparia, whereas 46,920 adults emerged from infested mangos. DISCUSSIONFruit y host status determination lies at the basis of trade and can help connect small-scale fruit producers in the developing world to lucrative export markets. To aid developing nations in the process of assessing whether a given fruit is a host to a particular fruit y species, well-dened protocols and experimental guidelines have been dened (FAO 2005; Hennessey 2007; Aluja & Mangan 2008; NAPPO 2008). Natural eld infestation trials and a set of screen-house or laboratory experiments all help determine whether a given fruit crop is natural host, non-host or conditional host (e.g., Jenkins & Goenaga 2008; De Graaf 2009). These protocols have been adopted for a wide range of fruit crops, such as mamey sapote (Pouteria sapota (Jacq.)), litchi ( Litchi chinensis Sonn.), rambutan ( Nephelium lappaceum L.), avocado Hass (Persea americana (Mill.) Hass), highbush blueberry ( Vaccinium corymbosum L.), green mango ( Mangifera indica L. Tommy Atkins and Keitt), and others. Although data from natural eld infestation trials provide the most accurate assessment of host status of a given fruit (NAPPO 2008), a key limitation of these trials is that one cannot control variability in fruit y abundance. In our experiments, no C. capitata adults were reared from eld-collected passionfruit in the principal production regions of Colombia. However, McPhail trapping in orchards and surrounding habitats also did not encounter any wild C. capitata populations in any of these zones. Purple passionfruit crops are located at 2016.1 250.9 m (mean SD) above sea level (Wyckhuys et al. in press), while C. capitata has not been reported above 1,600 m (ICA 2009). Thus, under the current altitudinal and geographic distribution of C. capitata in Colombia it is very unlikely that this species affects purple passionfruit orchards. Climate change could eventually bring C. capitata into actual cropping regions and equally shift current passionfruit production zones to higher altitudes (Hill et al. 2011). At present however, natural eld infestation data remain inconclusive with respect to passionfruit host status. Forced infestation trials under laboratory conditions proved critical in delineating purple passionfruit host status to C. capitata Even though C. capitata females oviposited in intact fruit (maturity degree 0) as in punctured fruit of different maturity degrees, larval development was very poor and no adults emerged. No adult emergence from fruit under laboratory conditions is either indicative of its character as non-host under experimental conditions (NAPPO 2008) or as nonhost overall (FAO 2005). Nevertheless, we need to indicate that adult development from purple passionfruit could have been affected by dissecting infested fruit 15 d after oviposition. On less suitable hosts, C. capitata likely develop slow and take longer to complete larval development. However, fruit was dissected according to its deterioration status (see FAO 2005; NAPPO 2008), while taking into account an upper C. capitata egg-larval development time of 15 d (EPPO 2010). In conclusion, even though early dissection of purple passionfruit may have affected pupation and adult eclosion, the poor larval development and lack of emergence of adults from 18 C capitata puparia clearly indicate the poor suitability of this fruit. For intact fruit, maturity level 0 was preferred, while fruit of more advanced maturity were not accepted for oviposition by C. capitata Fruit maturity state can greatly affect its acceptability as an oviposition substrate by certain y species (Armstrong 2001; Willink & Villagran 2007). Certain physical stimuli determined by fruit maturity level (e.g., color) inuence C. capitata acceptance or rejection of fruit of particular maturity levels (Prokopy et al. 1984; Suarez et al. 2007). Also, fruit maturity level can affect physical resistance to oviposition and interfere with successful C. capitata oviposition (Gould & Hallman 2001). To circumvent such, C. capitata tend to oviposit in existing oviposition holes, bird pecks or crevicesTABLE 1.OVIPOSITION AND SUBSEQUENT DEVELOPMENT OF C. CAPITATA ON MANGO AND PURPLE PASSIONFRUIT (PPF)OF 2 MATURITY LEVELS UNDER LABORATORY CONDITIONS. DATA REPRESENT CONSOLIDATED NUMBER OF INDIVIDUALS WITHIN EACH C. CAPITATA DEVELOPMENT STAGE ON A TOTAL OF 990 PPF FRUIT OR 495 MANGO FRUIT. C. capitata development stages Tested commodity Eggs Larvae Puparia Adults Mango 139,410*64,990 53,854 46,920 PPF maturity degree 0 84,410 194 18 0 PPF maturity degree 2 84,250 0 0 0*The total number of eggs was determined by counting the number of C. capitata eggs on 10% of (dissected) fruits, and extrapolating this for all tested fruits.

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Rengifo et al.: Host Status of Purple Passionfruit for Ceratitis capitata 95(Aluja & Mangan 2008). This could further explain high degrees of oviposition in punctured fruits and low acceptability of intact fruit, more so at advanced maturity degrees at which purple passionfruit has an exceptionally rm epicarp. Fruit y oviposition in hosts that are inadequate for larval development is commonly observed (Joachim-Bravo et al. 2001). Especially for highly polyphagous species such as C. capitata, behavioral adaptations cause oviposition in a wide range of fruit crops (Aluja & Mangan 2008). Additionally, under highly articial conditions, time-limited gravid females may accept a broad range of substrates for oviposition (see Robacker & Fraser 2002). A high level of acceptance for oviposition of intact and punctured fruit does not necessarily imply suitability of the infested fruit for further larval development or adult emergence. Increased mortality, poor larval development and reduced puparia size or weight all are indicative of antibiosis and biochemical defenses (Greany et al. 1983) that cannot be detected by ovipositing females. Passiora species are cyanogenic and liberate hydrogen cyanide in fruits or leaves when under (insect) attack (Spencer & Seigler 1983). Possibly, these compounds disrupt larval development in passionfruit. As presence of low numbers of larvae in fruit is not indicative that it is an acceptable host (Gould & Hallman 2001; Jenkins & Goenaga 2007; Willink & Villagran 2007), we can conclude that Colombia-grown purple passionfruit is a non-host under the experimental conditions used in these tests and may be a non-host in the eld. Since C. capitata is currently not established in the principal growing areas in Colombia, it is very unlikely that this pest will infest purple passionfruit under natural conditions. There may therefore be signicant potential for the establishment of pest free areas to allow exports to the United States or a systems approach based upon low C. capitata prevalence and poor host status. ACKNOWLEDGMENTSWe are grateful to Catherine Varela, Maribel Hurtado, Jaime Abell, Juan Camilo Rodrguez, Paola Urn, Liliana Crdenas, Nubia Esmeralda Guerrero, Rubn Molina, Jos Rey, Gloria Palma, Oscar Menjura, Jorge Enrique Arias, and Yaneth Bernal at the ICA Quarantine Treatment Laboratory for help in fruit dissections and fruit y colony maintenance. We are grateful to Gloria Marlene Vidal (USDA-APHIS) for help with data interpretation and research conceptualization. This research was nanced by the Colombian Ministry of Agriculture and Rural Development, with project grant MADR 2008L6772-3445 to KW. REFERENCES CITEDAGUIAR-MENEZES, E. L., MENEZES, E. B., CASSINO, P. C., AND SOARES, M. A. 2002. Passion Fruit, pp. 361, 390 In J. E. Pea, J. L. Sharp, and M. Wyoski [eds.], Tropical Fruit Pests and Pollinators: Biology, Economic Importance, Natural Enemies and Control. CABI Publishing, Wallingford, UK. AKAMINE, E. K., HAMILTON, R. A., NISHIDA, T., SHERMAN, G. D., AND STOREY, W. B. 1954. Passion Fruit Culture. University of Hawaii, Extension Circular 345. 23 p. ALUJA, M., AND MANGAN, R. L. 2008. Fruit y (Diptera: Tephritidae) host status determination: critical conceptual, methodological and regulatory considerations. Annu. Rev. Entomol. 53: 473-502. ALUJA, M., DIAZ-FLEISCHER, F., AND ARREDONDO, J. 2004. Nonhost status of commercial Persea americana Hass to Anastrepha ludens Anastrepha obliqua, Anastrepha serpentina and Anastrepha striata (Diptera: Tephritidae) in Mexico. J. Econ. Entomol. 97: 293-309. ARMSTRONG, J. W. 2001. Quarantine security of bananas at harvest maturity against Mediterranean and Oriental fruit ies (Diptera: Tephritidae) in Hawaii. J. Econ. Entomol. 94: 302-314. COWLEY, J. M., BAKER, R. T., AND HARTE, D. S. 1992. Denition and determination of host status for multivoltine fruit y (Diptera: Tephritidae) species. J. Econ. Entomol. 85: 312-317. DE GRAAF, J. 2009. Host status of avocado (hass) to Ceratitis capitata Ceratitis rosa and Ceratitis cosyra (Diptera; Tephritidae) in South Africa. J. Econ. Entomol. 102: 1148-1459. EPPO. 2010. Ceratitis capitata Datasheet on Quarantine Pests. Accessed online from www.eppo.org on November 22, 2010. FAO. 2005. The Asia and Pacic Plant Protection Commission (APPPC RSPM No 4). Regional Standards for Phytosanitary Measures; Guidelines for the Conrmation of non-host Status of Fruit and Vegetables to Tephritid Fruit Flies; Rap Publication 2005/27. FOLLETT, P. A., ARMSTRONG, J. W., AND ZEE, F. T. 2009. Host status of blueberry to invasive tephritid fruit ies in Hawaii. J. Econ. Entomol. 120: 1859-1863. FOLLETT, P. A., AND HENNESSEY, M. K. 2007. Condence limits and sample size for determining nonhost status of fruits and vegetables to tephritid fruit ies as a quarantine measure. J. Econ. Entomol. 100: 251-257. GOULD, W. P., AND HALLMAN, G. 2001. Host status of mamey sapote to Caribbean fruit y (Diptera: Thephritidae). Florida Entomol. 84: 370-375. GREANY, P. D., STYER, S. C., DAVIS, P. L., SHAW, P. E.,AND CHAMBERS, D. L. 1983. Biochemical resistance of citrus to fruit ies. Demonstration and elucidation of resistance to the Caribbean fruit y, Anastrepha suspensa Ent. Exp. Appl. 34: 40-50. HENNESSEY, M. K. 2007. Guidelines for the Determination and Designation of Host Status of a Commodity for Fruit Flies (Tephritidae). USDA-CPHST, Orlando, Florida. HILL, J. K., GRIFFITHS, H. M., AND THOMAS, C. D. 2011. Climate Change and Evolutionary Adaptations at Species Range Margins. Annu. Rev. Entomol. 56: 143-159. ICONTEC, 1999. Frutas Frescas. Mango. Especicaciones. NTC 5210. Bogota Colombia. ICA (INSTITUTO COLOMBIANO AGROPECUARIO). 2009. Mosca del Mediterrneo ( Ceratitis capitata ) en Colombia Ao 2008-2009. Plan Nacional Mosca de las Frutas PNMF. Boletin Epidemiologico.

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96 Florida Entomologist 94(1)March 2011JENKINS, D. A., AND GOENAGA, R. 2007. Host status of mamey sapote, Pouteria sapota (Sapotaceae), to the West Indian fruit y, Anastrepha obliqua (Diptera: Tephritidae) in Puerto Rico. Florida Entomol. 90: 384-388. JENKINS, D. A., AND GOENAGA, R. 2008. Host status of litchi and rambutan to the West Indian fruit y (Diptera: Tephritidae) Florida Entomol. 91: 228-231. JOACHIM-BRAVO, I. S., FERNANDES, O. A., DE BORTOLI, S. A., AND ZUCOLOTO, F. S. 2001. Oviposition behavior of Ceratitis capitata Wiedemann (Diptera: Tephritidae): association between oviposition preference and larval performance in individual females. Neotropical Entomol. 30: 559-564. LIQUIDO, N. J., SHINODA, L. A., AND CUNNINGHAM, R. T. 1991. Host Plants of the Mediterranean Fruit Fly (Diptera: Tephritidae): An Annotated World Review. Misc. Publ. No. 77, Entomol. Soc. America. NAPPO. 2008. Guidelines for the Determination and Designation of Host Status of Fruits and Vegetables for Fruit Flies (Diptera: Tephritidae). RSPM No. 30. OCAMPO, J. A. 2007. Study of the Diversity of the Genus Passiora L. (Passioraceae) and its Distribution in Colombia. Ph.D Dissertation, Centre International dtudes Suprieures en Sciences Agronomiques Montpellier Sup. Agro. 268 pp. PEA, J. E., GOULD, W. P., HENNESSEY, M. K., HALLMAN, G. J., AND CRANE, J. H. 2006. Laboratory and eld infestation studies on immature green Tommy Atkins and Keitt mangoes to determine host status to the Caribbean fruit y (Diptera: Tephritidae). Proc. Florida State Hort. Soc. 119: 16-20. PINZON, I. M., FISCHER, G., AND CORREDOR, G. 2007. Determinacin de los estados de madurez del fruto de gulupa (Passiora edulis Sims). Agronoma Colombiana 25: 83-95. PROKOPY, R. J., MCDONALD, P. T., AND WONG, T. T. Y. 1984. Inter-population variation among Ceratitis capitata ies in host acceptance pattern. Ent. Exp. Appl. 35: 65-69. ROBACKER, D. C., AND FRASER, I. 2002. Attraction of Mexican fruit ies (Diptera: Tephritidae) to grapefruit: enhancement by mechanical wounding of and experience with grapefruit. J. Ins. Behavior 15: 399413. ROBINSON, A. S., AND HOOPER, G. 1989. Fruit Flies: Their Biology, Natural Enemies and Control. World Crop Pests, Vol. 3A. Elsevier Amsterdam. SPENCER, K. C., AND SEIGLER, D. S. 1983. Cyanogenis of Passiora edulis J. Agric. Food Chem. 31: 794796. STAUB, C. G., DE LIMA, F., AND MAJER, J. D. 2008. Determination of host status of citrus fruit against the Mediterranean fruit y, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). Australian J. Entomol. 47: 184-187. SUAREZ, L., MOLINA, A., MURUA, F., ACOSTA, J. C., MOYANO, B., AND ESCOBAR, J. 2007. Evaluacin de colores para la oviposicin de Ceratitis capitata (Diptera, Tephritidae) en Argentina. Revista Peruana de Biologa 14: 291-293. THOMAS, M. C., HEPPNER, J. B., WOODRUFF, R. E., WEEMS, H. V., STECK, G. J., AND FASULO, T. R. 2001. Mediterranean fruit yCeratitis capitata http://entnemdept.u.edu/creatures/fruit/mediterranean_fruit_ y.htm, revised on July 24, 2010. VIDAL, C. G., ABELLO, S. J., AND AREVALO, P. E, 2005. Multiplicacin de la mosca del Mediterrneo Ceratitis capitata W. En condiciones controladas de laboratorio. Revista ICA 32: 25-30. WILLINK, E., AND VILLAGRAN, M. E. 2007. Evaluacin del riesgo cuarentenario de introduccin de Ceratitis capitata en la palta Hass de Argentina. Actas VI Congreso Mundial del Aguacate. Via Del Mar, Chile. 12-16 Nov. 2007. WYCKHUYS, K. A. G., LOPEZ ACOSTA, F., ROJAS, M., ANDOCAMPO, J. D. 2011. Do farm surroundings and local infestation pressure relate to pest management in cultivated Passiora species in Colombia? International J. Pest Management, in press

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Salas et al.: Population Dynamics of Greenidea on Guava and Ficus97 POPULATION DYNAMICS OF TWO SPECIES OF GREENIDEA (HEMIPTERA: APHIDIDAE) AND THEIR NATURAL ENEMIES ON PSIDIUM GUAJAVA (MYRT ACEAE) AND FICUS BENJAMINA (MORACEAE) IN CENTRAL MEXICO M ANUEL D ARO S ALAS -A RAIZA 1 R OBERT W. J ONES 2 A LEJANDRO P EA -V ELASCO 1 O SCAR A LEJANDRO M ARTNEZ -J AIME 1 AND E DUARDO S ALAZAR -S OLS 1 Departamento de Agronoma, Divisin Ciencias de la Vida, Universidad de Guanajuato, Mexico dariosalasaraiza@hotmail.co Facultad de Ciencias Naturales, Universidad Autnoma de Quertaro, Juriquilla, Queretaro, Mexico A BSTRACT Greenidea psiidi van der Goot and Greenidea ficicola Takahashi (Hemiptera: Aphididae), are Asiatic species that feed on guava, Psidium guajava and Ficus spp.; both of these aphids were reported as exotic pests in Florida in 2002 and in Mexico in 2003. The present study characterized the population dynamics of both aphid species and their natural enemies on guava and ornamental figs in the Bajio region of Central Mexico. This report represents the first record of G. psiidi on Ficus sp. in Mexico and the first report of the presence of both species in the state of Guanajuato. Greenidea psiidi and G. ficicola were detected on guava in Mar 2007 and on fig trees during the same year in Apr near Irapauto, Guanajuato. Populations of both alate and apterous forms of G. psiidi in Apr were greater on guava than on fig trees ( W = 119.0; P = 0.0122), which coincided with new vegetative growth after leaf loss in winter on guava. In Apr populations of apterous forms of both species were significantly greater than winged forms on both guava and figs. No correlation was found between temperature changes and population densities of aphids. The indigenous predators, Chrysoperla comanche Banks, Chrysoperla exotera (Navs) and Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae), fed readily on the aphids and were found on both guava and fig trees, although densities of all 3 species were in greater numbers on Ficus. The combined population densities of the 3 predators had a positive correlation with that of G. ficicola ( r = 0.74), with a best fit found with a quadratic model of simple regression: y (densities of Chrysoperla spp.) = 1.2479x 2 4.3073x + 9.6493, and R 2 = 0.703. Nine species of coccinelid beetles (Coleoptera: Coccinellidae) were identified, the most common being of the genus Scymnus Results suggest that non-deciduous ornamental fig trees may serve as reservoirs of beneficial insects for deciduous guava trees. Results from the present study provide basic biological data to aid in management of these 2 exotic species of Greenidea on guava in central Mexico. K ey Words: guava, aphids, Greenidea Psidium guajava Ficus benjamina R ESUMEN Greenidea psiidi van der Goot y Greenidea ficicola Takahashi (Hemiptera: Aphididae), son de origen asitico, en 2002 fueron reportados como plaga extica en Florida y en Mxico en 2003. En el presente estudio, se describe la dinmica poblacional de estas dos especies de fidos y sus enemigos naturales en guayaba y Ficus en la regin de El Bajo en el centro de Mxico. Es el primer reporte de G. psiidi en Ficus spp. en Mxico y el primer reporte de ambas especies en el estado de Guanajuato. Greenidea psiidi y G. ficicola fueron observadas en guayaba desde marzo y en Ficus desde mediados de abril, en el rea de estudio. Las poblaciones de fidos alados y pteros de G. psiidi fueron ms altas en abril en guayaba que en Ficus ( W = 119.0; P = 0.0122), esto coincide con los nuevos brotes despus de que el rbol de guayaba pierde las hojas. Las poblaciones de las formas pteras de ambas especies fueron significativamente mayores que las formas aladas tanto en guayaba como en Ficus No se encontr correlacin entre la temperatura y las poblaciones de fidos. Los enemigos naturales Chrysoperla comanche Banks, Chrysoperla exotera (Nvas) y Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae), se alimentan de estos fidos y fueron encontrados tanto en guayaba como en Ficus aunque la densidad de las tres especies fueron ms altas en Ficus Las poblaciones combinadas de las tres especies de depredadores presentaron una correlacin positiva con la especie G. ficicola ( r = 0.74), lo cual se explica con la ecuacin cuadrtica de regresin simple: y (densidades de Chrysoperla spp.) = 1.2479x 2 4.3073x + 9.6493, donde el coeficiente R 2 = 0.703. Se identificaron nueve especies de Coccinellidae, siendo el ms comn el gnero Scymnus Estos resultados sugieren que los rboles de Ficus cuyo follaje es perenne, puede servir como refugio de insectos benficos para los rboles de guayaba, cuyo h-

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98 Florida Entomologist 94(1) March 2011 bito es caducifolio. Los resultados de esta investigacin aportarn conocimientos sobre la biologa de estas especies exticas de fidos lo que ayudar en el manejo de la plaga en huertas de guayaba en la regin central de Mxico. Translation provided by the authors. Mexico contributes 25% of the world production of guava ( Psidium guajava L.; Myrtaceae), of whic h the states of Michoacan, Aguascalientes, and Zacatecas of the central Altiplano region are the principal producers (Gonzlez-Gaona et al. 2002). In 2008, the exotic aphid pests, Greenidea psidii van der Goot and Greenidea cicola Takahashi were observed on gua va and ornamental g trees, Ficus benjamina (L.), in the central Altiplano state of Guanajuato (Salas-Araiza, unpublished data). There is concern among producers and agricultural researchers of central Mexico that these new pests may pose a threat to guava production in Mexico. The genus Greenidea Schouteden belongs to the subfamily Greenideinae within the Aphididae and includes approximately 45 species (Prez Hidalgo et al. 2009). The natural distribution of the genus is Asiatic and species are found to favor young foliage of plants of the families, Fagaceae, Betulaceae, Juglandaceae, Myrtaceae, Rosaceae, and Rubiaceae (Blackman & Eastop 1994). The presence of Greenidea in the New World may be the result of importation of infested ornamental g trees whic h are widely commercialized throughout the world, and they have been implicated as vehicles for the introduction of exotic pests (ODonnell & Parrella 2005). Greenidea psidii was rst discovered in the New World in1916 on guava and its relative, Psidium cattleianum Sabine in Brazil (NoembergLazzari et al 2006). Halbert (2004) reported that G. cicola was collected in Florida in 2002, and together with G. psidii were found to feed on guava and ornamental g trees ( Ficus benjamina (L.)). Prez Hidalgo et al. 2009 reported the presence of G. psidii in Costa Rica in 2009. The rst reports of Greenidea in Mexico come from Pea-Martnez et al. (2003), who recorded G. psidii feeding on gua va in the states of Hidalgo, Morelos, Guerrero, and the Federal District, and G. cicola feeding on ornamental g trees in the state of Guanajuato Mexico. Trejo-Loyo et al. (2004) reported G. cicola occurs on Myrtaceae in Cuernavaca, Morelos Although further data is lacking, it is probable that both G. psidii and G. cicola are widely distributed in Mexico The biology of species of Greenidea on guava and other host plants is poorly known (Halbert 2004; Sousa-Silva et al. 2005; Noemberg-Lazzari et al. 2006). Northeld et al. (2008) noted that many insect pests feed on alternative host plants and that the understanding of the relationships of pest populations on wild hosts with those on cultivated hosts is crucial in the development of management strategies. The objective of the present study was to determine the population dynamics of Greenidea psidii G. cicola and their natural enemies on Psidium guajava and Ficus benjamina in the state of Guanajuato, Mexico. The data generated from this study will help evaluate the importance of these aphids within the pest complex attac king guava in the Bajo Region, as well as aid in the development of integrated pest management strategies. M ATERIALS AND M ETHODS The study site was conducted at the experimental eld station of the Division of Life Sciences of the University of Guanajuato at the ExHacienda El Copal (101N, 20O) at 1,750 masl in the municipality of Irapuato, Guanajuato, Mexico. The region has a mean annual precipitation of 750 mm, a mean temperature of 19C and mean relative humidity of 56% (INEGI 2009). Two orchards, 1 guava ( P. guajava ), and the other ornamental gs ( F. benjamina ) were c hosen for the study sites. All trees were 10 years old and the 2 orchards were separated by approximately 1 km. Weekly samples were made of aphids (alate and apterous forms) and predators (adults of Coccinellidae and larvae of Chrysopidae) from 10 trees of each host species. For each tree, samples consisted of 20 beats of a 1-m wooden rod on the branches of the trees, from which insects fell onto a 1-m 2 beating sheet. All insects on the sheet were collected and placed in a labeled vial with 70% alcohol. The sample period w as from 23 Mar to 22 Jun 2007 which corresponded to the early spring growth period of leaves of guava and to the reproductive activity of aphids. Samples were not taken on subsequent dates because aphids had ceased reproductive growth and individuals were virtually undetectable. All material from each tree was preserved in individual vials containing 70% alcohol, with a corresponding label and brought to the laboratory. The collected specimens were identied in the Entomology Laboratory of University of Guanajuato, with a compound and stereo microscope. Aphids were mounted for species determination following techniques given by Pea-Martnez (1995). The keys of Blackman & Eastop (1994, 2000) were used for species identication of aphids. For the neuropteran and coccinellid predators, the keys of Lpez-Arroyo et al. (2008) and Gordon (1985) were used, respectively. Meteoro-

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Salas et al.: Population Dynamics of Greenidea on Gua va and Ficus99logical data was obtained from a weather station maintained at the study site located 200 m from the orchards. All identied specimens were deposited in the Entomological Collection Leopoldo Tinoco Corona of the University of Guanajuato. Data were analyzed with the statistical software program SAS (SAS 1995). Abundance measures were calculated for G. psiidi and G. cicola and the various predator species on guava and gs. Due to non-linearity of data determined by the Shapiro-Wilks test, mean comparisons of aphid and predator numbers between hosts and among samples dates were made with MannWhitney non-parametric procedures. In addition, correlation analyses were conducted between population numbers of aphids with those of the various predators and also with climatic variables (temperature and precipitation). RESULTS AND DISCUSSIONSpecies of Greenidea in Guava and FigsBoth G. psidii and G. cicola were present on guava and the ornamental g, F. benjamina (Figs. 1, A, B, C). This is the rst report of G. psiidi on the widely planted F. benjamina from Mexico, and for this species for the state of Guanajuato. Although the habitual host for G. cicola is Ficus spp (Noemberg-Lazzari et al. 2006), Halbert (2004) reports that this aphid species also occurs on guava, as conrmed in the present study. Although the apparent habitual hosts of G. psidii is guava, and that of G. cicola is Ficus spp., it is unclear how populations on the habitual and other infrequent host plants interact and which of the infrequent hosts can maintain viable, reproductive populations in the absence of the habitual host plants. Both species of Greenidea were aggregated on new shoots and leaves of their hosts, a feeding preference previously reported by Prez Hidalgo et al. (2009) On guava, G psiidi and G. cicola were present on new leaf buds and on either side of young leaves, whereas on F. benjamina they were found principally on the underside of young leaves. The preference by aphids for new plant growth is a common behavior in aphids. Gould et al. (2007) state that certain stages of aphids have preferences for specic tissues of host plants. For example, Chaitophorus populicola Thomas, preferably feeds on new growth with diverse and high levels of amino acids These authors also note that high sugar levels and low amino acid concentration in leaves increases the production of winged individuals.Population Dynamics of Greenidea psidii and Greenidea cicolaBoth G. psiidi and G. cicola were rst recorded on guava trees in late Mar 2007, although the abundance of G. psiidi was notably greater during Mar and early Apr. This appearance and population growth coincided with the emergence of new shoots and leaves on guava, the timing of which corresponds to that described by DaminNava et al. (2004). Colonizing alate aphids are often attracted to specic volatiles of host plants (Chapman et al. 1981; Nottingham et al. 1991; Powell & Hardie 2001). Because G. psiidi and G. cicola are found initially on young shoots and leaves, it is probable that initial alate colonizers are attracted to volatiles associated with new growth of guava plants. It important to note that the 2 tree species studied have marked differences in leang patterns and the appearance of new growth. Guava is deciduous, with complete leaf loss occurring generally in Nov with new growth beginning in Mar in the study area. This is in contrast to F. benjamina which is a non-deciduous tree that produces new growth apparently in response to environmental factors. However, both G. psiidi and G. cicola appeared rst on guava, and then on F. benjamina (Fig. 1, A, B, C), although foliage was available on the latter host throughout winter months. These data suggest that G. psiidi and G. cicola rst colonize and establish on guava and then later move to F. benjamina. This behavior was expected for G. psiidi for which guava is considered a habitual host, but not for G. cicola which Ficus is considered the habitual host. Further study is needed to establish the initial colonization behavior of these 2 species and whether the behaviors are the result of greater attractiveness of guava in comparison to F. benjamina and/ or the leaf quality of F. benjamina is inadequate until late Apr. The peak abundance of both species (alate and apterous forms) occurred in mid and late Apr (7.5 and 6.0 aphids/sample, for G. psiidi and G. cicola, respectively) (Fig. 1, A, B). The densities of G. psiidi were greater on guava than on F. benjamina during the rst 5 sample periods, and signicantly so at peak densities during the third week of Apr (w = 119.0; P = 0.0122; Fig. 1, A). By mid May, densities of G. psiidi and G. cicola were barely detectable, and remained at very low densities through late Jun. These low densities are presumably the result of the maturation of leaves and the deterioration of the physical and nutritional requirements for these species (Fig. 2, A). The appearance and abundance of alate forms in relation to apterous forms of G. psiidi and G. cicola followed patterns expected from observations reported by Noemberg-Lazzari et al. (2006). Alates were found in very low numbers initially on both guava and F. benjamina and differences between the 2 forms were not signicantly different until the third week of Apr (Fig. 2, B, C). At that time, the abundance of apterous forms in-

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100 Florida Entomologist 94(1)March 2011 Fig. 1. Population dynamics of apterous and alate forms of Greenidea on 14 samples dates in Irapuato, Guanaj uato, Mxico. Means with the same letter not signicantly different based on Mann-Whitney non parametric test (A: 04/20/07, w = 119.0, P =0.0122). Mean number aphids/sample

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Salas et al.: Population Dynamics of Greenidea on Guava and Ficus101 Fig. 2. Population dynamics of apterous and alate forms of Greenidea on 14 samples dates in Irapuato, Guanaj uato, Mexico. 2007. Means with same letter not signicantly different based on Mann-Whitney non parametric test (B: 04/13/07, w = 138.5, P = 0.0179; 04/20/07, w = 100.5, P = 0.002; 04/27/07, w = 35.0, P = < 0.001; 05/04/07, w = 88.0, P = < 0.0003; 05/11/07, w = 118.0, P = 0.0037; C: 04/20/07, w =127.5, P = 0.016; 04/27/07, w = 17, P = < 0.0001). Mean number aphids/sample

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102 Florida Entomologist 94(1)March 2011creased considerably (16-18 fold) for both species. Signicant differences between the abundance of alate and apterous forms of G. psiidi on guava and F. benjamina were found from mid Apr through mid May (Fig. 2, B). Alate forms were also most abundant during this period, and continued to be found into Jun. It is assumed that many of these alate individuals dispersed from guava and F. benjamina when leaf and shoot quality declined after Apr. No correlation was found between the mean ambient temperature nor mean precipitation with the population changes of G. psiidi and G. cicola on guava and F. benjamina ( r = 0.15 for temperature; r = -0.21 for precipitation for G. psidii ; r =0.16 for temperature; and r = -0.18 for precipitation for G. cicola), This was expected given that there was little change in either temperature or precipitation during the sample period (Fig. 3). Even with greater temperature variation, however, little correlation would be expected given that temperature is often not a principal factor in aphid population growth. For example, Desneux et al (2006) reported that temperature was not positively correlated with population densities of Aphis glycines Matsumura on soybeans and some species, such as Schizaphis graminum (Rondan) on wheat produce more nymphs at lower temperatures (Pendleton et al. 2009). Differences in precipitation also were probably not an important factor in the changes in populations that were observed because the only noteworthy change in precipitation occurred between the last two samples. Although precipitation has been observed to physically dislodge aphids from plants and increase infection by pathogens (Nielson & Barnes 1961), in our study precipitation occurred primarily in Jun, and thus well after populations had already declined in late May. It could be argued that the life cycle of G. psiidi and G. cicola is adapted to avoid the unfavorable effects of the seasonal rains by completing their life cycle during the dry season. However, Noemberg-Lazzari et al. (2006) reported that in Brazil both species are present year round ( G. psiidi on guava and G. cicola on Ficus sp.), suggesting that the life cycle of the 2 insects is facultative and probably dependant more on host quality than on specic seasonal variations in climate.Natural EnemiesChrysopidae. Three species of Chrysopidae (Neuroptera) were identied on guava and F. benjamina: (1) Chrysoperla comanche (Banks), (2) Chrysoperla exotera (Nvas), and (3) Chrysoperla carnea (Stephens). The most abundant species was C. comanche and the present work represents the rst report of this species for the state of Guanajuato. This chrysopid is cited by Ramrez-Delgado et al. (2006) as the predominant species in pecans and grapes in the states of Coahuila and Durango, suggesting that it is one of the most important aphid predators of the central highlands of northern Mexico. The combined populations of the 3 species of Chrysoperla were present from 20 Apr 2007 to 15 Jun 2007 in guava and cus. Chrysoperla was rst recorded on F. benjamina on 20 Apr of 2007 (Fig. 4, A). In both guava and F benjamina, an increase of individuals of Chrysoperla spp. coincided with an apparent increase in the populations of the apterous forms for both aphid species on 20 Apr 2007 (Fig. 1, A, B, C). Populations of the Fig. 3. Mean temperature (C) and cumulative precipitation (mm) at El Copal, Irapuato, Guanajuato, Mxico, 2007.

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Salas et al.: Population Dynamics of Greenidea on Guava and Ficus103combined densities of Chrysoperla spp. peaked rst on F. benjamina on 27 Apr 2007 (0.32 individuals/sample) and later on guava on 11 May 2007 (0.15 individuals/sample). Densities of these predators gradually decreased from mid-May to mid Jun. Combined populations of the 3 species of Chrysoperla were positively correlated with populations of G. cicola (r = 0.74) on guava and F. benjamina and the relationship dened by y (densities of Chrysoperla spp.) = 1.2479x2 4.3073x + 9.6493, grade 2 polynomial with R2 = 0.703 (Fig. 5). This suggests that there was a numerical response in the populations of Chrysoperla species in conjunction with density changes in G. cicola. There was no apparent correlation found between populations of G. psiidi with Chrysoperla on either guava or F. benjamina. Coccinellidae. Species of Coccinellidae (Coleoptera) on guava and F. benjamina were: HippoFig. 4. Population dynamics of the combined totals of 3 species of Chrysoperla and Scymnus spp on 14 sample dates in Irapuato, Guanajuato, Mexico. 2007. Means with the same letter are not signicantly different based on the Mann-Whitney non parametric test ( B: 05/11/07, w =77.5, P = 0.016; 05/25/07, w = 92.0, P = < 0.001; 06/01/07, w = 95.0, P = < 0.001; 06/15/07, w = 83.0, P = < 0.006; 06/22/07, w = 83.0, P = < 0.006). Mean number predators/sample

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104 Florida Entomologist 94(1)March 2011damia convergens Gurin-Mneville, Harmonia axyridis Pallas, Zoglobra spp., Coccinella spp., Olla v-nigrum (Mulsant), Cycloneda sanguinea L., Stethorus spp., Hyperaspis quadrioculata (Motschulsky), and Scymnus spp. On F. benjamina, Scymnus spp. were the most abundant with 167 total individuals found compared with 126 captured in guava. Individuals of Scymnus spp. rst appeared in F. benjamina on 20 Apr 2007 and began increasing by 11 May 2007, reaching a maximum abundance on 1 Jun 2007 of approximately 6 individuals/sample and maintained densities of 2.5-4 individuals/sample until the end of the sample period. The overall population densities of Scymnus spp. was higher on F. benjamina than on guava (11.9 vs 9 individuals/sample, respectively), and on 5 of the 10 sample dates, and these predators had signicantly higher densities on F. benjamina than on guava (Fig. 4, B). Results suggest that other prey present on F. benjamina were more important in maintaining Scymnus spp. than the Greenidea species because these predators appeared after major populations peaks of Greenidea and increased after mid-May in the virtual absence of these aphids. Aphid mummies were not observed in the present study and there are no reports of parasitism of Greenidea suggesting that predators are the principal insect biological control agents of Greenideae in the region. Although F. benjamina was a host plant of Greenidea, both aphid species were found rst on guava, and then reached higher densities on guava than on F. benjamina. However, following aphid colonization, chrysopid and coccinelid predators developed greater densities on F. benjamina. These data suggest that F. benjamina is more important as a refuge for natural enemies of the pest aphids in the region, than as an alternative host plant from which Greenidea colonizes guava. The movement of predators from one host to another is well documented, in particular for aphidophagous generalists such as chrysopids and coccinellids (Sloggett et al. 2008). The following study lays the groundwork for the management of Greenidea psidii and G sicola in guava in the region. Indigenous predators were found to readily feed on G. psiidi and G. cicola and are apparently an important component in reducing densities of the pests. Further work is needed to study in detail the colonization behavior of G. psiidi and G. cicola in relation to guava and other hosts, and to investigate methods to optimize the action of native predators on these aphids. REFERENCED CITEDBLACKMAN, R. L., AND EASTOP, V. F.1994. Aphids on the Worlds Trees. An Identication and Information Guide CAB International, Oxon. 8+988 pp and 16 plates BLACKMAN, R. L., AND EASTOP, V. F. 2000. Aphids on the Worlds Crops: An Identication Guide. John Wiley & Sons. New York. 466 pp. CHAPMAN, R. F., BERNAYS, E. A., AND SIMPSON, S. J. 1981. Attraction and repulsion of the aphids Cavariella aegopodii, by plant odors. J. Chem. Ecol. 7: 881889. DAMIN-NAVA, A., GONZLEZ-HERNNDEZ, V. A., SNCHEZ-GARCA, P., PEA-VALDIVIA, C. B., LIVERA-MUOZ, M., AND BRITO-GUADARRAMA, T. 2004. Crecimiento y fenologa del guayabo (Psidium guajava L.) cv. Media China en Iguala, Guerrero. Rev. Fitotec. Mexicana 27(4): 349-358. DESNEUX. N., ONEIL. R. J., AND YOO, H. J. S. 2006. Suppression of population growth of the soybean aphid, Aphis glycines Matsumura, by predators: The identication of key predators and the effects of prey dispersion, predators abundance, and temperature. Environ. Entomol. 35(5): 1342-1349. GORDON, R. D. 1985. The Coccinellidae (Coleoptera) of America North of Mexico. J. New York Entomol. Soc. 93(1): 1-912 GOULD, G. G., JONES, C. G., RIFLEMAN, P., PREZ, A.,AND COLEMAN, J. S. 2007. Variation in eastern cottonwood (Populus deltoides Bartr.) phloem sap content caused by leaf development may affect feeding site selection behavior of the aphid, Chaitophorus populicola Thomas (Homoptera: Aphididae). Environ. Entomol. 36(5): 1212-1225. HALBERT, S. E. 2004. The genus Greenidea (Rhynchota: Aphididae) in the United States. Florida Entomol. 87(2): 159-163. INEGI. 2009. Instituto Nacional de Estadstica Geografa e Informtica. Informacin Geogrca. Mapa de Climas. (3 January 2009) (http://mapaserver.inegi.org.mx/geografa/espaol/ estados/gto/ clim.cfm?c=444&e=07) LPEZ-ARROYO, J. I., DE LEN-HERNNDEZ, T., RAMREZ-DELGADO, M., AND LOERA-GALLARDO, J. 2008. Especies de Chrysoperla (Neuroptera: Fig. 5. Correlation and simple regression model between number of individuals of Greenidea cicola and number of individuals of 3 species of Chrysoperla on 14 sample dates in Irapuato, Guanajuato, Mxico, 2007.

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Salas et al.: Population Dynamics of Greenidea on Guava and Ficus105Chrysopidae) presentes en Mxico, pp. 69-80 In M. D. Salas-Araiza y E. Salazar-Sols [eds.], Entomfagos en el Control de Plagas Agrcolas en Mxico. Primera Edicin. Universidad de Guanajuato, Guanajuato. Gto. 90 pp. NIELSON, M. W., AND BARNES, O. L. 1961. Population studies of the spotted alfalfa aphid Arizona in relation to temperature and rainfall. Ann. Entomol. Soc. America. 54(3): 441-448. NOEMBERG-LAZZARI, S. M., ZONTA-DE CARVALHO, R. C., CARDOSO, J. T., AND CALADO, D. C. 2006. First record of Greenidea psidii van der Goot and comparison with Greenidea cicola Takahashi (Hemiptera: Aphididae) in Brazil. Zootaxa 1235: 63-68. NORTHFIELD, T. B., PAINI, D. R., FUNDERBURK, J. E.,AND REITZ, S. R. 2008. Annual cycles of Frankliniella spp. (Thysanoptera: Thripidae) thrips abundance on North Florida uncultivated reproductive hosts: predicting possible sources of pest outbreaks. Ann. Entomol. Soc. America 101(4): 769778. NOTTINGHAM, S. F., HARDIE, J., DAWSON, G. W., HICK, A. J., PICKETT, J. A., WADHAMS, L. J., AND WOODCOCK, C. M. 1991. Behavioral and electrophysiological responses of aphids to host and nonhost plant volatiles. J. Chem. Ecol. 17: 1231-1242 ODONNELL, CH. A., AND PARRELLA, M. P. 2005. Host suitability of selected Ficus species for Thrips palmi (Thysanoptera: Tripidae). Florida Entomol. 88(1): 97-98. PADILLA-RAMREZ, J. S., GONZLEZ-GAONA, E., REYESMURO, L., AND MAYEK-PREZ, N. 2007. Produccin de fruto e ndices productivos en rboles de guayabo. Agric. Tc. Mxico 33(2): 191-196. PENDELTON, B. B., PALOUSEK COPELAND, A. L., ANDMICHELS, JR., G. J. 2009. Effect of biotype and temperature on tness of greenbug (Hemiptera: Aphididae) on sorghum. J. Econ. Entomol. 102(4): 1624-1627. PEA MARTNEZ, R. 1995. Colecta y montaje de dos (Homoptera: Aphididae). Curador Entomolgico y Acarolgico No. 2:9-14. PEA-MARTNEZ, R., VILLEGAS-JIMNEZ, N., REYESBARRN, L., PADILLA-BADILLO, G., AND HALBERT, S. 2003. Dos nuevas plagas potenciales en Mxico (Homoptera: Aphididae: Grenideinae). Entomologa Mexicana 3: 411-415. PREZ HIDALGO, N., VILLALOBOS-MULLER, W., AND MIER-DURANTE, P. 2009. Greenidea psidii (Hemiptera: Aphididae: Greenideinae) new invasive aphid in Costa Rica. Florida Entomol. 92(2): 396-398. POWELL, G., AND HARDIE, J. 2001. The chemical ecology of aphid host alternation: How do return migrants nd the primary host plant? Appl. Entomol. Zool. 36: 259-267. RAMREZ-DELGADO, M., LPEZ-ARROYO, J. I., NAVACANBEROS, U., GONZLEZ-HERNNDEZ, A., AND MERCADO-HERNNDEZ, R. 2006. Diversidad y uctuacin poblacional de especies de Chrysopidae en frutales del norte de Mxico. Entomologa Mexicana 5 (1): 480-485. SAS. 1995. Users guide for linear models. SAS Institute Inc. Cary, North Carolina. SLOGGETT, J. J., ZEILSTRA, I., AND OBRYCKI, J. J. 2008. Patch residence by aphidophagous ladybird beetles: Do specialists stay longer? Biol. Control 47: 199-206. SOUSA-SILVA, C. R., BROMBAL, J. C., AND ILHARCO, F. A. 2005. Greenidea cicola Takahashi (Hemiptera: Greenideidae), a new aphid in Brazil. Neotrop. Entomol. 34(6): 1023-12024. TREJO-LOYO, A. G., PEA-MARTNEZ, R. AND VILLEGASJIMNEZ, N. 2004. Adofauna (Hemiptera: Aphididae) de Cuernavaca, Morelos, Mxico. Folia Entomol. Mexicana 43(2): 191-202.

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106 Florida Entomologist 94(1) March 2011 TRANSMISSION OF THE MYCOPATHOGEN, HIRSUTELLA SPP., TO NYMPHS AND ADULTS OF THE GLASSY-WINGED SHARPSHOOTER, HOMALODISCA VITRIPENNIS (=COA GULATA) IN THE GREENHOUSE V ERENA -U LRIKE L IETZE 1 R USSELL F. M IZELL III 2 AND D RION G. B OUCIAS 1 1 Entomology and Nematology Department, University of Florida, Gainesville, FL 32611 2 North Florida Research and Education Center, University of Florida, Quincy, FL 32351 The mycopathogen Hirsutella spp. produces regular epizootics in Florida populations of the glassy-winged sharpshooter (GWSS), Homalodisca vitripennis a polyphagous, xylem-feeding cicadellid native to the southeastern U .S. and northeastern Mexico (Boucias et al. 2007). The GWSS transmits a lethal phytopathogenic bacterium, Xylella fastidiosa and thereby threatens the production of economically important plants (Redak et al. 2004). Recent introduction of GWSS into southern California, French Polynesia, Tahiti and Hawaii has stimulated interest in identifying potential biocontrol agents against invasive populations (Boucias et al. 2007; Hunnicutt et al. 2008). During a eld survey in Quincy, FL, a new species of Hirsutella H. homalodiscae nom. prov., w as detected in ~50% of mycosed sharpshooters (Boucias et al. 2007). Virtually nothing is known about the transmission of Hirsutella within GWSS populations Our eld observations have shown that Hirsutellainfected sharpshooters attac h via rhizoids to the plant bark in the summer and often remain attached throughout the winter. Placed in a moist environment, overwintered cadavers produce conidiospores suggesting they serve as microhabitats protecting Hirsutella and facilitating transmission to the new generation of GWSS This study examined potential transmission routes of Hirsutella under greenhouse conditions Experimental treatments included topical spore application and choice-exposure to sporulating cadavers on host plants. For insect rearing, eld-collected, healthy adults without hyphal bodies (HBs) in their hemolymph were identied by non-destructive antennal bleeding (Breaux 2005) and maintained on caged, potted plants (soybean, Glycine max cotton, Gossypium hirsutum and cowpea, Vigna unguiculata ) in a greenhouse (temperature 2630C 14:10 h light:dark photoperiod). Leaves with egg masses were removed and hatching neonates were transferred to caged lemon basil ( Ocimum basilicum ). Soil was watered to saturation once daily In each bioassay replicate, individual cotton or basil plants housing groups of 10-20 adults or nymphs, respectively, were covered with clear, gauze-covered acryl cylinders (15 cm diameter 45 cm high). Mortality and infection were recorded daily by removing dead individuals from the soil surface and examining their hemolymph for HB propagation. Cadavers were maintained on water agar to record mycosis. After 3 weeks, each plant was examined for mycosed cadavers, and surviving GWSS were subjected to hemolymph examination. Statistical analyses were conducted using the Proc Genmod procedure and Ls means statement of the Statistical Analysis System (SAS) for Windows to compare mortality or infection responses by logistic regression (Neter et al. 1990; SAS 2004). For topical application of Hirsutella spores, healthy adults were treated in three replicate assa ys by touching their ventral surface to either sporulating in vitro colonies of strain 3A (maintained at the UF Entomology/Insect P athology lab, Gainesville, FL) (total n = 46 adults), to GWSS cada vers displaying spores of Hirsutella ( n = 33), or to a nutrient agar plate (control, n = 35). Mortality in control, in vitro and cadaver treatments w as similar with 56 11%, 74 12%, and 60 8%, respectively ( 2 = 2.48, P = 0.1155). Contact with agar in vitro colonies or cadavers produced 0%, 13 6% and 42 5% infection, respectively, and only the latter treatment produced mycosis (28 6%). Infection transmitted from cadavers was signicantly higher than that transmitted from in vitro colonies ( 2 = 8.02, P = 0.0046). T o examine whether co-existence of healthy and mycosed GWSS on a plant would result in disease transmission, either overwintered cadavers collected in Jan (stored at -80C) or new cadavers collected in Jul/Aug were pinned to plants (10 per plant) and groups of healthy nymphs or adults were maintained on each plant for up to 3 weeks. Controls were conducted on plants without cadavers. The majority of pinned cadavers displayed sporulating Hirsutella mycelium within one week. New cadavers developed an unusually thick, white mycelium overgrowing the entire insect (Fig. 1A). Disease transmission was observed to varying degrees (Table 1). Dead exposed nymphs and adults (Fig. 1B and C), attached to the plant and displaying Hirsutella -induced mycosis were seen as early as 7 and 12 d after exposure, respectively. Hemolymph-borne HBs were detected as early as 8 d after exposure. New cadavers were more efcient in disease transmission when compared with overwintered cadavers (Table 1). When nymphs were exposed to new cadavers, no survivors were found and all dead insects were overgrown with thick mycelium

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Scientic Notes 107 (Fig. 1A). Exposure of nymphs to overwintered cadavers induced 13 8% mortality and 8 5% infection producing a light and at mycelium (Fig. 1B). Adult mortality after exposure to new and overwintered cadavers was 97 2% and 76 8%, respectively, and signicantly higher than in the controls (47 14%) (Table 1). Infection of adults induced by new cadavers (60 9%) was signicantly higher than that induced by overwintered cadavers (13 9%). Potentially, Hirsutella is one of the major mortality factors in endemic GWSS populations (Boucias et al. 2007). Our results clearly demonstrated that disease transmission from mycosed cadavers to healthy conspecics efciently occurs in undisturbed, small host populations. Identifying attractive cues that would cause foraging sharpshooters to contact infectious spores will be the next step for developing this pathogen as microbial control agent against H. vitripennis Fig. 1. Cadaver exposure experiments with Homalodisca vitripennis (A) Part of a lemon basil plant spiked with new cadavers 3 weeks after introduction of nymphs. Note the white, thick mycelium overgrowing the pinned cadaver (pc) in the center and the introduced, mycosed nymphs (ic). (B and C) Mycosed nymph and adult, respectively, displaying thin Hirsutella mycelium (m) 3 weeks after release on a plant spiked with overwintered cadavers. T ABLE 1.M EAN (SE) PERCENT MORTALITY AND INFECTION OF H OMALODISCA VITRIPENNIS 3 WEEKS AFTER INTRODUCTION TO CAGED PLANTS HARBORING H IRSUTELLA MYCOSED CADAVERS Life stage exposedCadavers Mortality a,b Infection a Induced mycosis c Nymphs None (control)0 0 a (139)0 0 a No Overwintered 13 8 b (68) 8 5 b Yes (2) New 100 0 f (53)100 0 d Yes (53) Adults None (control)47 14 c (103)0 0 a No Overwintered 78 6 d (61)13 5 b Yes (1) New 97 2 e (61)60 9 c Yes (30) a Numbers followed by different letters indicate significant differences (SAS proc genmod and lsmeans statement, P < 0.05). b Numbers in parentheses indicate the total number of GWSS exposed in 4 replicate assays. c Numbers in parentheses indicate the number of GWSS displaying mycosis within the 3-week observation period.

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108 Florida Entomologist 94(1) March 2011 S UMMARY For the rst time, transmission of Hirsutella from mycosed, sporulating H. vitripennis to healthy conspecics w as demonstrated. In addition, methods were established to amplify infectious material in vivo for potential inoculative release. R EFERENCES C ITED B OUCIAS D. G., S CHARF F. W., B REAUX S. E., P URCELL D. H., AND M IZELL R. E. 2007. Studies on the fungi associated with the glassy-winged sharpshooter Homalodisca coagulata with emphasis on a new species Hirsutella homalodiscae nom. prov. Biocontrol 52: 231-258. B REAUX S. E. 2005. Fungi associated with the glassy winged sharpshooter, Homalodisca coagulata in its native range University of Florida (Masters thesis), Gainesville, 108 pp. H UNNICUTT L. E., M OZORUK J., H UNTER W. B., C ROSSLIN J. M., C AVE R. D., AND P OWELL C. A. 2008. Prevalence and natural host range of Homalodisca coagulata virus-1 (HoCV-1). Arch. Virol. 153: 61-67. N ETER J. W ASSERMAN W., AND K UTNER M. H. 1990. Applied linear statistical models: regression, analysis of variance, and experimental designs. Richard Irwin Inc., Illinois. R EDAK R. A., P URCELL A. H., L OPES J. R. S., B LUA M. J., M IZELL R. F., III, AND A NDERSEN, P. C. 2004. The biology of xylem uid-feeding insect vectors of Xytella fastidiosa and their relation to disease epidemiology. Annu. Rev. Entomol. 49: 243-270. SAS. 2004. Users guide, version 9.1. SAS Institute, Cary, North Carolina.

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Scientic Notes 109 EXAMINATION OF METHODS FOR FORMOSAN SUBTERRANEAN TERMITE (ISOPTERA: RHINOTERMITIDAE) FECES RECOVERY T IMOTHY J. A RQUETTE 1 AND J OSE M. R ODRIGUEZ 2 1 Department of Entomology, Mississippi State University, Mississippi State, MS 39762 2 State Chemical Laboratory, Mississippi State, MS 39762 Chemical assay of undigested food from termite fecal material has been reported for Formosan subterranean termites ( Coptotermes formosanus Shiraki) and other species (Itakura et al. 1995; Hyodo et al. 1999; Mishra & Sen-Sarma 1979; Katsumata et al. 2007). Unlike drywood termites, which produce solid fecal pellets with a conspicuous appearance (Lewis et al. 2010), subterranean termites produce liquid feces (Gouge et al. 2001) that is potentially difcult to recognize and obtain. The current study describes the outcome of attempts at recovery of C. formosanus feces from laboratory arenas in whic h no food had been placed, and from arenas containing lter paper or wood. Termites used for this study were collected at Audubon Park, New Orleans, LA. Arenas without food were maintained and processed 2 different ways. For the rst set of arenas, 18 groups of approximately 50 workers that had been fed southern yellow pine ( Pinus sp.) for at least 72 h were transferred to pre-weighed bowls shaped from aluminum foil. Bowls containing termites were placed in polystyrene boxes (32 cm 25 cm 10 cm) (Tri-State Plastics Latonia, KY) lined with saturated paper towels. The boxes were covered and maintained at 23C. At 3, 6, 20, 24, 40, and 48 h, workers were removed from three randomly selected bowls by allowing them to crawl up a lter paper disc. After removal of dead insects and body parts, bowls were dried 15 min at 70C, and folded closed immediately upon removal from heat. Fecal dry weight was determined to 0.01 mg from the difference between the initial and nal weight of the bowl. For the second set of arenas, 3 groups of approximately 500 workers fed southern yellow pine for at least 72 h were transferred to polystyrene boxes lined with aluminum foil over saturated paper towels. Containers were covered and maintained at 23C. After 4, 8, 12, 28, and 38 h, termites were transferred to clean polystyrene boxes. Following each transfer, dead termites were removed, and air-dried feces transferred to pre-weighed aluminum foil bowls with a watercolor brush. Feces were further dried 15 min at 70C before weighing. For arenas containing lter paper, 3 groups of 50 workers were placed in plastic screw-top vials (40 mm 15 mm diameter) along with a piece of moistened Whatman 1 lter paper (Whatman Inc., Piscataway, NJ). Vials were loosely capped and maintained sideways at 28C in the dark. After 1 week insects were removed and collection of fecal material attempted by scraping vials and paper with a razor blade and spatula. For laboratory arenas containing wood blocks, 5 groups of approximately 200 to 500 workers and soldiers were placed in clear polystyrene containers lined with a piece of aluminum foil over saturated paFig. 1. Mean (+SEM) dry weight of C. formosanus feces recovered from laboratory arenas without food. Graph A: mean feces recoveries from 3 randomly selected foil bowls holding 50 workers. Graph B: repeated feces collection from containers that each held 500 workers and soldiers.

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110 Florida Entomologist 94(1) March 2011 per towels. Southern yellow pine blocks (either 50 mm 20 mm 5 mm, or 20 mm 3 ) that had been soaked in distilled water for 1 h were added to arenas. Containers were covered and stored at 28C. Accumulated debris on foil and wood was periodically collected from 4 arenas with a spatula and razor blade, while a fth arena was left undisturbed for 6 weeks. The weight of fecal material recovered from arenas that had not contained food averaged approximately 0.03 mg to 0.2 mg feces (dry weight) from foil bowls, and about 1 mg total feces (dry weight) after repeated collections from the same groups of insects (Fig. 1). Feces in these arenas were apparent either as liquid droplets that dried to dark solid specks, or collections of small, sticky black piles mixed with dead termites, body parts, and trapped live termites. No feces were recovered from arenas containing lter paper, although liquid fecal material was apparent from dark spots on the paper. For arenas with wood, solid debris was apparent on wood blocks and arena surfaces within 72 h. Debris was formed into branched structures in the single undisturbed arena (Fig. 2). The structures were soft and friable immediately upon removal from arenas, and when pulverized dried to a ne powder. Considering the minute quantity of feces recovered from arenas without food (Fig. 1), little fecal material appears to have been available for use in construction of the structures. Feces were not visually discernible from any arena containing wood blocks. Observations from the current study underscore the need for caution in collection of C. formosanus fecal material for assay because it can easily be contaminated by foreign partic les or unrecoverable if food is present. Furthermore, collection of feces for assay from arenas without food may be impractical because of low yield (Fig. 1). For instance, this species digests wood carbohydrate very efciently (Yoshimura 1995), so large numbers of termites would be needed to produce fecal material with enough undigested sugars for detection by conventional chromatographic methods. We appreciate the helpful assistance and advice from Ms. S. Parikh and Dr. J. Sun (Mississippi State University), and thank Dr. B. Layton and Dr. G. Baker (Mississippi State University) for reviewing an early draft of this manuscript. S UMMARY The current study attempted different methods for collection of Formosan subterranean terFig. 2. Cemented structures constructed by C. formosanus termites in a laboratory arena containing southern yellow pine blocks.

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Scientic Notes 111 mite feces. When food was present, the liquid fecal material was not recoverable either due to absorption into food, or occlusion by food particles that were used instead for construction purposes. Uncontaminated fecal material was recoverable from arenas without food only in minute quantity. R EFERENCES C ITED G OUGE D. H., S MITH K. A., O LSON C., AND B AKER P. 2001. Drywood termites. Cooperative Extension, College of Agriculture & Life Sciences, The University of Arizona. Available from: http://ag.arizona.edu/pubs/ insects/az1232/ H YODO F., A ZUMA J.-I., AND A BE T. 1999. Estimation of effect of passage through the gut of a lower termite, Coptotermes formosanus Shiraki, on lignin by Solid-State CP/MAS 13 C NMR. Holzforschung 53: 244-246. I TAKURA S., U ESHIMA K., T ANAKA H., AND E NOKI A. 1995. Degradation of wood components by subterranean termite, Coptotermes formosanus Shiraki. Mokuzai Gakkaishi 41: 580-586. K ATSUMATA K. S., J UN Z., H ORI K., AND I IYAMA K. 2007. Structural changes in lignin of tropical woods during digestion by the termite, Cryptotermes brevis. J. Wood Sci. 53: 419-426. L EWIS V. R., N ELSON L. J., H AVERTY M. I., AND B ALDWIN J. A. 2010. Quantitative changes in hydrocarbons over time in fecal pellets of Incisitermes minor may predict whether colonies are alive or dead. J. Chem. Ecol. (in press). M ISHRA S. C., AND S EN -S ARMA P. K. 1979. Studies on deterioration of wood by insects III. Chemical composition of faecal matter, nest material and fungus comb of some Indian termites. Mater. Organismen 14: 1-14. Y OSHIMURA T. 1995. Contribution of the protozoan fauna to nutritional physiology of the lower termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae). Wood Res. 82: 68-129.

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112 Florida Entomologist 94(1) March 2011 TRACHYAPHTHONA NIGRITA AND TRACHYAPHTHONA SORDIDA (COLEOPTERA: CHRYSOMELIDAE) REJECTED AS POTENTIAL BIOLOGICAL CONTROL AGENTS OF PAEDERIA FOETIDA L. (RUBIACEAE), AN INVASIVE WEED IN HA WAII AND FLORIDA R OBERT W. P EMBERTON 1 AND G LORIA L. W ITKUS United States Department of Agriculture, Agricultural Research Service, Invasive Plant Research Laboratory, 3225 College Ave., Ft. Lauderdale, FL 33314 1 Rpemberton5@gmail.com Skunk vine, Paederia foetida L. (Rubiaceae), is an invasive weed native to Asia but a problem in Florida and other areas of the southern USA, and in Hawaii (Pemberton & Pratt 2002). In the southern states of the USA it is mostly a problem in natural areas, whereas in Hawaii the plant is a problem in both natural areas and agriculture. Skunk vine is both a Category I weed (Florida Exotic Pest Plant Committee 2009) and a Florida Noxious Weed (USDA, Natural Resource Conservation Service). Biological control research began with eld surveys in Asia in 1997 (Pemberton & Pratt 2002). A lace bug, Dulinius conchatus Distant (Tingidae), and a leaf beetle, Sphenoraia rutilans Hope (Chrysomelidae) have undergone host specicity testing but neither demonstrated the nar rowness of host range needed in a skunk vine biocontrol agent (Pemberton et al. 2005; Pemberton & Witkus unpublished data). Two Trachyaphthona spp. ea beetles were identied as promising biological control candidates during surveys in J apan (Pemberton & Pratt 2002). Both species have larvae that feed on roots and adults that consume leaves. The biology and preliminary host ranges of Trachyaphthona sordida (Baly) and T. nigrita Ohno were studied in Japan (Okamato et al. 2008). Adult host range testing on T. nigrita was conducted in our F ort Lauderdale quarantine laboratory during 2007 and 2008, with eld-collected beetles shipped by Japanese cooperators during each Jun of those years. Adult host range testing on eld-collected T. sordida beetles was done in 2009 after J apanese cooperators shipped the beetles during Jun of that year. Adults of both species were set up in wooden screened sleeve cages, 50 cm 50 cm 53 cm, and fed cut vines of the control plant, Paederia foetida, and its invasive relative Paederia cruddasiana Pain Paederia cruddasiana, native to Nepal, Burma, China, and Thailand (Puff 1991), is a naturalized weed in Miami-Dade County, Florida. The beetles mated readily and laid eggs throughout the sleeved rearing cages. The most likely potential non-target plants of these beetles are the many native and economic members of the Rubiaceae in Florida. The host range testing for T. nigrita and T. sordida initially concentrated on 3 subfamilies within the Rubiaceae Test plants were native and ornamental members of this family selected to represent different taxonomic tribes. Host range testing on plants outside of the Rubiaceae consisted of species selected to represent related orders including the Cornales, Gentianales, Lamiales, and the Solonales. In total, 34 species in 7 families and 5 orders were tested. Host range tests on both beetles were conducted with a no-c hoice test design, and each individual test plant was exposed to 3 beetles for 1 month. Control plants of P. foetida and P. cruddasiana were exposed to 3 beetles for 1 month. Because researc h on T. nigrita spanned 2 seasons, eac h experiment was replicated 5 times. Research on T. sordida was conducted for 1 season so each test w as replicated 3 times. Host range testing for T. nigrita and T. sordida was conducted on branc hes of whole potted plants in the quarantine greenhouse. Clear acrylic tubes, 15.2 cm long 3.8 cm diameter (6 1.5 diameter), each with 2 mesh ventilation holes were placed on the end of a small branch or stem of each test plant. A slit sponge bung was placed around the base of the stem and up into the bottom of the tube. The stem and bung juncture was wrapped tightly with paralm. Another bung was used to seal the top of the tube. Due to the saltatory nature of the ea beetles, an aspirator was used to gently transfer 3 tiny beetles through the bung slit and into the tubes and onto each test branch. The 3 beetles consisted of 1 mating pair and another beetle of undetermined gender due to the difculty in determining gender in this ea beetle. Beetles were monitored for 1 month, and at the end of each week, all leaves with feeding damage were clipped, pressed and then scanned into jpeg les to enable feeding to be measured with SigmaScan Pro. The most important nding of the research was the nearly equal or greater feeding by both Trachyaphthona species on the native Florida test plant Diodia virginiana L. (Table 1). Trac hyaphthona nigrita adults ate a mean of 3.75 1.1 mm 2 leaf material of D. virginiana, compared to 4.76 1.53 mm 2 of leaf material on P. foetida

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Scientic Notes 113 T ABLE 1.H OST RANGE TESTING OF T RACHYAPHTHONA NIGRITA AND T RACHYAPHTHONA SORDIDA ADULT FLEA BEETLES SHOWING PLANTS THAT RECEIVED FEEDING T HREE ADULT BEETLES WERE TESTED FOR 1 MONTH FOR EACH SPECIES D IFFERENCES IN TOTAL AMOUNTS OF FEEDING ARE DUE TO 5 REPLICATES CONDUCTED OVER 2 SEASONS WITH T. NIGRITA VERSUS 3 REPLICATES WITH T. SORDIDA F LORIDA NATIVES ARE IN BOLD OrderFamily Tribe Species Trachyaphthona nigrita Trachyaphthona sordida total (mm 2 )mean (mm 2 )SDSEtotal (mm 2 )mean (mm 2 )SDSE Cornales Garryaceae Acuba japonica 0.01* 1 **0.000.000.000.00 Rubiales Rubiaceae Hedyotideae Pentas lanceolata 0.03* **0.110.040.040.02 Morindeae Morinda citrifolia 0.11* **0.000.000.000.00 Paederieae Paederia cruddasiana 16.533.310.570.334.151.380.630.36 Paederieae Paederia foetida 23.824.761.530.884.841.610.820.47 Paederieae Serissa foetida 3.330.670.370.220.450.150.210.12 Spermacoceae Diodea virginiana 18.763.751.100.649.703.230.500.29 Spermacoceae Ernodia littoralis 0.02* **0.040.010.010.01 Spermacoceae Mitchella repens 1.150.230.180.100.670.220.280.16 Spermacoceae Spermacoce assurgens 0.01* **0.01 ** Spermacoceae Spermacoce tetraquetra 2.790.560.160.090.12 ** Spermacoceae Spermacoce verticillata 0.000.000.000.000.06 ** 1 Test feeding was documented in only 1 replicate of each species.

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114 Florida Entomologist 94(1) March 2011and 3.31 0.57 mm2 on P. cruddasiana Trachyaphthona sordida adults consumed a mean of 3.23 0.5 mm2 leaf material on D. virginiana, compared to 1.61 0.82 mm2 on P. foetida, and 1.38 0.63 mm2 on P. cruddasiana Moreover, D. virginiana belongs to a taxonomic group within the Rubiaceae different than that of skunk vine, i.e., the tribe Spermacoceae rather than the tribe Paederieae for skunk vine. In addition, T. nigrita caused what is considered to be more than test feeding on Spermacoce tetraquetra A. Rich. another member of the Spermacoceae. Trachyaphthona nigrita adults consumed a mean of 0.56 0.16 mm2 of S. tetraquetra, compared to 1.61 0.82 mm2 on P. foetida, and 1.38 0.63 mm2 on P. cruddasiana. To prevent harm to native and important economic members of the Rubiaceae, the feeding of these ea beetles would need to be limited to plants in the skunk vine tribe, the Paederieae. It may be possible that the larvae of these beetles could have greater specicity than the adults. Two factors inuenced our decision to cease research on these Trachyaphthona species and to shift to other candidate insects of skunk vine. Most importantly, we have not successfully reared these univoltine beetles, which are, like many root-feeding ea beetles, difcult to culture in captivity. Secondly, our previous testing with the leaf beetle Sphenoraia rutilans found that it could complete its development both on Diodia virginiana and Spermacoce tetraquetra, indicating that these plants have an attractiveness and vulnerability to some Paederia feeding insects. SUMMARYTrachyaphthona nigrita and Trachyaphthona sordida, 2 potential biological control agents of Paederia foetida L. (Rubiaceae), were collected from Japan and brought into quarantine for adult host specificity testing. Testing indicated that they fed significantly on Florida native plants in the tribe Spermacoceae, a different tribe from that to which the target weed Paederia foetida belongs (Paederieae). These beetles, therefore, lack the appropriate level of host specificity, and this eliminates them from further consideration as potential biological controls of Paederia foetida, skunk vine. ACKNOWLEDGMENTSWe thank Junichi Yukawa, Fukuoka University (retired), and his students for collecting the Trachyaphthona beetles in Japan and shipping them to Florida. Rachel Taylor provided technical assistance and Luke Kasarjian grew the test plants. The South West Florida Water Management District and the Florida Department of Environmental Protection funded this research. REFERENCES CITEDFLORIDA EXOTIC PEST PLANT COMMITTEE. 2009. Florida Exotic Pest Plant Councils 2009 list of invasive species. Wildland Weeds 12: 13-16. OKAMOTO, C., TUDA, K., YAMAGUCHI, D., SATO, S., PEMBERTON, R. W., AND YUKAWA, J. 2008. Life history traits and host specicity of Japanese Trachyaphthona species (Coleoptera: Chrysomelidae), candidates as biological control agents against skunk vine, Paederia foetida (Rubiaceae), in Southeastern United States and Hawaii. Entomol. Sci. (Japan) 11: 143-152. PEMBERTON, R. W., AND PRATT, P. D. 2002. Skunk vine (Paederia foetida), pp. 343-351 In R. Van Driesche, B. Blossey, M. Hoddle, S. Lyon, and R. Reardon [eds.], Biological Control of Invasive Plants in the Eastern United States U.S. Forest Service, Forest Health Technology Enterprise Team-2002-04, Morgantown, West Virginia. PEMBERTON, R. W., MURAI, K., PRATT, P. D., AND TERAMOTO, K. 2005. Dulinius conchatus (Hemiptera: Tingidae), considered and rejected as a potential biological control agent of Paederia foetida, an invasive weed in Hawaii and Florida. Proc. Hawaiian Entomol. Soc. 37: 81-83. PUFF, C. 1991. Revision of the genus Paederia in Asia, pp. 207-289 In C. Puff [ed.], The Genus Paederia L. (RubiaceaePaederieae): A Multidisciplinary Study, Opera Botanica Belgica 3. National Botanic Garden of Belgium. Meise, Belgium. USDA NATURAL RESOURCE CONSERVATION SERVICE. 2010. http://plants.usda.gov/java/noxious?rptType= State&stateps=12. Last accessed on 29 Oct 2010.

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Scientic Notes 115 RICH MICROBIAL COMMUNITY ASSOCIATED WITH THE NEST MATERIAL OF RETICULITERMES FLAVIPES (ISOPTERA: RHINOTERMITIDAE) T HOMAS C HOUVENC 1* M ONICA L. E LLIOTT 2 AND N AN -Y AO S U 1 1 Department of Entomology and Nematology, Ft. Lauderdale Research and Education Center, University of Florida, Institute of Food and Agricultural Sciences, 3205 College Ave, Ft. Lauderdale, FL 33314, USA 2 Department of Plant Pathology, Ft. Lauderdale Research and Education Center, University of Florida, Institute of Food and Agricultural Sciences, 3205 College Ave, Ft. Lauderdale, FL 33314, USA *Corresponding author; E-mail: tomchouv@u.edu Subterranean termites live in a soil environment that can contain a rich community of microorganisms and the nest material can be a site of enhanced microbial activity (Holt & Lepage 2000). By using feces and saliva as building materials for nest construction and gallery systems, termites can alter the qualitative and quantitative composition of the resident soil microorganism community from adjacent soils (Jouquet et al. 2005). Keya et al. (1982) suggested that a signicant part of the altered microorganism community was mostly composed of cellulose decomposers, but function and origin of the microbial community associated with termite nests remains unclear. In this report we evaluate whether subterranean termites have the ability to import various microorganisms from distant soils or from their own gut microbial community into a microorganism-free soil environment. In Chouvenc et al. (2008), we investigated the survivorship of groups of 960 Reticulitermes avipes (Kollar) from large two-dimensional arenas lled with sterile sand (w ashed with 70% ethanol, rinsed with sterile deionized water and oven dried at 60C for 24 h). At the end of the 90-d experiment, 4 arenas from 2 different termite colonies were dismantled to obtain 2 types of nest material: (1) gallery material including sand mixed with termite fecal material from termites galleries, and (2) non-gallery material including undisturbed sand (not tunneled by the termite) at least 3 cm away from any termite gallery (Fig. 1). A total of 12 soil samples per arena (10 g per sample) including 6 samples for each type of nest material were collected. In order to estimate the microbial communities in the 2 types of nest material, 1 g sub-sample from each original sample was processed following the general procedure described in Elliott & Des Jardin (1998). Serial dilutions of the sub-samples were made (10 -1 to 10 -7 ) with sterile water and the diluted sub-samples were plated on 5 different selective media to isolate various groups of microbes including (1) overall aerobic bacteria, (2) uorescent pseudomonads, (3) actinobacteria, (4) overall fungi and, (5) entomopathogenic fungi. All selective media were previously described in Elliott & Des Jardin (1998) except the medium for entomopathogenic fungi (Veen & Ferron 1966). Soil samples collected from a control arena were established with only sand and no termites to conrm the absence of microbes in the sterile sand. Growth of microbes for each dilution and each selective medium was quantied to estimate the overall microbial structure of the 2 nest materials (Table 1). Individual microbe colonies were selected and transferred on 1/5 strength potato dextrose agar. Based on colony morphology of more than 2,000 isolates, we found 432 morphologically distinct isolates including 399 aerobic bacterial isolates, 11 actinobacterial isolates, and 22 fungal isolates. The molecular identication of these isolates and their potential function will be presented elsewhere. Samples from the control arena with no termites showed no microbial growth. Our results showed that when groups of 960 termites are allowed to forage and build a complex gallery system in sterile sand, a large microbial community establishes in the sand wall galleries mixed with fecal material. Because the sampling method cannot take into account the non-culturable microbes, our results suggest that the actual microbial diversity within the termites gallery system is probably greater than what we could observe. After 90 d in the presence of termites in the galleries, this microbial community moved away from the tunnels into the undisFig. 1. A group of termites in a two-dimensional arena for 90 d. Arrows show sampling sites for microbioal isolation. Note the dark shade in the gallery wall showing the presence of deposited fecal material. Bar = 1 cm.

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116 Florida Entomologist 94(1) March 2011 turbed sand, although the overall amount of bacteria remains 100-fold less than the fecal-sand mix. Previous studies have shown that actinobacteria can be isolated from the gut of various termites (Knig et al. 2006). Here we demonstrate that actinobacteria are also part of the microbe community that can colonize the termite nest structure. Fungi represented a very small fraction of the overall microbes enumerated, supporting the observation by Chouvenc et al. (2008, 2009) that termite gut contents, i.e., fecal material, used as building material for the gallery system possess highly fungistatic properties, and do not allow fungi to colonize the termite nest environment. The apparent absence of entomopathogenic fungi support that termites have mechanisms to prevent the spread of harmful fungi within the termite nest (Chouvenc et al. 2010). Since all the arenas were constructed of sterile sand, it is assumed all the microorganisms originated from the termite guts or from the surface of the cuticle of the termites. This indicates that a complex microbial community is associated with the termites themselves before they were introduced into the arena. We assume that most of the cuticular microbes were acquired from the eld nest where they were originally collected, although some could be strict termite-associated microbes and therefore ectosymbionts transmitted vertically. In addition, the gut can be a reservoir that temporarily hosts various microbes, which are later inoculated into various part of the gallery system. Because fungi were sampled in very small quantities, we hypothesize that the termite fecal material within the tunnel wall acts as an inhibitor against specic types of organisms. A broader sampling of termite colonies from diverse origin and molecular identication of the microbes will provide a better understanding of the origin and the function of some of these microbes. S UMMARY Field-collected subterranean termites were held in groups of 960 individuals for 90 d in twodimensional arenas lled with sterile sand. After 90 d, the tunnel walls made of sand mixed with fecal material, and sand from areas not disturbed by termites were compared for the presence of microbes. We show that a rich microbial community associated with the termites can colonize the termite nest environment and is primarily associated with the tunnels. R EFERENCES C ITED C HOUVENC T., AND S U N.-Y. 2010. Apparent synergy among defense mechanisms in subterranean termites (Rhinotermitidae) against epizootic events The limits and potential for biological control. J. Econ. Entomol. 103: 1327-1337. C HOUVENC T., S U N.-Y., AND E LLIOTT M. L. 2008. Interaction between the subterranean termite Reticulitermes avipes (Isoptera: Rhinotermitidae) and the entomopathogenic fungus Metarhizium anisopliae in foraging arenas. J. Econ. Entomol. 101: 885-893. C HOUVENC T., S U N.-Y., AND R OBERT A. 2009. Inhibition of Metarhizium anisopliae in the alimentary tract of the eastern subterranean termite Reticulitermes avipes J. Invertebr. Pathol. 101: 130-136. EL LIOTT M. L., AND D ES J ARDIN E. A. 1998. Comparison of media and diluents for enumeration of aerobic bacteria from bermudagrass golf course putting greens. J. Microbiol. Methods 34: 193-202. H OLT J. A., AND L EPAGE M. 2000. Termites and soil properties, pp. 389-407 In T. Abe, D. E. Bignell, and M. Higashi [eds.], Termite: Evolution, Sociality, Symbioses, Ecology. Kluwer Academic Publishers, Dordrecht. J OUQUET P., R ANJARD L., L EPAGE M., AND L ATA J. C. 2005. Incidence of fungus-growing termites (Isoptera, Macrotermitinae) on the structure of soil microbial communities. Soil Biol. Biochem. 37: 18521859. K EYA S. O., M URERIA N. K., AND A RSHAD M. A. 1982. Population dynamics of soil microorganisms in relation to proximity of termite mounds in Kenya. J. Arid Environ. 5: 353-359. K NIG H., F RHLICH J., AND H ERTEL H. 2006. Diversity and lignocellulolytic activities of cultured microorganisms, pp. 271-301 In H. Knig and A. Varma [eds.], Intestinal Microorganisms of Termites and Other Invertebrates. Springer, New York. V EEN K. H., AND F ERRON P. 1966. A selective medium for the isolation of Beauveria tenella and Metarrhizium anisopliae J. Inverteb. Pathol. 8: 268-269. T ABLE 1.C OLONY FORMING UNITS (CFUS) OF VARIOUS MICROBE GROUPS GROWN ON DIFFERENT SELECTIVE MEDIA PER GRAM OF SAMPLE OF TERMITE GALLERY MATERIAL AND NON GALLERY MATERIAL ( MEAN SE ). Microbe Group n a Gallery materialNon-gallery material t -test b Overall aerobic bacteria243.5 10 7 7.9 x 10 6 3.9 10 5 5.2 x 10 4 P < 0.001 Fluorescent Pseudomonads 247.2 10 2 3.0 x 10 2 8 6 P < 0.001 Actinobacteria 241.4 10 5 2.8 x 10 4 1.1 10 2 5.2 x 10 1 P < 0.001 Overall fungi 246.1 10 2 1.1 x 10 2 4.3 10 1 1.6 x 10 1 P < 0.001 Entomopathogenic fungi 12 0 0 0 0 N/A a Total number of samples plated on a given selective media per concentration from serial dilutions. b For a given microbe group based on selective media, comparison of the colony forming units between gallery material and nongallery material.

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Scientic Notes 117 GUT CONTENT ANALYSIS OF SOUTHERN AND TAWNY MOLE CRICKETS (ORTHOPTERA: GRYLLOTALPIDAE: SCAPERTISCUS ) D. E. S ILCOX AND R. L. B RANDENBURG Department of Entomology, North Carolina State University, Campus Box 7613, Raleigh, NC 27695 Two introduced species of mole crickets, Scapteriscus borellii Giglio-Tos, the southern mole cric ket and S. vicinus Scudder, the tawny mole cric ket cause economic damage to turfgrass in North Carolina and throughout the southeastern U.S. Previous studies indicate that Scapteriscus borellii primarily feed on insect material and Scapteriscus vicinus feed on plant material (Ulagaraj 1975; Taylor 1979; Matheny 1981; Fowler et al. 1985). There have been no additional research efforts to examine the gut content of these 2 pests as their range has expanded over the past 25 years to their northern-most habitat (North Carolina) with changes in ecology, host range, or diet. This research was conducted to determine the current feeding preferences of North and South Carolina populations of these 2 mole cricket species. Nymph and adult S. borellii and S. vicinus were collected during the spring and summer of 2009 with soapy w ater ushing (Short & Koehler 1979) and individual crickets were immediately preserved in 70% ethyl alcohol. Scapteriscus borellii nymphs were collected from Belvedere Country Club (P ender Co., NC, 34.3675, 77.710833) on 15 Sep and adults were collected from Olde Fort Golf Course (Brunswick Co., NC, 34.0857, 78.0536) on 5 May. Scapteriscus vicinus nymphs were collected from Scotch Meadows Country Club (Scotland Co ., NC, 34.4553, 79.2813) on 8 Oct and adults were collected from High Tech Turf (Horry Co., SC, 33.5097, 79.322) on 6 May and 12 May. All collected individuals were examined for species characteristics to ensure proper identication (Potter 1998). The alimentary canals (crop, proventriculous, and hindgut) of 25 late instars (large nymphs) and 25 adults for each species were removed and examined. Each cricket was placed in a petri dish (8.89 cm diameter, Fisher Scientic, Pittsburgh, PA) ventral side up, and an X-Acto knife (x3201, Elmers Products Inc., Columbus, OH) was used to remove the sterna to expose the alimentary canal. A pair of forceps was used to remove the alimentary canal. The cricket body cavity was placed into a plastic vial lled with 70% ethyl alcohol and labeled to identify each specimen. The alimentary canal remaining in the petri dish was covered with 70% ethyl alcohol to prevent desiccation. Forceps were used to tease open the crop, proventriculous, and hindgut. All gut content was noted for each cricket. The content was examined under a binocular microscope used in the 7X-30X power range and the contents were categorized as presence of plant material only, presence of insect material only, or presence of plant and insect material. Content was determined to be plant material if it was brous, green, or light brown in color and if it had a blade-like appearance (Castner & Fowler 1984). Content was determined to be animal material if it was dark brown or black, obviously sclerotized, or if they were a recognizable structure such as, tarsi, antennae, legs, etc (Castner & Fowler 1984). The gut content was thoroughly examined and analyzed until all pieces were identied and categorized. The contents were then removed from the petri dish with a bulb-pipette and placed in the vial with the cricket from which it was extracted. Data were analyzed by Chi-square analysis through use of Statistical Analysis System version 9.1 program (SAS Institute 2003). Of the 25 S. borellii nymph alimentary canals examined 28% 0.46% contained only plant material, 4% 0.20% contained only insect material, and 68% 0.48% contained plant material and insect material (Fig. 1). Of the 25 S. borellii adult alimentary canals examined 56% 0.51% contained only plant material, 20% 0.41% contained only insect material, and 24% 0.44% contained plant material and insect material (Fig. 1). There is signicant difference in the overall gut contents between S. borellii nymphs and S. borellii adults (Chi-square = 10.2609, P = 0.0059). Of the 25 S. vicinus nymph alimentary canals examined 60% 0.5% contained only plant material, 4% 0.2% contained only insect material and 36% 0.49% contained plant material and insect material (Fig. 1). Of the 25 S. vicinus adult alimentary canals examined, 96% 0.2% contained only plant material, 0% contained only insect material, and 4% 0.2% contained plant material and insect material (Fig. 1). There is signicant difference in the overall gut content between adult S. vicinus and nymph S. vicinus (Chi-square = 9.4769, P = 0.0088). There is Fig. 1. The frequency of various gut contents for S. borellii and S. vicinus nymphs and adults.

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118 Florida Entomologist 94(1) March 2011 also signicant difference ( P < 0.05) in the gut content between S. borellii (nymphs and adults) and S. vicinus (nymphs and adults) (Chi-square = 14.8027, P = 0.0006). W e determined that the gut content of North and South Carolina populations of S. borelli and S. vicinus is similar to previous ndings for South America and Florida populations (T able 1). Modest changes in feeding preferences were apparent as the southern mole cricket in North Carolina appeared to be more herbivorous than previous studies in other locations indicated in the past. The differences in length between S. borelli and S. vicinus alimentary canals could be related to differences in diet. The alimentary canal of S. vicinus is signicantly longer than in S. borellii (Nation 1983). The longer alimentary length seen in S. vicinus may reect the greater difculty of digesting plant cells; more area is needed for the chemical digestion to occur (Nation 1983). The differences in gut content seen in both species between nymphs and adults could be attributed to the different developmental needs of the life stages (Forrest 1987). We found a higher percentage of herbivory in both species as adults as compared to nymphs. We also observed greater herbivory in the southern mole cricket as compared to previous reports. Nymphs of the European mole cricket Gryllotalpa gryllotalpa Linnaeus (Ulagaraj 1975) that were fed on insect food completed development in 2 years while nymphs fed on vegetable matter completed development in over 4 years (Ulagaraj 1975). The protein found in animal/insect components of diet may be important in development. S UMMARY The alimentary canals of 25 nymph and 25 adult S. borellii and S. vicinus were dissected to determine the gut contents in North and South Carolina mole cric ket populations as compared to previous studies in other locations S borellii nymphs were found to primarily consume plant and insect materials while S. borellii adults were found to primarily consume only plant materials Scapteriscus vicinus nymphs and adults were found to primarily consume only plant material. The results of this study were similar to those of previous studies conducted more than 25 years ago in Brazil and Florida. R EFERENCES C ITED C ASTNER J. L., AND F OWLER H. G. 1984. Gut content analysis of Puerto Rican mole crickets (Orthoptera: Gryllotalpidae: Scapteriscus ). Florida Entomol. 67(3): 479-484. F ORREST T. G. 1987. Insect size tactics and developmental strategies. Oecologia 73: 178-184. F OWLER H. G., V IEIRA D E C AMARGO M. T., AND C RESTANA L. 1985. Feeding habits of Brazilian mole crickets (Orthoptera: Gryllotalpidae: Scapteriscus spp. and Neocurtilla sp.). J. Econ. Entomol. 78: 1076-1078. M ATHENY J R ., E. L. 1981. Contrasting feeding habits of pest mole cricket species. J. Econ. Entomol. 74: 444445. N ATION J. L. 1983. Specialization in the alimentary canal of some mole crickets (Orthoptera: Gryllotalpidae). Intl. J. Insect Morphol. & Embryol. 12(4): 201-210. P OTTER D. A. 1998. Destructive turfgrass insects: biology, diagnosis, and control. Ann Arbor Press, Chelsea, Michigan. SAS I NSTITUTE 2003. PROC Users Manual, version 9.1 SAS Institute, Cary, North Carolina. S HORT D. E., AND K OEHLER P. G. 1979. A sampling technique for mole crickets and other pests in turfgrass and pasture. Florida Entomol. 62: 282-283. T AYLOR T. R. 1979. Crop contents of two species of mole crickets, Scapteriscus acletus and S. vicinus (Orthoptera: Gryllotalpidae). Florida Entomol. 62(3): 278279. U LAGARAJ S. M. 1975. Food habits of mole crickets (Orthoptera: Gryllotalpidae: Scapteriscus ). J. Georgia Entomol. Soc. 10(3): 229-231. T ABLE 1.T HE PERCENT HERBIVORY CARNIVORY AND OMNIVORY FOR S. BORELLII AND S. VICINUS POPULATIONS IN B RAZIL AND F LORIDA ( OTHER STUDIES ) AND N ORTH AND S OUTH C AROLINA ( THIS STUDY ). Species Location % Herbivory% Carnivory% OmnivoryReference S. borellii Florida 4% 44% 8%Ulagaraj (1975) S. vicinus Florida 53% 15% 21%Ulagaraj (1975) S. borellii Florida 14% 90% Taylor (1979) S. vicinus Florida 50% 61% Taylor (1979) S. borellii Florida 6% 70% 43%Matheny (1981) S. vicinus Florida 72% 1% 10%Matheny (1981) S. borellii Brazil 18% 32% 12%Fowler et al. (1985) S. vicinus Brazil 44% 6% 10%Fowler et al. (1985) S. borellii North Carolina 42% 12% 46%NC 2009 S. vicinus North and South Carolina78% 2% 20%NC/SC 2009

PAGE 119

Book Reviews 119 G OLDSMITH M. R., AND M AREC F. (E DS .). 2010. Molecular Biology and Genetics of the Lepidoptera (Contemporary Topics in Entomology Series). CRC Press, Boca Raton. xv + 362 pp. ISBN 978-1-42006014, hardback, $129.95. With more than 150,000 described species, butteries and moths constitute one of the most diverse orders of insects. The number of actual species of Lepidoptera may be much greater, possibly as high as 500,000. Lepidoptera are thought to have radiated contemporaneously with owering plants, and they are prominent in the literature on co-evolution. Caterpillars, both as external and internal feeders, are important components of terrestrial ecosystems worldwide, and many are pests to agriculture while also serving as pollinators. As perhaps one of the most conspicuous taxa of terrestrial invertebrates, Lepidoptera are also of central utility in ecosystem assessment and in educating the public in environmental conservation. Lepidoptera include numerous model organisms that have played central roles in biology, for studies on physiology, development, evolution, and genetics. Molecular Biology and Genetics of the Lepidoptera edited by Goldsmith & Marec, is a book of eighteen chapters that highlights recent developments on the genetics of Lepidoptera model systems. The book is in many ways a follow up to Goldsmith & Wilkins (1995). With the advancement of molecular tools and methods, this timely work serves as an updated compilation, highlighting lepidopteran model systems that have become prominent in recent years. The book begins with an overview of lepidopteran relationships written by thirteen authors (Chapter 1). The chapter is divided into several sections, each representing a separate superfamily, outlining the role of model organisms in these superfamilies (Bombycoidea, Hesperoidea + Papilionoidea, Noctuoidea, Pyraloidea, and Tortricoidea). Each of these superfamily sections includes phylogenies compiled from various studies, and the trees are useful for identifying the phylogenetic position of models in each superfamily. The chapter also includes a backbone phylogeny of the Lepidoptera based on Kristensen & Skalskis (1998) tree from the Handbook of Zoology. Readers should be aware that two key studies on the molecular phylogeny of Lepidoptera have appeared very recently in the literature, namely Regier et al. (2009) and Mutanen et al. (2010) that are updates to Kristensens tree. Researchers interested in understanding the phylogenetic position of model organisms should refer to these publications for an updated backbone phylogeny of the Lepidoptera. Subsequent chapters focus on particular model species or genera, such as Bombyx (Chapter 2), Heliconius (Chapter 6) and Helicoverpa (Chapter 12). There are also chapters on sex c hromosome systems and genetics underlying sexual dimorphism (Chapters 3, 4), development of buttery wing patterns and the genetics underlying them (Chapters 5, 6), molecular innovations of buttery eyes (Chapter 7), and pheromone production and perception (Chapter 10). Many of these chapters contain fascinating facts, and information that is highly specialized. Of interest to me were the chapters on lepidopteran circadian clocks (Chapter 8) and the genetics that inuence host ranges in Lepidoptera (Chapter 11). The former chapter includes numerous diagrams illustrating the mechanisms of circadian rhythms. This chapter also includes a section on how the circadian clock plays a role in the migration of monarch butteries, which I found to be especially compelling. The latter chapter on host range (Chapter 11) reviews model taxa ( Euphydryas, Helicoverpa, Heliothis, Papilio ) in which the genetics underlying host specicity has been examined. As someone interested in host-use evolution in Lepidoptera, I found this chapter to be an especially informative source. Latter chapters focus on another dominant theme, insect control. For instance, there are chapters on the genetics underlying insecticide resistance (Chapter 13), innate insect immune response (Chapter 14), interhemocoelic toxins for pest management (Chapter 16), and polydnaviruses in lepidopteran parasitoids (Chapter 17). These chapters are written in great detail to explain various aspects of insect/pest control, which also contain quite a bit of specialized terminology, and acronyms such as PBLEs, pro-PAP-3, and V421M mutation might overwhelm the average Lepidoptera enthusiast. In conclusion, Goldsmith & Marec do an excellent job compiling a book written by a broad range of leading researchers studying various aspects of lepidopteran genetics. The compilation is not a coffee-table book, and the average reader may nd it a bit hard to follow without a background in molecular biology. However, the book is an excellent summary of the recent progress in the eld, and will be an invaluable resource for researchers and driven enthusiasts that are interested in learning more about the genetics of Lepidoptera. R EFERENCES C ITED G OLDSMITH M. R., AND W ILKINS A. S. 1995. Molecular Model Systems in the Lepidotera. Cambridge University Press, Cambridge, England.

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120 Florida Entomologist 94(1) March 2011 K RISTENSEN N. R., AND S KALSKI A. W. 1998. Phylogeny and palaeontology, pp. 7-25 In N. P. Kristensen (ed.), Lepidoptera, Moths and Butteries. Volume 1: Evolution, Systematics and Biogeography. Walter de Gruyter, New York. M UTANEN M., W AHLBERG N., AND K AILA L. 2010. Comprehensive gene and taxon coverage elucidates radiation patterns in moths and butteries. Proc. Royal Soc. of London, Series B 277: 2839-2848. R EGIER J. C., Z WICK A., C UMMINGS M. P., K AWAHARA A. Y., C HO S., W ELLER S., R OE A., B AIXERAS J., B ROWN J. W., P ARR C., D AVIS D. R., E PSTEIN M., H ALLWACHS W., H AUSMANN A., J ANZEN D. H., K ITCHING I. J., S OLIS M. A., Y EN S. H., B AZINET A. L., AND M ITTER C. 2009. Toward reconstructing the evolution of advanced moths and butteries (Lepidoptera: Ditrysia): an initial molecular study. BMC Evolutionary Biology 9: 280. Current address Akito Y. Kawahara Department of Plant and Environmental Protection Sciences University of Hawaii, Manoa 3050 Maile Way Honolulu, HI 96822 USA ak43@hawaii.edu Future address Akito Y. Kawahara McGuire Center for Lepidoptera and Biodiversity Florida Museum of Natural History University of Florida Gainesville, FL 32611

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Corporate and Sustaining Members 121 MAR 2011 FLORIDA ENTOMOLOGICAL SOCIETY CORPORATE AND SUSTAINING MEMBERS C ORPORATE Atlantic Turf andDow Agrosciences Ornamental ConsultingAttn: Ellen Thoms Attn: Scott Ferguson7257 NW 4th Blvd. #20 7145 58th AvenueGainesville, FL 32607 Vero Beach, FL 32967 Bayer Crop Science Craig Shelton Lake Placid, FL S USTAINING Bayer Crop ScienceMark B. Sivic Attn: John H. Paige16744 W. Brighton Dr. 328 Indian Lilac Rd.Loxahatchee, FL 33470 Vero Beach, FL 32963 Best Termite & Pest ControlSengenta Crop Protection Attn: Frank A. MongioviAttn: Nancy Rechiegl 8120 N. Armenia Ave.11601 Erie Road Tampa, FL 33604Parrish, FL 34219 Dow AgrosciencesTaylor Pest Management Attn: Joe EgerAttn: James Taylor 2606 S. Dundee Blvd.851 NE Jensen Beach Blvd. Tampa, FL 33629Jansen Beach, FL 34957 E.O. Painter Printing Co. Attn: Dick Johnston P.O. Box 877 DeLeon Springs, FL 32130


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Christensen et al.: Acquired Natural Enemies of Oxyops vitiosa


ACQUIRED NATURAL ENEMIES OF THE WEED BIOLOGICAL CONTROL
AGENT OXYOPS VITIOSA (COLEPOTERA: CURCULIONIDAE)

ROBIN M. CHRISTENSEN, PAUL D. PRATT, SHERYL L. COSTELLO, MIN B. RAYAMAJHI AND TED D. CENTER
USDA/ARS, Invasive Plant Research Laboratory, 3225 College Ave., Ft. Lauderdale, FL 33314

ABSTRACT

The Australian curculionid Oxyops vitiosa Pascoe was introduced into Florida in 1997 as a
biological control agent of the invasive tree Melaleuca quinquenervia (Cav.) S. T. Blake. Pop-
ulations of the weevil increased rapidly and became widely distributed throughout much of
the invasive tree's adventive distribution. In this study we ask if 0. vitiosa has acquired nat-
ural enemies in Florida, how these enemies circumvent the protective terpenoid laden exu-
dates on larvae, and what influence 1 of the most common natural enemies has on 0. vitiosa
population densities? Surveys of 0. vitiosa populations and rearing of field-collected individ-
uals resulted in no instances of parasitoids or pathogens exploiting weevil eggs or larvae. In
contrast, 44 species of predatory arthropods were commonly associated (>5 individuals when
pooled across all sites and sample dates) with 0. vitiosa. Eleven predatory species were ob-
served feeding on 0. vitiosa during timed surveys, including 6 pentatomid species, 2 formi-
cids and 3 arachnids. Species with mandibulate or chelicerate mouthparts fed on adult
stages whereas pentatomids, with haustellate beaks, pierced larval exoskeletons thereby by-
passing the protective larval coating. Observations of predation were rare, with only 8% of
timed surveys resulting in 1 or more instances of attack. Feeding by the pentatomid Podisus
mucronatus Uhler accounted for 76% of all recorded predation events. Podisus mucronatus
numerically responded to fourth instars but no response was observed for other life stages.
Damage to M. quinquenervia plants from feeding by 0. vitiosa, however, was not influenced
by P. mucronatus densities, indicating that predation does not alter plant suppression.

Key Words: biological control, biotic resistance, predation, Oxyops vitiosa, Melaleuca Quin-
quenervia, Podisus mucronatus

RESUME

El curculi6nido australiano Oxyops vitiosa Pascoe fue introducido a la Florida en 1997 como
un agent de control biol6gico para el arbol invasor, Melaleuca quinquenervia (Cav.) S. T
Blake. Poblaciones del gorgojo aumentaron rapidamente y se distribuyeron ampliamente
por much de la distribuci6n del arbol invasor adventivo. En este studio, preguntamos si O.
vitiosa han adquerido enemigos naturales en la Florida, como estos enemigos evitan las se-
creciones de turpenoides que protejen las larvas, y que influencia tiene uno de los enemigos
naturles mas comunes sobre la densidad de la poblaci6n de 0. vitiosa? La inspecci6n de la po-
blaci6n de 0. vitiosa y la cria de individuos recolectados en el campo result en no caso de pa-
rasitoides y pat6genos usando los huevos o larvas de los gorgojos. En contrast, 44 species
de artr6podos depredadores fueron comunmente asociadas (>5 individuos cuando se agrega-
dos por todos los sitios y fechas de muestreo) con 0. vitiosa. Se observaron once species de
depredadores alimentandose sobre 0. vitiosa durante los sondeos, incluyendo 6 species de
pentat6midos, 2 formicidos y 3 aracnidos. Especies con parties bucales mandibuladas y que-
liceradas se alimentaron sobre los estadios adults mientras que los pentat6midos, con su
pico chupador, puncharon los exo-esqueletos de las larvas asi pasando el cubertura protec-
tiva de las larvas. Observaciones de depredaci6n fueron raras, con solamente 8% de los es-
tudios que llevaron el tiempo resultaron en 1 6 mas instancias de ataque. La alimentaci6n
del pentat6mido Podisus mucronatus Uhler cont6 con 76% de los events de depredaci6n re-
gistrados. Podisus mucronatus respondio numericamente al los instares de cuarto estadio
pero ninguna respuesta fue observada en los otros estadios de vida. El dano a las plants de
M. quinquenervia debido a la alimentaci6n por 0. vitiosa, sin embargo, no fue influenciado
por la densidad de P. mucronatus, que indica que la depredaci6n no altera la supresi6n de la
plant.


Acquisition of novel natural enemies may in- Partland & Nicholson 2003; Norman et al. 2009;
fluence the successful establishment, spread, and Paynter et al. 2010). Of the arthropods introduced
impact of introduced weed biological control for control of invasive plants world wide, approx-
agents in their adventive range (Goeden & Louda imately 50% suffer sufficient mortality from
1976; Semple & Forno 1987; Simberloff 1989; higher trophic levels to significantly limit sup-
Cornell & Hawkins 1993; Hill & Hulley 1995; Mc- pression of target weeds (Goeden & Louda 1976).







Florida Entomologist 94(1)


The spider mite Tetranychus lintearius (Dufour),
for example, was introduced into New Zealand,
Australia, and the United States as a biological
control agent of the invasive plant Ulex europaeus
L. (Fabaceae) (Hill & Stone 1985; Hill et al. 1991).
Although successfully established and widely dis-
tributed, mites in each country rarely sustained
sufficient population densities to provide perma-
nent control of the target weed (Rees & Hill 2001).
Subsequent studies demonstrated that a complex
of native and introduced predators suppressed T.
lintearius populations and limited control of the
invasive weed (Peterson 1993; Peterson et al.
1994; Pratt et al. 2003).
Considering the ecological risks (Carvalheiro
et al. 2008) and expense of biological control, in-
creased attention in the scientific literature has
focused on predicting susceptibility of introduced
biological control agents to natural enemies in the
adventive range (Kuhlmann et al. 2006). Hill &
Hulley (1995), for instance, demonstrated that
variation in susceptibility of introduced herbi-
vores to parasitoids is related, in part, to evolu-
tionary strategies that render the prey less acces-
sible, apparent, or palatable to the attacker.
Along this continuum of use by natural enemies
lie those species that are highly apparent yet ex-
perience relatively less attack due to the expres-
sion of chemical deterrents that render them less
palatable or even toxic to prospective natural en-
emies. The introduced weevil Oxyops vitiosa Pas-
coe sequesters terpenoids from leaves of its host
plant Melaleuca quinquenervia (Cav.) S. T. Blake
and larvae excrete these compounds through
their integument (Wheeler et al. 2002). The con-
sumption and expression of these terpenoids re-
pels the red imported fire ant (Solenopsis
invicta Buren) and red wing blackbird (Agelaius
phoeniceus L.) under controlled feeding tests
(Wheeler et al. 2002). It remains unclear, how-
ever, if this acquired repellency confers protection
from the suite of potential novel natural enemies
that exist in the herbivore's adventive range.
Oxyops vitiosa is native to eastern Australia
and is a specialist herbivore of the invasive tree
M. quinquenervia (Balciunas et al. 1994). Based
on its narrow host range, the weevil was permit-
ted for release in Florida in 1997 and readily es-
tablished in M. quinquenervia dominated habi-
tats (Center et al. 2000; Pratt et al. 2003). Adult
weevils feed on M. quinquenervia foliage whereas
larvae consume only newly-developed leaves that
are ephemerally produced in seasonal flushes at
branch apices (Purcell & Balciunas 1994).
Following its introduction, 0. vitiosa popula-
tions increased rapidly and became widely dis-
tributed throughout much of the invasive tree's
geographic distribution in Florida (Pratt et al.
2003; Balentine et al. 2009). When considering
the large densities of these herbivores in the envi-
ronment, we questioned (1) whether 0. vitiosa


had acquired natural enemies in Florida, (2) how
these enemies mitigated the defensive strategies
of the herbivore, and (3) what impact the most
abundant of these natural enemies has on O. vi-
tiosa population densities?

MATERIALS AND METHODS

Surveys for natural enemies associated with
0. vitiosa were conducted at 4 locations in south
Florida. Site 1 was located near Ft. Lauderdale,
Broward Co., FL. The site was a 0.5-ha field con-
sisting of 2- to 5-m tall trees occurring at a den-
sity of 21,560 trees/ha. In general, M. quinquen-
ervia trees were growing in organically rich soils
typical of reclaimed 'glades' systems. Melaleuca
quinquenervia trees at site 2 occurred under a
power line right of way near Weston, Broward
Co., FL. Prior to 1997 land managers cut M. quin-
quenervia trees near their bases, resulting in
multi-stemmed coppices. The study area was 0.5
ha and trees were 2-5 m tall, occurring at a den-
sity of 2,517 trees/ha. Site 3 was located near Es-
tero, Collier Co., FL and consisted of an 8-ha area
of drained wetland converted to pasture. To sup-
press M. quinquenervia growth, land managers
mowed trees at 6-month intervals, resulting in
coppices 0.5-2 m in height. These coppicing
clumps formed a dense, nearly continuous canopy
of leaves with 4,406 stumps/ha. In contrast to the
previous sites, the soil type was primarily sand,
consistent with an invaded pine flatwoods. Site 4
consisted of a 1-ha area within the historically
mesic flatwoods of the Picayune Forest near Belle
Meade, Collier Co., FL. A fire burned much of the
M. quinquenervia dominated areas in 1998 re-
sulting in recruitment of 129,393 trees/ha of pri-
marily small 1-2 m tall saplings, with an occa-
sional large, mature tree interspersed (Table 1).
Surveys were conducted monthly at each site
from Nov 2000 through Jun 2001 and sampling
occurred between 10 AM and 2 PM on days without
precipitation. Sampling consisted of sweeping M.
quinquenervia foliage, and occasionally trunks,
with a 90-cm diameter sweep net. One sample
consisted of 100 sweeps of the net in a sweeping
motion of 180 with sweeps spaced ca. 1.0 m apart
along a randomly selected 100-m transect. Four
samples along separate transects were collected
each month. The content of the net after 100
sweeps was emptied into a 4-liter sealable bag
and frozen at minus 20 (1) C until processed.
Arthropods were then separated from plant mate-
rial, sorted by morphological types, and pinned or
stored in 70% ethanol.
One limitation of our sweep sampling method
included collecting arthropods that were not
closely associated with 0. vitiosa, but were tran-
sients, merely resting on the plant foliage or dis-
turbed from understory vegetation while sam-
pling. Additionally, this method was biased to-


March 2011







Christensen et al.: Acquired Natural Enemies of Oxyops vitiosa


TABLE 1. RESEARCH SITE DESCRIPTION AND SUMMARY OF SURVEYS CONDUCTED FOR O. VITIOSA IN SOUTH FLORIDA.

Site GPS Coordinates Surveys conducted Habitat Hydro-period

1 N 26.05605 W -80.25168 1, 2, 3, 4 Swale Short
2 N 26.03548 W -80.43495 1, 2, 3, 4 Swale Medium
3 N 26.42550 W -81.81033 1, 2, 3, 4, 5 Mesic flatwoods Short
4 N 26.10478 W -81.63392 1, 2, 3,4 Mesic flatwoods Short
5 N 26.46017 W -81.70186 4 Mesic flatwoods Short
6 N 26.54698 W -81.79820 4 Wet flatwoods Short
7 N 28.47323 W -81.33632 4 Upland lake Long
8 N 25.71341 W -80.47949 4 Swale Medium
9 N 25.81208 W -80.41780 4 Swale Medium
10 N 26.16227 W -80.36269 4 Swale Long

1= Arthropods associated with 0. vitiosa, 2 = 0. vitiosa population density, 3 = 0. vitiosa egg parasitism, 4 = Entomopathogens
ofO. vitiosa, 5 = Impacts ofP. mucronatus on 0. vitiosa populations.


wards poor fliers. All study sites possessed
smaller trees that facilitated sampling but may
have biased collections to lower rather than
higher canopy dwelling species. Therefore, cau-
tion should be used when drawing inferences
from these data due to the unknown relationships
between sampled arthropods and 0. vitiosa. For
this reason, a minimum of 2 observers also
searched for direct predation or parasitism for 30
min/survey at each site monthly.
All specimens, except formicids, were submit-
ted to and deposited at the Florida State Collec-
tion of Arthropods (FSCA, Division of Plant In-
dustries, Gainesville, FL) for identification and
incorporated into their taxonomic database (Cos-
tello et al. 2003). Most formicids were identified
and retained by L. Davis of the Fire Ant Unit, Ag-
ricultural Research Service, USDA, Gainesville,
FL. A few formicids were identified by M. Deyrup
of the Archbold Biological Station, Lake Placid,
FL. Several dipteran specimens were identified at
the Systematic Entomology Laboratory, Agricul-
tural Research Service, USDA, Beltsville, MD.
Population densities of 0. vitiosa at sites 2 and
3 were monitored by delineating a 0.5-ha study
site within the existing M. quinqueneruia stands,
respectively. Within these plots, transects were
arranged in a grid pattern with 8 transects ori-
ented east to west at 10-m intervals and points on
each transect spaced 10 m apart. Beginning in
Nov 2000 through Jun 2001, M. quinqueneruia
trees were sampled monthly at 20 randomly se-
lected transect points. Plants at each sampling
point were selected based on the quarter method
of vegetation sampling (Smith 1966). The area
was divided into 4 quarters at each sampling
point based on the 4 cardinal directions. The
nearest tree to the sample point in each quarter
was examined to determine the number of O. vi-
tiosa per plant. The ordered distance method was
used to quantify weevil population densities over
time at sites 1 and 4 (Krebs 1999). In total, 30
points were randomly selected at each sampling


interval and the nearest tree to each point was in-
ventoried. At all sites, 0. vitiosa life stages were
counted along with plant resource availability.
Resource availability was assessed on a 5-point
scale based on visual estimation of percentage of
suitable foliage for feeding by 0. vitiosa: 0 = no
suitable foliage; 1 = less than 25%; 2 = 26 to 50%;
3 = 51 to 75%; 4 = 76 to 100%.
A partial correlation analysis was used to iden-
tify those predators positively associated with O.
vitiosa (PROC CORR), after controlling for the in-
fluence of site by the PARTIAL statement (SAS
1999). For all tests, a P-value <0.05 was consid-
ered significant evidence for association among
predators and 0. vitiosa. However, caution should
be used when interpreting these data because as-
sociation is not sufficient evidence to suggest that
a trophic relationship exists among the species.
To determine if 0. vitiosa had acquired egg or
larval parasitoids in its adventive range, 50 eggs
were collected at random from sites 1-4 at
monthly intervals. Eggs were examined under a
dissecting microscope (10-50X) to detect presence
of larval exit holes indicating larval eclosion; eggs
with exit holes were discarded. The remaining
eggs were left attached to leaf material and
placed in gelatin capsules. These capsules were
transferred into a Petri dish (10 x 1.5 cm) that
was then sealed with Parafilm to retain leaf mois-
ture. Petri dishes were placed in an environmen-
tal chamber at 25 (+1) C, with a photoperiod of
16:8 (L:D) and a relative humidity of 65% 10%.
Hatching of eggs was monitored once a week.
Eggs that did not hatch after 1 month were dis-
sected.
To detect larval parasitoids, 50 third or fourth
instars of 0. vitiosa were reared to the pupal
stage. Each larva was placed individually in 1
Petri dish (10 x 1.5 cm) with moistened filter pa-
per and M. quinqueneruia leaves. Petri dishes
were sealed with Parafilm and kept in an environ-
mental chamber under the same conditions as de-
scribed earlier. Host leaves were replaced every







Florida Entomologist 94(1)


other day until the prepupal stage, when leaves
were removed for the remainder of the study.
Surveys for entomopathogens of 0. vitiosa
were conducted at 10 sites between Jun 2003 and
Jan 2004 (Table 1). Late instars and adults were
collected, packaged in an ice-cooler, transported
to the laboratory, and examined by USDA/ARS
insect pathologists at the Center for Medical, Ag-
ricultural, and Veterinary Entomology, Gaines-
ville, FL. All live individuals originating from the
same location were homogenized in 3-5 mL
of deionized water and a sample of the crude sus-
pension was examined with a phase-contrast mi-
croscope to search for pathogens, such as mi-
crosporidia, fungal spores, or occluded viruses.
During initial surveys, the pentatomid bug Po-
disus mucronatus Uhler was commonly associ-
ated with 0. vitiosa and observed feeding on lar-
val stages of the biological control agent at each of
the 4 study sites. Therefore, we quantified the
population dynamics of P mucronatus and 0. vi-
tiosa at site 3. Sampling was conducted as de-
scribed earlier except 20 transects were oriented
east to west at 20-m intervals with 9-10 points on
each transect spaced 20 m apart. Melaleuca quin-
quenervia plants were sampled at 50 randomly
selected transect points every 6 weeks (approxi-
mate generation time; Purcell & Balciunas 1994)
beginning in Dec 2000 and continuing through
Oct 2002. As before, the nearest plant to the sam-
ple point in each quadrant was examined to deter-
mine the number and life stage of each 0. vitiosa
and P mucronatus individual. In addition to
these data, we also noted the amount of damage
due to herbivory, plant resource availability (as
described earlier), and the number of dead larvae.
Herbivory damage was assessed on a 5-point
scale based on visual estimation of the percentage
of suitable foliage destroyed by 0. vitiosa feeding:
0 = no damage; 1 = less than 25% destroyed; 2 =
26 to 50%; 3 = 51 to 75%; 4= 76 to 100% destroyed.
Linear regression was used to test for a numerical
response of predators to life stages of 0. vitiosa (n
= 6), dead larvae, food availability, and herbivory
damage (i.e., aggregation of predators to patches
of high prey density; Schenk & Bacher 2002).

RESULTS AND DISCUSSION

Biotic resistance describes the collective influ-
ence of parasitoides, predators, pathogens, and
competitors on the establishment and prolifera-
tion of non-indigenous species, including intro-
duced biological control agents (Simberloff & Von
Holle 1999). Historically, native predators, para-
sitoids, and pathogens have interfered with half
of the published case histories involving insect in-
troductions for weed control (Goeden & Louda
1976). Considering this high rate of interference,
we questioned if 0. vitiosa had acquired natural
enemies in its adventive range.


We observed no instances of parasitoids (egg
and larval) or pathogens exploiting 0. vitiosa. Of
the 1138 0. vitiosa eggs collected from the study
sites (Table 1), 782 hatched and developed nor-
mally while the remaining 356 did not hatch. Of
the 1266 fourth-instars collected from study sties,
913 survived to become adults. Dissection of both
unhatched eggs and dead larvae yielded no evi-
dence that mortality was due to parasitism. Sim-
ilarly, no pathogens were found in the late instars
and adults collected from sampled sties (Table 1).
These results indicate that despite the herbi-
vore's high population densities and large geo-
graphic distribution (Pratt et al. 2003; Balentine
et al. 2009), native parasitoids and pathogens
have failed to exploit these lifestages of 0. vitiosa.
One explanation for the lack of 0. vitiosa para-
sitization may be that native parasitoids require
more than the 4 years allotted in this study to ad-
just behaviorally and physiologically to exploit
the new host as well as produce sufficient densi-
ties to be discovered through our sampling proto-
cols. In contrast, Hill & Hulley (1995) determined
that 16 of the 40 established weed biological con-
trol agents in South Africa had acquired native
parasitoids within 3 years of release. Similarly,
the biological control agent Neomusotima con-
spurcatalis Warren acquired a suite of parasitoids
within months of its release in Lygodium micro-
phyllum (Cav.)-dominated habitats of Florida
(Kula et al. 2010). These and other examples of
rapid parasitoid acquisition by biological control
agents (Carvalheiro et al. 2008; Paynter et al.
2010) suggest that the timing of our study was
not premature but that future parasitoid (or
pathogen) surveys may yield new discoveries as
the region continues to recruit exotic species
(Klassen et al. 2002; Dobbs & Brodel 2004;
Childers & Rodrigues 2005).
Surveys of 0. vitiosa populations resulted in
the collection of 154 species of predatory arthro-
pods, yet only 44 had an overall abundance
greater than 5 individuals when pooled across all
sites and dates (Table 2). Species positively corre-
lated with 0. vitiosa (all stages) included the sal-
ticid Eris flava (Peckham & Peckham), the crab
spiders Misumenops bellulus (Banks) and Mis-
umenops sp., and the pentatomid bug Podisus
mucronatus Uhler (Table 2). Although these data
indicate that predators are associated with the in-
troduced herbivore, direct observation of preda-
tion provides conclusive evidence of these novel
trophic interactions. Eleven predatory species
were observed feeding on 0. vitiosa during timed
surveys, including 6 pentatomid species (Euthy-
rhynchus floridanus (L.), P mucronatus (Say), Po-
disus jole (Stal), Podisus maculiventris (Say), Po-
disus sagitta (F.), Stiretrus anchorage (F.)), 2 for-
micids (Pseudomyrmexgracilis (F.), Solenopsis in-
victa) and 3 arachnids (Peucetia viridans (Hentz),
Latrodectus mactans (F.), Latrodectus geometri-


March 2011








Christensen et al.: Acquired Natural Enemies of Oxyops vitiosa


TABLE 2. THE ABUNDANCE OF PREDACEOUS ARTHROPODS COLLECTED FROM THE INVASIVE TREE MELALEUCA QUIN-
QUENERVIA AND THEIR CORRELATION WITH THE INTRODUCED BIOLOGICAL CONTROL AGENT OXYOPS VITIOSA.

Site Abundance Partial Correlation

Family Species 1 2 3 4 PCC P-value


Alydidae
Anyphaenidae

Araneidae



Cixiidae
Clubionidae
Formicidae








Lycosidae

Lygaeidae

Membracidae
Mimetidae
Miridae
Miturgidae
Oxyopidae
Pentatomidae
Pisauridae
Reduviidae
Salticidae





Scelionidae
Tetragnathidae
Theridiidae



Thomisidae


Hyalymenus sp.A
Hibana sp.
Hibana sp.
Acacesia hamata (Hentz)
Eriophora ravilla (C.L. Koch)
Neoscona sp.
Neoscona arabesca (Walckenaer)
Bothriocera sp.
Clubiona sp.
Camponotus planatus (Roger)
Camponotus floridanus (Buckley)
Dolichoderus pustulatus Mayr
Paratrechina longicornis (Latreille) *
Paratrechina guatemalensis (Forel) *
Pseudomyrmex pallidus (F. Smith) *
Pseudomyrmex gracilis (Fabricius)
Solenopsis invicta Buren
Pardosa sp.
Pirata sp.
Oedancala crassimana (Fabricius)
Paromius longulus (Dallas)
Spissistilus festinus (Say)
Mimetus sp.
Z. ... .... pallidulus (Blanchard)
Cheiracanthium inclusum (Hentz)
Peucetia viridans (Hentz)
Podisus mucronatus Uhler
Pisaurina sp.
Zelus longipes Linnaeus
Eris flava (Peckham & Peckham)
Hentzia palmarum (Hentz)
Pelegrina galathea (Walckenaer)
Phidippus sp.
Thiodina peurpera (Hentz)
Trissolcus sp.
Tetragnatha sp.
Anelosimus studiosus (Hentz)
Chrysso pulcherrima (Mello-Leitao)
Theridion glaucescens (Becker)
Theridion flavonotatum (Becker)
Misumenops sp.
Misumenops bellulus (Banks)
Misumenops sp.
Tmarus sp.


cus C.L. Koch). The formicids and arachnids were counted for 76% of all recorded predation events
observed feeding exclusively on adult weevils and the remaining species each represented <5%
whereas the pentatomids attacked larvae of O. vi- of the events, respectively.
tiosa; E. floridanus was the only species observed Ecological theory suggests that host range ex-
exploiting all active stages of the introduced her- pension is influenced in part by host phylogeny,
bivore. Observing predation was rare, with only with close relatives more readily adopted than
8% of timed surveys resulting in 1 or more in- distant ones (Paynter et al. 2010). Therefore, an
stances of attack. Feeding by P mucronatus ac- alternative explanation for the lack of acquired


1 12
0 0
4 0
15 14
5 1
27 15
6 0
0 6
0 6
75 46
14 0
6 0
85 102
16 2
15 11
2 1
8 16
1 0
18 2
13 2
7 4
0 0
1 0
2 2
96 140
137 9
6 1
7 5
25 0
4 5
72 45
12 5
0 1
0 19
0 8
33 8
67 0
1 0
12 0
24 0
2 3
26 37
10 0
2 0


-0.23098
0.04519
-0.06653
0.15511
-0.08230
-0.08867
-0.13606
-0.11674
0.01856
0.05757
0.09479
-0.14389
-0.14078
-0.08637
-0.13190
-0.17915
0.02664
0.33110
0.33916
0.33885
0.06829
0.29864
0.07110
0.31276
-0.01003
0.06787
0.52700
0.13199
-0.05046
0.36503
0.00540
0.18343
-0.14807
-0.13863
-0.10483
0.11216
0.08956
0.26530
0.13018
-0.01400
0.30519
0.35739
0.39715
-0.24154


0.2112
0.8093
0.7222
0.4047
0.6599
0.6353
0.4655
0.5317
0.9211
0.7584
0.6120
0.4400
0.4500
0.6441
0.4794
0.3349
0.8869
0.0688
0.0620
0.0622
0.7151
0.1027
0.7039
0.0867
0.9573
0.7168
0.0023
0.4791
0.7875
0.0435
0.9770
0.3233
0.4267
0.4570
0.5746
0.5480
0.6318
0.1492
0.4852
0.9404
0.0950
0.0484
0.0269
0.1905







Florida Entomologist 94(1)


parasitoids and pathogens may be due to the pau-
city of closely related species in the biological con-
trol agent's adventive range. The Australian wee-
vil 0. vitiosa belongs to the tribe Goniopterini,
which has no representatives in the New World
(Alonso-Zarazaga & Lyal 1999). Similarly, inva-
sion by M. quinquenervia markedly alters com-
munity structure in ways that are likely to repel
habitat specialists. Therefore, the acquisition of
parasitoids will likely require evolutionary rather
than ecological time scales (Hill & Hulley 1995).
With the exception of E. floridanus, the exclu-
sive use of adult versus larval prey observed
herein may be explained by mouthpart morpholo-
gies and the antipredatory activity of the viscous
coating that covers immature stages of 0. vitiosa
(Purcell & Balciunas 1994). Larvae of the intro-
duced weevil sequester terpenoids from M. quin-
queneruia leaves and excrete these compounds
through their integument (Wheeler et al. 2003).
This larval coating has been shown to repel the
red imported fire ant (S. invicta) and likely con-
fers protection against other mandibulate preda-
tors (Wheeler et al. 2002). However, adults and
pupae lack the coating and are susceptible to pre-
dation by a range of predator types. The larval
coating does not confer protection against pen-
tatomid species observed herein. The haustellate
mouthparts of pentatomid species pierce the lar-
val integument and largely bypass the terpenoid-
laden coating to access the internal contents of
the larval prey. Yet, mouthpart type alone does
not facilitate exploitation of the abundant novel
resource as other predators with haustellate
mouthparts (i.e., Zelus longipes L.) occurred at
the study sites but were not common or observed
directly feeding on 0. vitiosa larvae.
Increased densities of 0. vitiosa eggs, early in-
stars, and adults did not influence patch coloniza-
tion by P mucronatus (Table 3). A numerical re-
sponse by P. mucronatus was observed, however,
on plants harboring fourth instars (Table 3), indi-
cating a preference for larger larval stages of the
introduced weevil. These findings are consistent

TABLE 3. LINEAR REGRESSION OF PODSUS MUCRONATUS
DENSITIES ON OXYOPS VITIOSA STAGE SPECIFIC
DENSITIES, PLANT QUALITY, AND FOLIAGE
AVAILABILITY.

Life stage df Estimate t-value Pr > t

Egg 1 -0.00097 -0.29 0.7752
1st instar 1 -0.00185 -0.24 0.8075
2nd instar 1 0.01394 1.66 0.0966
3rd instar 1 0.01175 1.27 0.2025
4th instar 1 0.0251 2.88 0.004
Adults 1 0.00818 1.06 0.2874
Dead 1 0.13755 16.16 <.0001
Damage 1 -0.00265 -1.31 0.1893
Plant foliage 1 -0.00176 -0.94 0.3462


with Hawkins et al. (1997), who reported that in-
sect predation is higher in late developmental
stages due, in part, to resource concentration and
handling time. Not surprisingly, a positive rela-
tionship between P. mucronatus and larval
corpses also was observed.
While it is clear that P. mucronatus attacks O.
vitiosa larvae and numerically responds to the
single most damaging stage of the herbivore, does
this predation disrupt biological control of M.
quinqueneruia? We hypothesized that increases
in P mucronatus densities results in concomitant
increases in predation and ultimately decreases
in plant damage caused by 0. vitiosa. Damage
levels observed herein, however, were not influ-
enced by P. mucronatus densities (Table 3), indi-
cating that predation does not alter plant sup-
pression within the sampled patch. Similarly, the
amount of plant resource availability for con-
sumption by 0. vitiosa does not vary based on
predator loads, which suggest that predation does
not result in a corresponding increase in undam-
aged plant material (Table 3). These results are
supported by independent studies that also were
conducted at site 3 and reported marked reduc-
tions in M. quinqueneriva growth and survival
despite the presence of these predators (Center et
al. 2000; Pratt et al. 2002; Pratt et al. 2004). The
limited influence of P mucronatus on 0. vitiosa
population growth and herbivory is likely related
to low predation rates (mean = 9.5%, SE = 0.5).
The introduction of 0. vitiosa has resulted in
marked reductions in growth and survivorship of
the invasive tree M. quinqueneriva (Pratt et al.
2003, 2005; Rayamajhi et al. 2008; Tipping et al.
2008, 2009; Balentine et al. 2009), with no direct
non-target impacts to plant species in the weevil's
adventive range (Pratt et al. 2009). The acquisi-
tion of higher trophic levels by 0. vitiosa, how-
ever, suggests that indirect effects of apparent
competition may exist as predators are subsi-
dized by the introduced weevil and their result-
ant increased population densities may exert
asymmetrical predation on their historical prey
species (Carvalheiro et al. 2008). In the absence of
pre-introduction food web analyses, it remains
unclear how the exploitation of 0. vitiosa by na-
tive predators affects apparent competition on
shared prey densities. The limited predation by
generalists suggests that the strength of appar-
ent competition is weak but additional research is
needed to quantify interactions among intro-
duced and native prey species as mitigated by
common predators.

ACKNOWLEDGMENTS

We thank 2 anonymous reviewers for comments on
earlier versions of the manuscript. We also thank Scott
Wiggers, Willey Durden, Kirk Tonkel, Andrea Kral, Carl
Belnavis, Tafana Fiore, and Stacey Grassano for assis-


March 2011







Christensen et al.: Acquired Natural Enemies of Oxyops vitiosa


tance with data collection and site maintenance. Men-
tion of trade names or commercial products in this
publication is solely for the purpose of providing specific
information and does not imply recommendation or en-
dorsement by the U.S. Department of Agriculture. This
research was supported, in part, by grants from the
South Florida Water Management District, the Florida
Department of Environmental Protection Bureau of In-
vasive Plant Management, and the USDA Areawide
TAME Melaleuca Program (tame.ifas.ufl.edu).


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Florida Entomologist 94(1)







Yang et al.: Mating Strategy of Ectropis oblique


COMPARATIVE MATING STRATEGIES OF MALE AND FEMALE
ECTROPIS OBLIQUE (LEPIDOPTERA: GEOMETRIDAE)


YUN-QIU YANG', XU-HUI GAO1, YAN-ZHUO ZHANG3, LONG-WA ZHANG AND XIAO-CHUN WAN'*
1Key Laboratory of Tea Biochemistry & Biotechnology, Ministry of Education and Ministry of Agriculture,
Anhui Agricultural University, Hefei, Anhui 230036, P.R. China

2Provincial Key Laboratory of Microbial Pest Control, Anhui Agricultural University, Hefei,
Anhui230036, P.R. China

'State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology,
Chinese Academy of Sciences, Beijing 100101, P.R. China

*Corresponding author; E-mail: tealab@ahau.edu.cn

ABSTRACT

The mating strategies of male and female Ectropis oblique Prout were investigated with the
aid of male antennae as an electroantennogram (EAG) detector and capillary-GC analysis.
Each male was capable of mating with several females, but females that had received a sper-
matophore mated only once. Antennae dissected from males 0, 1, 2, 3, and 4 d post-mating
and antennae from virgin males of corresponding ages displayed similar EAG responses to
sex pheromone extracts from sexually active females. Pheromone extracts of mated females
elicited significantly weaker male EAG responses than the pheromone extracts of virgin fe-
males. EAG responses of males to sex pheromone extracts taken from mated females at 0,
1, 2, 3, and 4 d post-mating were consistently weak. Pheromone production in the phero-
mone glands of mated females was strongly suppressed and declined during each of 4 suc-
cessive nights after they had mated.

Key Words: EAG, GC, mating, sex pheromone emission, spermatophore

RESUME

Las estrategias de apareamiento de machos y hembras de Ectropis oblique Prout fueron in-
vestigadas con el uso de la antena del macho como un detector electro-antenogramatico
(EAG) y de CG capilar andlisis. Cada macho fue capaz de aparearse con varias hembras,
pero las hembras que han recibido un espermat6foro se aparearon solamente una vez. Las
antenas diseccionadas de los machos 0, 1, 2, 3, y 4 dias despu6s de aparearse y las antenas
de machos virgenes de edades corespondientes mostraron respuestas de EAG similares a los
extractos de feromonas sexuales de hembras sexualmente activas. Los extractos de feromo-
nas de hembras apareadas provocaron una respuesta del EAG de los machos significativa-
mente mas debil que los extractos de feromonas de las hembras virgenes. Las respuestas de
EAG de machos hacia los extractos de feromonas sexuales tomados de hembras apareadas
a los 0, 1, 2, 3 y 4 dias despu6s de aparear fueron consistentemente mas debiles. La produc-
ci6n de feromonas en las glandulas de feromonas de hembras apareadas fue fuertemente su-
primida y decline durante cada una de las noches consecutivas despu6s de que las hembras
se aparearon.


Male insects typically mate many times during
their lifetime, while females display diverse mat-
ing strategies (Arnqvist & Nilsson 2000). In some
species, females need to mate once or only a few
times to produce an optimal number of viable
eggs; in many other species females mate fre-
quently to maximize reproductive potential (Rad-
wan & Rysinska 1999). No matter what strate-
gies females use, they tend to discontinue sex
pheromone production after mating, either tem-
porarily or permanently; and this avoids prob-
lems associated with excessive male sexual ha-
rassment (Giebultowicz et al. 1991). This phe-


nomenon was demonstrated by bioassay or chem-
ical analysis in many studies (Webster & Carde
1984; Coffelt & Vick 1987; Ahn et al. 2002). In
contrast to the numerous studies on changes in
female reproductive behavior after mating, few
studies have focused on male response to dimin-
ished pheromone production of mated females.
Recently, the electroantennogram (EAG) has
been widely used in studies on semiochemical in-
volvement in sex pheromones (Park et al. 2001;
GAkge et al. 2007). An EAG response profile is
thought to represent the sensitivity and relative
abundance of olfactory receptor neurons on the







Florida Entomologist 94(1)


antennae that are tuned to the compounds tested.
The EAG response amplitudes are thought to rep-
resent the quantity of semiochemicals (Pouzat &
Ibeas 1989). Thus, the EAG may be used as a tool
to investigate the sensitivity of males to variable
amounts of sex pheromone.
Ectropis oblique Prout (Lepidoptera:
Geometridae) is an important tea bush pest in
Southeast China. Population outbreaks can com-
pletely defoliate leaves on the bushes (Hu et al.
1994). The sex pheromone components of the fe-
male E. oblique were identified as (Z,Z,Z)-3,6,9-
octadecatriene and 6,7-epoxy-(Z,Z)-3,9-octadeca-
diene (Yao et al. 1991). Most efforts to control E.
oblique populations are focused on the develop-
ment of methodologies to disrupt reproduction.
Therefore, understanding the behavior of E. ob-
lique is necessary. In this study, the mating fre-
quency and longevity of E. oblique males and fe-
males was investigated. At the same time, the dif-
ferent mating strategies of the two genders were
examined using male antennae as EAG detectors.
To verify the results of the electrophysiological
analysis, changes in sex pheromone titers pro-
duced by mated and virgin females were also in-
vestigated using capillary-GC analysis.

MATERIALS AND METHODS

Insect Culture

The E. oblique insects were obtained from Qian-
shan County (31.5N0, 116.3E0), Anhui Province,
China, and maintained for many generations in the
laboratory. Larvae were reared on tea leaves. Adults
and larvae were maintained in controlled condi-
tions at 22 + 3C, 60-70% relative humidity, and a
photoperiod of 14L:10D, with scotophase and photo-
phase reversed from a natural light cycle to permit
scotophase observations during normal working
hours. Pupae were sexed based on the morphology
of the 8th-10th abdominal segments, and main-
tained in moist sand for eclosion. Adults were kept
individually in 240-mL plastic jars and fed a 10%
honey solution soaked in cotton.

Effect of Mating Frequency on Longevity of Females
and Males

In preliminary observations of behavior, both
females and males copulated during the first sc-
otophase after emergence, and were sexually ac-
tive at the second scotophase. Therefore, a 1-d-old
female (0 d after emergence) and a 2-d-old male
were paired. Copulation occurred in the sc-
otophase lasted approximately 6 h. After mating,
the female insects did not resume calling during
the same scotophase (Yang et al. 2008). Thus, each
female in this experiment was replaced daily with
another 1-d- old virgin female. When the female
died, the number of times it had mated was ascer-


trained by counting the spermatophores present in
her bursa copulatrix. The experiment was re-
peated 40 times. The number of times a male E. ob-
lique mated was determined by keeping a record of
the number of females mated during successive sc-
otophases. The experiment was also repeated 40
times. The longevity of each male and each female
was recorded in order to determine whether mat-
ing affected longevity of either gender.

Extraction of Sex Pheromones

Active sex pheromones were extracted from
the glands of the virgin females. The terminal sec-
tion of the abdomen, which included the phero-
mone gland, was excised from the virgin female
moth 6 h after the onset of the second scotophase,
when the virgin female had been calling for 1 h.
Experimental procedures were performed under
a red light to facilitate observation without dis-
turbing the insects. Each excised abdominal tip
was immersed in 10 pL of redistilled hexane for 4-
6 h at room temperature. Then, the tip was re-
moved and the extract without any purification
was submitted for EAG or GC analysis. The pro-
cedure for extracting sex pheromones from either
mated or virgin females was the same.

Electroantennographic Analysis of the Effect of Mating
on Pheromone Production and on Male Responsiveness
at Various Days after Mating

Electroantennograms (EAG) were obtained
with an EAG apparatus (Syntech Co., 79199
Kirchzarten, Germany). The antennae of either
mated or virgin males were excised at the bases
and a few distal segments were cut off to facilitate
conductivity. The antennae were then attached to
the electrodes of the EAG probe with Spectra 360
Electrode Gel (Parker Laboratories Inc., Orange,
New Jersey). Antennal preparations were ex-
posed to a stream of humidified and charcoal-fil-
tered air emitted at 4 mL s-lafter having flowed
through a 35-cm long glass tube (inner diameter,
8 mm; outer diameter, 10 mm). To facilitate inser-
tion of the Pasteur pipette used to administer the
pheromone test stimulus, a 3-mm hole was bored
5 cm from the outlet of the glass tube. Ten pL of
the extract (test stimulus) was applied to a piece
of filter paper (1 x 5 cml x 5 cm). The filter paper
was placed in a Pasteur pipette after the solvent
(hexane) had been allowed to evaporate for 5 min.
Each test stimulus was delivered within a 0.5 s
pulse of 4 mL s-1 of air with a stimulus controller
(type CS-55) to transport the volatiles to the an-
tenna. The EAG signal was amplified 10x through
an intelligent data acquisition controller (type
IDAC-2) and viewed on an oscilloscope. A period
of at least 30 s was allowed between 2 successive
stimuli for the recovery of antennal responsive-
ness. Redistilled hexane (10 pL) was used as a


March 2011







Yang et al.: Mating Strategy of Ectropis oblique


control stimulus in every test. The absolute EAG
amplitude (mV) minus the solvent response was
used for data analysis.
First, the influence of the male's mating status
on the male's EAG responses to active sex phero-
mones was studied. To obtain mated males and fe-
males, the insects were paired in the first sc-
otophase and allowed to mate. Mating pairs were
then removed. After mating, the mated females
were used for the next experiment. With the same
procedure as described above, the antennae of the
mated male were dissected at 0, 24, 48, 72, and 96
h after mating for use as EAG test detectors. Sex
pheromone extracts from sexually active females
were used as stimuli. The EAG responses of an-
tennae dissected from virgin males at each of
these times post-mating were compared to the
EAG responses of antennae obtained from mated
males at corresponding times post-mating. Each
treatment was repeated 6-8 times.
Secondly, the influence of the female's mating
status on the EAG responses of a male was stud-
ied with sex pheromone extract of a mated female
as the stimulus to elicit an EAG response from a
2-d-old virgin male. As described above, the sex
pheromone of the mated females was extracted at
0, 24, 48, 72, and 96 h after mating. The EAG re-
sponses of antennae of 2-d-old virgin males ex-
posed to pheromone extract obtained from virgin
females at each of the above times post-mating
were compared to the EAG responses to phero-
mone extract obtained from mated females at cor-
responding times post mating. Each treatment
was also repeated 6-8 times.

Pheromone Titer Analysis

To assess the effect of mating on pheromone
production, the sex pheromone titers of the mated
and virgin females were analyzed by the proce-
dure described above. Thus sex pheromone ex-
tract was analyzed in a gas chromatograph (Agi-
lent 6890) equipped with a capillary column (DB-
5, 60m x 0.5mm i.d x 0.25 pm film). The oven tem-


perature was programmed at 50C for 2 min, then
15C min-1 to 250C and held for 5 min. The tem-
peratures of the injector and detector were 200C
and 250C, respectively. Nitrogen with a flow ve-
locity of 40 mL min-' was used as the carrier gas.
To quantify the pheromones in the female gland,
only the amount of epo3,Z6,Z9-19:H, the major
sex pheromone component of E. oblique, was de-
termined. Each treatment was repeated 6-8
times.

Statistical Analysis

The data were analyzed by one-way ANOVA,
followed by a LSD multiple comparison test at P <
0.05 (SPSS 11.0 for Windows, 2002; SPSS Inc.,
Chicago, IL).

RESULTS

Mating Frequencies of Females and Males

The results of mating frequency are shown in
Table 1. When 40 females were paired individu-
ally with 2-d-old virgin males on successive days
until they died, only 1 spermatophore was de-
tected in the abdomens of 31 females (77.5%),
while no spermatophore was detected in the abdo-
mens of the rest of the females (9, 22.5%). Thus
any female that had received a spermatophore in
1 mating did not copulate again. When males
were repeatedly offered 2-d-old virgin females,
the numbers of males that mated various times
during their lifespan and the corresponding per-
centages were as follows: 0 matings (10; 25.0%); 1
mating (9;22.5%); 2 matings (8; 20.0%), 3 matings
(7; 17.5%), 4 matings (5; 12.5%); 5 matings (0; 0%)
and 6 matings (1; 2.5%).

The Effect of Mating on the Longevity of Adults

Males lived significantly longer than females
(Table 1), whether mated or unmated (P < 0.05).


TABLE 1. MATING FREQUENCY OF ECTROPIS OBLIQUE AND ITS EFFECT ON FEMALE AND MALE LONGEVITY.

No. of Female Female No. of Male Male
females mating rates Longevity males mating rates longevity
Mating time observed (%) (days) observed (%) (days)

0 9 22.5 *11.36 + 2.67 a 10 25.0 14.2 0.85 b
1st 31 77.5 10.07 + 2.87 a 9 22.5 19.6 1.82 c
2nd 0 0 ** 8 20.0 16.0 1.47 b
3rd 0 0 7 17.5 15.5 2.50 b
4th 0 0 5 12.5 15.5 1.50 b
5th 0 0 0 0
6th 0 0 1 2.5 15

*Values are mean + SE. Different letters indicate significant difference (P < 0.05) by LSD test.
** not tested.







Florida Entomologist 94(1)


In addition, the males that mated only once lived
significantly longer than unmated males or males
that had mated more than once (P < 0.05). How-
ever males that had mated 2 times did not live
significantly longer than males that had mated
either 3 or 4 times. The life spans of mated and
unmated females did not differ significantly.

Electroantennographic Analysis of the Effect of Mating
on Pheromone Production and on Male Responsiveness
at Various Days after Mating

The effects of mating and lapsed time after
mating of males on their EAG responses to sex
pheromone extracts from sexually active females
are shown in Fig. 1. No significant (P > 0.05) dif-
ferences in EAG responses to sex pheromone ex-
tracts from sexually active females were observed
between the mated and virgin males. The anten-
nae of males that had been amputated 0, 1, 2, 3,
and 4 d post-mating exhibited the same magni-
tude of the EAG responses to sex pheromone ex-
tracts from sexually active females as those of an-
tennae of the corresponding virgin males.
The male EAG responses to female sex phero-
mone extracts from virgin females and mated fe-
males (Fig. 2) differed profoundly (P < 0.05) with
the former evoking much stronger responses than
the latter. Moreover the male EAG responses to
sex pheromones extracted from the females at 0,
1, 2, 3, and 4 d post mating remained at very low
levels (data not shown).

Pheromone Titer Analysis

The virgin females began to produce sex pher-
omones during the first scotophase after emer-


14 *matedmate
a
12 Ovirgn male
0 b b

C c C C
S- 4


2
1-


0 1 2 3 4
Daw after matin

Fig. 1. Influence of the mating status of males on
their EAG response to female sex pheromone extracts
from sexually active females. Antennae of unmated
males (solid bar) and virgin males (open bar) amputated
0, 1, 2, 3, and 4 d post-mating were used as EAG detec-
tors. Data were presented as mean values SE (n = 6-
8) and analyzed by one-way ANOVA, followed by an
LSD multiple range test (P < 0.05). Significant differ-
ences among various dosages of the same stimulant are
indicated with different letters.


0 1 2
Days after mating


Smatred male
Dvinfm*e

b





d d


3 4


Fig. 2. Influence of differential mating status of fe-
males on male EAG responses to their pheromones. Sex
pheromone extracts of mated females (solid bar) and
virgin females (open bar) obtained 0, 1, 2, 3, and 4 d
post-mating were used as stimuli. Data were presented
as mean values SE (n = 6-8) and analyzed by one-way
ANOVA, followed by an LSD multiple range test (P <
0.05). Significant differences among various dosages of
the same stimulant are indicated with different letters.



gence (Fig. 3). Maximal pheromone titers were
present in the glands during the second and third
scotophase after emergence. Thereafter the pher-
omone titer decreased gradually. When the fe-
males mated on the first night after eclosion, the
sex pheromone titers decreased strongly and sig-
nificantly compared with the titers of virgin fe-
males. Pheromone production in mated females
remained suppressed during each of 4 successive
nights after they had mated. These results are
consistent with the weak male antennographic
responses coinciding with the male response to
mated female sex pheromones.


a e

tr t
Oaysafter mating


Smaed fenmle
vnini female


d




3 4


Fig. 3. Influence of mating on the production of 6,7-
epoxy-(Z,Z)-3,9-octadecadiene, the major female sex
pheromone. The female sex pheromone was extracted
with hexane from pheromone glands of mated females
(solid bar) and virgin females (open bar). Pheromone
glands of mated females were extracted 0, 1, 2, 3, and 4
d post-mating.


__ __ __ __ __


March 2011


id







Yang et al.: Mating Strategy of Ectropis oblique


DISCUSSION

Our data suggest that the females in our labo-
ratory colony of E. oblique are monandrous. Two
different views exist concerning female mating
strategies. Polyandry presents a variety of bene-
fits to females, including full fertilization of their
egg complement, increased genetic diversity of
offspring, receipt of non-sperm nutrients, and re-
duced chances of fertilization by sperm that are
genetically defective due to age. Conversely, poly-
andry may decrease female fitness due to the eco-
logical cost of mating, including energy costs, and
risks of physical injury and sexually transmitted
pathogens and parasites (Arnqvist & Nilsson
2000). We observed that E. oblique females after
having mated fended off males and did not accept
a second mating partner. Mated females began
laying eggs during the first scotophase and laid
nearly all of their eggs before the fourth day (data
not shown). The life spans of females ranged from
about /2 to 23 of the lengths of the life spans of
males. The lifespan of the female in the wild is
likely to be even shorter than in the laboratory.
Thus, a single mating is sufficient to fertilize
nearly all eggs and minimize the above men-
tioned risks associated with multiple matings.
Males, on the other hand, are potentially po-
lygynous. Male polygyny is to be expected because
most evolutionary theories contend that the con-
tributions and consequences of mating are much
greater for females than for males (Thornhill &
Alcock 1983).
This study revealed that multiple matings re-
duced the longevity of males but not of females.
This result is in agreement with the results of
studies on several other species (Proshold et al.
1982; Svensson et al. 1998). It is thought that al-
location of nutritional reserves for egg develop-
ment and maturation after mating may be re-
sponsible for causing the lifespan of mated fe-
males to be shorter than that of virgin females.
In most species, there is a causal relationship
between male calling behavior and female phero-
mone emission. Only when the female pheromone
gland becomes exposed to emit pheromone, may
the male display calling behavior. Permanent or
even temporary reductions in the emissions of sex
pheromones caused loss of attraction and sexual
receptivity in males (Kingan et al. 1995). The
present study showed that E. oblique male EAG
responses to mated female pheromone gland ex-
tracts were significantly diminished, which could
be the result of reduced pheromone release. The
results support the hypothesis that mating con-
siderably suppressed pheromone production in fe-
males. Indeed according to our capillary-GC anal-
ysis, pheromone titers in pheromone gland ex-
tracts did not increase at all up to 4 d after mating.
Because the male EAG response to pheromone
gland extract of already mated females did not


show any increase up to 4 d after mating, it may
be deduced that females may mate only once. This
deduction is in accordance with the observation
that each female actually mates only 1 time. Even
though a few females were observed to copulate
twice, but no more than 1 spermatophore was
ever detected in a bursa copulatrix, possibly be-
cause the first copulation was an unsuccessful
mating.
Mating-induced termination of sex pheromone
production has been investigated in several moth
species (Raina et al. 1994; Ando et al. 1996). The
inactivation of pheromone production after copu-
lation, which reduces the ability of females to
elicit a sexual response in males, may be due to
the secretion of pheromonostatic peptide (Kingan
et al. 1995), or the presence of viable sperm in the
spermatheca (Giebultowicz et al. 1991). The
mechanisms involved in pheromone suppression
after copulation in E. oblique are currently un-
known. Thus, it is suggested that further studies
must be conducted on this area.
The present results show that mating status
did not appear to have a significant effect on the
responses of male antennae to sex pheromone ex-
tracts of sexually active females. This suggests
that a past mating does not cause the male to be
unresponsive to the sex pheromone, and such a
male can be expected to continue to seek females
for additional matings. This deduction is also in
accordance with direct observations of multiple
matings by males. The same mating system de-
scribed for E. oblique has been observed in some
other species (Royer & McNeil 1993; Foster& Ay-
ers 1996; Svensson et al. 1998).

ACKNOWLEDGMENT

The authors express gratitude to Professor M. Z.
FAN for assistance during this study.

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Florida Entomologist 94(1)







Farnum & Loftin: Distribution of P. curvatus and P. tricuspis in Arkansas



DISTRIBUTION OF PSEUDACTEON CURVATUS AND PSEUDACTEON
TRICUSPIS (DIPTERA: PHORIDAE) IN ARKANSAS

JAKE M. FARNUM AND KELLY M. LOFTIN
Department of Entomology, University of Arkansas, 319 Agriculture Bldg., Fayetteville, AR 72701

ABSTRACT

From 1995 to 2009, four Pseudacteon species were released in the U.S. with 3 species, P. cur-
vatus, P. tricuspis, and P. obtusus released in Arkansas. To determine Pseudacteon establish-
ment and expansion, sticky traps were used to monitor phorid fly species at and near 10
release sites, in counties bordering neighboring states, and along regional transects. Pseu-
dacteon flies were captured in 16 Arkansas counties: Ashley, Chicot, Clark, Desha, Drew,
Hempstead, Howard, Little River, Montgomery, Nevada, Perry, Phillips, Pike, Polk, Sevier,
and Union. Pseudacteon curvatus was found in areas far from release sites, suggesting dis-
persal from neighboring states. The range of P. tricuspis evidently also expanded from its ini-
tial release sites in southern Arkansas.

Key Words: biological control, phorid fly, Solenopsis invicta, parasitoid

RESUME

De 1995 hasta el 2009, cuatro species de Pseudacteon fueron liberadas en los Estados Uni-
dos con 3 de ellas, P. curvatus, P. tricuspis y P. obtusus, liberadas en el estado de Arkansas.
Para determinar el establecimiento y expansion de Pseudacteon, se usaron trampas pegajo-
sas para monitorear las species de f6ridos en y alrededor de los 10 sitios donde fueron libe-
radas, en condados fronterizos con los estados vecinos, y a lo largo de las lines regionales.
Las moscas Pseudacteon fueron capturadas en los siguientes 16 condados de Arkansas: As-
hley, Chicot, Clark, Desha, Drew, Hempstead, Howard, Little River, Montgomery, Nevada,
Perry, Phillips, Pike, Polk, Sevier y Union. Pseudacteon curvatus fue encontrada en areas le-
jos de los sitios done fue liberada, que indica que se disperse de los estados vecinos. Eviden-
temente, el rango de P. tricuspis tambien se expandio de los sitios donde fue liberada
inicialamente en el sur de Arkansas.


Pseudacteon phorid flies parasitize and kill
their fire ant host (Solenopsis invicta Buren, S.
richteri Forel and their hybrid). Due to their high
host specificity (Folgarait et al. 2002; Gilbert &
Morrison 1997; Morrison & Gilbert 1999; Porter
1998a, 2000; Porter & Alonso 1999; Porter & Gil-
bert 2004; Vazquez et al. 2004), several Pseudac-
teon spp. have been introduced from South Amer-
ica as classical biological control agents against
imported fire ants. While internal development of
the fly larvae eventually decapitates and kills in-
dividual ants (Porter et al. 1995a; Consoli et al.
2001), perhaps the most important effect is the
disruption of the foraging behavior of the ants
(Feener & Brown 1992; Orr et al. 1995; Porter et
al. 1995b; Mehdiabadi & Gilbert 2002), which
leads to a decrease in food uptake and a decline in
colony health (Folgarait & Gilbert 1999). Four
Pseudacteon species were released in the U.S.
from 1995 to 2009: P curvatus Borgmeier, P. lito-
ralis Borgmeier, P. obtusus Borgmeier, and P tri-
cuspis Borgmeier. Each species fills a different
niche, in terms of diurnal activity (Pesquero et al.
1996), seasonal occurrence (Fowler et al. 1995;
Folgarait et al. 2003) and preferred size of host
(Campiolo et al. 1994); all complementing traits
for control of S. invicta and S. richteri (Morrison


et al. 1997; Porter 2000; Folgarait et al. 2002,
2005).
Pseudacteon flies use semiochemicals to locate
their ant hosts (Orr et al. 1997; Vander Meer &
Porter 2002; Morrison & King 2004; Chen & Fad-
amiro 2007) and then hover over the ants before a
rapid aerial attack (Morrison & Porter 2005).
Within a period of an hour a female fly can make
up to 120 oviposition attempts (Morrison et al.
1997), before either tiring or being captured and
killed by the ants (Porter 1998b). A single egg laid
in the thorax develops through 3 instars before
decapitating the ant's head, which is then used as
a pupal case (Pesquero et al. 1995; Porter et al.
1995a).
The first release of P tricuspis occurred in
Texas in 1995 (Gilbert 1996), and was unsuccess-
ful due to unfavorable conditions (Vazquez et al.
2006). The first successful release of P tricuspis in
northern Florida was in 1997 (Porter et al. 1999).
Pseudacteon spp. have been released in 11 south-
ern states: Alabama, Arkansas, Florida, Georgia,
Louisiana, Mississippi, North Carolina, Okla-
homa, South Carolina, Tennessee, and Texas
(Porter et al. 1999; Graham et al. 2003; Williams
& deShazo 2004; Parkman et al. 2005; Thead et
al. 2005; Henne et al. 2007; Weeks & Callcott







Florida Entomologist 94(1)


2008). Pseudacteon curvatus, P. obtusus, and P.
tricuspis have been released in Arkansas and in
adjacent states except Missouri (Clemons et al.
2003; Weeks & Callcott 2008).
Dispersal rates for Pseudacteon spp. are vari-
able and the majority of flies disperse only a few
hundred meters (Morrison et al. 1999). However,
some flies in each generation are known to travel
2 to 4 km or more (Porter et al. 2004) and popula-
tions of flies expanding on average 74 km over a
period of 3.5 years (Porter 2010).
Monitoring of Pseudacteon spp. is achieved
with a variety of methods: actively through direct
collections of flies at disturbed ant colonies with
either manual or electrical stimulation (Barr &
Calixto 2005; Morrison & Porter 2005), or pas-
sively by trapping with a sticky trap (Puckett et
al. 2007). Until the current study, monitoring of
these species in Arkansas had been concentrated
at and near release sites to determine Pseudac-
teon establishment. The objective of this study
was to determine the distribution of Pseudacteon
spp. in Arkansas through wider-scale monitoring.

MATERIALS AND METHODS

Trap Design

To determine presence or absence, passive
trapping was used based on a modified version of
a PTS (pizza tri-stand) sticky trap (Puckett et al.
2007). The modified Puckett trap (Fig. 1) con-
sisted of a pizza tri-stand (Polyking No. 20431)
covered in Tanglefoot, glued to the flat side of an
inverted portion cup (Dart No. 100PC), which
was hot glued to the underside of a plastic cup lid
(Dart No. 8JL). This device was placed in the
center of the bottom half of a plastic Petri dish
(150 by 15 mm). The surfaces of the portion cup
and the inner lip of the Petri dish were coated


Fig. 1. A modified Puckett trap.


with Fluon, to prevent ants from climbing up
the trap or out of the Petri dish.
One trap was placed per location by first locat-
ing a mound of substantial size and activity. The
mound was then disturbed, by kicking it over cre-
ating a flat surface on which the Petri dish was
placed. As ants climbed into the Petri dish, a few
ants were crushed by hand to induce alarm pher-
omone release and the trap was placed in the cen-
ter of the Petri dish. A brightly colored pin flag (91
cm long) was positioned alongside the trap. Glo-
bal positioning system (GPS) coordinates were re-
corded for each location, and a corresponding
number written on the lid of the trap.
Traps were retrieved 20 to 24 h after place-
ment. At time of retrieval, an 8 oz expanded poly-
styrene foam cup (Dart No. 8J8) was placed over
the trap, and the lid was snapped in place. The
cup prevented damage and contamination to the
sticky portion of the trap.

Sampling at Release Locations

Fourteen releases of Pseudacteon spp. were
made in Arkansas from 1998 to 2009 (Table 1).
Sampling along transects in the cardinal direc-
tions from the release sites began in 2002 (Pike
Co.) and 2004 (Miller Co.). In 2009, the Miller,
Perry, Pike, and Sevier County release sites were
revaluated for this study to confirm establishment
ofPseudacteon spp. A 1.6-km interval was used be-
tween traps along each transect placed in Pike,
Miller, and Sevier Counties, and at 0.8-km inter-
vals in Perry County. Transects were determined
by locating roads and highways on aerial maps
that radiated out from the release site in north,
south, east, and west directions. One modified
Puckett trap was placed at each sampling location.

Sampling Bordering Counties

Sampling was intended to monitor spread of
established populations of Pseudacteon spp. in
bordering counties/parishes of neighboring states
of Louisiana (Henne et al. 2007), Mississippi
(Thead et al. 2005), and Tennessee (Graham et al.
2003; Parkman et al. 2005; Weeks & Callcott
2008), and sampling packages were sent to Uni-
versity of Arkansas Cooperative Extension Ser-
vice County Agents in imported fire ant infested
counties in eastern and southern Arkansas in the
early summer of 2009. Each package contained
four modified Puckett traps, latex gloves, and an
information sheet. Instructions were to place the
traps during the summer months, as previously
described, at 4 locations within the county. De-
ployed traps were returned from 4 bordering
counties: Columbia, Lafayette, Phillips, and St.
Francis. Geographic coordinates were recorded on
the instruction sheet with the number of the cor-
responding trap.


March 2011







Farnum & Loftin: Distribution of P. curvatus and P. tricuspis in Arkansas 17


TABLE 1. HISTORY OF PSEUDACTEON SPP. RELEASES IN ARKANSAS, 1998-2009.

Date County Species Number Released

1998, Jul-Aug Drew P. tricuspis 1,350
2002, May2 Pike P. tricuspis 3,000
2002, Oct2 Bradley P. tricuspis 1,200
2003, Sep2 Bradley P. tricuspis 1,500
2004, May2 Miller P. tricuspis 2,580
2005, May2 Sevier P. tricuspis 4,300
2005, Oct2 Clark P. curvatus 8,5003
2006, Sep2 Perry P. curvatus 15,8164
2007, Sep2 Perry P. tricuspis 1,900
2008, Jun2 Jefferson P. tricuspis 120
2008, Oct2 Polk P. obtusus 3,2005
2009, May2 Grant P. curvatus 25,2966
2009, Jun2 Jefferson P. tricuspis 120
2009, Sep2 Garland P. tricuspis 360

'Release made by Lynne Thompson, University of Arkansas Monticello.
Release made by Kelly Loftin, University of Arkansas Cooperative Extension Service.
'Number released based on 35.3 g S. invicta workers, 800/gram and 30% parasitism.
'Number released based on 65.9 g S. invicta workers, 800/gram and 30% parasitism.
"Number released based on 23.7 g S. invicta workers, 450/gram and 30% parasitism.
'Number released based on 105.4 g S. invicta workers, 800/gram and 30% parasitism.


Sampling Along Regional Transects

Due to possible expansion ofPseudacteon spp.
from neighboring states, sampling transects were
identified and mapped in 3 regions: western,
southeastern, and southwestern Arkansas
(Fig. 2). Transects began at the state line of the
adjacent state and traveled inward, with modi-
fied Puckett traps placed at 3-mile intervals and
GPS coordinates recorded for each trap. Sampling
was conducted for 36 h along transects from each
region between 19 Sep and 3 Oct 2009.


Fig. 2. Regional transects in Arkansas (SE1, SE2,
SE3, SW1, SW2, SW3, SW4, W1, W2, W3, W4) (ESRI
Inc. 2009).


Phorid Fly Identifications

Upon retrieval, modified Puckett traps were
taken to the University of Arkansas laboratory
(Fayetteville, AR) and GPS coordinates logged
into Google EarthTM. Traps were examined under
a dissecting microscope for presence of Pseudac-
teon spp. The sticky portion of the trap was
sprayed with liquid degreaser (Goo GoneTM) if fly
removal was necessary. Species were identified
based on the morphology of the female ovipositor
(Porter & Pesquero 2001). Voucher specimens of
P. curvatus, P. tricuspis, and P. obtusus were ob-
tained from laboratory specimens and the phorid
fly rearing facility in Gainesville, FL for identifi-
cation of male and female flies. Male Pseudacteon
spp. are currently difficult to identify through use
of keys (Morrison et al. 1997; Porter & Pesquero
2001).


RESULTS AND DISCUSSION

Release Locations

Traps placed along transects radiating from
the phorid fly release sites yielded surprising re-
sults. Of the 20 traps retrieved from Sevier
County, no Pseudacteon spp. were caught. Traps
from Miller, Perry, and Pike Counties captured
Pseudacteon spp. Along the Miller County release
site transects, P. curvatus was captured at 3 loca-
tions north of Texarkana, in the southeastern
part of Little River County (Fig. 3). However, this
was unexpected because the phorid fly species re-
leased in Miller County in 2004 was P. tricuspis







Florida Entomologist 94(1)


Fig. 3. Pseudacteon curvatus release sites in (A)
Clark Co., (B) Grant Co., (C) Perry Co., and 2009 cap-
ture locations including (D) Phillips Co., Arkansas.
Pseudacteon tricuspis release sites in (E) Bradley Co.,
(F) Drew Co., (G) Garland Co., (H) Jefferson Co., (I)
Miller Co., (J) Perry Co., (K) Pike Co., (L) Sevier Co., Ar-
kansas and 2009 capture locations (ESRI Inc. 2009).


(Fig. 3, I on map). While possible, it is unlikely
that these captures resulted from cross contami-
nation of the 2 species at the rearing facility. Sev-
eral factors are in place to prevent cross contami-
nation including parasitization of the ants in sep-
arate rooms and lack of available ants of pre-
ferred host size. Furthermore, voucher sampling
is conducted to insure parasitization by the cor-
rect species (Amy Croft, Florida Department of
Agriculture and Consumer Services, personal
communication). A plausible explanation could be
the expansion ofP. curvatus from released and es-
tablished populations in the bordering states of
Louisiana, Oklahoma, and Texas (Weeks &
Callcott 2008; Anne-Marie Callcott, USDA-
APHIS, personal communication). Similar results
were observed along the northeast transect of the
Pike County release site; capture of a Pseudac-
teon fly (P. curvatus) different from the species
originally released (P. tricuspis). Due to the prox-
imity (5 km) with the Clark County release site
(Fig. 3A), it is possible that this capture was from
movement of P. curvatus from Clark County, al-
though revaluation of Clark County was not con-
ducted in 2009. In Perry County, capture of P.
curvatus was recovered at the initial release site
and at locations to the south (0.5 km) and east
(2.2 km).

Bordering Counties

Puckett traps from Columbia, Lafayette, and
St. Francis Counties were devoid of Pseudacteon
flies, although all traps returned from Phillips
County on the Mississippi border captured P. cur-


vatus. Phillips County is the one of 2 counties in
Arkansas (Crittenden Co. the other) currently
known to have only S. richteri and no record of S.
invicta (Robert Vander Meer, USDA-ARS, per-
sonal communication). Sampling locations in
Phillips Co. were located on the western levee of
the Mississippi River, northwest of Friars Point,
MS (Fig. 3D), adjacent to counties in Mississippi
and Tennessee with known S. richteri popula-
tions (Streett et al. 2006; Oliver et al. 2009). The
high number ofP. curvatus found on 1 trap (~172)
implies sufficient colonies of S. richteri to support
P. curvatus populations.

Regional Transects

A total of 176 modified Puckett traps were
placed along transects in western, southeastern,
and southwestern Arkansas (Fig. 2). In the west-
ern region of transects, 28 traps contained P cur-
vatus, and 2 contained P tricuspis (Fig. 3). Of the
traps that captured P tricuspis, one was located
29.5 km west of the P tricuspis release in Pike
County, and the other was 46 km northwest of the
release site. All 4 transects in this region included
captures of P curvatus. On the 2 northerly routes
(W1 and W2), traps with P curvatus were found
at regular intervals with the most easterly cap-
ture 54 km from the Arkansas/Oklahoma state
line. The remaining 2 transects (W3 and W4) also
had captures. Transect (W3) picked up P curva-
tus 13 km from the Arkansas/Oklahoma border.
The 3 traps from the most southerly transect
(W4) that contained P. curvatus were located near
the northern section of the Miller County release
site transect. This directional pattern of recover-
ies supports the hypothesis of immigration from
confirmed P curvatus populations in Le Flore
County, Oklahoma (Weeks & Callcott 2008) ap-
proximately 24 km northwest of Mena, Arkansas.
Prevailing winds generally do not correlate with
dispersion patterns (Morrison et al. 2000), as flies
move close to the ground where wind is reduced.
Pseudacteon curvatus but not P tricuspis was
trapped along the southeastern region transects.
P curvatus was found at 1 location in Union
County, north of El Dorado, and on 12 traps along
each of the other transects (SE2 and SE3) to the
east (Fig. 3). From the 2 easterly transects (SE2,
SE3), traps with P curvatus were found 91 km
north of the Arkansas/Louisiana border and 65
km west of the Arkansas/Mississippi border. This
distribution in Arkansas is expected based on col-
lections of P curvatus in bordering counties along
the western side of Mississippi (Adams, Bolivar,
Claiborne, Coahoma, Desoto, Jefferson, Tunica,
Warren, Washington, Wilkinson), and along Loui-
siana's northern side (Claiborne, East Carroll,
Morehouse, Union, West Carroll) (Anne-Marie
Callcott, USDA-APHIS, personal communica-
tion).


March 2011







Farnum & Loftin: Distribution of P. curvatus and P. tricuspis in Arkansas


Pseudacteon tricuspis was captured at 2 loca-
tions along a transect (SW3) of the southwestern
region of Arkansas (Fig. 3). One of the traps was
located in the northern part of Nevada County, in
the city of Prescott, and the other was 18 km to
the southeast. Their proximity to the Pike County
release site (Fig. 3K), 20 km northwest, may sug-
gest their origin, although P curvatus appeared
in no samples taken from the Pike County release
site transect. Weather conditions may have
played a role in the lack of Pseudacteon spp.
present on the remaining 55 traps in this region.
Temperatures for the region on 3 Oct 2009 ranged
from a low of 8C to a high of 26C. Temperatures
below 20C inhibit activity of Pseudacteon flies
(Morrison et al. 1999; Wuellner et al. 2002). On 3
Oct 2009 temperatures were above 20C for a 7-h
period midday (11:00 AM to 6:00 PM CST) and be-
low 20C for the entire day of 4 Oct 2009, when
the traps were retrieved. However, 2 traps located
on the transect southeast of Prescott, AR collected
P tricuspis. Weather data suggests similar condi-
tions for the Prescott, AR area, although slightly
warmer temperatures (17C) were recorded for an
additional 3.5 h on the morning of 4 Oct 2009.
Of the traps that collected Pseudacteon spp.,
no traps contained both species. Perhaps because
P tricuspis is reliant on larger ants, P. curvatus is
able to establish more readily where P. curvatus
and P tricuspis overlap, and thus were not de-
tected if present in low densities (Gilbert et al.
2008). Another factor for the lack of both species
in the trap may be due to the modification of the
Puckett trap. The 2 differences in the traps design
were the attractant used and the placement of the
trap. Traps were placed on a disturbed mound
with live ants whereas Puckett traps were placed
in an open area with midden (Puckett et al. 2007).
The Puckett trap as originally designed captured
more P. tricuspis than P. curvatus, although sea-
sonal fluctuations could be a variable (Puckett et
al. 2007).

CONCLUSION

Passive Pseudacteon trapping with the modi-
fied Puckett trap provided advantages over direct
collection from disturbed mounds. Because it is
deployed quickly, multiple traps can be placed
over a large area which allows continuous and si-
multaneous sampling (Puckett et al. 2007). The
manpower needed to achieve similar coverage us-
ing observational sampling is cost prohibitive.
The addition of the protective cup to the original
design allowed longer storage time between col-
lection and examination of the trap, protection of
the sticky portion, and reduced contamination.
The modification based on disturbed mounds and
fluon-coated petri dishes rather than fire ant mid-
den was advantageous in that maintenance of a
fire ant colony for collection of midden is no longer


necessary. With this modification, trapped fire
ants rather than fire ant midden serve as the
Pseudacteon fly attractant.
The results suggest establishment and expan-
sion of P curvatus from the release site in Perry
County, and P. tricuspis from the release site in
Pike County. While limited, the current range of
P tricuspis in Arkansas appeared to be along a
narrow 86-km band stretching from northwest
Pike County to south central Nevada County, and
25 km west of the release site. The current distri-
bution ofP. curvatus in Arkansas suggested natu-
ral movement from surrounding states. Despite
extensive sampling across southern Arkansas,
many areas remained unsampled. Additional
trapping would provide a better understanding of
the distribution of Pseudacteon spp. in Arkansas.

ACKNOWLEDGMENTS

We thank Anne-Marie Callcott of the USDA APHIS
lab in Gulfport, MS for approving Pseudacteon spp. for
release, Amy Bass, Amy Croft, and Deborah Roberts of
the Florida Department of Agriculture and Consumer
Services for assistance in supplying phorid flies, Ed
Brown, Jerry Clemons, Randy Forst, Rex Herring, Mike
McCarter, Shawn Payne, Doug Petty, Amy Simpson,
Rebecca Thomas, Shaun Rhodes, Carla Vaught, Joe Ves-
tal, and Danny Walker of the University of Arkansas
Cooperative Extension Service for assistance placing
phorid fly traps, Michael Hamilton and Robert Goodson
for collecting imported fire ants for identification, and
Ricky Corder of the University of Arkansas Cooperative
Extension Service for assistance.

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Florida Entomologist 94(1)


March 2011


MINI-ASPIRATOR: A NEW DEVICE FOR
COLLECTION AND TRANSFER OF SMALL ARTHROPODS TO PLANTS

MAHMUT DOGRAMACI1', JIANJUN CHEN2, STEVEN P. ARTHURS' CINDY L. MCKENZIE3, FABIELI IRIZARRY,'
KATHERINE HOUBEN1, MARY BRENNAN' AND LANCE OSBORNE1
'University of Florida, Department of Entomology and Nematology, Mid-Florida Research and Education Center,
Apopka, FL 32703, USA

2University of Florida, Department of Environmental Horticulture, Mid-Florida Research and Education Center,
Apopka, FL 32703, USA

3U.S. Horticultural Research Laboratory, ARS-USDA, Fort Pierce, FL 34945, USA

ABSTRACT

The process of collecting and/or infesting plants with a designated number of small arthro-
pods in biological experiments is tedious and laborious. We developed a modified mini-aspi-
rator, powered with a vacuum pump and fitted with a specially adapted (removable)
collection vial to reduce the handling effort. The efficiency of the mini-aspirator was tested
with the chilli thrips, Scirtothrips dorsalis Hood (Thysanoptera: Thripidae), a predatory
mite,Amblyseius (= Neoseiulus) cucumeris (Oudemans) (Acari: Phytoseiidae), and the insid-
ious flower bug, Orius insidiosus (Say) (Heteroptera: Anthocoridae). Using the mini-aspira-
tor, operators collected 10A. cucumeris mites and 10 S. dorsalis thrips and transferred them
onto pepper plants in 43 s and 37 s, respectively, compared with 639 and 229 s, respectively,
using a camel's hair brush as a conventional method. The use of the mini-aspirator for col-
lectingA. cucumeris predatory mites and S. dorsalis thrips and infesting pepper plants with
them represents a 15-fold and 6-fold time saving, respectively. Collection of 10 0. insidiosus
flower bugs took 20 s with the mini-aspirator compared with 30 s when an unmodified aspi-
rator was used. Proportionally, the amount of time saved with the mini-aspirator for the
handling of 0. insidiosus flower bugs was minimal compared with the timesavings when
handling S. dorsalis thrips and the A. cucumeris predatory mites with the mini-aspirator.
Additionally, the mini-aspirator can be fitted with a battery-powered Mini-Vac, which makes
it portable for field applications, such as in sampling field populations when screening for
pesticide resistant individuals.

Key Words: mini-aspirator, Neoseiulus cucumeris, Scirtothrips dorsalis, Orius insidiosus,
manual infestation

RESUME

El process de recolectar y/o infestar plants con un cierto numero de artr6podos pequeios en
experiments biol6gicos es tedioso y laboroso. Desarrollamos una mini-aspiradora modifi-
cada, que funciona mediante una bomba de succi6n especialmente montada con un frasco de
recolecci6n especialmente adaptado (desmontable) para reducir el esfuerzo de manejo. La
eficiencia de la mini-aspiradora fue probada con el trips de chile, Scirtothrips dorsalis Hood
(Thysanoptera: Thripidae), un acaro depredador, Amblyseius (=Neoseiulus) cucumeris
(Oudemans) (Acari: Phytoseiidae), y el chinche pirata diminuto, Orius insidiosus (Say) (He-
teroptera: Anthocoridae). Usando la mini-aspiradora, los operadores recolectaron y transfe-
rieron 10 acaros depredadores y trips de chile a plants de chile en 43 y 37 segundos,
comparado con 638 y 229 segundos usando el metodo conventional con una brocha de pintar.
El uso de la mini-aspiradora para mover e infestar plants con acaros depredadores y trips
represent un ahorro de 15 y 6 veces, respectivamente. La recolecci6n de 10 chinches pirates
diminutos tom6 20 segundos con la mini-aspiradora comparada con 30 segundos cuando se
uso una aspiradora no modificada. El tiempo ahorrado proporcionalmente fue minimo com-
parado con la recolecci6n de trips de chile y los acaros depredadores con la mini-aspiradora.
Ademas, la mini-aspiradora puede ser mantada con una mini-bomba de succionar de bate-
rfa, que la hace portable para aplicaciones en el campo, como en la evaluaci6n de resistencia
de plaguicidas en poblaciones de campo.


Manual collection and infestation of small ar- thropods. Thrips and predatory mites are exam-
thropods (<2 mm) can be labor intensive and cum- ples of small arthropods used by scientists in nu-
bersome and result in injury to the handled ar- merous experiments (Mound & Palmer 1981;







Dogramaci et al.: Mini-aspirator for Handling Small Arthropods


Chiu et al. 1991; Tatara & Furuhushi 1992; Ts-
chuchiya et al. 1995; Bournier 1999; Seal et al.
2006; Arthurs et al. 2009). Scirtothrips dorsalis
Hood, chilli thrips, is one of the smallest thrips
species with adults ranging from 1.5-2.0 mm. The
S. dorsalis adult moves rapidly, and may jump or
fly when disturbed. Immature stages of S. dorsa-
lis, especially first instars, are very small (<1 mm)
and have fragile easily injured bodies. Thus, an
important requirement for studying small arthro-
pods is to have a reliable method for collecting
and releasing designated numbers of individuals
without harm.
Three methods are used for infesting plants
with thrips. One is manual infestation of arthro-
pods with a soft-bristled camel's hair brush
(Cloyd & Sadof 1998). This method has been
used widely but is labor-intensive and time-con-
suming. For instance, Cloyd et al. (2001) re-
ported that infesting 50 plants each with 10
adult western flower thrips (WFT), Frankliniella
occidentals (Pergande), required a technician 3.5
h. In addition, the process of mechanical transfer
involved a risk of injury to the specimen. The
second method involves placement of plants in a
location where thrips are known to occur in or-
der to allow a natural population of the pest to
build-up on the test plants. Although this
method is less cumbersome than manual infes-
tation, the number of thrips transferred onto
test plants cannot be accurately regulated,
which introduces variation among test plants.
The third method is the use of a commercial
mouth operated aspirator purchased from Bio-
Quip, Rancho Dominquez, CA. An improvement
to the mouth-operated aspirator was reported by
Cloyd et al. (2001), who developed a "Small In-
sect Aspirator" for collecting WFT. The latter in-
volved the use of an aspirator from BioQuip at-
tached to a battery operated 'Mini-Vac' (MV In-
strument, Glendale, CA 91205; http://www.mini-
vac.com/index02.html). Use of the small insect
aspirator is attended with operational difficul-
ties similar to those encountered with the use of
regular aspirators, in particular the collection of
extraneous materials. In our studies, we found
that the wide suction tubes of the aspirators col-
lect too many extraneous materials along with
thrips (e.g., other organisms and plant debris)
and sometimes causes physical damage to
thrips. Additionally, in our experience the design
of the collection vial does not allow the thrips to
be released easily following collection.
In order to overcome some of the design limita-
tions of previous small arthropod handling de-
vices, we developed a "mini-aspirator" that can be
powered either with a laboratory vacuum pump
or with a small portable vacuum pump and fitted
with a specially adapted and removable collection
vial that allows rapid transfer of the collected ar-
thropods onto plants. We compared the efficiency


of the mini-aspirator with a paintbrush and com-
mercial aspirator for collecting and releasing the
chilli thrips, Scirtothrips dorsalis Hood and 2 of
its natural enemies, a predatory mite, Neoseiulus
cucumeris (Oudemans), and the flower bug, Orius
insidiosus (Say).

MATERIALS AND METHODS

Mini-aspirator

The mini-aspirator was built from clear 6.35-
mm diam vinyl tubing fitted with a 1-mL filtered
pipette tip (VWR International, West Chester,
PA). The intake tubing opening was reduced by
using an adaptor to attach a 200-pL pipette tip,
which facilitated the collection of individual
small arthropods (Figs. 1A and 1B). The modi-
fied mini-aspirator was powered by an electrical
laboratory vacuum pump (Rocker vacuum pump,
Rocker Scientific Co., Ltd., Kaohsiung, Taiwan)
(Fig. 1A). To collect S. dorsalis, an infested leaf
was placed under a stereomicroscope (Fig. 1C)
and the desired number of thrips was captured
in the collection vial for transfer onto plants. The
collection vial was removed from the mini- aspi-
rator and attached to a plant with a hair clip to
allow the voluntary dispersal of the thrips onto
the plant (Fig. 1D). To make the mini-aspirator
portable, we integrated the mini-aspirator with
a Mini-Vac (MV Instrument, Glendale, CA
91205; http://www.mini-vac.com/index02.html)
(Fig. 1E). However, to compensate for the re-
duced suction power of the Mini-Vac, the pipette
tip filter was replaced with fine woven nylon fab-
ric. The assembly of the pipette tip, collection
vial, filter and vacuum tube is shown in Fig. 2.
Such integration of the "mini-aspirator" with the
battery powered Mini-Vac made the system por-
table for field use.

Arthropods

Scirtothrips dorsalis specimens were obtained
from a colony that originated from rose plants in
Winter Park, FL. The colony was maintained on
cotton plants, Gossypium hirsutum, 'Deltapine
493 Conventional'. The health of the colony was
maintained by periodically introgressing thrips
from naturally infested plants. Commercially
available thrips predators (a predatory mite, N.
cucumeris (Oudemans), and the insidious flower
bug, Orius insidiosus (Say)) were obtained from
Koppert Biological Systems, Berkel en Rodenrijs,
The Netherlands.

Plant Material
The infestation methods were tested on sweet
pepper plants, Capsicum annum L. Pepper seeds
were germinated on moist filter papers inside







Florida Entomologist 94(1)


March 2011


Fig. 1. Novel minute arthropod infestation apparatus and its use. A. The complete system in use; B. The mini-
aspirator small arthropod collector; C. Collecting S. dorsalis from a leaf; D. Mini-aspirator along with collected ar-
thropods attached to a plant to allow dispersal; E. The portable mini-aspirator consisting of clear 6.35 mm diam vi-
nyl tubing fitted with a 1-mL pipette tip and nylon cloth filter connected with an adaptor to a 200-jL pipette tip;
with suction provided by a Mini-Vac.

Petri dishes. Germinated seeds were transferred Collection and Infestation of Small Arthropods with the
to seedling trays. Seedlings each with 4-6 fully ex- Conventional Camel's Hair Brush
panded leaves were planted into 15 cm diam pots.
Pepper plants at >10 leaf stage were used for the Collecting chilli thrips and infesting plants
arthropod infestation experiments, with them. Either 10 or 20 chilli thrips were cap-







Dogramaci et al.: Mini-aspirator for Handling Small Arthropods


u r, ube



R.: ,n gfilter
Cp11:: :lllgtube


S 00 p, pipette tip-
:: Ii r _.lle ting
' .ric, lualsmall
.. .. ." ar hroDods


Fig. 2. Assembly of the pipette tip, collection vial, fil-
ter and vacuum line of the mini-aspirator.


turned with a moistened camel's hair brush. Each
of these thrips was then placed onto a pepper
plant leaf. The number of seconds needed to cap-
ture and transfer the above designated number of
thrips was recorded. This was repeated 8 times
each for groups of 10 or 20 chilli thrips.
Sorting and releasing N. cucumeris. The entire
content of the package containing N cucumeris
and substrate was emptied into a Petri dish (15
cm diam) lined with a filter paper. To separate N.
cucumeris predatory mites from the substrate,
the closed Petri dish was agitated gently several
times. Under a stereomicroscope each predatory
mite was collected individually from the filter pa-
per and placed onto a pepper plant with a camel's
hair brush. Groups of either 10 or 20 N. cucumeris
were placed on a plant. The number of seconds re-
quired to collect and release either 10 or 20 pred-
atory mites was recorded. This was repeated 8
times each for groups of 10 and 20 mites.
Collecting and releasing 0. insidious flower
bugs on pepper plants. Orius insidious flower
bug adults in groups of either 10 or 20 were col-
lected from a purchased colony with a BioQuip as-
pirator. The contents of the package (vermiculite
substrate and bugs) were emptied onto a board
and bugs that crawled on the board were cap-
tured with the BioQuip aspirator. Each collection
vial containing the 0. insidiosus was placed at the
base of a pepper plant that had been infested with
thrips to allow the bugs to exit the vial and dis-
tribute onto the plant. The number of seconds re-
quired to collect and to release either 10 or 20
bugs was recorded. This was repeated 8 times
each for groups of 10 or 20 0. insidiosus.

Use of the Mini-aspirator to Collect Small Arthropods
and Transfer Them onto Plants

Collecting S. dorsalis thrips and infesting plants
with them. Ten or 20 S. dorsalis were collected with


the mini-aspirator as described above. The mini-as-
pirator along with the collected S. dorsalis was at-
tached to a pepper plant leaf with a hair clip in a
manner that allowed thrips to distribute them-
selves on plant leaves (Fig. 1D). The time required
for collection and release (the latter being the time
required to attach the mini-aspirator along with
the collected S. dorsalis to a pepper plant leaf) S.
dorsalis was recorded. This was repeated 15 times
each for groups of 10 and 20 S. dorsalis.
Collecting N. cucumeris and releasing them
onto plants. The entire contents of the package
with N. cucumeris predatory mites were emptied
into a Petri dish (15 cm diam). To separate N. cu-
cumeris mites from the packaging material, a fil-
ter paper was placed in the Petri dish and the
closed Petri dish was agitated gently several
times. Under a stereomicroscope, either 10 or 20
N. cucumeris mites on the filter paper were col-
lected with the mini-aspirator. The collection
tube with the N. cucumeris mites was attached
to a pepper leaf as described above (Fig. 1D). The
seconds required for collection and release (the
latter being the time required to attach the col-
lector containing the collected N. cucumeris to a
pepper plant leaf were recorded. This was re-
peated 15 times each for groups of 10 and 20 N.
cucumeris.
Collecting adult 0. insidiosus flower bugs and
releasing them onto plants. The mini-aspirator
was modified by enlarging the opening of the pi-
pette tip to accommodate the bugs. The bugs were
collected from among the vermiculate particles
scattered on a board. Orius insidiosus in groups of
either 10 or 20 were collected with the mini-aspi-
rator. The collection tube containing the bugs was
then attached to a leaf with a hair clip as de-
scribed previously. The number of seconds re-
quired to collect and release the bugs was re-
corded. This was repeated 15 times each for
groups of 10 and 20 0. insidiosus.

Statistical Analysis

The efficiency (time) of the mini-aspirator was
compared with the conventional method for col-
lecting and releasing each of the three arthro-
pods. The study was repeated 8 and 15 times for
the conventional and new method, respectively.
Data were analyzed by ANOVA procedure (PROC
GLM) and means were separated by Fisher's pro-
tected LSD test for all the experiments (SAS In-
stitute 1997).

RESULTS

The collection of 10 adult thrips from cotton
leaves and their release onto a pepper plant us-
ing the mini-aspirator took 37 s compared with
229 s with the camel's hair brush method. This
represented a 6-fold reduction in infestation







Florida Entomologist 94(1)


o0 AvSa*SuS


N ucumans


Fig. 3. Comparison of seconds needed
and infesting 10 0. insidiosus, S. dorsalis
eris onto plants with conventional met]
hair brush or commercial aspirator) and 1
mini-aspirator.


= 0.0001 and F,,21 = 379.00; P = 0.0001), respec-
tively. No predatory mite was found to be dam-
aged by the mini-aspirator.
The time required to collect and release adult
0. insidiosus with a BioQuip aspirator was less
than time needed for collecting and releasing by
the paintbrush method. However, collecting and
Releasing 10 and 20 0. insidious with the com-
mercial aspirator required 31 and 58 s, but only
s ," 20 and 32 s with the mini-aspirator (Figs. 3 and
for collecting 4). These time differences between the 2 methods
andN. cucum- of collection and release were also significantly
hods (camel's different (F,,, = 43.19; P= 0.0001) (F,,2 = 33.52; P
the developed = 0.0001), respectively.


time when the mini-aspirator method was used
(Fig. 3). The difference between the 2 methods
was highly significant (F1,, = 325.66;P < 0.0001).
The time required to place 20 thrips on a plant
with the mini-aspirator was 42 s, compared with
461 s by using the camel's hair brush method,
which represented an 11-fold reduction in infes-
tation time when the mini-aspirator was used
(F,,, = 974.68; P < 0.0001) (Fig. 4). With the new
method, the amount of time to collect and re-
lease per thrips decreased as number of in-
creased. Such a reduction was not observed with
the paintbrush method. Some thrips adults were
also observed to be injured by the commercial as-
pirator (Fig. 5-A). We also observed some dam-
aged thrips and reduced thrips activity when
they were handled with a commercial aspirator
(Fig. 5-B).
The collection and release of 10 and 20 pred-
atory mites required 639 and 1154 s with a
moistened camel's hair brush compared with 43
and 90 s, respectively, with the mini-aspirator
(Figs. 3 and 4). The use of the mini-aspirator
saved 15-fold and 13-fold more time in collect-
ing and releasing 10 and 20 predatory mites, re-
spectively, compared with the use of the paint-
brush method. The difference between the two
methods was highly significant (F,,, = 284.86; P


0 mdlosus


J


-U


S orsais


Fig. 4. Comparison of seconds needed for collecting
and infesting 20 0. insidiosus, S. dorsalis and N. cucum-
eris onto plants with conventional methods (camel's
hair brush or commercial aspirator) and the developed
mini-aspirator.


DISCUSSION
The mini-aspirator reduces the time required
to collect and release a designated number of
small arthropods to the plants. The camel's hair
brush method in addition to being very slow can
cause injury, especially to soft-bodied small ar-
thropods. An operator using the mini-aspirator
can collect and transfer small arthropods using
controlled air intake velocity, which minimizes in-
jury to the collected arthropods. To avoid injury to
thrips, the vacuum was adjusted to the minimum
sufficient to collect thrips. Although injury to
thrips was not investigated in detail, thrips col-
lected with the mini-aspirator were checked un-
der a stereomicroscope and no serious thrips in-
jury was observed.
The mini-aspirator is different from the com-
monly available commercial aspirators, which
employs larger diameter removable glass or
plastic collecting vials. Initially, we used the
commercial aspirators but experienced difficul-
ties in transferring the designated numbers of
arthropods. The commercial aspirator available
to us has a 4-mm diam collection tube that could
not be adjusted for the selective collection of in-
dividual thrips; and this is a significant limita-
tion when working with mixed colonies of in-
sects. Another advantage of the mini-aspirator is
that unlike the traditional collection vials, the
smaller removable and disposable collection
tubes can easily be attached to small leaves or
plant stems without disturbing the insects in-
side the tube.
The mini-aspirator developed in this study can
be powered with a Mini-Vac, which makes the
technique portable for field applications. The
technique may be used to quickly census wild
populations for laboratory testing or for use in in-
secticide efficacy trials. The mini-aspirator could
also be adapted for quick pesticide resistance or
efficacy trials in the field (Rueda and Shelton
2003). The inside of the collection tube of the
mini-aspirator could be treated with pesticides of
interest or a treated leaf disk could be placed in
the mini-aspirator before collecting small arthro-


f) A


I


March 2011


N ct"nrs







Dogramaci et al.: Mini-aspirator for Handling Small Arthropods


Fig. 5. Illustration of an injured S. dorsalis when collected with a regular made aspirator. A, Injury to abdomen
of S. dorsalis. B, Injury to wing of S. dorsalis.


pods, and then collected arthropods would be held
for a fixed exposure period to quantify pesticide
efficacy. This kind of monitoring would be helpful
to confirm pest susceptibility to pesticides before
their wide area applications.

ACKNOWLEDGMENTS

We are grateful to Kenneth E. Savage, Russell D.
Caldwell, and Younes Belmourd for help during this
study. The study was supported by the USDA Tropical
and Subtropical Agricultural Research (T-STAR) Pro-
gram, American Floral Endowment and the USDA-ARS
Floriculture and Nursery Research Initiative.

REFERENCES CITED

ARTHURS, S., MCKENZIE C. L., CHEN, J., DOGRAMACI,
M., BRENNAN, M., HOUBEN, K., AND OSBORNE, L.
2009. Evaluation ofNeoseiulus cucumeris andAmbl-
yseius cucumeris (Acari: Phytoseiidae) as Biological
Control Agents of Chilli Thrips, Scirtothrips dorsalis
(Thysanoptera: Thripidae) on Pepper. Biol. Cont. 49:
91-96
BOURNIER, J. P. 1999. Two Thysanoptera, new cotton
pests in Cote d'Ivorie. Annales de la Societe Ento-
mologique de France 34: 275-281.
CHIU, H. T., SHEN, S. M., AND WU, M. Y. 1991. Occur-
rence and damage of thrips in citrus orchards south-
ern Taiwan. Chinese J. Entomol. 11: 310-316.


CLOYD, R. A., WARNOCK, D. F., AND HOLMES, K. 2001.
Technique for collecting thrips for use in insecticide
efficacy trials. Hort. Sci. 36: 925-926.
CLOYD, R. A., AND SADOF, C. S. 1998. Flower quality,
flower numbers, and Western flower thrips density
on transversal daisy treated with granular insecti-
cides. Hort. Tech. 8: 567-570.
MOUND, L. A., AND PALMER, J. M. 1981. Identification,
distribution and host plants of the pest species of
Scirtothrips (Thysanoptera: Thripidae). Bull. Ento-
mol. Res. 71: 467-479.
RUEDA, A., AND SHELTON, A. M. 2003. Development and
evaluation of a thrips insecticide bioassay system for
monitoring resistance in Thrips tabaci. Pest. Man-
age. Sci. 59: 553-558.
SAS INSTITUTE. (1997) SAS User's Guide. SAS Institute
Cary, North Carolina.
SEAL, D. R., CIOMPERLIK, M. A., RICHARDS, M. L., AND
KLASSEN, W. 2006. Distribution of chilli thrips, Scir-
tothrips dorsalis (Thysanoptera: Thripidae), in pep-
per fields and pepper plants on St. Vincent. Florida
Entomol. 89: 311-320.
TATARA, A., AND FURUHASHI, K. 1992. Analytical study
on damage to Satsuma mandarin fruit by Scirto-
thrips dorsalis Hood (Thysanoptera: Thripidae),
with particular reference to pest density. Japanese J.
Appl. Entomol. 36(4): 217-223.
TSCHUCHIYA, M., MAUI, S., AND KUBOYAMA, N. 1995.
Color attraction of yellow tea thrips (Scirtothrips
dorsalis Hood). Japanese J. App. Entomol. Zool. 39:
299-303.







Florida Entomologist 94(1)


TAXONOMY OF KOREAN LESTEVA WITH A DESCRIPTION OF A NEW
SPECIES (COLEOPTERA: STAPHYLINIDAE: OMALIINAE)

TAE-KYU KIM AND KEE-JEONG AHN
Department of Biology, Chungnam National University, Daejeon 305-764, Republic of Korea

ABSTRACT

A taxonomic study of the genus Lesteva Latreille in Korea is presented. Four species includ-
ing a new species, Lesteva coreana sp. nov., are recognized. Three species, L. cordicollis
Motschulsky, L. distinct Watanabe and L. miyabi Watanabe, are new to the Korean fauna,
and L. plagiata Sharp previously recorded from Korea is a misidentification of L. miyabi. A
key and a comparison of morphological features of Korean Lesteva species with illustrations
of the diagnostic features are provided.

Key Words: Staphylinidae, Omaliinae, Lesteva, new species, Korea

RESUME

Se present un studio taxon6mico del g6nero Lesteva Latreille en Corea. Se reconocen cua-
tro species incluyendo una nueva especie, Lesteva coreana sp. nov. Tres species, L. cordi-
collis Motschulsky, L. distinct Watanabe y L. miyabi Watanabe son nuevas para la fauna de
Corea y se determine que L. plagiata Sharp, anteriormente registrada en Corea, fue basado
sobre una identificaci6n equivocada de L. miyabi. Se proven una clave y una comparisi6n
de las caracteristicas morfol6gicas de las species de Lesteva en Corea con ilustraciones de
las caracteristicas diagn6sticas.


The genus Lesteva Latreille (tribe Anthoph-
agini Thomson) is composed of 104 species dis-
tributed in the Holarctic and Oriental regions
(Watanabe 1990, 2004, 2005; Herman 2001;
Smetana 2004; Li 2005; Sharvrin et al. 2007). In
East Asia, 19 and 15 species of the genus are re-
ported in Japan and in China, respectively (Wa-
tanabe 1990, 2004; Smetana 2004; Li 2005).
Lesteva plagiata Sharp recorded by Cho et al.
(2002) in Korea is a misidentification ofL. miyabi
Watanabe.
Members of Lesteva occur in montane riparian
areas and are often found in moss or wet litter,
sometimes in caves. Adults and larvae are preda-
tors (Steel 1970; Newton et al. 2001). We have
studied 20 specimens of L. cordicollis Motschul-
sky, 28 specimens ofL. coreana sp. nov., 60 spec-
imens of L. distinct Watanabe, and 125 speci-
mens ofL. miyabi.
In this paper, we report 4 Lesteva species (L.
cordicollis, L. coreana sp. nov., L. distinct, and
L. miyabi) from Korea. A key, habitus photo-
graphs, and the illustrations of diagnostic fea-
tures are provided. All specimens are deposited in
the Chungnam National University Insect Collec-
tion (CNUIC), Daejeon, Korea.
Genus Lesteva Latreille, 1797
Lesteva Latreille, 1797: 75.
Tevales Casey, 1894: 398. Synonymized by Steel, 1952: 9.

Diagnosis. Body ovoid and flattened, densely
pubescent, covered with punctures. Head sub-


quadrate; eyes convex, large, with pubescence be-
tween facets; ocelli distinct; temple round; vertex
with 2 longitudinal depressions; gular sutures
separated, divergent posteriorly; mandibles sub-
triangular, curved inwardly with distinct internal
teeth, mola distinct; maxillary palpomere 4 as
wide and about 4.0 times as long as palpomere 3;
antenna extending to near middle of elytra.
Pronotum convex, widest at anterior third or
fourth, more narrowed posteriorly than anteri-
orly; mesoventrite with longitudinal carina along
midline and several foveae on each side; elytra
flat, broader than pronotum, expanded posteri-
orly; legs long and slender, protarsus thin in both
sexes. Abdomen broad, flat and abruptly nar-
rowed posteriorly, tergites IV-V with a pair of
wing folding patches.

Lesteva cordicollis Motschulsky, 1860

(Figs. 1, 5, 9, 13, 17, 19, 21-22)

Lesteva cordicollis Motschulsky, 1860: 549; Sharvrin,
2001: 191.
Description. Body (Fig. 1) length 3.6-4.0 mm
(head to abdominal end), covered with fine
punctures and pubescence, brown to dark
brown and glossy; head and pronotum black,
mouthparts, antennae and legs light brown.
Head about 1.4 times as wide as long; eye about
3.3 times as long as temple; antennae (Fig. 5)
pubescent, reaching middle of elytra, 4th an-
tennomere 2.1 times as long as wide, 8th anten-


March 2011







Kim & Ahn: Korean Lesteva Species


1 2


Figs. 1-4. Habitus. 1. Lesteva cordicollis, length 3.8
mm; 2. L. coreana sp. nov., length 3.4 mm; 3. L. dis-
tincta, length 4.0 mm; 4. L. miyabi, length 3.9 mm.



nomere 1.8 times as long as wide. Pronotum
slightly convexed with fine punctures, widest
near anterior fourth with ambiguous U-depres-
sion near middle, 1.3 times as wide as long,
about 1.2 times as wide and 1.4 times as long as
head; scutellum (Fig. 9) subtriangular, prescu-
toscutellar suture gently curved, scutellar pro-
cess broad subtriangular; elytra bicolor, hu-
meral region with large yellow patch and fine
punctures, posterior margin truncated, 1.1
times as wide as long, 1.5 times as wide and 1.7
times as long as pronotum (Figs. 1, 13); apex of
metaventral process round (Fig. 17); external
surface of metatibia with 3-4 long golden setae
(Fig. 19). Abdominal segments III-VIII with mi-
crosculpture. Median lobe of aedeagus elongate,
parallel-sided, apical process triangular, apical
middle area elevated, internal sac backbone-
shaped; parameres slender, slightly longer than
median lobe, four setae present with two at
apex (Figs. 21-22).
Materials Examined. KOREA: Gangwon
Prov.: Chuncheon-si, Nam-myeon, Mt. Bongh-
wasan (N3746'1.2" E 12735'59.0" 186m) 17 IX
2008, TK Kim ex under stone near stream (1649,
CNUIC); Chungnam Prov.: Daejeon, Yuseong-
gu, Sutong-gol, 9 V 1998, KR You, HJ Lim, HJ
Kim, ex near stream (29, CNUIC); Jeonbuk
Prov.: Muju-gun, Anseong-myeon, Mt. Deokyu-
san, Chilyeon-fall, 27 V 2005, TK Kim, ex under
stone near stream (8619, CNUIC); Jinan-gun,
Jeongcheon-myeon, Mt. Unjangsan, V 19 1998,
YB Cho (2 CNUIC);
Distribution. Korea (South), Russia (East Si-
beria).


Lesteva coreana Kim and Ahn sp. nov.
(Figs. 2, 6, 10, 14, 23-24)

Description. Body (Fig. 2) length 3.1-3.5 mm
(head to abdominal end), covered with fine punc-


i' ~


N .1 IV


tures and pubescence, brown to dark brown and
glossy; mouthparts, antennae and legs light
brown. Head about 1.4 times as wide as long; eye
about 3.1 times as long as temple; antennae (Fig.
6) pubescent, reaching middle of elytra, 4th an-
tennomere 2.6 times as long as wide, 8th antenno-
mere 2.4 times as long as wide. Pronotum slightly
convexed with fine punctures, widest near ante-
rior fourth with obscure U-depression near mid-
dle, 1.3 times as wide as long, about 1.2 times as
wide and 1.3 times as long as head; scutellum
(Fig. 10) subtriangular, prescutoscutellar suture
arcuate, scutellar process narrow triangular;
elytra bicolor, humeral region with indistinct yel-
low patch and fine punctures, posterior margin
truncated, 1.1 times as wide as long, 1.4 times as
wide and 1.7 times as long as pronotum (Figs. 2
and 14); apex of metaventral process round; ex-
ternal surface of metatibia with 3-4 long golden
setae. Abdominal segments III-VIII with micros-
culpture. Median lobe of aedeagus narrowed api-
cally, lateral margin weakly arcuated; basal re-
gion of parameres broad, narrowed apically, api-
cal third constricted, slightly longer than median
lobe, four setae present with two at apex (Figs. 23-
24).
Type Series: Holotype, 6: 'KOREA: Jeonbuk
Prov.: Muju-gun, Anseong-myeon, Mt. Deokyu-
san, Chilyeon-fall, 27 V 2005, TK Kim, ex under
stone near stream; Holotype, Lesteva coreana
Kim and Ahn, Desig. T.-K. Kim and K.-J. Ahn
2010.' Deposited in CNUIC, Daejeon. Paratypes,
same data as holotype (10619, CNUIC),
Paratype, Lesteva coreana Kim and Ahn, Desig.
T.-K. Kim and K.-J. Ahn 2010. Other materials:
Mt. Deokyusan, Chilyeon-fall, 22-23 V 1998, HJ
Kim, ex near stream (1 CNUIC); same data as
holotype (2 Y, CNUIC); Chungnam Prov.: Dae-
jeon, Mt. Gyeryongsan, Keumsubong, 21 V 2000,
SJ Park, ex near stream (5659, CNUIC); Yu-
seong-gu, Sutong-gol, 5 IX 1998, SJ Baek (16,
CNUIC); Sutong-gol, 9 V 1998, KR You, HJ Lim,
HJ Kim, ex near stream (1 CNUIC).
Distribution. Korea (South).
Remarks. The species is similar to L. cordicol-
lis but can be distinguished by the shape and
structures of antennomeres, scutellum, and me-
dian lobe of aedeagus (Table 1).

Lesteva distinct Watanabe, 1990
(Figs. 3, 7, 11, 15, 18, 25-26)

Lesteva distinct Watanabe, 1990: 178; Herman, 2001:
315; Smetana, 2004: 247.

Description. Body (Fig. 3) length 3.5-4.1 mm
(head to abdominal end), covered with coarse
punctures and pubescence, reddish brown to
black and glossy; mouthparts, antennae and legs
brown. Head about 1.2 times as wide as long; eye





Florida Entomologist 94(1)


5

6




7
___ 5


10


13 14


11 12


15


16


Figs. 5-16. 5-8. Antenna, ventral aspect. 5. Lesteua cordicollis; 6. L. coreana sp. nov.; 7. L. distinct; 8. L. miyabi.
9-12. Scutellum, dorsal aspect. 9. L. cordicollis; 10. L. coreana sp. nov.; 11. L. distinct; 12. L. miyabi. 13-16.
elytron, ventral aspect. 13. L. cordicollis; 14. L. coreana sp. nov.; 15. L. distinct; 16. L. miyabi. Scales = 0.1 mm
(Figs. 9-12); 0.3 mm (Figs. 5-8, 13-16).


March 2011







Kim & Ahn: Korean Lesteva Species


0




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about 1.7 times as long as temple; antennae
(Fig. 7) pubescent, reaching middle of elytra, 4th
antennomere 2.1 times as long as wide, 8th an-
tennomere 1.9 times as long as wide. Pronotum
much convexed with coarse punctures, about 1.2
times as wide as long, 1.2 times as wide and 1.3
times as wide as head, widest near anterior
fourth with distinct U-depression near middle;
scutellum (Fig. 11) subtriangular, prescutoscutel-
lar suture arcuate, scutellar process broad pen-
tagonal; elytra bicolor, humeral region with red-
dish brown patch and somewhat coarse punc-
tures, posterior margin round, 1.1 times as long
as wide, about 1.8 times as wide and 1.9 times as
long as pronotum (Figs. 3 and 15); apex of
metaventral process notched (Fig. 18); external
surface of metatibia with 10-14 long dark brown-
ish setae. Abdominal segments III and VIII with
microsculpture. Median lobe of aedeagus broad,
basal two third parallel-sided, apical third nar-
rowed suddenly, apical process triangular, mid-
line area elevated with longitudinal carina, lat-
eral margin rolled dorsally; parameres robust,
symmetrical, as long as median lobe, lateral mar-
gin rolled ventrally, apical region coiling ventrally
and inwardly, four setae present (Figs. 25 and 26).
Materials Examined. KOREA: Gangwon
Prov.: Chuncheon-si, Sabuk-myeon, Jiam-ri, 15
IV 2001, SI Lee (16, CNUIC); Chungbuk Prov.:
Danyang-gun, Danyang-eup, Mt. Sobaeksan,
Cheondong-area, 8-9 V 1999, US Hwang, HJ Kim,
sifting (2 9, CNUIC); Yeongdong-gun, Sangchon-
myeon, Mulhan-ri, Mt. Minjujisan, Mulhan-
stream (N363'15" E12752'31"), 16 VI 2006, TK
Kim, ex under stone near stream (16, CNUIC);
Mt. Manloi, 30 V 1998, HJ Lim, sifting (19,
CNUIC); Chungnam Prov.: Daejeon-si, Yu-
seong-gu, Gung-dong, Chungnam National Uni-
versity (N3622'38.7" E12720'43.5"), 18 IV 2007,
HW Kim, ex near pond (3629, CNUIC); Chung-
nam National University (N3622'38.7"
E127020'43.5"), 7 V 2007, YH Kim, ex near pond
(7649, CNUIC); Chungnam National University
(N36022'38.7" E127020'43.5"), 14 V 2007, HW
Kim, ex near pond (1268 9, CNUIC); Yuseong-gu,
Deokmyeong-dong, Sutonggol, 9 V 1998, KR You,
HJ Lim, HJ Kim, ex near stream (16, CNUIC);
Buyeo-gun, Naesan-myeon, Mt. Wolmyeongsan,
Geumgisa, 3 V-1 VI 2000, US Hwang, HJ Kim,
FIT (19, CNUIC); Geumgisa, 1 VI 2000, US
Hwang, HJ Kim, sifting (19, CNUIC); Jeonbuk
Prov.: Buan-gun, Byeonsan-myeon, Mt. Nae-
byeonsan, Jikso-fall, 30 V 2001, YB Cho, sifting
(1629, CNUIC); Jinan-gun, Jeongcheon-myeon,
Mt. Unjangsan, V 19 1998, YB Cho (2 9, CNUIC);
Jeonnam Prov.: Gurye-gun, Mt. Jirisan, Toji-
myeon, Piagol, 24 V 2000, HJ Kim, ex near stream
(1612, CNUIC); Piagol, 24-27 V 2000, KJ Ahn,
SJ Park, US Hwang, FIT (19, CNUIC); Jindo-
gun, Uisin-myeon, Sacheon-ri, Mt. Cheomchilsan
(N34027'53.7" E12618'42.6" 115m), 23 II 2007







Florida Entomologist 94(1)


17 18


19


20


Figs. 17-20. 17-18. Metaventrite, ventral aspect. 17. Lesteva cordicollis; 18. L. distinct. 19-20. Metatibia (pu-
bescence omitted), anterior aspect. 19. L. cordicollis; 20. L. miyabi. Scales = 0.3 mm.


TK Kim, sifting, leaf litter (1 CNUIC); Yeongg-
wang-gun, Hongnong-eup, Sangha-ri (N3523'24.9"
E126025'57.9"), 2 V 2007, KJ Ahn, TK Kim, YH
Kim, ex near stream (29, CNUIC); Hadong-gun,
Hwagye-myeon, Ssanggyesa, 25 V 2000, HJ Kim, ex
near stream (16, CNUIC); Gyeongbuk Prov.:
Cheongsong-gun, Budong-myeon, Mt. Juwangsan,
29 VI 1987, YB Cho, ex under moss (16, CNUIC);
Gyeongnam Prov.: Geoje-si, Yeoncho-myeon, Mt.
Aengsan (N34056'17.3" E128036'6.6" 85m), 21 I
2009, DH Lee, JH Song, ex under stone near mount
stream (2 1 2, CNUIC).
Distribution. Korea (South), Japan.

Lesteva miyabi Watanabe, 1990
(Figs. 4, 8, 12, 16, 20, 27-28)

Lesteva miyabi Watanabe, 1990: 175; Herman, 2001:
324; Smetana, 2004: 247.
Lesteua plagiata: Cho et al., 2002: 36. Misidentification.

Description. Body (Fig. 4) length 3.8-4.5 mm
(head to abdominal end), covered with coarse
punctures and pubescence, dark brown to black
and glossy; mouthparts, antennae and legs brown
to reddish brown. Head about 1.3 times as wide as
long; eye about 1.7 times as long as temple; anten-
nae (Fig. 8) pubescent, reaching middle of elytra,
4th antennomere 1.9 times as long as wide, 8th
antennomere 1.8 times as long as wide. Pronotum


mostly convexed with coarse punctures, widest
near anterior third with distinct U-depression
near middle, about 1.2 times as wide as long,
about 1.1 times as wide and 1.2 times as long as
head; scutellum (Fig. 12) subtriangular, prescu-
toscutellar suture round, scutellar process broad
pentagonal; elytra unicolor with coarse punc-
tures, posterior margin round, 1.04 times as long
as wide, 1.8 times as wide and 2.0 times as long as
pronotum (Figs. 4 and 16); apex of metaventral
process notched; external surface of metatibia
with 10-14 long dark brownish setae (Fig. 20). Ab-
dominal segments III and VIII with microsculp-
ture. Median lobe of aedeagus broad, narrowed
apically with longitudinal carina, lateral margin
almost straight; parameres robust, symmetrical,
as long as median lobe, lateral margin rolled ven-
trally, apical region coiling ventrally and in-
wardly, four setae present (Figs. 27 and 28).
Materials Examined. KOREA: Jeju Prov.:
Jeju-si, Arail-dong, Gwaneumsa, 26 V 2003, SJ
Park, ex near stream (10699, CNUIC); Jeju-si,
Bonggae-dong, Muljang-oreum, 23 V 1998, YB
Cho (3659, CNUIC); Jeju-si, Nohyeong-dong,
Cheonwangsa (N3324'25.4" E12629'42.7" 395
m), 8 XI 2006, TK Kim, ex under stone near
stream (16, CNUIC); Jeju-si, Orai-dong, Eorimok
(N3323'26.0" E12629'41.1" 1000 m), 31 V 2007,
TK Kim, ex under stone near stream (29,
CNUIC); Seoguipo-si, Hawon-dong, Seoguipo
Natural Recreation Forest (N3318'54.2"


March 2011






Kim & Ahn: Korean Lesteva Species


22 23


25 26


27 28


Figs. 21-28. Aedeagus. 21-22. Lesteva cordicollis. 21. dorsal aspect; 22. lateral aspect. 23-24. L. coreana sp. nov.
23. dorsal aspect; 24. lateral aspect. 25-26. L. distinct. 25. dorsal aspect; 26. lateral aspect. 27-28. L. miyabi. 27.
dorsal aspect; 28. lateral aspect. Scales = 0.3 mm.


E126027'56.0" 735 m), 30 V 2007, TK Kim, sifting,
flood debris (1 CNUIC); Seoguipo Natural Rec-
reation Forest (N3318'36" E126028'9.2" 665 m),
31 V 2007, DH Lee, YH Kim, sifting, leaf litter
(1 CNUIC); Bukjeju-gun, Aewol-eup, 1100-goji,
28 v 2003, CW Shin, ex near stream (26, CNUIC);
1100-goji (N33021'40.6" E126027'44.6" 1097 m),
12 x 2006, TK Kim, sifting, wet leaf litter (162$2,
CNUIC); 1100-goji (N33021'37.5" E126027'45.8"
1110 m), 31 V 2007, TK Kim, sifting, leaf litter
(9669, CNUIC); Bukjeju-gun, Jocheon-eup,


Goepyeongi-oreum, 23 V 2006, SJ Park, DH Lee,
SI Lee, YH Kim, leaf litter (16, CNUIC); Goepyeo-
ngi-oreum (N33025'2.7" E126038'32.6" 530 m), 8 IX
2006, DH Lee, ex leaf litter (1659, CNUIC);
Goepyeongi-oreum (N33025' 1.8" E126038'32.2" 539
m), 8 IX 2006, TK Kim, ex wet grit near pond (16,
CNUIC); Namjeju-gun, Namwon-eup, Dongsu-
bridge (N33022'8.4" E126037'30.7" 640 m), 8 XI
2006, TK Kim, ex under stone near stream (36,
CNUIC); Dongsu-bridge, 1 III 2007, TK Kim, ex un-
der stone near stream (2619, CNUIC); Dongsu-


21


24


\\\







Florida Entomologist 94(1)


bridge (N3322'8.5" E12637'30.5" 635 m), 29 V
2007, TK Kim, ex under stone near stream
(186129, CNUIC); Namjeju-gun, Namwon-eup,
Goepyeongi-oreum, 28 V 2003, SJ Park, CW Shin,
MJ Jeon, sifting (166119, CNUIC); Goepyeongi-
oreum, 28 V-27 VI 2003, YB Cho, SJ Park, CW Shin,
FIT (16, CNUIC); Mt. Hallasan, 900 m alt., Jejudo
Is., 17 VII 1994, G. Sh. Lafer leg (16, CNUIC).


Distribution. Korea (South), Japan.
Remarks. Cho et al. (2002) reported L. pla-
giata in Korea. However, we have determined
that this was a misidentification of L. miyabi,
based on our examination of their voucher speci-
men (16: Mt. Hallasan, 900 m alt., Jejudo Is., 17.
VII 1994, G. Sh. Lafer leg). The species was col-
lected only in Jeju-do island.


KEY TO THE KOREAN SPECIES OF THE GENUS LESTEVA LATREILLE

1. Pronotum slightly convexed with fine punctures; prosternal process without carina; posterior margin of elytra
truncated with fine punctures; apex of metaventral process round (Fig. 17); metatibia without long dark
brownish setae (3-4 long golden setae present) (Fig. 19). ........................................ 2

- Pronotum distinctly convexed with coarse punctures; prosternal process with short, sinuous longitudinal car-
ina; posterior margin of elytra round with coarse punctures; apex of metaventral process notched (Fig. 18);
metatibia with 10-14 long dark brownish setae (Fig. 20) ........................................ 3

2. Fourth antennomere 2.1 times as long as wide, 8th antennomere 1.8 times as long as wide (Fig. 5); scutellar pro-
cess broad (Fig. 9); median lobe of aedeagus elongate, in basal three fourth parallel-sided, and in apical
fourth abruptly narrowed in dorsal view (Figs. 21 and 22). ............................ L. cordicollis

- Fourth antennomere 2.6 times as long as wide, 8th antennomere 2.4 times as long as wide (Fig. 6); scutellar pro-
cess narrow (Fig. 10); median lobe of aedeagus narrowed apically, lateral margin weakly arcuated in dorsal
view (Figs. 23 and 24) ..................................................... L. coreana sp. nov.

3. Pronotum widest at anterior fourth; elytra bicolor with reddish patch around humeral region, moderately
broad and long (Fig. 3) .......................................................... L. distinct

Pronotum widest at anterior third; elytra unicolor, broad and long (Fig. 4) ..................... L. miyabi


ACKNOWLEDGMENTS
We thank Dr. Watanabe (Tokyo University of Agri-
culture, Japan) for the loan of specimens. This research
was supported by the project on survey and excavation
of Korean indigenous species of the National Institute of
Biological Resources (NIBR) under the Ministry of En-
vironment, Korea.

REFERENCES CITED

CASEY, T. L. 1894. Coleopterological notices. V. Ann.
New York Acad. Sci. 7: 281-606.
CHO, Y. B., LAFER, G. S., PAIK, J. C., AND PARK, J. K.
2002. Contribution to the staphylinid fauna (Co-
leoptera, Staphylinidae) of Korea. Korean J. Soil
Zool. 7(1-2): 35-44.
HERMAN, L. H. 2001. Catalog of the Staphylinidae (In-
secta: Coleoptera). 1758 to the End of the Second
Millennium. I. Introduction, History, Biographical
Sketches, and Omaliine Group. Bull. American Mus.
Nat. Hist. 265: 309-333.
LATREILLE, P. A. 1797. Pr6cis des Caracteres
Generiques des Insectes, Dispos6s dans un Ordre
Naturel. xiv + 201 + 7 pp. Brive: F. Bourdeaux.
LI, X.-J., LI, L.-Z., AND ZHAO, M.-J. 2005. A new species
of the genus Lesteva (Coleoptera: Staphylinidae:
Omaliinae) from China. Entomotaxonomia 27(2):
111-113.
MOTSCHULSKY, V. 1860. Enum6ration des nouvelles es-
peces de coleopteres rapport6es de ses voyages. 3e
parties. Bull. Soc. Imper. Nat. Moscou 33(2): 539-588.
NEWTON, A. F., THAYER, M. K., ASHE, J. S., AND CHAN-
DLER, D. S. 2001. 22. Staphylinidae, pp. 272-342 In
R. H. Arnett and M. C. Thomas [eds.], American Bee-


tles. Vol. 1. Archostemata, Myxophaga, Adephaga,
Polyphaga: Staphyliniformia. CRC Press, Boca Ra-
ton, Florida.
SHARP, D. 1889. The Staphylinidae of Japan. Ann. Mag.
Nat. Hist. (6)3: 463-476.
SHAVRIN, A. V. 2001. New and little-known species of
Omaliinae from the Baikal-Transbaikal area (Co-
leoptera: Staphylinidae). Zoosyst. Rossica 9: 189-193.
SHAVRIN, A. V., SHILENKOV, V. G., AND ANISTSCHENKO,
A. V. 2007. Two new species and additional records
of Lesteva Latreille, 1797 from the mountains of
South Siberia (Coleoptera: Staphylinidae: Omalii-
nae: Anthophagini). Zootaxa 1427: 37-47.
SMETANA, A. 2004. Staphylinidae, subfamily Omalii-
nae, pp. 237-268 In I. L6bl and A. Smetana [eds.],
Catalogue of Palaearctic Coleoptera. Volume 2, Hy-
drophiloidea Histeroidea Staphylinoidea. Applo
Books, Steustrup.
STEEL, W. O. 1952. Notes on the Omaliinae (Col., Sta-
phylinidae). 4. On the genera Lesteva Latr., Paraleste-
va Casey and Tevales Casey, with a key to the British
species of Lesteva. Entomol. Mon. Mag. 88: 8-9.
STEEL, W. O. 1970. The larvae of the genera of the Oma-
liinae (Coleoptera: Staphylinidae) with particular
reference to the British fauna. Trans. R. Entomol.
Soc. London 122(1): 1-47.
WATANABE, Y. 1990. A taxonomic study on the subfami-
ly Omaliinae from Japan (Coleoptera, Staphylin-
idae). Mem. Tokyo Univ. Agric. 31: 149-179.
WATANABE, Y 2004. Two new species of the genus Lesteva
(Coleoptera, Staphylinidae) from the Island of D6go of
the Oki Islands, West Japan. Elytra 32(1): 71-77.
WATANABE, Y. 2005. A new species of the genus Lesteva
(Coleoptera, Staphylinidae) from Taiwan. Elytra
33(1): 30-33.


March 2011







Goyal et al: Corn-infesting Ulidiidae of Florida


DISTRIBUTION OF PICTURE-WINGED FLIES
(DIPTERA: ULIDIIDAE) INFESTING CORN IN FLORIDA

GAURAV GOYAL1, GREGG S. NUESSLY', DAKSHINA R. SEAL2, JOHN L. CAPINERA3, GARY J. STECK4
AND KENNETH J. BOOTE'
'Everglades Research and Education Center, University of Florida (UF),
Institute of Food and Agricultural Sciences (IFAS), 3200 E. Palm Beach Rd., Belle Glade, FL 33430

2Tropical Research and Education Center, UF, IFAS, 18905 S.W. 280 St., Homestead, FL 33031

3Department of Entomology and Nematology, UF, IFAS, P.O. Box 110620, Gainesville, FL 32611

4Division of Plant Industry, Florida Department of Agriculture and Consumer Services,
P.O. Box 147100, Gainesville, FL 32614

5Department of Agronomy, UF, IFAS, P.O. Box 110500, Gainesville, FL 32611

ABSTRACT

The picture-winged fly Euxesta stigmatias Loew (Diptera: Ulidiidae) has been a serious pest
of sweet corn (Zea mays L.) in Florida since 1930. Several other species in the family are
known to infest corn grown in the Caribbean, Central America, and South America. Surveys
were conducted throughout Florida to evaluate species richness and distribution of corn-in-
festing Ulidiidae. Adults were sampled with sweep nets and reared from fly larvae-infested
corn ears collected from representative corn fields in 16 and 27 counties in 2007 and 2008,
respectively. Four Ulidiidae species were found in corn fields using both sampling tech-
niques. Euxesta eluta Loew and Chaetopsis massyla (Walker) were found throughout the
state on field and sweet corn. Euxesta stigmatias was only found in Martin, Miami-Dade,
Okeechobee, Palm Beach, and St. Lucie Counties on field and sweet corn. Euxesta annonae
(F.) was found in sweet corn in Miami-Dade, Okeechobee, and Palm Beach Counties, but field
corn was not sampled in these counties. Euxesta eluta, E. stigmatias, and C. massyla were
collected from corn throughout the corn reproductive stage. Raising adults from fly larvae-
infested ears provided the best method for assessing rates of ear infestation and species rich-
ness. Sweep netting did not provide reliable information on the presence or species compo-
sition of ulidiid species infestation. We report for the first time E. annonae and E. eluta as
pests of corn in Florida and the USA.

Key Words: Euxesta annonae, Euxesta eluta, Euxesta stigmatias, Chaetopsis massyla, maize

RESUME

La mosca de alas pintadas, Euxesta stigmatias Loew (Diptera: Ulidiidae), ha sido una plaga
seria de maiz dulce (Zea mays L.) en la Florida desde 1930. Varias species de la familiar Uli-
diidae son conocidas de infestar maiz sembrado en el Caribe y el Centroam6rica y Sudam6-
rica. Se realizaron sondeos por todo la Florida para evaluar la diversidad de species y
distribuci6n de moscas de la familiar Ulidiidae que infestan maiz. Se muestrearon los adults
con redes de recolecci6n y criandolos de mazorcas infestadas con larvas de moscas de campos
representatives de maiz en 16 y 27 condados en 2007 y 2008, respectivamente. Se encontra-
ron Euxesta eluta Loew y Chaetopsis massyla (Walker) por todo el estado en maiz de campo
y maiz dulce. Euxesta stigmatias fue encontrada solamente en los condados de Martin,
Miami-Dade, Okeechobee, Palm Beach y St. Lucie sobre maiz de campo y maiz dulce.
Euxesta annonae (F.) fue encontrada en maiz dulce en los condados de Miami-Dade, Okee-
chobee y Palm Beach, pero no se muestrearon maiz de campo en estos condados. Se recolec-
taron Euxesta eluta, E. stigmatias y C. massyla durante toda la etapa reproductive del maiz.
Euxesta annonae fue criada de mazorcas solamente de 8 a 21 dias de edad, pero los campos
con mazorcas de s 8-dias no fueron muestreados en los condados donde esta especie fue en-
contrada. El criar los adults de mazorcas infestadas con larvas de moscas fue el mejor me-
todo para evaluar la taza de infestacion de las mazorcas y la diversidad de species. Las
recolecciones con redes no dieron un estimado confiable para identificar infestaciones de es-
pecies de ulidiidos. Reportamos por primera vez E. annonae y E. eluta como plagas de maiz
en la Florida y EEUU.







Florida Entomologist 94(1)


There are 671 species of Ulidiidae worldwide,
but less than 10 species in 2 genera are known to
damage corn (Allen & Foote 1992; Anonymous
2008c; Goyal et al. 2010; Van Zwaluwenburg
1917). Van Zwaluwenburg (1917) first reported
the pest nature of Euxesta stigmatias Loew
(Diptera: Ulidiidae) (Figs. 1 g, lh) in Puerto Rico
where it damaged up to 100% of untreated corn. It
was first discovered damaging corn in Miami,
Florida in 1938 (Barber 1939) and had moved
north into central Florida by 1951 (Hayslip 1951).
This species has become a serious pest of Florida
sweet corn (Zea mays L.) requiring multiple in-
secticide applications during the ear stage to
maintain a marketable crop (Mossler 2008;
Nuessly & Hentz 2004; Seal 1996, 2001; Seal &
Jansson 1994). Sweet corn is an important crop in
Florida with 22.8% of the total USA fresh market
sweet corn production (Anonymous 2009). Eux-
esta stigmatias also has been reported infesting
sweet corn in Georgia (Daly & Buntin 2005),
Texas (Walter & Wene 1951), California (Fisher
1996), Guatemala (Painter 1955) and Brazil
(Franca & Vecchia 1986). The insect deposits its
eggs primarily on silks (styles) in the tips of ears.
The larvae feed on silks, kernels, and cobs. Bailey
(1940) observed disruption of pollination due to
larval feeding on silks. Larvae enter through the
soft pericarp of milk stage kernels to completely
consume the developing embryo and endosperm
(Seal & Jansson 1989). App (1938) observed lar-
val feeding on cobs followed by mold development
resulting in significant reduction in market value.
Several other ulidiid species are known maize
pests in the Caribbean and in the Americas south
of Texas (Arce de Hamity 1986; Barbosa et al.
1986; Chittenden 1911; Diaz 1982; Evans & Zam-
brano 1991; Gossard 1919; Painter 1955; Wyck-
huys & O'Neil 2007), but only 1 other species is
currently recognized as a pest in the USA. Cha-
etopsis massyla (Walker) (Figs. 1 a, b) was recently
determined to be a primary pest of sweet corn in
Florida (Goyal et al. 2010). Evidence suggesting
the possibility of additional picture-winged species
attacking corn in Florida include a picture of Eux-
esta eluta Loew (Diptera: Ulidiidae) (Figs. 1 e, fl on
the cover of Hayslip's (1951) paper entitled "Corn
silk fly control on sweet corn" misidentified as E.
stigmatias. Examination of the Ulidiidae collection
at the Division of Plant Industry in Gainesville,
Florida revealed that E. eluta and E. annonae (F.)
(Diptera: Ulidiidae) (Figs. 1 c, d) have been col-
lected in several Florida counties since at least
1948, but these specimens were not labeled as be-
ing collected or reared from corn. These later 2 spe-
cies are recognized pests of corn in South America
(Diaz 1982; Frias-L 1978). Therefore, it is possible
that additional Ulidiidae species may be feeding on
corn in Florida. The objective of this study was to
evaluate species richness and distribution of corn-
infesting ulidiids throughout Florida.


MATERIALS AND METHODS

Corn grown throughout Florida was sampled
for ulidiid species. Extension personnel and re-
searchers from all 67 Florida counties provided
information on corn types and growing season
needed to select representative fields. Corn fields
were visited with the assistance of extension
agents. One to 2 corn fields were sampled for Uli-
diidae in each of 16 counties from Jul through Oct
2007 (Table 2). One to 4 corn fields were sampled
for Ulidiidae in each of 27 counties during Feb
through Jun 2008 (Table 3), including 10 counties
visited in 2007.
Adult ulidiids can be elusive and difficult to re-
liably observe and collect. They frequently avoid
direct sunlight and walk or fly away from the di-
rect line of sight of workers approaching them.
They are more easily collected from the tassels
and upper leaves of corn plants in the hour just
after sunrise and just before sunset, but it was
not possible to sample all fields at these times.
Adults can also be killed after ovipositing on a
plant host before they are sampled, particularly
within crops that are frequently treated with in-
secticides, such as sweet corn. Therefore, fields
were sampled for both adults and immatures to
determine whether the plants served as develop-
mental hosts for ulidiid species and to determine
the feasibility of using adult collection records for
determining ear infestation. Preference was
given to sampling corn that was between the silk-
ing and dough stages because both the adult and
immature stages of flies can best be collected dur-
ing the first 3 weeks of corn reproduction. Neither
adults nor immatures in ears were found in fields
sampled before silking in Lake County (sweet
corn) in 2007 and in Jefferson (field corn) and
Walton (sweet corn) Counties in 2008, therefore;
data from these 3 fields were not included in the
results. Sweet corn fields were preferred over
field corn for sampling because the flies cause less
damage in field corn than in sweet corn (Scully et
al. 2000). Corn type (i.e., field, sweet, Bt-en-
hanced, and standard corn) and variety, number
of days before or after first silk, and locations of
the field were recorded. Visual observations were
taken for the presence of ulidiid adults.
Flies were collected from corn fields with a
sweep net (37.5 cm diameter). The sample size
was adjusted depending on the estimated field
size. In fields <4 ha, 3 pairs of corn rows were se-
lected for sampling: 1 pair from each side of the
field and 1 pair in the middle of the field. In fields
>4 ha, 9 pairs of rows were selected for sampling:
1 on each side of the field, 1 in the middle of the
field, and 6 pairs of rows randomly selected from
between the field margins. Sweep net sampling
for flies was done while walking the length of the
field swinging the net 100 times between 2 rows
in each pair of selected rows. Flies were preserved


March 2011




Goyal et al: Corn-infesting Ulidiidae of Florida


Fig. 1. Chaetopsis massyla male (a) and female (b); Euxesta annonae male (c) and female (d); E. eluta male (e)
and female (f); E. stigmatias male; and (g) and female (h).


...........


9rj


a







Florida Entomologist 94(1)


in 70% ethyl alcohol for later identification and
counting with a dissecting microscope. Identified
Ulidiidae specimens housed at the Division of
Plant Industry, Gainesville, FL and keys of Eux-
esta (Ahlmark & Steck unpublished, Curran
1928, 1934, 1935) and Chaetopsis (G. Steyskal un-
published) were used to confirm identifications.
Corn ears were examined for the presence of fly
larvae in the same fields sampled with sweep nets.
Ears found to contain larvae were collected and held
for adult emergence to confirm species infestation.
The number of ears sampled per field was adjusted
depending on the number of planted rows in each
field. Fifty-six ears were examined in fields with
<90 rows and 88 ears were examined in fields with
>90 rows. In a field with <90 rows, 10 groups of 4
plants each were randomly selected for ear inspec-
tion. In a field with 90 rows, ears were examined in
every tenth row starting from the first row and con-
tinuing to the other side of the field (total of 10
rows). In a field with >90 rows, 6 rows were sampled
from each side of the field (each sampled row sepa-
rated by 10 rows), and 6 additional rows were ran-
domly selected and sampled in the middle of the
field. One ear on each of 4 plants in the middle of
each selected row was examined for fly larvae (40
and 72 ears per field for <90 and >90 rows, respec-
tively). An additional 4 plants in each corner of the
field were examined for larvae-infested ears (16
ears per field). The top third of each infested ear was
removed with a knife and placed individually in a
Ziploc bag (1.83 L, S.C. Johnson & Son, Inc., Ra-
cine, WI). Two paper towels were added to each bag
to reduce moisture accumulation. Bags were stored
in portable coolers in the field and during transpor-
tation back to the laboratory.
Infested ears kept in the Ziploc bags were held in
an air conditioned room maintained at 26.0 + 1C
and L14:D10 h photoperiod to collect pupae for
adult identification. To reduce the accumulation of
moisture and associated fungus growth, bags with
corn were left partially open, paper towels were
changed frequently, and the air was constantly cir-


culated by box fans. Corn ears collected on Mar 6,
2007 were placed collectively in 3.78 L Ziploc bags
and then transferred to plastic containers with
mesh tops. Pupae were removed from the bags and
plastic containers, and placed on moistened filter
paper (Whatman@ 3, Whatman International Ltd.,
Maidstone, England) in covered Petri dishes for
adult emergence. The dishes were sealed with Para-
film (Pechiney Plastic Packaging, Chicago, IL) to
reduce moisture loss. Adults that emerged were pre-
served in 70% ethyl alcohol for later identification
and counting as above.

Statistical Analysis

The results were tested by analysis of variance
to examine the effects of sample technique, corn
type (field and sweet), corn ear age day and
month of sampling (1-7, 8-14, 15-21 d) and sample
year on the mean numbers of each species col-
lected (Proc GLM, Version 9.0; SAS Institute
2008). Year was used as a random variable in the
model. The mean number of flies sweep netted per
pair of rows used in the data analysis was calcu-
lated for each field by dividing the total number of
flies caught in sweep nets by the number of pairs
of rows sampled in that field. The mean number of
flies per infested ear was calculated for each field
by dividing the total number of flies reared from
infested corn ears by the number of infested ears
in each field. Different numbers of ears and plant
rows were sampled in each field and more fields
were sampled in 2008 than in 2007; therefore the
results were presented as least square means
rather than arithmetic means of flies caught per
row and reared per corn ear.

RESULTS

The mean number of ulidiid adults caught in
sweep nets was significantly affected by fly spe-
cies, corn type, survey year, and the species x year
interaction (Table 1). Significantly more E. eluta


TABLE 1. ANALYSIS OF VARIANCE FOR FLIES CAPTURED BY SWEEP OR REARED FROM CORN EAR ON SPECIES, CORN TYPE,
AGE OF CORN AND YEAR

Sweep net Corn ears

Source df F P F P

Species of Ulidiidae 3 8.12 <0.0001 3.73 0.0120
Corn type 1 6.77 0.0099 6.53 0.0113
Age of corn 2 1.82 0.1638 1.09 0.3368
Year 1 33.17 <0.0001 13.82 0.0003
Species x corn type 3 0.68 0.5642 1.52 0.2091
Species x age of corn 6 0.31 0.9304 0.67 0.6739
Species x year 3 5.50 0.0011 2.00 0.1145

ANOVA (Proc GLM, SAS Institute 2008); denominator df= 236.


March 2011







Goyal et al: Corn-infesting Ulidiidae of Florida


(least squares mean SEM; 3.80 + 0.63) and C.
massyla (3.62 0.63) were caught in sweep nets
per row than E. stigmatias (1.33 0.63) and E.
annonae (0.14 0.63). More adults were caught
per row with sweep nets in sweet corn (2.95 +
0.34) than in field corn (1.49 0.49). Sweep net
counts per row were greater in 2007 (3.89 0.51)
than in 2008 (0.55 0.32).
The mean number of adults emerged per ear
were significantly affected by fly species, corn
type, and survey year (Table 1). Significantly
more E. eluta (1.41 0.30) were reared from each
corn ear than E. stigmatias (0.40 0.30) and E.
annonae (0.05 0.30). The mean number of C.
massyla per ear (0.82 0.30) was not significantly
different than the other species. More adults were
reared from each corn ear in 2007 (1.19 0.25)
than in 2008 (0.15 0.15). Significantly more
adults per ear were reared from sweet corn (1.02
+ 0.16) than from field corn (0.33 0.24). Results
for species by county and reared from fields were
presented separately for 2007 (Table 2) and 2008
(Table 3) due to significant differences in mean
counts between years.
The correlation between adults caught in
sweep nets and those reared from ears varied by
species. Correlation coefficients were as follows:
0.79 (P < 0.0001) for E. stigmatias, 0.62 (P <
0.0001) for C. massyla, 0.58 for (P < 0.0001) E. an-
nonae, and 0.51 (P < 0.51) for E. eluta.

2007 Field Survey

Four Ulidiidae species were caught in sweep
nets and reared from fly larvae-infested ears in
Florida corn during the first survey year (Table
2). Chaetopsis massyla was collected in more
counties throughout the state than other species
and was netted in 100% of the sampled fields.
This was followed by E. eluta, which was netted in
88% of sampled fields in all counties except Lake
and Lee Counties (Table 2). Euxesta annonae and
E. stigmatias were netted from only 3 counties in
central and southern Florida, i.e., Miami-Dade,
Okeechobee, and Palm Beach Counties. As a re-
sult of the more limited distribution, both E. an-
nonae and E. stigmatias were netted in only 18%
of fields sampled. The species netted and reared
varied by corn type. Adults of E. annonae and E.
stigmatias were netted only from sweet corn
fields in Miami-Dade, Okeechobee, and Palm
Beach Counties, but field corn was not sampled in
these counties (Table 2). Adults ofE. eluta and C.
massyla were netted from both field and sweet
corn fields throughout the state. Euxesta eluta
and C. massyla were netted from 50 and 100% of
field corn fields, respectively, while both species
were netted from 100% of sweet corn fields.
The percentage of ulidiid-infested ears ranged
from 5% in Escambia to 38% in Santa Rosa
County (Table 2). Euxesta eluta and C. massyla


were reared from ears collected from all but Lee,
Lake and St. Johns Counties. These 2 species
were reared from infested ears in 82% of corn
fields statewide. Euxesta annonae and E. stigma-
tias were reared only from corn ears collected
from Miami-Dade, Okeechobee, and Palm Beach
Counties, amounting to only 18% of fields sam-
pled. Adults of E. eluta and C. massyla were
reared from both field and sweet corn ears. Eux-
esta annonae and E. stigmatias emerged from
sweet corn ears in fields from Miami-Dade,
Okeechobee, and Palm Beach Counties, but field
corn fields were not sampled in these counties.
Euxesta eluta and C. massyla were each reared
from 50% of field corn and 92% of sweet corn
fields. Euxesta annonae and E. stigmatias were
reared from 100% of the sweet corn fields in above
mentioned Counties.
The age of sampled corn in 2007 ranged from 4
to 21 d after first silk (Table 2). Chaetopsis mas-
syla was sweep netted in fields of all ages sam-
pled. Euxesta eluta was sweep netted in fields 7-
21 d after first silk. Euxesta annonae and E. stig-
matias were sweep netted from fields 8 to 21 d af-
ter first silk, but no fields <8 d after first silk were
sampled in counties infested with these 2 species.
Euxesta eluta and C. massyla emerged from corn
ears collected from fields 4 to 21 d after first silk,
while E. annonae and E. stigmatias from ears col-
lected 8 to 21 d after first silk (Table 4). Corn ears
were not collected from fields <8 d after first silk
in counties with E. annonae and E. stigmatias.

2008 Field Survey

The same 4 species were again collected in
sweep nets and reared from fly-larvae infested
ears in Florida corn during the second study year
(Table 3). Ulidiid adults were netted in 23 of 27
counties sampled in 2008. No adult picture-
winged flies were captured in corn in Dixie, Jack-
son, Sumter, Taylor or Volusia Counties. Chaetop-
sis massyla was collected from more counties
than other species throughout the state and was
netted in 66% of the fields sampled. Euxesta eluta
was netted in 49% of the fields sampled. Chaetop-
sis massyla was the only species collected from
corn in Alachua, Jefferson and Marion Counties,
while E. eluta was the lone species collected from
corn in Okaloosa County. Euxesta stigmatias was
netted only in Martin, Okeechobee, Palm Beach,
and St. Lucie Counties amounting to only 11% of
the fields sampled. Euxesta annonae was not col-
lected in sweep samples in 2008. Euxesta eluta, E.
stigmatias and C. massyla were netted from field
and sweet corn fields (Table 3). Euxesta eluta
were netted from 27 and 59%, and C. massyla
from 40 and 78% of the field and sweet corn fields
throughout the state, respectively. Euxesta stig-
matias was caught from 100 and 67% of the field
and sweet corn fields, respectively, in Martin,
















TABLE 2. ULIDIIDAE SPECIES COLLECTED IN FIELDS OR REARED FROM INFESTED EARS IN FLORIDA, 2007.

No. rows Mean no. adults captured per 100 sweeps3 Mean no. adults emerged per ear (per infested ear)
sampled No. ears
County-field Corn Sample Ear with sampled
no. type' date age (d)2 sweep net E. annonae E. eluta E. stigmatias C. massyla (no. infested) E. annonae E. eluta E. stigmatias C. massyla

Alachua Swt 16 Aug 15-21 3 0.0 1.8 0.0 4.2 56(6) 0.0 (0.0) 0.9 (8.3) 0.0 (0.0) 1.4 (13.5)
Bradford Swt 17 Oct 15-21 9 0.0 0.9 0.0 2.9 88 (14) 0.0 (0.0) 0.7 (4.4) 0.0 (0.0) 1.6 (9.9)
Miami-Dade Swt 6 Mar 15-21 3 2.8 26.0 11.2 11.7 56 (16) 0.4(1.3) 17.5(61.3) 5.6(19.4) 5.9 (20.7)
Escambia 1 Fld 2 Aug 8-14 3 0.0 11.5 0.0 3.8 56 (5) 0.0 (0.0) 0.2 (2.2) 0.0 (0.0) 0.3 (3.2)
Escambia 2 Bt swt 2 Aug 7 9 0.0 5.8 0.0 6.9 56 (3) 0.0 (0.0) 0.3 (6.3) 0.0 (0.0) 0.6 (10.3)
Gadsden Swt 17 Sep 7 3 0.0 6.3 0.0 4.8 56 (6) 0.0 (0.0) 1.1(10.3) 0.0(0.0) 1.0 (9.3)
Holmes Swt 16 Oct 15-21 3 0.0 3.7 0.0 2.3 56 (11) 0.0 (0.0) 0.6 (3.2) 0.0 (0.0) 0.6 (2.8)
Lake Fld 14 Sep 4-5 9 0.0 0.0 0.0 3.2 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Lee Fld 17 Oct 15-21 3 0.0 0.0 0.0 1.5 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Liberty Fld 13 Sep 15-21 3 0.0 2.3 0.0 1.8 56 (4) 0.0 (0.0) 0.3 (4.3) 0.0 (0.0) 0.5 (7.3)
Marion Swt 4 Sep 8-14 3 0.0 16.5 0.9 26.0 56 (11) 0.0 (0.0) 4.2(21.2) 0.0(0.0) 7.1(36.3)
Okeechobee Swt 18 Sep 8-14 3 1.1 0.9 1.0 2.9 88 (6) 0.1 (1.3) 0.6 (8.3) 0.3 (4.3) 0.8 (12.3)
Palm Beach Swt 14 Nov 8-14 3 1.7 21.3 33.5 18.3 56 (17) 1.7 (5.5) 5.8(19.1) 8.8(29.1) 1.8 (5.9)
Santa Rosa Swt 3 Aug 8-14 3 0.0 3.7 0.0 8.3 56(21) 0.0(0.0) 4.5(12.1) 0.0(0.0) 1.1(3.0)
St. Johns Swt 13 Sep 15-21 3 0.0 10.7 0.0 11.3 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Suwannee Swt 13 Sep 15-21 3 0.0 11.5 0.0 2.7 56 (12) 0.0 (0.0) 2.8(13.2) 0.0(0.0) 4.6 (21.3)
Washington Swt 2 Aug 15-21 3 0.0 4.3 0.0 3.3 88 (16) 0.0(0.0) 11.3(62.3) 0.0(0.0) 3 (16.3)

Corn type: Fld = field corn; Swt = sweet corn; Bt swt = Bacillus thuringiensis-enhanced sweet corn.
'Estimated days after first silk at time of sampling.
'0 = no flies were detected in sweep nets.








TABLE 3. ULIDIIDAE SPECIES COLLECTED IN FIELDS OR REARED FROM INFESTED EARS IN FLORIDA, 2008.

No. rows Mean no. adults captured per 100 sweeps3 Mean no. adults emerged per ear (per infested ear)
sampled No. ears
County-field Corn Sample Ear with sampled
no. type' date age (d)2 sweep net E. annonae E. eluta E. stigmatias C. massyla (no. infested) E. annonae E. eluta E. stigmatias C. massyla

Alachua Swt 4 Jun 7 3 0.0 0.0 0.0 1.3 56 (8) 0.0 (0.0) 0.2 (1.4) 0.0 (0.0) 0.0 (0.0)
Bradford 1 Swt 23 Jun 18-21 3 0.0 2.3 0.0 8.7 56 (15) 0.0 (0.0) 3.3 (12.3) 0.0 (0.0) 1.1(4.3)
Bradford 2 Swt 23 Jun 10-14 3 0.0 3.3 0.0 5.7 56 (17) 0.0(0.0) 10.2(33.5) 0.0 (0.0) 0.4 (1.5)
Columbia Swt 24 Jun 8-14 9 0.0 1.1 0.0 2.2 88 (2) 0.0 (0.0) 0.1(2.5) 0.0 (0.0) 0.05 (2.0)
Dixie Fld 4 Jun 2-3 9 0.0 0.0 0.0 0.0 88 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) o
Gilchrest 1 Swt 23 Jun 15-21 9 0.0 0.7 0.0 1.3 88 (6) 0.0 (0.0) 0.6 (9.3) 0.0 (0.0) 0.8 (11.3)
Gilchrest 2 Bt fid 23 Jun 7 9 0.0 0.0 0.0 2.1 88 (4) 0.0 (0.0) 0.1(2.3) 0.0 (0.0) 0.2 (5.3)
Gilchrest 3 Bt fid 23 Jun 7 9 0.0 1.8 0.0 0.8 88 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Hamilton 1 Swt 24 Jun 7 3 0.0 1.3 0.0 1.3 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Hamilton 1 Fld 24 Jun 14 3 0.0 0.0 0.0 0.0 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Hendry Swt 26 Feb 15-21 9 0.0 4.6 0.0 1.4 88 (18) 0.0 (0.0) 2.7 (13.3) 0.0 (0.0) 4.4 (21.4)
Holmes 1 Swt 26 Jun 21 3 0.0 8.3 0.0 0.7 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Holmes 2 Fld 26 Jun 21 9 0.0 2.1 0.0 2.9 88 (9) 0.0 (0.0) 0.4 (3.4) 0.0 (0.0) 0.1(1.4)
Jackson 1 Bt fid 5 Jun 2-3 9 0.0 0.0 0.0 0.0 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Jackson 2 Bt fid 5 Jun 2-3 9 0.0 0.0 0.0 0.0 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Jefferson -1 Fld 25 Jun 8-14 9 0.0 0.0 0.0 0.0 88 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Jefferson 1 Swt 25 Jun 14 3 0.0 0.0 0.0 0.7 56 (10) 0.0 (0.0) 0.4 (2.4) 0.0 (0.0) 0.0 (0.0)
Lafayette 1 Swt 24 Jun 14 3 0.0 1.0 0.0 1.3 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Lafayette 2 Swt 24 Jun 14 3 0.0 0.0 0.0 1.7 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Lafayette 3 Fld 24 Jun 14 9 0.0 0.0 0.0 0.0 88 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Lake 1 Swt 6 Jun 21 9 0.0 0.0 0.0 5.2 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) o
Lake 2 Swt 6 Jun 21 9 0.0 2.0 0.0 1.8 56 (22) 0.0 (0.0) 1.7 (4.4) 0.0 (0.0) 4.1(10.4)
Marion Bt fid 3 Jun 14 3 0.0 0.0 0.0 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Martin 1 Swt 11Mar 7 3 0.0 3.0 1.0 1.3 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Martin 2 Swt 11 Mar 1-2 3 0.0 0.0 0.0 0.0 56 (9) 0.0 (0.0) 0.4 (2.3) 3.1(19.0) 0.5 (3.3)
Martin 3 Swt 11 Mar 1-2 9 0.0 0.0 0.0 0.9 88 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Martin -4 Swt 11 Mar 14 3 0.0 0.0 14.3 4.7 56 (8) 0.0 (0.0) 0.0 (0.0) 0.5 (3.4) 0.0 (0.0)
Nassau 1 Swt 23 Jun 15-21 3 0.0 0.0 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Nassau 2 Fld 23 Jun 15-21 9 0.0 0.0 0.0 0.0 88 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Okaloosa 1 Swt 5 Jun 5 3 0.0 2.0 0.0 0.0 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Okaloosa 2 Swt 5 Jun 10 3 0.0 0.0 0.0 0.0 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)

Corn type: Fld = field corn; Swt = sweet corn; Bt swt = Bacillus thuringiensis-enhanced sweet corn.
'Estimated days after first silk at time of sampling.
0 = no flies were detected in sweep nets; = fly species was observed only, not collected.
















TABLE 3. (CONTINUED) ULIDIIDAE SPECIES COLLECTED IN FIELDS OR REARED FROM INFESTED EARS IN FLORIDA, 2008.

No. rows Mean no. adults captured per 100 sweeps3 Mean no. adults emerged per ear (per infested ear)
sampled No. ears
County-field Corn Sample Ear with sampled
no. type' date age (d)2 sweep net E. annonae E. eluta E. stigmatias C. massyla (no. infested) E. annonae E. eluta E. stigmatias C. massyla

Okeechobee Swt 19 Apr 14 9 0.0 3.6 7.2 1.9 88 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Palm Beach Swt 10 Apr 15-21 3 0.0 11.3 20.7 12.7 56 (15) 0.9 (3.2) 3.3 (12.3) 1.1(4.3) 2.5 (9.2)
Santa Rosa 1 Bt swt 5 Jun 10 3 0.0 0.0 0.0 1.7 56 (16) 0.0(0.0) 16.1(56.4) 0.0 (0.0) 0.4 (1.4)
Santa Rosa 2 Swt 5 Jun 14 3 0.0 0.0 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Santa Rosa 3 Bt swt 5 Jun 15-21 3 0.0 2.3 0.0 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
St. Johns 1 Bt swt 6 Jun 8-14 3 0.0 6.3 0.0 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
St. Johns 2 Bt swt 6 Jun 14 3 0.0 0.0 0.0 0.0 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
St. Lucie Bt fld 29 May 14 9 0.0 0.0 4.1 5.7 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Sumter Swt 6 Jun 15-21 3 0.0 0.0 0.0 0.0 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Suwannee Fld 4 Jun 15-21 3 0.0 1.0 0.0 1.0 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Taylor Fld 25 Jun 15-21 3 0.0 0.0 0.0 0.0 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Volusia Swt 6 Jun 21 3 0.0 0.0 0.0 0.0 56 (1) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.02 (1.0)
Walkulla Swt 25 Jun 14 3 0.0 2.0 0.0 56 (6) 0.0 (0.0) 0.3 (3.2) 0.0 (0.0) 0.3 (2.3)
Walton Fld 26 Jun 14 9 0.0 1.8 0.0 0.0 88 (4) 0.0 (0.0) 0.1(2.3) 0.0 (0.0) 0.0 (0.0)
Walton Swt 26 Jun 15-21 3 0.0 0.0 2.0 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)
Walton Swt 26 Jun 15-21 9 0.0 0.0 0.0 0.0 88 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)

'Corn type: Fld = field corn; Swt = sweet corn; Bt swt = Bacillus thuringiensis-enhanced sweet corn.
Estimated days after first silk at time of sampling.
0 = no flies were detected in sweep nets; = fly species was observed only, not collected.







Goyal et al: Corn-infesting Ulidiidae of Florida


TABLE 4. PERCENTAGE OF FIELDS WITH ULIDIIDAE SPECIES SWEEP NETTED OR REARED FROM CORN EARS BY EAR AGE

Sweep netted Reared from corn ears
Ear age (d) Ear age (d)

Species 0 to 7 d 8 to 14 d 15 to 21 d 0 to 7 d 8 to 14 d 15 to 21 d

2007
E.annonae 0 100 100 0 100 100
E. eluta 67 100 89 67 100 78
E. stigmatias 0 100 100 0 100 100
C. massyla 100 100 100 67 100 78

2008
E.annonae 0 0 0 0 0 100
E. eluta 36 42 64 27 32 35
E. stigmatias 33 100 100 33 33 100
C. massyla 54 68 71 18 21 41


Okeechobee, Palm Beach, and St. Lucie Counties.
No E. annonae adults were netted in field or
sweet corn fields.
Ulidiid-infested ears were found in 13 of 27
counties sampled (Table 3). The percentage of ul-
idiid infested ears ranged from 2% in Volusia to
39% in Lake County. Only E. eluta were reared
from corn ears collected from Alachua, Jefferson,
and Walton Counties. Chaetopsis massyla was the
only species reared from corn ears collected from
Volusia County. Euxesta eluta and C. massyla
were reared from 32% and 28% of the corn fields
sampled throughout the state. Euxesta eluta were
reared from 20 and 38% and C. massyla from 13
and 34% of the field and sweet corn fields, respec-
tively throughout the state. Euxesta stigmatias
was only reared from infested sweet corn ears in
Martin and Palm Beach Counties amounting to
6% of the total fields sampled. Adults of E. an-
nonae were only reared from infested sweet corn
ears collected from Palm Beach County amount-
ing to approximately 2% of the total fields sam-
pled. Field corn fields were not sampled in the
counties where E. stigmatias and E. annonae
were reared from ears.
The age of corn ears in surveyed fields ranged
from 1 to 21 d after first silk (Table 3). More E.
eluta and C. massyla were caught in sweep nets
and reared from corn ears 15 to 21 d post-silking
compared to 0 to 14 d post-silking. More E. stig-
matias were caught in sweep nets and reared
from corn ears in fields with 15 to 21 d post-silk-
ing ears than in younger fields. In counties where
E. annonae was found, it was only reared from
fields sampled 15 to 21 d after first silk.

DISCUSSION

The results of this 2-year study confirmed that
several species of Ulidiidae flies were infesting
corn in Florida. Ulidiidae flies were found infest-


ing both sweet and field corn fields across the
Florida panhandle from Escambia to Nassau
Counties and south through the peninsula to Mi-
ami-Dade County. Flies were collected in sweep
nets or reared from corn ears from 29 out of 33
sampled counties during the 2 survey years
(Fig. 2). Flies were more common in the 2007
compared to 2008 surveys probably due to differ-
ences in sampling times. Corn fields in 2007 were
largely sampled from Aug to Oct, except for Mi-
ami-Dade County that was sampled in Mar. In
contrast, surveys were conducted from Feb to Jun
in 2008. The flies may be more common in mid-
summer through fall months in northern Florida.
While more research has been conducted on E
stigmatias than the other species, it was found to
be much less common than C. massyla and E.
eluta in this survey. Euxesta eluta and C. massyla
were distributed in most fields sampled through-
out the state in both years, while E. stigmatias
and E. annonae were found in only several coun-
ties of southern Florida (Martin, Miami-Dade,
Okeechobee, Palm Beach, and St. Lucie).
The distribution of alternate host plants and
differences in acceptable temperature ranges for
each species may explain some of the variation
present in the distribution of ulidiids infesting
corn in Florida. Euxesta eluta, E. stigmatias, and
C. massyla were collected from both field and
sweet corn, while E. annonae was collected only
from sweet corn fields. Sweet corn is mostly grown
in southern Florida in comparison to northern
Florida where field corn predominates (Anony-
mous 2008a). However, E. stigmatias was not col-
lected or reared from sweet corn fields in northern
Florida. Frias-L (1978) in Chile found that higher
temperature and lower relative humidity led to
greater numbers of E. annonae while the reverse
led to greater numbers ofE. eluta.
Sampling with both sweep nets and collecting
infested corn ears gave a more complete picture of







Florida Entomologist 94(1)


WASHINGTON
Y GADSDEN JEFFERSON


WALTON

CALHOUN


E. annonae 0
E. eluta A
E. stigmatias *
C. massyla *


BRADFORD


FLAGLER


GILCHREST


SEMINOLE


.BREVARD
OKEECHOBEE


S( NI INIANRIVER
IAnTEE PDEE A ST.LUCIE
HANDS E A
SARASO A E STO Ar T MARTIN

GLADES PALM BEACH
LEE HEN A'
CHARLOTTE A *

BROWARD
COLLIER
DADE

MONROE 0


Fig. 2. Distribution of Ulidiidae species infesting corn in Florida by county during the 2007-2008 surveys. Sym-
bols in figure represent species collected using sweep nets and reared from corn ears in each of the sampled (shaded)
counties.


fly distribution in Florida corn fields than either
sampling technique alone. Low correlation values
indicate that sweep netting is not an efficient
method to estimate ulidiid species infesting corn
ears. The relationship between sweep nets and fly
species that emerged from infested ears ac-
counted for >60% of the variation for E. stigma-
tias and C. massyla, but <60% for E. eluta and E.
annonae. There were also a few locations where
flies were observed but not collected with sweep
nets. These were the places where flies were un-
common (1 or 2 per site) and netting was not the
best sampling technique for insects at low densi-
ties. Seal et al. (1996) found that E. stigmatias
congregated on the top of plants late in the
evening. Fly species in our study may have been
more active or more accessible with nets at times
of the day other than when sampling was con-
ducted. Therefore, sweep netting can be used to
indicate the potential for ear infestation, but the
identification of adults reared from infested ears


is currently the only method available for differ-
entiating the species developing within ears. The
external physical characteristics of the immature
stages of Ulidiidae infesting Florida corn are cur-
rently being examined by the authors to deter-
mine the possibility of using eggs, larvae or pupae
for the identification of species of flies infesting
corn.
Euxesta eluta, E. stigmatias and C. massyla were
collected from corn throughout the reproductive
stage of corn. Adult E. annonae may be present in
fields during the first week of silking, but only fields
>8 d after first silk were sampled in counties where
this species was found. In general, there was a ten-
dency for greater infestation by all 4 species as
sweet corn ears neared harvest and as field corn
ears approached the dough stage. The authors also
have frequently reared E. eluta, E. stigmatias, and
C. massyla from tassels and stems of corn plants.
Therefore, the potential host period on this crop is
longer than just the reproductive stage.


March 2011







Goyal et al: Corn-infesting Ulidiidae of Florida


This is the first report of E. annonae infesting
corn in Florida and the USA. This species was not
common in any location but was always netted
from fields and reared from ears along with other
Ulidiidae species. Euxesta annonae was the least
collected species in sweep nets and it was reared
from corn ears collected only from the southern
end of the Florida peninsula (Fig. 2). Euxesta an-
nonae is also reported as a pest of corn in Chile
(Frias-L 1978). The authors have frequently ob-
served E. annonae on Annona spp. (Magnoliales:
Annonaceae) and Chinese long bean, Vigna un-
guiculata ssp. sesquipedalis (L.) Verdc. (Fabales:
Fabaceae) in southern Florida and reared E. an-
nonae adults from field collected Annona spp.
fruit (Magnoliales: Annonaceae). Plants of An-
nona spp. are recorded in several southern and
central Florida counties (Brevard, Broward, Col-
lier, De Soto, Glades, Hendry, Highlands, Indian
River, Lee, Manatee, Martin, Miami-Dade, Mon-
roe, Palm Beach, and St. Lucie) (Wunderlin &
Hansen 2008) where they may provide alterna-
tive food resources for this species. The authors
have reared this species from decaying corn
stalks and from spiny amaranth, Amaranthus
spinosus L. (Caryophyllales: Amaranthaceae)
roots collected from the field at Belle Glade, Flor-
ida.
Euxesta eluta was widely collected in this
study from fields sampled throughout Florida
(Fig. 2). These flies were commonly observed in
fields and as many as 62 were reared from an in-
dividual ear. While this is the first known record
of E. eluta being a pest of corn in Florida and the
USA, its image in Hayslip (1951) suggests that it
was present in Florida corn fields >50 yr ago, but
incorrectly identified as E. stigmatias. The wide
distribution of E. eluta in Florida and its discov-
ery on both sweet and field corn indicates this fly
is a much greater threat to corn than E. stigma-
tias, which is found in a much smaller portion of
Florida. Euxesta eluta was recognized as infesting
corn in Puerto Rico >60 yr ago (Wolcott 1948) and
has been recorded as an ear pest in Ecuador
(Evans & Zambrano 1991), Chile (Frias-L 1978;
Olalquiaga 1980), Peru (Diaz 1982), Argentina
(Arce de Hamity 1986), and Brazil (Franca & Vec-
chia 1986). Euxesta eluta is a pest of loquat, Erio-
botrya japonica (Thumb.) Lindl. (Rosales: Ro-
saceae) in Alachua County, Florida (Anonymous
2008b). Loquat is grown as a dooryard plant and
is distributed in several counties throughout the
state (Wunderlin & Hansen 2008).
Euxesta stigmatias was found in sweep net col-
lections and reared from corn ears from southern
and central Florida counties only (Fig. 2).
Weather differences in southern and northern
Florida may explain part of the variation in dis-
tribution of the species. Adult E. stigmatias have
been reared from damaged or decayed inflores-
cences of sorghum, Sorghum bicolor (L.) Moench


(Cyperales: Poaceae), tomato fruit, Lycopersicon
esculentum L. (Solanales: Solanaceae) (Seal &
Jansson 1989), and decaying carrot roots, Daucus
carota L. (Apiales: Apiaceae) (Franca & Vecchia
1986).
Chaetopsis massyla was caught in sweep nets
and reared from corn ears in the majority of sur-
veyed counties (Fig. 2). This fly was common in
field and sweet corn fields throughout the year in
southern Florida counties. The relative abun-
dance and development range across corn types
indicates this species is a much greater threat to
Florida corn than previously recognized. Its habit
of feeding on a range of monocots may help ex-
plain its widespread distribution throughout
Florida. Allen & Foote (1992) reported it to be a
secondary invader of wetland monocots. Chaetop-
sis massyla has been reared from cattail, Typha
spp. (Typhales: Typhaceae) in California (Keiper
et al. 2000). Typha spp. are found in most Florida
counties except Flagler, Gadsden, Glades, Her-
nando, and Suwannee (Wunderlin & Hansen
2008). The authors made several personal obser-
vations of C. massyla plant associations during
the course of this statewide survey. Chaetopsis
massyla was frequently observed by the authors
on cattail plants on ditch banks and feeding on
sugary exudates from sugarcane plants (a com-
plex hybrid of Saccharum spp.) in Belle Glade
(Palm Beach County). Chaetopsis massyla adults
were reared from sugarcane stems that were ac-
tively infested with the sugarcane borer, Diatraea
saccharalis (F). (Lepidoptera: Crambidae) col-
lected by the authors in November 2009 from sug-
arcane fields at Clewiston (Hendry County) and
Sebring (Highlands County), Florida. Chaetopsis
massyla was also successfully reared by the au-
thors from otherwise healthy sugarcane stems ex-
posed to colonies in the laboratory in which 0.5 cm
diam holes were drilled in billets to simulate
emergence and frass evacuation holes produced
by D. saccharalis. Other plants from which C.
massyla has been reared include hairy sedge,
Carex lacustris Willd. (Cyperales: Cyperaceae)
(Allen & Foote 1992), Narcissus spp. (Liliales: Lil-
iaceae) (Blanton 1938) and onions, Allium cepa L.
(Asparagales: Alliaceae) (Merrill 1951). Carex
spp. are found throughout the state while the dis-
tribution of Narcissus spp. is considered to be lim-
ited to Alachua, Calhoun, Escambia and Leon
Counties (Wunderlin & Hansen 2008).
Two additional Chaetopsis spp. have been re-
ported feeding on corn, but neither was found in
this 2-year survey of Florida corn fields. Large
populations of fly larvae that were discovered in
corn stalks within tunnels likely produced by Eu-
ropean corn borer, Ostrinia nubilalis Hiibner
(Lepidoptera: Pyralidae) in Ohio were reared to
adults and identified as Chaetopsis aenea (Wiede-
mann) by Gossard (1919). Larvae of C. fulvifrons
(Macquart) were reared from within the tunnels







Florida Entomologist 94(1)


of southwestern corn borer, Diatraea grandiosella
Dyar (Lepidoptera: Crambidae), in the Texas high
plains (Knutson 1987). Langille (1975) reported
that Chaetopsis spp. larvae were commonly asso-
ciated with diapausing D. grandiosella within
corn stalks in Missouri and hypothesized that ul-
idiid larvae feed on the decaying stalks or micro-
bial growth within the bored stalks.
In conclusion, 4 species of picture-winged flies
were found infesting corn in Florida. Evidence
presented herein is the first known documenta-
tion for E. annonae and E. eluta as pests of corn in
Florida and the USA. The 4 species were not uni-
formly distributed throughout Florida corn grow-
ing regions. Euxesta eluta and C. massyla were
found infesting field and sweet corn throughout
Florida. Euxesta stigmatias was only found in-
festing corn in Martin, Miami-Dade, Okeechobee,
Palm Beach, and St. Lucie Counties. Euxesta an-
nonae (F.) was found in sweet corn in Miami-
Dade, Okeechobee, Palm Beach Counties, but
field corn was not sampled in these counties. Eu-
xesta eluta, E. stigmatias and C. massyla were
collected from corn throughout the corn reproduc-
tive stage. Euxesta annonae was reared from 8-21
d old ears only, but fields with ears <8 d old were
not sampled in the counties where this species
was found. The relative abundance ofE. eluta and
C. massyla in Florida field and sweet corn indi-
cates the need for more research into their biology
and ecology. The discovery ofE. eluta and C. mas-
syla attacking corn ears in many of the northern-
most Florida counties suggests that further sur-
veys of corn growing areas across the borders into
neighboring states is warranted to determine the
extent of corn infesting picture-winged fly infesta-
tions in the southern U.S. The statewide distribu-
tion of E. eluta and C. massyla in reproducing
corn also suggests that studies should be con-
ducted to evaluate additional food sources that
support these species in the absence of corn.


ACKNOWLEDGEMENTS

We thank Harsimran K. Gill and Bijayita Thapa for
assistance in rearing Ulidiidae from collected ears, and
Nicholas A. Larsen for help contacting extension agents
and field navigation. We acknowledge Jaya Das for help
with the Florida county maps. The photographic assis-
tance of Lyle Buss was instrumental in producing the
fly images used in this report. We are also thankful to
University of Florida Cooperative Extension Agents and
researchers for their help in selecting corn fields and ar-
ranging the visits, and corn growers of Florida for allow-
ing us to survey their fields. This research was made
possible by a Hand Fellowship awarded by the Dolly
and Homer Hand Group.

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Florida Entomologist 94(1)


March 2011


A CHECKLIST AND KEY TO SPECIES OF THE GENUS BETACIXIUS
MATSUMURA (HEMIPTERA: FULGOROMORPHA: CIXIIDAE) WITH
DESCRIPTIONS OF TWO NEW SPECIES FROM GUIZHOU PROVINCE, CHINA

PEI ZHANG1'2'3 AND XIANG-SHENG CHEN1'2'*
'The Provincial Key Laboratory for Agricultural Pest Management of Mountainous Region, Guizhou University,
Guiyang, Guizhou 550025, P.R. China

2Institute of Entomology, Guizhou University, Guiyang, Guizhou 550025, PR. China

3Xingyi Normal University for Nationalities, Xingyi, Guizhou 562400, P.R. China

*Corresponding author; E-mail: chenxs3218@163.com

ABSTRACT

Two new species of Betacixius Matsumura, 1914 (Hemiptera: Fulgoromorpha: Cixiidae:
Cixiini), B. bispinus Zhang and Chen sp. nov. (China: Guizhou) and B. flagellihamus Zhang
and Chen sp. nov. (China: Guizhou), from southwest China, are described and illustrated.
A key for identifying 23 known species of Betacixius is provided.

Key Words: Hemiptera, Fulgoroidea, Cixiidae, Oriental region, Betacixius, new species,
China

RESUME

Se described e ilustran dos nuevas species de Betacixius Matsumura, 1914 (Hemiptera:
Fulgoromorpha: Cixiidae: Cixiini), B. bispinus Zhang y Chen sp. nov. (China: Guizhou) y B.
flagellihamus Zhang y Chen sp. nov. (China: Guizhou) del suroeste de China. Se provee una
clave para identificar las 23 species conocidas de Betacixius.


The cixiid planthopper genus Betacixius (Cixi-
inae: Cixiini) was established by Matsumura
(1914) for the type species, B. ocellatus Mat-
sumura, 1914, from Japan. To date, 21 species
with 2 subspecies have been recorded worldwide
and all species occur in the southern region (Mat-
sumura 1914; Schumacher 1915; Metcalf 1936;
Jacobi 1944; Fennah 1956; Hori 1982; Chou et al.
1985, 1988; Tsaur et al. 1991; Hua 2002).
During the course of studying species biodiver-
sity of the suborder Auchenorrhyncha in south-
west China, 2 specimens belonging to unde-
scribed species of the genus Betacixius were
found. The purpose of this paper is to describe
these 2 new species and to provide an identifica-
tion key to all species of Betacixius.

MATERIALS AND METHODS

Morphological terminology follows L6cker et
al. (2006). Dry specimens were used for the de-
scription and illustration. External morphology
was observed under a stereoscopic microscope
and characters were measured with an ocular mi-
crometer. The genital segments of the examined
specimens were macerated in 10% KOH and
drawn from preparations in glycerin jelly with
the aid of a Leica MZ 12.5 stereomicroscope. Illus-


trations were scanned with Canon CanoScan
LiDE 200 and imported into Adobe Photoshop 8.0
for labeling and plate composition. Specimens ex-
amined are deposited in the Institute of Entomol-
ogy, Guizhou University, Guiyang, Guizhou Prov-
ince, China (IEGU).

DESCRIPTIVE TAXONOMY
Betacixius Matsumura, 1914 (Figs. 1-25)

Betacixius Matsumura 1914: 412; Chou et al. 1985: 23;
Tsaur et al. 1991: 27.

Type species: Betacixius ocellatus Matsumura
1914, by original designation.
Description. This is a redescription incorporat-
ing the descriptions previously published by Chou
et al. (1985) and Tsaur et al. (1991) as follows.

Body Size and Coloration. Small to medium-
size cixiids (4.3-7.3 mm). Body coloration varying
from green, brown to fulvous, mostly bearing spe-
cial markings on anteclypeus; lateroapical parts
of frons, otherwise unicolorous throughout.

Head and Thorax. Head including eyes
slightly narrower than pronotum. Vertex much
wider than long in midline, widest basally or






Zhang & Chen: New Species of Betacixius from China


7


4 6
Figs. 1-13. Betacixius bispinus Zhang and Chen sp. nov. 1. Head and thorax, dorsal view; 2. Frons; 3. Forewing;
4. Male genitalia, lateral view; 5. Pygofer and genital styles, ventral view; 6. Anal segment, dorsal view; 7. Anal seg-
ment, caudal view; 8. Connective, caudal view; 9. Right genital style, ventral view; 10. Aedeagus, left side; 11. Aede-
agus, right side; 12. Aedeagus, dorsal view; 13. Aedeagus, ventral view. Scale bars = 0.25 mm (Figs. 6, 7), 0.5 mm
(Figs. 1, 2, 4, 5, 8-13), 1 mm (Fig. 3).







Florida Entomologist 94(1)


apically, lateral carinae moderately elevated,
disc shallowly hollowed on each side of median
carina. Frons rounded at base, usually with in-
complete median carina not reaching anterior
margin of vertex, lateral carinae slightly ele-
vated below level of eyes, with small median
ocellus and semicircular frontoclypeal suture.
Clypeus tricarinate, convex to midline. Prono-
tum small, with distinct median carina, inter-
median carinae curving laterad, angularly
rounded posteriorly. Mesonotum tricarinate,
convex between lateral carinae, flattened pos-
teromedially. Forewings broadest at apical
third, rounded at apex, with 4-5 subapical cells
and 8-9 apical cells, hyaline, sometimes with an
oblique band or ocellated stripe. Hind tibia with
2-4 lateral spines and 6 apical spines. Chaetot-
axy of hind tarsus 7/7.


Male Genitalia. Pygofer symmetrical, U-
shaped, with thumb-shaped dorsolateral angles
in ventral view. Medioventral process triangular
or subtriangular in ventral view, generally wider
at base than long in midline. Anal segment tubu-
lar. Genital styles symmetrical in ventral view.
Aedeagus slender in lateral view.
Distribution. Oriental and Palaearctic regions.
Remarks. This genus may be easily distin-
guished from other genera of Cixiini by the presence
of 4-5 subapical cells and 8-9 apical cells on the
forewing, vertex much wider than long in midline,
frons with incomplete median carina distinct near
frontoclypeal suture, and chaetotaxy of hind tarsus
7/7. The 2 new species, B. bispinus Zhang and Chen
sp. nov. (China: Guizhou) and B. flagellihamus
Zhang and Chen sp. nov. (China: Guizhou), fit into
the genus by the presence of features as above.


WORLD CHECKLIST OF SPECIES OF BETACIXTUS MATSUMURA

B. bispinus Zhang and Chen sp. nov.; southwestern China (Guizhou).
B. brunneus Matsumura (1914); China (Taiwan), Japan.
B. clypealis Matsumura (1914); China (Taiwan).
B. clypealis vittifrons Matsumura (1914); China (Taiwan).
B. delicatus Tsaur & Hsu (1991); China (Taiwan).
B. euterpe Fennah (1956); China (Guangdong).
B. flagellihamus Zhang and Chen sp. nov.; southwestern China (Guizhou).
B. flavovittatus Hori (1982); China (Taiwan).
B. fuscus Tsaur & Hsu (1991); China (Taiwan).
B. herbaceus Tsaur & Hsu (1991); China (Taiwan).
B. .... .. Y...,. Matsumura (1914); South China, Japan (Okinawa).
B. maculosus Tsaur & Hsu (1991); China (Taiwan).
B. michioi Hori (1982); China (Taiwan).
B. nelides Fennah (1956); China (Zhejiang, Guangdong).
B. nigromarginalis Fennah (1956); China (Hubei).
B. obliquus Matsumura (1914); South China, Japan (Gifu).
B. obliquus pallens Matsumura (1914); Japan (Tokyo, Harima, Kumanoto)
B. ocellatus Matsumura (1914); China (Taiwan), Japan.
B. pallidior Jacobi (1944); South China (Fujian).
B. rinkihonis Matsumura (1914); China (Taiwan), Japan.
B. robustus Jacobi (1944); South China (Fujian).
B. shirozui Hori (1982); China (Taiwan).
B. sparsus Tsaur & Hsu (1991); China (Taiwan).
B. tonkinensis Matsumura (1914); South China, Vietnam.
B. transversus Jacobi (1944); South China (Fujian).

KEY TO SPECIES OF THE GENUS BETACIXTUS MATSUMURA

1. Forewings with m arkings (Figs. 3, 16, 24, 25) ..................................................... 2
- Forewings without any markings ............................................................ 9
2. Forewings with a large ocellate marking in apical half (Figs. 16 and 25) ............................... 3


March 2011






Zhang & Chen: New Species of Betacixius from China


23


20


17


Figs. 14-23. Betacixius flagellihamus Zhang and Chen sp. nov. 14. Head and thorax, dorsal view; 15. Frons; 16.
Forewing; 17. Male genitalia, lateral view; 18. Pygofer and genital styles, ventral view; 19. Anal segment, dorsal
view; 20. Anal segment, caudal view; 21. Genital styles, ventral view; 22. Aedeagus, dorsal view; 23. Aedeagus, right
side. Scale bars = 0.25 mm (Figs. 18-21), 0.5 mm (Figs. 14, 15, 17, 22, 23), 1 mm (Fig. 16).


-- -c 18







Florida Entomologist 94(1)


March 2011


- Forewings without such a marking in apical half (Figs. 3 and 24) .................................... 6

3. Forewings with an oblique brown band extending from clavus across middle of corium........ B. tonkinensis

- Forewings without such a band (Figs. 16 and 25) .................................................. 4

4. Flagellum of aedeagus with 1 spine, hook-shaped (Figs. 22 and 23) ............... B. flagellihamus sp. nov.

- Flagellum of aedeagus with 2 spines, not hook-shaped. ............................................. 5

5. Aedeagus with 2 L-shaped processes apically ........................................... B. maculosus

- Aedeagus with 1 nearly straight and 1 arched process apically. .............................. B. ocellatus

6. Forewings with an oblique band extending from stigma passing through its middle part ................. 7

- Forewings without such a band (Figs. 3 and 24) .................................................. 12

7. Frons with median carina distinct on apical third; hind basitarsus much longer than the 2nd and 3rd segment
put together. ................................................................. B. michioi

- Frons and hind basitarsus without features as above. .............................................. 8

8. Forewings with apical cells of M and Cu strongly infuscate. .................. ........... B. transversus

- Forewings with apical cells not infuscate. ........................................................ 9

9. Forewings with apical margin black or distinctly darkened. ......................................... 10

- Forewings with apical margin fuscous or not distinctly darkened .................................... 11

10. Frons with a pallid spot at centre of lateral margins, clypeus dark, mesonotum testaceous. .... B. .. . ... Y .

- Frons without such spots; mesonotum, except scutellum, castaneous-piceous ................... .B. euterpe

11. Forewings with an oblique dark band extending from clavus into centre of corium, slightly distad of level of
union of claval veins ............................................................. B. obliquus

- Forewings with a spot near sutural margin of clavus near union of claval veins, no oblique dark band at this level
extending into corium ........................................................... B. pallidior

12. Forewings with a long black stripe from base, along clavus extending to Rs ..................... .B. fuscus

- Forewings without such a stripe (Figs. 3 and 24) ................................................. 13

13. Fore tibiae with black, longitudinal stripes ...................................................... 14

- Fore tibiae without such stripes ............................................................... 15

14. Mid- and hind- tibiae with black, longitudinal stripes ..................................... B. delicatus

- Mid- and hind- tibiae without such stripes ............................................... B. sparsus

15. Forewings infuscated at base, extending along clavus to end of Cu,; mesonotum with a large, very distinct brown
marking between lateral carinae ................................................... B. shirozui

- Forewings and mesonotum without spots as above (Figs. 3 and 24) .................................. 16

16. Forewings with apical margin black or very dark (Figs. 3 and 24) .................. ................ 17

- Forewings with apical margin not particularly dark. .............................................. 18

17. Medioventral process of pygofer in ventral view right-angled triangular, pointed at apex (Fig. 5); on ventral mar-
gin, periandrium with lobate processes, 2 broad processes basally, bending forward, directed ventroceph-
alad, flagellum semisclerotised, with several serrated processes near apex (Figs. 10 and 12)
.................. ................................................ B. bispinus sp. nov.

- Medioventral process in ventral view subtriangular, rounded at apex; periandrium without lobate processes, but
with serrated processes basally, flagellum, without any processes ...................... B. rinkihonis

18. First apical cell of forewing piceous .................................................... .B. robustus

- A dark suffusion over all apical cells and across base of forewing .............................. B. nelides







Zhang & Chen: New Species of Betacixius from China


19. Aedeagus with a curved spine on left near apex and a short ledge in a similar position on right; flagellum arising
above left margin, sides parallel for most of length, distally a short curved spine directed cephalad, and a
subquadrate plate with a stout spine directed ventrad .......................... B. nigromarginalis
- Aedeagus and flagellum without features as above ............................................. 20
20. Frons without median carina ......................................................... B. clypealis
- Frons with median carina ................. ...................................... ........ 21
21. Frons with white or yellowish lateroapical parts; anteclypeus entirely black ............... B. flavovittatus
- Frons and clypeus unicolorous ................................... .......................... 22
22. Body pale brown; flagellum of aedeagus with 2 processes on right side ....................... B. brunneus


Body green; flagellum of aedeagus with one process on each side ....


Betacixius bispinus Zhang and Chen sp. nov.
(Figs. 1-13, 24)

Description. Body length (from apex of vertex
to tip of forewings): male 5.0-6.1mm (n = 2), fe-
male 5.5-6.8mm (n = 7); forewing length: male
4.0-4.9mm (n = 2), female 4.5-5.7mm (n = 7).
Coloration. General color brown. Body slightly
covered with powdery wax. Eyes yellowish brown
to blackish brown; ocelli reddish yellow. Vertex
yellowish brown. Pronotum blackish brown ex-
cept median carina yellowish brown. Mesonotum
blackish brown, with yellowish brown area pos-


...... B. herbaceus


teromedially, carinae concolorous. Frons yellow-
ish brown except lateral carinae blackish brown.
Postclypeus yellowish brown, anteclypeus black-
ish brown. Rostrum generally yellowish brown,
blackish brown near tip. Forewings pale brown,
hyaline; veins brown, tubercles dark brown;
stigma black; clavus with a short transversal
brown band, just distad of fork PCu+A1. Hind-tib-
iae yellowish brown, lateral- and apical- spines
yellowish brown basally, black apically; platellae
of tarsi dark brown. Abdomen black ventrally.
Head and Thorax. Vertex narrowing to apex as
shown in Figs. 1 and 24, wider than distance be-


U 24 L VW25

Figs. 24-25. Body of adult in dorsal view. 24. Betacixius bispinus Zhang and Chen sp. nov.; 25. Betacixius flagel-
lihamus Zhang and Chen sp. nov.







Florida Entomologist 94(1)


tween eyes, 2.5 times wider than long in midline;
anterior margin arched convex, with small emar-
gination at midpoint, posterior margin arched
concave; median carina indistinct, lateral carinae
without branches, subapical carina absent. Me-
dian ocellus very small, located the centre of the
frontoclypeal suture. Pedicel of antenna 1.6 times
longer than wide. Frons broad, with small spots
on middle area, widest at level of lateral ocelli,
narrowing to both ends, 1.25 times longer than
wide in midline; median carina indistinct, extend-
ing from slightly above level of lateral ocelli to
median ocellus, lateral carinae distinct and
ridged, arched convex; anterior margin arched
concave. Clypeus with median carina distinct and
elevated throughout, widest at level of endpoints
of frontoclypeal suture; lateral carinae distinct
and elevated. Rostrum relatively short, reaching
hind coxae, apical and subapical segments
equally long. Pronotum short and narrow, collar-
like, twice as long as vertex in midline; median
carina distinct and complete; intermedian cari-
nae corrugated and curving into posterior margin
which is concave in obtuse angle. Mesonotum 1.89
times longer than pronotum and vertex com-
bined; 3 longitudinal carinae all reaching ante-
rior and posterior margins, median carina indis-
tinct on posteromedian area, which bears trans-
verse striations. Forewings 2.22 times longer
than wide, with sparse setae on surface, tubercles
along veins, claval veins without tubercles; 2 in-
distinct subapical lines of cross veins; fork Sc+RP
distad of fork CuAl+CuA2; r-m crossvein slightly
distad of fork MA+MP; RP apically bifid, MA api-
cally bifid, MP apically bifid; fork PCu+A1
slightly basad of centre of clavus; Sc+R and M
fused at superior-outside angle of basal cell; fork
MA1+MA2 distad of fork MP1+MP2. 2nd hind-
tarsus with 5 platellae; hind-tibia with 4 lateral
spines, 6 apical spines: 2 large, 1 medium, 3
small, divided into 2 groups.

Male Genitalia. In ventral view, pygofer stout,
slightly concave medially, widening laterally; dor-
sal margin caudad obliquely raised in lateral
view, inferior part with bristles; lateral lobes sym-
metrical, medium-inferior part arched convexly
in lateral view. Medioventral process right-angled
triangular in ventral view, relatively broad, 1.5
times wider than long, distance between tips of 2
lateral lobes 3.06 times as long as width of medio-
ventral process; narrow triangular in lateral view,
covered basally. Anal segment short and stout as
shown in Figs 4, 6 and 7; 2.13 times longer than
wide in dorsal view; incompactly connected with
pygofer, freely movable; anal style, finger-like api-
cally, not beyond anal segment; anal opening
nearly subelliptical in dorsal view. Genital styles
as shown in Figs. 4, 5 and 9, in ventral view, wid-
ening to apex, apical margin truncated, with
sharp angle outside, internal processes broad,


touching each other; in lateral view, ventral mar-
gin curving upward, outside slightly corrugated,
dorsal margin smooth and bent forming a right
angle approximately; incompactly connected with
connective, freely movable; apical margin with
bristles in ventral view. Aedeagus broad, short,
connected with anal segment by 2 points; each
side with broad spine arising at apex of aedeagal
shaft, curving upward; periandrium ventrally
with lobate processes, basally 2 broad processes,
bending forward, directed ventrocephalad. Con-
nective anchor-like, relatively long, the width of
aedeagal shaft 1.65 times as wide as width of con-
nective plus ventral arm. Flagellum semi-sclero-
tised, structure simple, generally curving left,
with several serrated processes near apex.

Type Material. Holotype: 6, Mayanghe Na-
tional Natural Reserve (600m), Yanhe County,
Guizhou Province, China, 5-12 June 2007, X.-S.
Chen. Paratypes: 16, 79 9, same data as holo-
type.

Etymology. The name is derived from the Latin
words bi- (double) and spinus (spine), which re-
fers to the 2 spines on ventral margin of perian-
drium.
Distribution: Southwest China (Guizhou Prov-
ince).
Remarks. This new species is similar in ap-
pearance to B. rinkihonis, but differs from the lat-
ter in the shape of the medioventral process and
the anal segment and by having 2 spines on the
ventral margin of the periandrium and several
serrated processes near apex of flagellum.
Betacixius flagellihamus Zhang and Chen sp. nov.
(Figs. 14-23, 25)
Description. Body length (from apex of vertex
to tip of forewings): male 5.0-5.8mm (n = 13), fe-
male 5.4-6.2mm (n = 15); forewing length: male
4.5-5.0mm (n = 13), female 4.6-5.1mm (n = 15).

Coloration. General color brown. Body covered
with powdery wax. Median area of eyes black, ven-
tral margin yellow, other part reddish brown to
blackish brown. Median ocellus pale yellow, semi-
hyaline; lateral ocelli red. Vertex pale yellow.
Pronotum pale yellow. Mesonotum brown. Frons
generally brown, bright yellow above frontoclypeal
suture; postclypeus yellow to brown, with oblique
streaks; anteclypeus black. Apical segment of ros-
trum brown, subapical segment yellow. Forewings
pale brown, semihyaline, with ocellated marking
as shown in Figs. 16 and 25; veins and tubercles
brown; stigma brown to dark brown; clavus with
dark brown stain on apical third, sometimes ex-
tending to end of clavus; clavus suture brown.
Hind-tibia brown, lateral- and apical-spines brown
at base, black apically; membranous tooth of tarsi
dark yellow. Abdomen black ventrally.


March 2011







Zhang & Chen: New Species of Betacixius from China


Head and Thorax. Vertex narrowing towards
apex as shown in Figs. 14 and 25, wider than dis-
tance between eyes, 2.3 times wider than long in
midline, separated into 2 hollow areolets by me-
dian carina; anterior margin generally arched
convex slightly, slightly concave at midpoint, pos-
terior margin concave in obtuse angle; median ca-
rina distinct and complete, subapical carina ab-
sent. Median ocellus indistinct, in the centre of fr-
ontoclypeal suture. Frons with well-distributed
small spots, widest at level of antennae, narrow-
ing to both ends, length equal to width; median
carina distinct and elevated near frontoclypeal
suture; lateral carinae slightly S-shaped, ele-
vated; apex of frons elevated and lobate; anterior
margin semicircle concave. Median carina of
clypeus distinct and elevated; lateral carinae of
postclypeus elevated. Rostrum reaching hind-
femora, apical and subapical segments equally
long. Pronotum short and narrow, collar-shaped,
2.25 times longer than vertex in midline; median
carina distinct and elevated; posterior margin
concave in obtuse angle. Mesonotum 1.44 times
longer than pronotum and vertex combined; three
longitudinal carinae elevated except for median
carina weakly elevated at base, all reaching ante-
rior and posterior margins. Forewings 2.2 times
longer than wide; surface of Forewings with se-
tae, basal part less, apical part more; veins with
distinct tubercles, C vein with 34 tubercles, claval
apically vein with tubercles; 2 indistinct subapi-
cal lines of cross veins; fork Sc+RP distad of fork
CuAl+CuA2; r-m crossvein distad of fork
MA+MP; RP apically bifid, MA apically bifid, MP
apically bifid; fork PCu+A1 slightly basad of cen-
tre of clavus; Sc+R and M fused at superior-out-
side angle of basal cell; fork MA1+MA2 distad of
fork MP1+MP2. 2nd hindtarsus with 3 platellae;
hind-tibia with 3 lateral spines, 6 apical spines,
being divided into 2 groups by a relatively wide
gap, one group with 3 equal spines, arranged
closely, the other group with 1 large and 2 small,
arranged sparsely.

Male Genitalia. In ventral view, pygofer stout,
shallowly U-shaped, slightly widening from base
to end, ventral margin slightly concave; dorsal
margin caudad obliquely upward in lateral view;
lateral lobes symmetrical, inferior half slightly
arched concave in lateral view. Medioventral pro-
cess mastoid in ventral view, with bristles, 1.25
times wider than long, distance between tips of 2
lateral lobes 2.4 times as long as width of medio-
ventral process; tongue-shaped in lateral view.
Anal segment short and stout as shown in Figs
17, 19 and 20; in dorsal view 2 times longer than
wide; compactly connected with pygofer, immov-
able; anal styles, not beyond anal segment; anal
opening pear-like in dorsal view. Genital styles as
shown in Figs 17, 18 and 21, in ventral view, wid-
ening to apex, internal processes broad, not


touching each other; in lateral view, ventral mar-
gin curving upward, dorsal margin strongly bend-
ing upward; compactly connected with connec-
tive, immovable. Aedeagus stout, structure sim-
ple; each side with a broad spine at apex of
aedeagal shaft, right one curving dorsad, left one
relatively straight, directed up-cephalad. Connec-
tive relatively slender, the width of aedeagal shaft
1.33 times as wide as width of connective plus
ventral arm. Flagellum strongly sclerotised,
freely movable, structure simple, with a barb-
shaped spine at apex.

Type Material. Holotype: 6, Leigongshan Na-
tional Natural Reserve, Leishan County, Guizhou
Province, China, 13 May 1985, Z.-Z. Li.
Paratypes: 766, 99 9, same data as holotype;
2S 39 9, Guiyang, Guizhou Province, China,
June 1983, Students of Grade 79, Major Plant-
protecting; 266, 19, Huaxi (1000m), Guiyang,
Guizhou Province, 20 May 2007, Q.-Z. Song; 16,
29 9, Forest Park (1000m), Guiyang, Guizhou
Province, China, 20 May 2007, X.-S. Chen.

Etymology. The name is derived from the Latin
words flagell (flagellate) and hamus (hook), which
refers to the hook-like spine of flagellum.
Distribution: Southwest China (Guizhou Prov-
ince).
Remarks. This new species is similar in ap-
pearance to B. ocellatus, but differs from the lat-
ter in the shape of the anal segment and the num-
ber of spines on the flagellum.

ACKNOWLEDGMENTS

We are grateful to Prof. Zi-Zhong Li, Ms. Qiong-
Zhang Song (Institute of Entomology, Guizhou Univer-
sity, China) for providing valuable specimens. We thank
Prof. Dr. Shun-Chern Tsaur (Research Center for Biodi-
versity, Academia Sinica, Taibei, Taiwan), Kun-Wei
Huang (National Museum of Natural Science, Tai-
chung, Taiwan) and Ms. Gail Charabin (Saskatoon Re-
search Centre, Agriculture and Agri-Food Canada) for
obtaining literature. This work was supported by the
National Natural Science Foundation of China
(No.30560020, 31060290), the China Postdoctoral Sci-
ence Foundation (No. 2005037111), the Program for
New Century Excellent Talents in University (NCET-
07-0220), the Specialized Research Fund for the Doc-
toral Program of Higher Education (No. 20060657001),
and the International Science and Technology Coopera-
tion Program of Guizhou (20107005).

REFERENCES CITED

CHOU, I., LU, J.-S., HUANG, J., AND WANG, S.-Z. 1985.
Homoptera, Fulgoroidea. Economic Insect Fauna of
China. Fasc. 36. Science Press, Beijing pp. 1-152.
CHOU, I. WANG, Y.-L, HUANG, B.-K., AND YUAN, X.-Q.
1998. Homoptera: Fulgoroidea: Cixiidae, pp. 379-382
In B.-K Huang [ed.], Insect Fauna of Fujian Prov-
ince, Volume II. Fuzhou, China, Science and Tech-
nology Press.







56 Florida Ento



FENNAH, R. G. 1956. Fulgoroidea from Southern China.
Proc. California Acad. Sci. 28: 441-527.
HORI, Y. 1982. The Genus Betacixius Matsumura, 1914
(Homoptera: Cixiidae) of Formosa, pp. 175-182 In M.
Sat6, Y. Hori, Y. Arita and T. Okadome [eds.], Special
issue to the memory of retirement of Emeritus Pro-
fessor Michio Chdj6. Association of the Memorial Is-
sue of Emeritus Professor M. Chdj6 C/O Biological
Laboratory, Nagoya Women's University, Nagoya,
Japan.
HUA, L.-Z. 2002. Catalogue of Insects of China I. Guang-
zhou, China: Sun Yat-Sen University Press.
JACOBI, A. 1944. Die Zikadenfauna der Provinz Fukien
in Stidchina und ihre tiergeographischen Beziehun-
gen. Mitteilungen der Mtinchner Entomologischen
Gesellschaft 34: 5-66.


1m


ologist 94(1) March 2011



LOCKER, B., FLETCHER, M. J., LARIVIERE, M.-C., AND
GURR, G. M. 2006. The Australian Pentastirini
(Hemiptera: Fulgoromorpha: Cixiidae). Zootaxa
1290: 1-138.
MATSUMURA, S. 1914. Die Cixiinen Japans. Annota-
tiones Zoologicae Japanenses, Tokyo 8: 393-434
METCALF, Z. P. 1936. General Catalogue of the Homoptera
Fascicle IV Fulgoroidea, Part 2, Cixiidae. pp 269.
SCHUMACHER, F. 1915. Der gegenwartige Stand unserer
Kenntnis von der Homopteren-Fauna der Insel For-
mosa unter besonderer Beriicksichtigung von Sau-
ter'schem Material. Mitteilungen aus dem Zoologis-
chen Museum in Berlin. Berlin 8: 73-134
TSAUR, S.-C., HSU, T.-C., AND VAN STALLE, J. 1991. Cixi-
idae of Taiwan, Part V. Cixiini except Cixius. J. Tai-
wan Museum 44: 1-78.







Scheffrahn & Crowe: Termites on Boats


SHIP-BORNE TERMITE (ISOPTERA) BORDER INTERCEPTIONS IN
AUSTRALIA AND ONBOARD INFESTATIONS IN FLORIDA, 1986-2009

RUDOLF H. SCHEFFRAHN AND WILLIAM CROWE2
'Fort Lauderdale Research and Education Center, University of Florida,
Institute of Food and Agricultural Sciences, 3205 College Avenue, Davie, Florida, 33314, U.S.A.
rhsc@ufl.edu

Australian Quarantine and Inspection Service, P.O. Box 222, Hamilton Central, Queensland, 4007, Australia
bill.crowe@aqis.gov.au

ABSTRACT

Alate termite flights from mature colonies infesting marine vessels is a primary mechanism
for anthropogenic transoceanic establishment of invasive termite species. A taxonomic re-
view is given of 133 recorded termite infestations onboard vessels in Australia and Florida
between 1986 and 2009. The differing governmental approaches to regulating entry by for-
eign boats appears to reflect the relative frequency of exotic termite establishments in Aus-
tralia and Florida.

Key Words: invasive species, biosecurity, overwater dispersal, Kalotermitidae, Rhinotermiti-
dae, Termitidae

RESUME

El vuelo de termitas aladas de colonies maduras que infestan barcos es el mecanismo prin-
cipal para el movimiento transocednico y establecimiento de species de termitas invasoras.
Se provee una revision taxon6mica de 133 infestaciones de termitas encontradas en barcos
en Australia y la Florida entire 1986 y 2009. Los diferentes enfoques gubernamentales en re-
gular la entrada de barcos extranjeros tienden a reflejar la frecuencia relative de estableci-
miento de termitas ex6ticas en Australia y la Florida.


Natural overwater dispersal of infested flot-
sam and anthropogenic dispersal by maritime
vessels are the 2 primary means by which ter-
mites are transported across distant sea barriers
(Scheffrahn et al. 2009). The cessation of rapid
late Pleistocene/Holocene sea level rise at about
7K years before present (ybp, Fleming et al. 1998)
predates the first known long-distance human
maritime voyages by some 3.5K ybp (Anderson et
al. 2006). Therefore, contemporary nonathropo-
genic termite distributions were established be-
tween these 2 periods. Distant termite dispersal
by flotsam can be presumed to be a very rare
event with a success rate inversely proportional
to distance. Establishment of the depauparate
native terrestrial faunas on distant oceanic volca-
nic islands such as Hawaii was the result of tran-
soceanic dispersal (Cowie & Holland 2006). Near-
shore islands like the Krakatau can be colonized
much more frequently by both flotsam transport
and cross-water termite dispersal flights (Gath-
orne-Hardy & Jones 2000).
Shipboard transport of termite colonies, where
success is not affected by travel distance, has
been suspected in recent (Gay 1967; Scheffrahn &
Su 2005) and early (Scheffrahn et al. 2009) trans-
oceanic termite establishments. Vessels can be
colonized during construction (usually only Kalo-


termitidae) or by alates (all taxa) flying onboard
during dockage, either on water or in dry dock.
Hochmair & Scheffrahn (2010) showed a strong
correlation between land-borne infestations of
Coptotermes spp. in Florida and their distance to
maritime boat dockage suggesting that marine
vessels are predominant vehicles for dissemina-
tion of this pest genus.
Within the last century, 6 exotic termite spe-
cies have become established in Florida (Schef-
frahn et al. 1988; Scheffrahn et al. 1992; Schef-
frahn et al. 2002; Scheffrahn & Su 1995; Su et al.
1997), more than any other state or territory in
America, followed by Hawaii with 5 species (or 4
species if Zootermopsis angusticollis Hagen is not
established, Woodrow et al. 1999; Yeap et al.
2007). Australia has a slightly greater human
population than Florida, a much larger tidal
coastline, and both share similar economies, cli-
mates, pleasure boating industries, and proximi-
ties to tropical nations to the north and south, re-
spectively. Yet, in the last century, a single exotic
termite, Cr. brevis (Walker), has become estab-
lished in Australia (Peters 1990). As for other sus-
pected exotic Cryptotermes, Gay & Watson (1982)
determined that Cr. cynocephalus Light and Cr.
domesticus (Haviland) are endemic to northeast
Australia. Gay (1967) reported that Cr. dudleyi







Florida Entomologist 94(1)


Banks, an exotic drywood termite from Southeast
Asia, was already established in Darwin by 1913.
In a further attempt to understand the dynam-
ics and taxa involved in exotic termite establish-
ments, we provide a summary of onboard termite
infestations in Florida and border interceptions
in Australia and we contrast the regulatory pro-
cedures used for boats arriving from foreign
ports.

MATERIALS AND METHODS

Termite specimens from Australia were found
during interceptive inspections by WC and other
Australian Quarantine and Inspection Service
(AQIS) personnel. Florida samples were collected
by or submitted to RHS by pest control profes-
sionals and boat owners or operators. In both
cases, onboard specimens were collected and
stored in ethanol. Identifications were made by
the authors using voucher specimens from their
respective collections. Other information sought
included date of collection, given in Table 1 only in
years, location collected (city), vessel origin (if
known), and vessel and/or infestation type. For
species identification, samples were required to
contain morphologically robust winged images
and/or soldiers. Workers were identified morpho-
logically only to genus.

RESULTS AND DISCUSSION

During 1986-2009, 74 and 59 termite incidents
onboard boats were recorded in Australia and Flor-
ida, respectively (Table 1). The Australian records
are comprehensive and represent all known AQIS
interceptions. The Florida incidents represent an
informal and very incomplete sampling of the ac-
tual number of boat infestations occurring around
the State. Three vessels were infested simulta-
neously by 2 species and each is recorded as a sep-
arate incident in Table 1. Unlike Australia where
only Cr brevis is established, most boats in Florida
are infested in their home waters where exotic spe-
cies abound. This phenomenon enhances the
spread of termites in Florida from boat-to-land or
land-to-boat by dockside dispersal flights and also
elevates the likelihood that boats voyaging from
Florida could spread termites to foreign ports. Al-
though long, open ocean voyages are not the norm
for Florida boaters, some will "island-hop"
throughout the West Indies. One yacht, suspected
of acquiring Co. gestroi Wasmann (= havilandi,
Kirton & Brown 2003) during a winter dockage in
the Turks and Caicos Islands (Scheffrahn & Su
2005) was simultaneously infested with Incisiter-
mes minor (Hagen). It was presumed that the lat-
ter species infested this boat while under construc-
tion in San Diego, California.
The subterranean genus Coptotermes (Family
Rhinotermitidae) was observed in 53% of all boat


infestations (Fig. 1) with Co. formosanus Shiraki
being the most common onboard pest in both
Florida (27 records) and Australia (13 records)
followed by Co. gestroi with 15 and 4 records, re-
spectively. Three other infestations by subterra-
nean termites were recorded including 2 by Reti-
culitermes virginicus (Banks) in Florida and 1by
Heterotermes sp. in a boat that sailed from Flor-
ida to Grand Cayman Island. The second most
prevalent genus, at 30% of boat infestations, was
Cryptotermes (Family Kalotermidae). Australian
interceptions yielding 8 infestations each of Cr
brevis (Walker) and Cr domesticus (Haviland), 3
infestations of Cr dudleyi Banks, 2 of Cr cyno-
cephalus Light, and 15 Cr. species undetermined.
Only 3 infestations of Cr brevis were recorded
from Florida; however, fumigations for this spe-
cies are so routine in Florida that samples are sel-
dom collected for identification. One pest control
company in Fort Lauderdale estimates that it is
contracted to fumigate about 15 boats a year for
drywood termites (read Cr brevis, Edwards, J. K.,
personal communication). On the other hand, 6
infestations of I minor were recorded in both
Australia and Florida. Alates of I minor are
much more robust and dark (reddish pronotum
and head) than Cr brevis and have a different
flight season and diel periodicity. Therefore, I mi-
nor flights prompt elevated identification re-
quests by the Florida pest control industry. Aus-
tralia recorded a single shipboard infestation
each of immigrants (Snyder) and I. sp., while in
Florida, a single infestation of I. snyderi (Light)
was observed on a houseboat in Key West. The
most unexpected find of this study was a mature
infestation in 2009 of Rugitermes panamae (Sny-
der) from an itinerant yacht intercepted while
visiting Bundaberg Australia. The yacht was ap-
parently infested by this "dampwood" species dur-
ing a voyage in 2003 to Central America. Colonies
of the predominantly arboreal genus Nasutiter-
mes (Termitidae) were found on boats 3 times
during the last 25 years. Nasutitermes acajutlae
(Holmgren) was found twice and N. nigriceps
(Haldeman) once in Florida. Although not re-
corded in Table 1, N. corniger (Motschulsky) was
found infesting 2 boats in dry dock in Dania
Beach, Florida, as part of a land-borne infestation
of this pest (Scheffrahn et al. 2002).
We suggest herein that the difference in the
number of exotic termite species established in
Florida versus Australia is attributable, at least
in part, to differing laws and regulations intended
to exclude exotic pests. The U.S. Customs and
Border Patrol (CBP) requires that pleasure ves-
sels arriving in the U.S. from a foreign port must
report their arrival by telephone and be directed,
with passengers and crew, to the nearest port of
entry or nearest designated reporting location for
a CBP face-to-face interview and/or vessel inspec-
tion (Anonymous 2009). Inspections focus on im-


March 2011








Scheffrahn & Crowe: Termites on Boats


TABLE 1. TERMITE INFESTATIONS BY GENUS AND SPECIES ONBOARD VESSELS DURING 1986-2009 (VOUCHER SPECIMENS
IN THE UNIVERSITY OF FLORIDA TERMITE COLLECTION OR AQIS RECORDS).


Vessel location
where termites found

Islamorada Key, FL
North Palm Beach, FL
Fort Lauderdale, FL
Jacksonville, FL
Lighthouse Point, FL
Hypoluxo, FL
Palm Beach, FL
Brunswick, Georgia
Palm Beach Gardens, FL
Hillsborough Beach, FL
Hallandale, FL
Tampa, FL
Pompano Beach, FL
Tampa, FL
Fort Lauderdale, FL
Fort Lauderdale, FL

Holmes Beach, FL
Dania Beach, FL
Hollywood, FL
Fort Lauderdale, FL
Fort Lauderdale, FL

Fort Lauderdale, FL
Jacksonville Beach, FL
Marathon Key, FL
Fort Lauderdale, FL
Volusia County, FL

Panama City, FL
Lake Park, FL
Fort Pierce, FL
Hollywood, FL
Fort Lauderdale, FL
Key West, FL
Key West, FL

Key West, FL
Tequesta, FL
Key Largo, FL
Key West, FL
Key West, FL
Miami Beach, FL

Stock Island Key, FL


Vessel origin'


Tennessee










Hong Kong


Hong Kong



Jamaica
Virgin Gordo, B.V.I.
Turks, Caicos






Cuba





Key West


St. Petersburg, FL
Boca Chica Key, FL Key West
Stock Island (Key West), FL
Franklin, Louisiana Florida
Marathon Key, FL
Cudjoe Key, FL
Grand Cayman, Cayman Is. Florida

Unknown if blank.
dead images only, no live infestation.


Genus


Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes

Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes

Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes

Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes

Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes

Coptotermes

Coptotermes
Coptotermes
Coptotermes
Cryptotermes
Cryptotermes
Cryptotermes
Heterotermes


Species Year Vessel comments


formosanus
formosanus
formosanus
formosanus
formosanus
formosanus
formosanus
formosanus
formosanus
formosanus
formosanus
formosanus
formosanus
formosanus
formosanus
formosanus

formosanus
formosanus
formosanus
formosanus
formosanus

formosanus
formosanus
formosanus
formosanus
formosanus

formosanus
formosanus
gestroi
gestroi
gestroi
gestroi
gestroi

gestroi
gestroi
gestroi
gestroi
gestroi
gestroi

gestroi

gestroi
gestroi
gestroi
breuis
breuis
breuis
sp.


1986 boat
1995 boat
1995 11 m boat
1997 cable ship
1998 boat
1998 26 m boat
1999 9 mboat
1999 10 m boat
2000 9 mboat
2000 large boat
2000 23 m boat
2001 10m boat
2001 16m boat
2002 10 m boat
2002 26 m speed boat
2002 15 m cabin
cruiser
2002 boat
2003 11 m boat
2004 boat
2004 boat
2004 18 m fishing
yacht
2005 15 m sailboat
2006 13 m boat
2006 small boat
2008 15 m boat
2008 18 mcabin
cruiser
2008 9 mboat
2008 8 mboat
1991 Boat
1995 boat in dry dock
2001 27m yacht
2003 15 m sailboat
2005 houseboat, nest
with queen
2005 8 mmotorboat
2005 9 m fishing boat
2006 Sailboat
2007 Sailboat
2007 Boat
2007 12 m fishing boat
transom
2007 15 m cabin
cruiser
2007 11 m Yacht
2007 sailboat
2007 Boat
2000 17 m boat
2005 Sailboat
2006 Boat
1995 Boat








Florida Entomologist 94(1)


March 2011


TABLE 1. (CONTINUED) TERMITE INFESTATIONS BY GENUS AND SPECIES ONBOARD VESSELS DURING 1986-2009
(VOUCHER SPECIMENS IN THE UNIVERSITY OF FLORIDA TERMITE COLLECTION OR AQIS RECORDS).


Vessel location
where termites found

Fort Lauderdale, FL
Miami, FL
Fort Lauderdale, FL
Marathon Key, FL
Dania Beach, FL
St. Augustine, FL
Key West, FL
Fort Lauderdale, FL
Jacksonville, FL
Fort Lauderdale, FL
Key West, FL
Jacksonville Beach, FL
Darwin, NT, AUS
Perth, WA, AUS
Brisbane, Qld, AUS
Sydney, NSW, AUS
Brisbane, Qld, AUS
Brisbane, Qld, AUS
Brisbane, Qld, AUS
Brisbane, Qld, AUS

Townsville, Qld, AUS
Bundaberg, Qld, AUS
Brisbane, Qld, AUS
Newcastle, NSW, AUS
Brisbane, Qld, AUS
Darwin, NT, AUS
Darwin, NT, AUS
Bundaberg, Qld, AUS
Brisbane, Qld, AUS
Brisbane, Qld, AUS
Cairns, Qld, AUS

Sydney, NSW, AUS
Brisbane, Qld, AUS

Brisbane, Qld, AUS
Brisbane, Qld, AUS

Mackay, Qld, AUS
Brisbane, Qld, AUS
Brisbane, Qld, AUS
Perth, WA, AUS

Townsville, Qld, AUS
Brisbane, Qld, AUS

Brisbane, Qld, AUS
Cardwell, Qld, AUS
Brisbane, Qld, AUS
Brisbane, Qld, AUS
Airlie Beach, Qld, AUS


Vessel origin'


Los Angeles, CA
San Diego, CA
Taiwan
western Mexico
FL Keys

St. Thomas U.S.V.I.
Puerto Rico



China
Hong Kong
China

Hong Kong
China
USA / Japan
USA

Hong Kong/Asia
Hawaii
Japan
China
USA
Thailand
Thailand
Marshall Islands
China
USA
AUS

Unknown
New Caledonia

Taiwan
Hong Kong

Taiwan
France
USA
Singapore


USA

USA

South Africa
USA
USA


Genus


Incisitermes
Incisitermes
Incisitermes
Incisitermes
Incisitermes
Incisitermes
Incisitermes
Nasutitermes
Nasutitermes
Nasutitermes
Reticulitermes
Reticulitermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes

Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes
Coptotermes


Species Year Vessel comments


minor
minor
minor
minor
minor
minor
snyderi
acajutlae
acajutlae
nigriceps
virginicus
uirginicus
formosanus
formosanus
formosanus
formosanus
formosanus
formosanus
formosanus
formosanus

formosanus
formosanus
formosanus
formosanus
formosanus
gestroi
gestroi
gestroi
gestroi
sp.
sp.


Coptotermes sp.
Coptotermes sp.


Coptotermes
Coptotermes

Coptotermes
Coptotermes
Coptotermes
Coptotermes

Cryptotermes
Cryptotermes

Cryptotermes
Cryptotermes
Cryptotermes
Cryptotermes
Cryptotermes


sp.
sp.

sp.
sp.
sp.
travians?

brevis
brevis

brevis
brevis
brevis
brevis
brevis


2000
2000
2001
2006
2007
2008
2000
2002
2002
1996
2000
2003
1994
2000
2002
2003
2003
2003
2005
2005

2005


Boat
26 m boat
27 m yacht
Boat
20 m boat
Boat
Houseboat
15 mboat
container on ship
Sailboat
Houseboat
Boat
boat (refugee)
Boat
boat, fibreglass
Yacht
boat
Yacht
9 m boat
boat
(with I. minor)
Boat


2006 itinerant yacht
2007 Boat
2008 Boat
2009 itinerant yacht
1986 Yacht
1994 Boat
1996 Yacht
2003 Boat
2002 Yacht
2005 dinghy fromTIto
Cairns
2006 navy boat
2008 boat (returning
AUS yacht)
2008 Boat
2005 yacht flybridgee
in lockers)
2005 Boat
2008 Yacht
2008 Boat
2002 boat, fibreglass &
wood
1989 Yacht
2003 wooden yacht
(with I. Minor)
2005 Superyacht
2006 Trimaran
2007 Boat
2008 Boat
2009 Catalina
400 MK II


Unknown if blank.
dead images only, no live infestation.








Scheffrahn & Crowe: Termites on Boats


TABLE 1. (CONTINUED) TERMITE INFESTATIONS BY GENUS AND SPECIES ONBOARD VESSELS DURING 1986-2009
(VOUCHER SPECIMENS IN THE UNIVERSITY OF FLORIDA TERMITE COLLECTION OR AQIS RECORDS).


Vessel location
where termites found

Mackay, Qld, AUS
Bundaberg, Qld, AUS
Darwin, NT, AUS

Darwin, NT, AUS

Darwin, NT, AUS
Darwin, NT, AUS
Brisbane, Qld, AUS
Sydney, NSW, AUS
Broome, WA, AUS

Gove, NT, AUS

Gove, NT, AUS

Broome, WA, AUS

Darwin, NT, AUS
Darwin, NT, AUS
Darwin, NT, AUS

Darwin, NT, AUS
Gove, NT, AUS

Darwin, NT, AUS
Gove, NT, AUS

Thursday Island, AUS

Darwin, NT, AUS
Darwin, NT, AUS

Broome, WA, AUS

Broome, WA, AUS

Bundaberg, Qld, AUS
Broome, WA, AUS

Broome, WA, AUS

Broome, WA, AUS

Darwin, NT, AUS

Broome, WA, AUS

Brisbane, Qld, AUS
Bundaberg, Qld, AUS
Brisbane, Qld, AUS
Brisbane, Qld, AUS

Brisbane, Qld, AUS


Vessel origin'

USA
USA
Indonesia

Indonesia

Indonesia
Indonesia
Vanuatu

Papela, Roti, Indonesia

Karja Sama, Indonesia

Indonesia

Indonesia

Indonesia
Philippines
Indonesia

Indonesia
Indonesia

Vietnam
Indonesia

Indonesia

Indonesia
Indonesia

Papela Roti, Indonesia

Indonesia

USA
Sulawesi, Indonesia

Indonesia

Indonesia

Indonesia

Indonesia

Thailand
Hawaii

USA

USA


Genus

Cryptotermes
Cryptotermes
Cryptotermes

Cryptotermes

Cryptotermes
Cryptotermes
Cryptotermes
Cryptotermes
Cryptotermes

Cryptotermes

Cryptotermes

Cryptotermes

Cryptotermes
Cryptotermes
Cryptotermes

Cryptotermes
Cryptotermes

Cryptotermes
Cryptotermes

Cryptotermes

Cryptotermes
Cryptotermes

Cryptotermes

Cryptotermes

Cryptotermes
Cryptotermes

Cryptotermes

Cryptotermes

Cryptotermes

Cryptotermes

Drepanotermes2
Incisitermes
Incisitermes
Incisitermes

Incisitermes


Species

brevis?
cavifrons'
cynocephalus

cynocephalus

domesticus
domesticus
domesticus
domesticus
domesticus

domesticus

domesticus

domesticus

dudleyi
dudleyi
dudleyi

sp.
sp.

sp.
sp.

sp.

sp.
sp.

sp.

sp.

sp.
sp.

sp.

sp.

sp.

sp.

sp.
immigrants
minor
minor

minor


Year

2009
2008
2005

2009

1986
1987
1999
2003
2005

2006

2006

2007

1994
2006
2008

1993
1993

2001
2004

2004

2004
2005

2006

2007

2007
2005

2005

2005

2005

2009

2008
2007
2001
2003

2005


Vessel comments

28 m super yacht
Yacht
foreign fishing
vessel
foreign fishing
vessel
Yacht
yacht
boat
yacht
foreign fishing
vessel
foreign fishing
vessel
foreign fishing
vessel
foreign fishing
vessel
boat
boat
foreign fishing
vessel
boat
foreign fishing
vessel
boat
foreign fishing
vessel
foreign fishing
vessel
boat
foreign fishing
vessel
foreign fishing
vessel
foreign fishing
vessel
boat
foreign fishing
vessel
foreign fishing
vessel
foreign fishing
vessel
foreign fishing
vessel
foreign fishing
vessel
boat
boat
yacht
wooden yacht
(with Cr. brevis)
boat (with Co.
formosanus)


Unknown if blank.
dead images only, no live infestation.







Florida Entomologist 94(1)


TABLE 1. (CONTINUED) TERMITE INFESTATIONS BY GENUS AND SPECIES ONBOARD VESSELS DURING 1986-2009
(VOUCHER SPECIMENS IN THE UNIVERSITY OF FLORIDA TERMITE COLLECTION OR AQIS RECORDS).

Vessel location
where termites found Vessel origin' Genus Species Year Vessel comments

Brisbane, Qld, AUS Fiji (made in USA) Incisitermes minor 2006 super yacht
Bundaberg, Qld, AUS USA Incisitermes minor 2006 trimaran
Cairns, Qld, AUS USA Incisitermes minor 2009 trimaran
Broome, WA, AUS Papela Roti, Indonesia Incisitermes sp. 2006 foreign fishing
vessel
Bundaberg, Qld, AUS Central Amer. Rugitermes panamae 2009 Yacht

Unknown if blank.
dead images only, no live infestation.




50



40


4-




20



10 --
(ft








0 3











Fig. 1. Frequency of termite genera collected on vessels in Australia and Florida. "Other" includes Drepanoter-
mes and Rugitermes from Australia and Heterotermes from Florida.


migration compliance by the passengers and
crew, possible illegal contraband, and agricul-
tural pests in cargo. Structural and household
pests, which are usually disassociated with cargo
and dwell within the vessel's own structure, are
not mandated for inspection. In contrast to Flor-
ida practices, passengers and crew aboard vessels
arriving to Australia from a foreign port must ob-
tain clearance by the Australian Customs and
Border Protection Service and the Australian
Quarantine and Inspection Service (AQIS). Ves-
sels with timber in their cargo or construction


must also be inspected by AQIS. The level of AQIS
inspection required will depend on the amount of
timber present and the construction/re-fit and
sailing history of the vessel. The inspection can be
conducted by an AQIS quarantine officer or AQIS
entomologist with or without a licensed pest con-
trol professional and approved termite detection
method. If termites are found upon inspection,
the vessel must be fumigated with methyl bro-
mide (AQIS method T9047) or sulfuryl fluoride
(AQIS method T9090) at the owner's expense
(Anonymous 2010).


March 2011







Scheffrahn & Crowe: Termites on Boats


ACKNOWLEDGMENTS

We thank boat owners and pest control companies in
Florida for submitting termite samples to RHS.


REFERENCES CITED

ANDERSON, A., CHAPPELL, J., GAGAN, M., AND GROVE, R.
2006. Prehistoric maritime migration in the Pacific
islands: An hypothesis of ENSO forcing. The Ho-
locene 16: 1-6.
ANONYMOUS. 2009. Code of Federal Regulations (United
States) 19CFR4.2.
ANONYMOUS. 2010. ICON Condition C9645. Australian
Quarantine and Inspection Service.
COWIE, R. H., AND HOLLAND, B. S. 2006. Dispersal is
fundamental to biogeography and the evolution of
biodiversity on oceanic islands. J. Biogeography 33:
193-198.
FLEMING, K., JOHNSTON, P., ZWARTZ, D., YOKOYAMA, Y.,
LAMBECK, K., AND CHAPPEL, J. 1998. Refining the
eustatic sea-level curve since the last glacial maxi-
mum using far- and intermediate-field sites. Earth
and Planetary Science Letters 163: 327-342.
GATHORNE-HARDY, F. J., AND JONES, D. T. 2000. The re-
colonization of the Krakatau islands by termites
(Isoptera), and their biogeographical origins. Biol. J.
Linnean Soc. 71: 251-267.
GAY, F. J. 1967. A world review of introduced species of
termites. CSIRO Bulletin Melbourne, Australia 286:
1-88.
GAY, F. J., AND WATSON, J. A. L. 1982. The genus Cryp-
totermes in Australia (Isoptera: Kalotermitidae).
Australian J. Zool. Supplementary Series 30, 88: 1-
64.
HOCHMAIR, H. H., AND SCHEFFRAHN, R. H. 2010. Spatial
association of marine dockage with land-borne infes-
tations of invasive termites in urban South Florida.
J. Econ. Entomol. 103: 1338-1346.


KIRTON, L. G., AND BROWN, V. K. 2003. The taxonomic
status of pest species of Coptotermes in Southeast
Asia: Resolving the paradox in the pest status of the
termites, Coptotermes gestroi, C. havilandi and C. tra-
vians (Isoptera: Rhinotermitidae). Sociobiol. 42: 43-
63.
PETERS, B. C. 1990. Infestations of Cryptotermes brevis
(Walker) (Isoptera: Kalotermitidae) in Queensland,
Australia. 1. History, detection and identification.
Australian Forester 53: 79-88.
SCHEFFRAHN, R. H., AND SU, N.-Y. 1995. A new subterra-
nean termite introduced to Florida: Heterotermes
Froggatt (Rhinotermitidae: Heterotermitinae) estab-
lished in Miami. Florida Entomol. 78: 623-627.
SCHEFFRAHN, R. H., AND SU, N.-Y. 2005. Distribution of
the termite genus Coptotermes (Isoptera: Rhinoter-
mitidae) in Florida. Florida Entomol. 88: 201-203.
SCHEFFRAHN, R. H., CABRERA, B. J., KERN JR., W. H.,
AND SU, N.-Y. 2002. Nasutitermes costalis (Isoptera:
Termitidae) in Florida: First record of a non-endemic
establishment by a higher termite. Florida Entomol.
85: 273-275.
SCHEFFRAHN, R. H., KRECK, J. RIPA, R., AND LUPPICHINI,
P. 2009. Endemic origin and vast anthropogenic dis-
persal of the West Indian drywood termite. Biol. Inva-
sions 11: 787-799.
SCHEFFRAHN, R. H., MANGOLD, J. R., AND SU, N.-Y 1988.
A survey of structure-infesting termites of peninsular
Florida. Florida Entomol. 71: 615-630.
Su, N.-Y., SCHEFFRAHN, R. H., AND WEISSLING, T. 1997. A
new introduction of a subterranean termite, Coptoter-
mes havilandi Holmgren (Isoptera: Rhinotermitidae)
in Miami, Florida. Florida Entomol. 80: 408-411.
WOODROW, R. J., GRACE, J. K., AND YATES III, J. R. 1999.
Hawaii's Termites: An Identification Guide. Honolulu
(HI): University of Hawaii. 6 p. (Household and Struc-
tural Pests; HSP-1).
YEAP, B.-K., OTHMAN, A. S., LEE, V. S., AND LEE, C.-Y.
2007. Genetic relationship between Coptotermes
gestroi and Coptotermes vastator (Isoptera: Rhinoter-
mitidae). J. Econ. Entomol. 100: 467-474.







Florida Entomologist 94(1)


March 2011


FIRST RECORD OF THE GENUS ADOXOMYIA
(DIPTERA: STRATIOMYIDAE) WITH FOUR SPECIES FROM TURKEY

TURGAY USTUNER1 AND ABDULLAH HASBENLI2
'Selcuk University, Faculty of Science, Department of Biology, Alaaddin Keykubat Kampuisii, 42100 Selcuklu,
Konya, Turkey
E-mail: turgayustuner@gmail.com

2Gazi University, Faculty of Arts and Sciences, Department of Biology, 06500 Teknikokullar, Ankara, Turkey.

ABSTRACT

Adoxomyia aureovittata (Bigot, 1879), A. cinerascens (Loew, 1873), A. obscuripennis (Loew,
1873) andA. sarudnyi (Pleske, 1903) are recorded from Turkey for the first time. Both sexes
of the first 3 species and the male ofA. sarudnyi are redescribed and photographs of all spe-
cies are provided. The distributions of all species are briefly discussed. The male genitalia
and some other diagnostic characters of all the examined species are illustrated. An identi-
fication key to all East-Mediterranean species was constructed and is included in this report.

Key Words: Adoxomyia aureovittata, A. cinerascens, A. obscuripennis, A. sarudnyi, new
records, distribution, Turkey

RESUME

Se registran por primera vez Adoxomyia aureovittata (Bigot, 1879), A. cinerascens (Loew,
1873),A. obscuripennis (Loew, 1873) yA. sarudnyi (Pleske, 1903) para Turquia. Se proven
redescripciones y fotos de ambos sexos de las primeras 3 species y del macho deA. sarudnyi.
Se discutan la distribuci6n de todas las species. Se ilustran las genitalias de los machos y
algunas de las caracteristicas diagn6sticas de las species examinadas. Una clave para la
identificaci6n de todas las species de la region Este del Mediterraneo es incluida.


The family Stratiomyidae belongs to the sub-
order Brachycera in Diptera (Rozkosny 1982).
This large family includes more than 2650 species
in 375 genera composed of 12 subfamilies world-
wide of which 426 species in 55 genera in 7 sub-
families occur in the Palaearctic region (Woodley
2001). So far 48 species in 14 genera and 7 sub-
families (Beridinae, Pachygastrinae, Clitellarii-
nae, Hermetinae, Sarginae, S' .,ri,.. i... Nem-
otelinae) have been recorded in Turkey (Rozkosny
& Nartshuk 1988; Woodley 2001; Ustiiner et al.
2002, 2003; Ustfiner & Hasbenli 2003, 2004).
The subfamily Clitellariinae contains 50 gen-
era worldwide, 10 genera in the Palaearctic re-
gion and 1 genus (Pycnomalla) in Turkey (Wood-
ley 2001; Ustfiner et al. 2002). The genus Adox-
omyia (Kertesz, 1907) belongs to the subfamily
Clitellariinae and includes 36 described species.
They are distributed in the Palaearctic region (16
species), the Nearctic region (13 species), the Neo-
tropical region (4 species), the Oriental region (3
species) and the Afrotropical region (2 species)
(Woodley 2001; Hauser 2002; Nartshuk 2004).
Palearctic species ofAdoxomyia are found mainly
in south-eastern Europe, Transcaucasus, Near
East, Central Asia and China (Rozkosny 1983;
Rozkosny & Nartshuk 1988; Nartshuk 2004).
Adoxomyia had not been recorded in Turkey be-
fore this report.


The larvae of Adoxomyia are known only for
some Nearctic and one Oriental species; they
were found in decaying cacti and nests of pack
rats (Neotoma sp.) (McFadden 1967; James & Mc-
Fadden 1969).
In addition to the 4 species recorded in Turkey at
least 5 additional species may occur here. A dahlii
(Meigen 1830) is known from southern Europe (incl.
Ukraine), Armenia and Israel. A. ruficornis (Loew
1873) occurs in Azerbaijan, Iran and Kyrgyzstan.A.
hermonensis Lidner, 1975 and A. paleastinensis
Lindner, 1937 were described from Israel and A
transcaucasica Nartshuk, 2004 is based on types
from Armenia and Azerbaijan. According to a recent
paper by Nartshuk (2004), A. portschinskii (Pleske)
is a mere synonym ofA. dahlii (Meigen).
MATERIALS
A total of 16 specimens (12 males and 4 fe-
males) were collected by hand net at Antalya, Is-
parta, and Konya in Turkey between 1999 and
2001. Some specimens ofAdoxomyia were caught
sunbathing on stones or on the ground in dry
creek beds. The specimens are deposited in the
collection of the Selcuk University Department of
Biology in Konya, Turkey. The fact that previ-
ously the genusAdoxomyia had not been recorded
from Turkey reflects the poor knowledge of the
Turkish Stratiomyidae fauna.







Ustiiner & Hasbenli: First Records of Adoxomyia Species in Turkey


THE GENUSADOXOMYIA Kert6sz 1907

The generic name Adoxomyia was proposed
by Kertesz (1907) and according to the catalog
published a year later (Kertesz 1908) this ge-
nus embraced 23 species. In 1923, Kertesz tried
to establish a new separate genus, Euclitellaria
Kertesz, but it was not accepted by Pleske
(1925) and subsequent authors. The last com-
prehensive key to the Palaearctic species is that
by Lindner (1937), who followed Pleske's con-
cept ofAdoxomyia in a broad sense. Besides the
work by Rozkosny (1983), which treated the Eu-
ropean species, some other relevant papers


were chiefly devoted to descriptions of separate
additional species (Ouchi 1938; Dusek & Rozko-
sny 1963; Lindner 1975; Nartshuk 2004).
In Turkey, the species belonging to Adoxomyia
are mid-sized (6-11 mm) with a predominantly
black body. The eyes, which are contiguous in
males and widely separated in females, are cov-
ered with dense hairs. The antennae are rela-
tively long, predominantly black, but can be red-
orange to dark brown in some species. Scape and
pedicel are of equal length. The flagellum consists
of 8 flagellomeres. No thoracic spines but two
scutellar spines are present. All of 4 M-veins
reach the wing margin.


KEY TO THE EAST MEDITERRANEAN SPECIES OF ADOXOMYIA

The following key is based partly on Lindner (1937). The male ofA. hermonensis and the female of
A. palaestinensis are unknown.

1. Legs completely black ..................................... ............................ 2
- Legs bicoloured or mainly ................ .............................................. 5
2. Antenna entirely black .............................................................. 3
- At least basal half of antennal flagellum red. ................................. A. ruficornis (Loew 1873)
3. Scutellar spines short, slender and bare, basal 3-4 flagellomeres in female unusually broad ............... 4
- Femora Scutellar spines longer, thickened and haired, basal 3-4 flagellomeres not as broad (Figs. 19 and 20)
................ .................... ........................ A. obscuripennis (Loew 1873)
4. Female eyes black haired, postocular band wider than scape is long; male unknown
............... ...................................... .......... herm onensis Lindner 1975
- Female eyes white haired, postocular band as wide as scape is long ........ A. transcaucasica Nartshuk 2004
5. Legs entirely yellow ................. ................... ................. A. sarudnyi (Pleske 1903)
- At least femora black....................................................................... 6
6. Antenna black ....................... .............. ............................ 7
- Abdomen with silverish white hair patches ............................. A. palaestinensis Lindner 1930
8. Abdomen with golden yellow hair patches ............................... .A. aureovittata (Bigot 1879) Y
- Abdomen with silverish white (rarely coppery) hair patches ...................................... 9
9. Male flagellum almost cylindrical, female apical flagellomere at base half as broad as scape at distal end
...................................................... .......... A dahlii (M eigen 1830)
- Male flagellum distinctly swollen in middle, female apical flagellomere broader, at most slightly narrower than
scape at distal margin (Figs. 17 and 18) ............................... .A. cinerascens (Loew 1873)


Redescription of the 4 Adoxomyia species recorded in
Turkey

1. Adoxomyia aureouittata (Bigot 1879);
see Figs. 1-4, 15-16 and 22-23.

Male: Head transversely oval, in dorsal view.
Eyes touching on frons. Hairs on eyes as long as
pedicel, dense and black. Black postocular area
swollen in lower half and narrowed in upper part,


covered with appressed yellowish hairs. Face,
cheeks and posteroventral part of head covered
with hairs as long as scape, erect, black and
partly brown. Antenna entirely black and more
slender in male than in female. Scape and pedicel
black with black and erect hairs being as long as
scape. Flagellum about four times as long as
scape and pedicel combined, the first 5 flagellom-
eres with dense yellowish pubescence. Scutum
black, with semi-erect and black hairs. Scutellum






Florida Entomologist 94(1)


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12 1


14\


Figs. 1-14. Adoxomyia aureovittata: 1- male in dorsal view, 2- female in dorsal view, 3- male in lateral view, 4-
female in lateral view; 5-8Adoxomyia cineracens 5- male in dorsal view, 6- female in dorsal view, 7- male in lateral
view, 8- female in lateral view; 9-12 Adoxomyia obscuripennis 9- male in dorsal view, 10- female in dorsal view, 11-
male in lateral view, 12- female in lateral view; 13-14. Adoxomyia sarudnyi 13- male in dorsal view, 14- male in lat-
eral view (Scale 1 mm).


and scutellar spines mainly black but tip of
scutellar spines brown. Scutellum covered with
semi-appressed sparse black hairs. Legs black
and yellow. Coxa, trochanter, and femur black ex-
cept for yellow bases of femora. Tibia mainly
black, its both ends narrowly yellowish brown.
Tarsi yellow, front tarsus darkened dorsally as
well as tarsomeres 3-5 of mid and hind legs. Coxa,
trochanter, and femur with semi-erect black
hairs. Tibia covered with appressed yellowish
hairs. Tarsi with appressed golden yellowish
hairs. Abdomen entirely black, abdominal pile
black except for golden yellow lateral markings


on tergite 4 and a transverse, golden yellow band
on tergite 5.
Female: Eyes densely black haired, hairs only
one-fourth as long as pedicel. Black postocular
area approximately as wide as fore tibia and cov-
ered with appressed yellowish hairs. Frons about
1/3 of head width, with fine longitudinal groove in
middle, black, densely punctate, with yellowish
hairs. Face black, with yellowish hairs on sides
below antennae. Remainder of face, cheeks and
posteroventral part of head covered with erect,
black hairs being as long as scape. Antenna in-


2


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13




I


March 2011


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Usttiner & Hasbenli: First Records of Adoxomyia Species in Turkey


serted at middle of head profile, partly black.
Scape black but reddish brown at tip. Pedicel and
first 5 flagellomeres dark red, last 3 flagellomeres
black. Scape and pedicel with erect, black hairs as
long as antenal scape. Flagellum about 4.5 times
as long as both basal antennal segments com-
bined, first 5 flagellomeres densely golden yellow
dusted. Thorax black, scutum with 2 golden yel-
low dusted longitudinal stripes which can be re-
duced to absent in some specimens (especially
smaller ones). Scutellum black with appressed
golden yellow hairs. Tips of scutellar spines
brown. Legs black and yellow, femora black ex-
cept for yellow base, tibiae mainly darkened, with
both ends broadly yellowish brown on fore and
mid legs. Hind tibia mainly black, with both ends
broadly yellowish brown. Tarsi yellow, with dor-
sally darkened tarsomeres 3-5.
All legs with yellowish hairs that are semi-
erect on femora and appressed on tibiae and tarsi.
Abdomen black but lateral markings on tergites
2-4 and a triangular apical spot on tergite 5 with
golden yellow hair patches.
Adoxomyia aureovittata (Bigot 1879) was de-
scribed from an unknown locality. Apparently,
this species is distributed in the eastern part of
the Mediterranean area. In addition to Turkey it
was also found in Greece (M. Hauser, personal
communication).
Material Examined: Turkey: Konya, Hadim,
between Tosmur and Gevne Village, Gevne Valley,
1450-2020 m, 10 June 1999, 1 male and 1 female;
Konya, Taskent, Begreli Village, Gevne Valley,
1570 m, 10 August 2001, 1 male; all T Ustiiner
leg.
Distribution: Greece, Turkey.

2. Adoxomyia cinerascens (Loew 1873); see Figs
5-8, 17-18 and 24-25.

Male: Hairs above compound eyes as long as
pedicel, densely black. Postocular area black, cov-
ered with pale yellowish, appressed hairs. Fron-
tal triangle black with dense, erect, pale yellow-
ish hairs being about 1.5 times as long as the
scape. Hairs on black face erect, as long as
pedicel, pale yellowish. Antenna as long as head
in lateral view. Scape, pedicel and first 3 basal
flagellomeres brownish orange, rest of flagellum
black. Hairs on scape and pedicel erect, as long as
scape, pale yellowish. Thorax including scutellum
black, with dense yellowish hairs. Scutellar
spines yellow. Wing veins brown. Legs bicoloured,
coxae black, trochanters brownish, femora and
tibiae mainly black, partly yellow at tips. Fore
and hind tarsi yellowish on inner surface and
darkened on outer surface, mid tarsi yellow but
basal 2 tarsomeres darkened on outer surface.
Femora with sparse semi-erect pale yellow hairs.
Tibiae and tarsomeres with dense appressed yel-
low hairs. Abdomen mainly black, with trans-


verse, pale yellow hair band on posterior margin
of tergite 4.

Female: Hairs on eyes black, about 0.3 times as
long as pedicel. Postocular area black, covered
with pale yellow and semi-appressed hairs. Frons
about 0.3 of head-width, shining black and
densely punctate, with fine longitudinal groove in
middle and with sparse pale yellow hairs. Face
black, with whitish, dense, erect hairs being as
long as pedicel. Antenna long, about 1.5 times as
long as head in lateral view, bicolored and in male
more slender than in female. Scape brownish or-
ange on lower surface but darkened on upper sur-
face. Pedicel and first 3 flagellomeres brownish
orange, rest of flagellum black. Flagellomeres 2-3
darkened on outer surface. Thorax black, with ap-
pressed dense pale yellow hairs, but tip of post-
pronotal callus brownish. Scutellum black with
pale yellow hairs, scutellar spines yellow. Wing
veins brown. Legs bicoloured. Coxae black, tro-
chanters yellow. Femora and tibiae mainly black
and partly yellow on tips. Fore and hind tarsi yel-
lowish on inner surface, darkened on outer sur-
face, basal 2 tarsomeres of mid tarsi yellow, other
tarsomeres darkened on outer surface. Femora
with semi-erect pale yellow hairs. Tibiae with ap-
pressed yellowish hairs. Tarsomeres with ap-
pressed yellow hairs. Abdomen mainly black but
with pale yellow hair patches at posterior margin
of tergite 4.
Material Examined: Turkey: Antalya, Giundog-
mus district, Glineycik Village, Topraktepe place,
elev. 200 m, 23 June 1999, 7 males, 12 July 2000,
1 male; Konya, Taskent, Begreli Village, Gevne
Valley, elev. 1585 m, 10 July 2000, 1 female; all T.
Ustiiner leg.
Distribution: Palaearctic: Iran, Israel, Kaza-
khstan, Kyrgyzstan, Tajikistan (Kertesz 1908;
Lindner 1937, 1974, 1975; Rozkosny & Nartshuk
1988; Woodley 2001). This is the first record for
the fauna of Turkey

3. Adoxomyia obscuripennis (Loew 1873); Figs 9-
12, 19-20 and 28-29.

Male: Head transverse, hemispherical, eyes
touching on frons. Hairs on eyes dense, black, as
long as scape. Frontal triangle black, with silver-
ish white pubescence. Face slightly produced in
lateral view, with dense, black hairs, as long as
scape. Cheeks and posteroventral part of head
with erect black hairs. Black postocular area
prominent but narrower than pedicel in upper
half and somewhat swollen in lower half, about as
wide as antennal scape is long, covered with ap-
pressed silverish white hairs. Antenna entirely
black, flagellomeres 1-3 thickened, following
flagellomeres small and slender, last flagellomere
longer than 4 preceding combined. Thorax com-
pletely black. Scutum covered with long erect







Florida Entomologist 94(1)


Imm.


i;J
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j
-.,--- i1


4_


15 16


i /7 ***.
** ''* "
I ,
S1`


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23 % ;25 27 '
0.1 mm. 0.1 mm.
0\1 mm. mm
I 0.1 mm.


Figs. 15-21.Adoxomyia Antennae: 15-A. aureovittata male, 16-A. aureovittata female, 17-A. cineracens male,
18-A. cineracens female, 19-A. obscuripennis male, 20-A. obscuripennis female, 21-A. sarudnyi male (Scala 1 mm.):
22-23Adoxomyia aureovittata male terminalia: 22- dorsal part of male genitalia, 23- ventral part of male genitalia;
24-25 Adoxomyia cineracens male terminalia: 24- dorsal part of male genitalia, 25- ventral part of male genitalia;
26-27Adoxomyia sarudnyi male terminalia: 26- dorsal part of male genitalia, 27- ventral part of male genitalia; 28-
29 Adoxomyia obscuripennis male terminalia: 28- dorsal part of male genitalia, 29- ventral part of male genitalia
(Scale 0.1 mm.).


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March 2011


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Ustiiner & Hasbenli: First Records of Adoxomyia Species in Turkey


black hairs and short appressed silverish white
hairs. Scutellum with very strong and thick,
black scutellar spines. Legs black, but knees
brown. Femora with semi-erect silverish white
hairs. Tibiae with semi-appressed, dense, silver-
ish white hairs. Tarsomeres black on outer sur-
face, brown on inner surface, with semi-ap-
pressed, dense, silverish white hairs. Halteres
pale yellow. Abdomen entirely black, tergites 1-2
with erect, moderately long, white hairs. White
hairs also distinct on distal halfoftergite 4 and on
entire tergite 5.

Female: Hairs above compound eyes dense and
black, as long as antennal scape. Frons black, as
wide as flagellum is long, with fine median
groove. Frontal hairs as long as antennal scape,
pale. Face black, covered below antennae with
long black hairs. Cheeks and posteroventral part
of head with erect, black and yellowish hairs. Pos-
tocular area as wide as flagellum, black and with
appressed, dense, silverish white hairs. Antenna
slender, about 1.1 times as long as head. Scape,
pedicel and flagellomeres 4-7 shining black, three
basal flagellomeres black, whitish grey dusted.
Last flagellomere about 1.5 times as long as 4 pre-
ceding. Thorax black, covered with appressed, sil-
verish white hairs. Scutellum, including strong
and short scutellar spines, black. Top of post-
pronotal callus and postalar callus brownish.
Legs black but knees brown, femora with semi-
erect white hairs, tibiae with semi-appressed,
dense, silverish white hairs. Tarsomeres black on
outer surface, brown on inner surface, with semi-
appressed, dense, silverish white hairs. Halteres
pale yellow. Wings transparent and partly brown-
ish, with brown veins. Abdomen entirely black,
with erect, moderately long, silverish white hairs
on sides of tergites 2-3. White hairs also devel-
oped on distal half of tergite 4 and on entire terg-
ite 5.
Material Examined: Turkey: Isparta, Yalvag,
The Sultan Mountains, elev. 1660 m, 29 May
2001, 1 male, 2 females, T. Ustiiner leg.
Distribution: Palaearctic: Azerbaijan, Kazakh-
stan, Russia, Tajikistan, Uzbekistan (Kertesz
1908, 1923; Lindner 1937; Nartshuk 2004; Pleske
1925; Rozkosny 1983; Rozkosny & Nartshuk
1988; Woodley 2001). Adoxomyia obscuripennis
(Loew 1873) is recorded from Turkey for the first
time.

4. Adoxomyia sarudnyi (Pleske 1903); Figs 13-14,
21 and 26-27.

Male: Head transverse, hemispherical. Eyes
touching on frons, dense, black eye hairs about
0.3 times as long as scape. Frontal triangle shin-
ing black, with fine median groove and white pile
in upper part. Face, cheeks and posteroventral
part of head black with whitish hairs. Postocular


area black, about 0.4 times as wide as length of
antennal scape, somewhat swollen in lower half,
about 0.75 times as long as scape, covered with
dense, silverish white hairs. Antenna relatively
long, about twice as long as head in lateral view.
Scape and pedicel orange but darkened on outer
surface, with strong, erect, black hairs. First 3
flagellomeres orange on inner surface but dark-
ened on outer surface, rest of flagellum black.
Thorax black, with short appressed yellowish
golden hairs and long semi-appressed black hairs
intermixed. Tops of postpronotal callus reddish
brown. Scutellum black and scutellar spines yel-
low. Wings transparent brownish, basal wing
veins bright orange and distal veins brown, wing
tip much darker and contrasting to clear wing
base. Legs entirely bright yellow to orange except
for black coxae. Fore tarsi yellowish on inner sur-
face, darkened on outer surface. Mid and hind
tarsi yellow, but last three tarsomeres darkened
on outer surface. Femora with semi-erect, sparse,
yellowish hairs. Tibia with appressed, dense, yel-
lowish hairs. Tarsi with thick, adpressed, dense,
yellowish hairs. Abdomen black with posterolat-
eral, silverish white lateral markings on tergirtes
3-4.
Material Examined: Turkey: Konya, Taskent,
Begreli Village, Gevne Valley, elev. 1570 m, 1 July
2001, 1 male, T. Ustiner leg.
Distribution: Palaearctic: Afghanistan, Iran
(Kertesz 1908, 1923; Lindner 1937; Pleske 1925;
Rozkosny & Nartshuk 1988; Woodley 2001).

DISCUSSION

The genus Adoxomyia was previously un-
known from Turkey. This fact is fairly surprising
because many species were recorded from adja-
cent countries and are known from southern, of-
ten arid parts of the Palaearctic region. Therefore
it was to be expected that at least some species of
this genus would be found in Turkey as well. That
is why in this report we constructed an actual
identification key to the all East-Mediterranean
species. Due to intense collecting efforts in many
different localities in Turkey, 4 species of this ge-
nus were collected. The most remarkable record is
Adoxomyia aureovittata which was described as
Euparyphus aureovittatus by Bigot in 1879 from
an unknown locality. The record from Turkey rep-
resents the first evidence that it is a Palaearctic
species and an unpublished record from Greece
(Hauser, personal communication) confirms that
this species probably has an East- Mediterranean
distribution.
Adoxomyia cineracens is distributed in Tran-
scaspia, in the Near East (Iran, Israel) and Cen-
tral Asia. The type locality is Kizilkum (Kazakh-
stan). Our record thus closes the distribution gap
between the known records from Israel and the
type locality.











Our record ofAdoxomyia obscuripennis repre-
sents the most southern and western point of its
range and the second evidence of its occurrence in
the western part of Asia (cf. a record from Azer-
baijan in Nartshuk 2004).
Adoxomyia sarudnyi was only known from Af-
ghanistan and Iran. Our record in Turkey repre-
sents the most western locality of this very rare
species.

ACKNOWLEDGMENTS

Our thanks to Dr. Martin Hauser and Prof. Dr. R.
Rozkosny for critically reviewing the manuscript.


REFERENCES CITED

DUSEK, J., AND ROZKOSNY, R. 1963. Revision mitteleu-
ropaischer Arten der Familie Stratiomyidae
(Diptera) mit besonderer Berticksichtigung der Fau-
na der CSSR I. Acta Societatis Entomologicae C'ech-
osloveniae 60(3): 202-221.
HAUSER, M. 2002. A new species ofAdoxomyia Kert6sz,
1907 (Diptera: Stratiomyidae) from Socotra, Yemen.
Fauna of Arabia 19: 463-466.
JAMES, M. T., AND MCFADDEN, M. W. 1969. The genus
Adoxomyia in America North of Mexico (Diptera:
Stratiomyidae). Journal of the Kansas Entomologi-
cal Society 42: 260-276.
KERTESZ, K. 1907. Eine neuer Dipteren-Gattungsname.
Annalen Historico-Naturales Musei Nationalis Hun-
garici 5(2): 499.
KERTESZ, K. 1908. Catalogus dipterorum hucusque de-
scriptorum. Volumen III. Stratiomyiidae, Erinnidae,
Coenomyidae, Tabanidae, Pantophthalmidae, Rha-
gionidae. Museum Nationale Hungaricum, Budap-
estini, 366 pp.
KERTESZ, K. 1923. Vorarbeiten zu einer Monographie
der Notacanthen. XLV-L. Annales Historico Natu-
rales Musei Nationalis Hungarici, 20: 85-129.
LINDNER, E. 1937. 18. Stratiomyiidae [part]. Lieferung
114. Pp. 145-176. In Lindner, E. [ed.], Die Fliegen


March 2011


der palaearktischen Region. E. Schweizert'sche Ver-
lagsbuchhandlung, Stuttgart, 218 pp.
LINDNER, E. 1974. On the Stratiomyidae (Diptera) of
the Near East. Israel J. Entomol. 9: 93-108.
LINDNER, E. 1975. On some Stratiomyidae (Diptera)
from the Near East. Israel J. Entomol. 10: 41-49.
MCFADDEN, M. W. 1967. Soldier fly larvae in America
north of Mexico. Proc. U.S. Natl. Mus. 121: 1-72.
NARTSHUK, E. P. 2004. New data on Adoxomyia Bezi
from the Caucasus and Eastern Europe (Diptera:
Stratiomyidae). Zoosystematica Rossica 12: 263-266,
St. Petersburg.
OUCHI, Y. 1938. On some stratiomyiid flies from eastern
China. J. Shanghai Sci. Inst., Section III 4: 37-61.
PLESKE, T. 1925. Etudes sur les Stratiomyiinie de la r6-
gion pal6arctique-III.-Revue des esp6ces pal6arc-
tiques de la sous-famille des Clitellariinae. Encyclo-
p6die Entomologique, S6rie B (II), Diptera 1(3-4):
105-119, 165-188.
ROZKOSNY, R. 1983. A Biosystematic Study of the Euro-
pean Stratiomyidae (Diptera), Volume 2: Clitellarii-
nae, Hermnnetiinae, Pachygasterinae and Bibliography.
Dr. W. Junk, The Hague, Boston, London, 431 pp.
ROZKOSNY, R., AND NARTSHUK, E. P. 1988. Family Stra-
tiomyidae, pp. 42-96 In A. So6s and L. Papp [eds.],
Catalogue of Palaearctic Diptera. Volume 5. Atheri-
cidae-Asilidae. Akad6miai Kiad6 Budapest, 446 pp.
USTUNER, T., AND HASBENLI, A. 2003. First record of
the subfamily Beridinae (Diptera: Stratiomyidae)
from Turkey. Studia Dipterologica 10(1): 186-188,
Halle (Saale).
USTUNER, T., HASBENLi, A., AND AKTUMSEK, A. 2002.
Contribution to subfamily 360 Clitellariinae
(Diptera, Stratiomyidae) in Fauna of Turkey. J. En-
tomol. Res. Soc. 4(1): 19-24.
USTUNER, T., HASBENLI, A., AND ROZKOSNY, R. 2003.
First record of Hermetia illucens (Linnaeus, 1758)
(Diptera, Stratiomyidae) from the Near East. Studia
Dipterologica 10(1): 181-185, Halle (Saale).
USTUNER, T., AND HASBENLi, A. 2004. A new species of
Oxycera Meigen (Diptera: Stratiomyidae) from Tur-
key. Entomol. News 115(3): 163-167.
WOODLEY, N. E. 2001. A World Catalog of the Stratiomy-
idae (Insecta: Diptera). Backhuys Publishers,
Leiden, 474 pp.


Florida Entomologist 94(1)







Wiesenborn: Nitrogen Contents in Riparian Arthropods


NITROGEN CONTENT IN RIPARIAN
ARTHROPODS IS MOST DEPENDENT ON ALLOMETRY AND ORDER

WILLIAM D. WIESENBORN
U.S. Bureau of Reclamation, Lower Colorado Regional Office, P.O. Box 61470, Boulder City, NV 89006

ABSTRACT

I investigated the contributions of body mass, order, family, and trophic level to nitrogen (N)
content in riparian spiders and insects collected near the Colorado River in western Arizona.
Most variation (97.2%) in N mass among arthropods was associated with the allometric effects
of body mass. Nitrogen mass increased exponentially as body dry-mass increased. Significant
variation (20.7%) in N mass adjusted for body mass was explained by arthropod order. Ad-
justed N mass was highest in Orthoptera, Hymenoptera, Araneae, and Odonata and lowest in
Coleoptera. Classifying arthropods by family compared with order did not explain signifi-
cantly more variation (22.1%) in N content. Herbivore, predator, and detritivore trophic-levels
across orders explained little variation (4.3%) in N mass adjusted for body mass. Within or-
ders, N content differed only among trophic levels of Diptera. Adjusted N mass was highest in
predaceous flies, intermediate in detritivorous flies, and lowest in phytophagous flies. Nitro-
gen content in riparian spiders and insects is most dependent on allometry and order and least
dependent on trophic level. I suggest the effects of allometry and order are due to exoskeleton
thickness and composition. Foraging by vertebrate predators, such as insectivorous birds, may
be affected by variation in N content among riparian arthropods.

Key Words: nutrients, spiders, insects, trophic level, exoskeleton, cuticle

RESUME

Se investiguo las contribuciones de la masa de cuerpo, orden, familiar y el nivel tr6fico al con-
tenido de nit6geno (N) en araias e insects riparianos (que viven en la orilla del rio u otro
cuerpo de agua) recolectadaos cerca del Rio Colorado en el oeste del estado de Arizona. La ma-
yoria de la variaci6n (97.2%) en la masa (N) entire los artr6podos fue asociado con los efectos
alom6tricos de la masa de cuerpo. La masa de nitr6geno aument6 exponencialmente con el au-
mento de masa-seca del cuerpo. La variaci6n significativa (20.7%) en la masa N ajustada por
la masa del cuerpo se explica segun el ord6n del artr6podo. La masa ajustada N fue mas alta
en Orth6ptera, Hymen6ptera, Araneae, Odonata y mas baja en Coleoptera. Al clasificar los ar-
tr6podos por familiar comparado con el ord6n no explica la variacion mayor significativa
(22.1%) en el contenido de N. Los niveles tr6ficos de los herbivoros, depredadores y detritivoros
en todos los ordenes explica la pequefia variaci6n (4.3%) en la masa N ajustada por la masa del
cuerpo. Entre los ordenes, el contenido N varia solamente entire los niveles tr6ficos de Diptera.
El valor ajustado de la masa de N fue mayor para las moscas depredadores, intermedio para
las moscas detritivoras y menor para las moscas fit6fagas. El contenido de nitr6geno en araias
e insects riparianos es mas dependiente sobre la alometria y ord6n y menos dependiente so-
bre el nivel tr6fico. Sugiero que los efectos de alometria y ord6n son debidos al grosor y la com-
posici6n del exo-esqueleto. El forraje por los depredadores vertebrados, como aves insectivoras,
puede ser afectado por la variaci6n del contenido N entire los artr6podos riparianos.


Nitrogen concentrations in organisms are de-
pendent on trophic level. This is most apparent
between plants and herbivores, because N com-
prises 0.03-7% of dry mass in plants compared
with 8-14% in animals (Mattson 1980). Variation
in N concentration among and within plants, and
its effects on abundances of herbivores including
arthropods, especially agricultural pests, has
been frequently examined (reviewed in Mattson
1980; Scriber 1984). Fewer studies have consid-
ered variation in N concentration among spiders
and insects. Bell (1990) and Studier & Sevick
(1992) tabulated measurements of %N in various
insects from different studies. Fagan et al. (2002)
compared %N between arthropod herbivores and


predators by analyzing data compiled from vari-
ous sources. Concentrations of N in spiders and
insects were dependent on trophic level after con-
trolling for body length, representing allometry,
and taxonomic group, representing phylogeny
(Fagan et al. 2002). Predators generally con-
tained higher %N than herbivores. Predaceous
arthropods may concentrate N from food similar
to phytophagous arthropods.
Variation in N concentration among spiders
and insects may affect foraging by arthropod-
feeding vertebrates and the qualities of food they
obtain. Diet protein has been implicated as affect-
ing egg production (Ramsay & Houston 1997) and
nestling growth (Johnston 1993) in insectivorous







Florida Entomologist 94(1)


birds. Identifying sources of variation in arthro-
pod N content may improve our understanding of
the prey composition required to support species
of insectivorous wildlife.
I examined variation in N content among spi-
ders and insects collected from trees and shrubs
established to restore riparian habitat for insec-
tivorous vertebrates, especially birds. Variation
in N mass was partitioned into various sources. I
first determined the allometric relationship be-
tween N mass and body dry-mass. After adjusting
N mass for this relationship, N contents of arthro-
pods were compared among orders and families
and among trophic levels across and within or-
ders. I interpreted N contents in relation to exosk-
eleton scaling and chemical composition and con-
cluded by applying the results to diets of insectiv-
orous birds.

MATERIALS AND METHODS

Arthropod Collections

Spiders and insects were collected next to the
Colorado River within Havasu National Wildlife
Refuge in Mohave County, Arizona. Most arthro-
pods were collected at an irrigated 43-ha riparian
restoration area (3446'N, 11431'W; elevation
143 m) of planted or volunteer trees and shrubs
12 km southeast and across the river from Nee-
dles, California. Plots were planted during 2003-
2005 with cuttings that were taken from nearby
areas along the river and rooted in containers.
The area is straddled by Topock Marsh (16 km2)
and Beal Lake (0.9 km2), 2 impoundments con-
taining mostly emergent cattails (7Fi. ..... sp.,
Typhaceae) and open water. Undeveloped areas of
the surrounding floodplain support mostly natu-
ralized tamarisk (Tamarix ramosissima Ledeb.,
Tamaricaceae) shrubs. The floodplain is flanked
by Sonoran desertscrub dominated by creosote
bush (Larrea tridentata (DC.) Cov., Zygophyl-
laceaae). Maximum temperatures average 42.7C
during Jul, and minimum temperatures average
5.6C during Jan at Needles (DRI 2010).
I collected arthropods from plants and trapped
insects in flight. Arthropods were swept with a
38-cm diameter muslin net from planted cotton-
wood (Populus fremontii S. Watson, Salicaceae)
and Goodding's black willow (Salix gooddingii C.
Ball, Salicaceae) trees, planted narrow-leaved
willow shrubs (Salix exigua Nutt.), volunteer
honey mesquite (Prosopis glandulosa Torrey, Fa-
baceae) and screwbean mesquite (Prosopis pubes-
cens Benth.) trees, and volunteer arrowweed
shrubs (Pluchea sericea (Nutt.) Cov., Asteraceae).
I also swept arthropods from T ramosissima bor-
dering the plots. Additional arthropods on S. ex-
igua were swept from plants growing along a dirt
irrigation canal 2 km northwest of the restoration
area. Plant species were swept separately except


for Prosopis spp., which grew together. Each spe-
cies was swept 10-15 min on 9 dates: 30 Apr, 14
May, 27 May, 08 Jun, 22 Jun, 30 Jun, 21 Jul, 4
Aug, and 18 Aug 2009. All plant species were in
flower or fruit except for P. fremontii. Arthropods
swept from plants were placed into plastic bags,
kept in a refrigerator, and killed in a freezer. Fly-
ing insects were trapped with a Malaise trap
(Santee Traps, Lexington, KY) that was placed in
the center of a plot supporting S. gooddingii and
P sericea and elevated 1 m aboveground with
fence posts. Trapped insects were collected into a
dry plastic bottle containing a nitrogen-free, di-
clorvos insecticide strip. Insects were trapped for
6.1-7.3 h during 0855-1640 PDT on each of the
above dates except 30 Apr, 14 May, and 18 Aug
2009.
Spiders and insects collected on each date were
sorted under a microscope into morphotypes (sim-
ilar-looking specimens). Representatives of each
morphotype were placed into 70% ethanol for
identification. I counted and split the remaining
specimens of each morphotype into samples each
with an estimated maximum dry mass of 10 mg.
Individual specimens with dry masses >10 mg
were placed into separate samples. Arthropod
samples for N analyses were cleaned by vortexing
in water, transferred to filter paper with a Biuch-
ner funnel, dried 2 h at 80'C, and stored in stop-
pered vials.

Arthropod Identifications and Trophic Levels

Spiders and insects were identified to the low-
est taxon possible, at least to family and typically
to genus. Vouchers of adult insects were deposited
at the Bohart Museum of Entomology, University
of California, Davis, and vouchers of spiders were
deposited at the California Academy of Sciences,
San Francisco. Arthropod taxa were classified
into the trophic levels of herbivore, predator, and
detritivore based on published descriptions (Ta-
ble 1). Holometabolous insects were classified by
larval diet. Herbivores included consumers of pol-
len, nectar, or honeydew homopterann egesta).
Predators included parasites and consumers of
already-dead animals.

Arthropod Nitrogen Estimates

The mass of N in each arthropod sample was
estimated with the Kjeldahl method adapted
from Isaac & Johnson (1976). Samples of dried ar-
thropods were weighed (+0.01 mg) with a mi-
crobalance (model C-30, Cahn Instruments, Cer-
ritos, CA) and ground into water with a 5-mL
glass tissue homogenizer. Homogenized samples
were poured and rinsed with water, to a total vol-
ume of 20 mL, into 100-ml digestion tubes. I
added 6 mL of concentrated sulfuric acid, contain-
ing 4.2% selenous acid, and 3 mL of 30% hydrogen


March 2011









TABLE 1. ADULT ARTHROPODS COLLECTED FROM RIPARIAN HABITAT NEAR THE COLORADO RIVER IN ARIZONA AND ANALYZED FOR NITROGEN CONTENT.

No. specimens Mean body Mean SD
Order or suborder Family Genus' Source2 No. Samples per sample Trophic level3 dry mass (mg) % N

Araneae Philodromidae Philodromus E.S 2 3-4 P 1.93 10.6 0.9


Salticidae

Thomisidae
2 families4'6
3 families5'6

Libellulidae

Acrididae
Tettigoniidae

Largidae
Lygaeidae
Pentatomidae

Reduviidae


Cicadellidae





Cixiidae
Flatidae
Membracidae

Chrysopidae

Myrmeliontidae


Habronattus
Metaphidippus
Misumenops



Pachydiplax

Acridinae7
Scudderia

Largus
Nysius
Brochymena
Thyanta
Pselliopus
Zelus

Cicadellinae
Gyponinae
Opsius6
Typhlocybinae

Oecleus
Ormenis


Chrysoperla

Myrmelion


S
S

S
S
F,G,P
E
P
F,P,S

E,F,G
G
T
F
S
S
G,T
G

F,G,S
G
F


1-3
1

1
67
1
1
1
1-3

1-3
2
28-41
19-22
5
4
2
2

2-14
11
1


9.3 1.2
13.0
12.1 1.8
14.3
13.8 0.1
12.3 1.0

13.9+ 2.7

14.6
9.2

9.0
11.0 1.5
11.6
13.3
10.5 2.0
10.1+ 2.2

8.6
11.2 1.5
11.4 0.0
14.6
10.1
8.9 1.2
10.6
9.1 1.5

11.8
12.5


13.0
115.0

49.2
0.46
55.2
17.1
14.1
7.20

6.62
3.36
0.68
0.35
4.37
1.24
5.72
5.22

1.51
1.37
8.99


Subfamily in Acrididae and subfamily or genus in Cicadellidae.
E, Salix exigua; F, Populus fremontii; G, Salix gooddingii; M, Malaise trap; P, Prosopis glandulosa or P. pubescens; S, Pluchea sericea; T, Tamarix ramosissima.
'D, Detritivore; H, Herbivore; P, Predator. Reference for all (Borror et al. 1981) except Apioceridae (Cole 1969) and Andrenidae, Formicidae, and Tettigoniidae (Essig 1926).
'Salticidae, Habronattus sp.; Thomisidae, Misumenops sp.
5Araneidae, Hypsosinga sp.; Salticidae, Metaphidippus sp. & Habronattus sp.; Thomisidae, Misumenops sp.
'Adults and immatures.
'Immatures.


Odonata


Orthoptera


Heteroptera







Homoptera









Neuroptera











TABLE 1. (CONTINUED) ADULT ARTHROPODS COLLECTED FROM RIPARIAN HABITAT NEAR THE COLORADO RIVER IN ARIZONA AND ANALYZED FOR NITROGEN CONTENT.

No. specimens Mean body Mean SD
Order or suborder Family Genus' Source2 No. Samples per sample Trophic level3 dry mass (mg) % N

Coleoptera Bruchidae Algarobius P 1 6 H 3.01
Coccinellidae Chilocorus F,P 3 2-4 P 4.75 9.8+ 1.2
Hippodamia F,S 3 2-8 P 6.26 6.6 2.8
Diptera Apioceridae Apiocera M 1 1 P 52.87 11.4
Asilidae Proctacanthus M 1 1 P 42.3 11.7
Dolichopodidae Asyndetus M 13 17-113 D 0.39 9.9 2.0
Lauxaniidae Homoneura F,G 2 4-5 D 1.31 7.8 1.0
Minettia F,G 2 2-6 D 2.37 8.1 4.6
Sarcophagidae Eumacronychia F,G 1 2 P 1.68 11.5
Tabanidae Apatolestes M 1 1 P 15.0 11.6
Tabanus M 13 2-3 P 13.8 10.9 2.2
Tachinidae Zaira M 2 1-2 P 7.66 9.2 2.3
Tephritidae Acinia F 2 7-9 H 1.01 5.1+ 1.5
9.8
Hymenoptera Andrenidae Perdita S 1 2 H 1.74
Formicidae Formica E,S 4 6-16 H 0.76 10.9 1.8
Halictidae Agapostemon E 1 1 H 7.42 11.7
Dieunomia S 1 3 H 5.57 14.1
Lasioglossum E 1 9 H 2.71 16.7
13.4
Sphecidae Bembix M 1 1 P 33.5
Cerceris M 1 1 P 10.6 8.8
Tachysphex M 1 1 P 7.23 8.5
21.2
Tiphiidae Myzinum E 1 6 P 4.54
Vespidae Polistes G 1 1 P 28.8 14.0

Subfamily in Acrididae and subfamily or genus in Cicadellidae.
'E, Salix exigua; F, Populus fremontii; G, Salix gooddingii; M, Malaise trap; P, Prosopis glandulosa or P. pubescens; S, Pluchea sericea; T, Tamarix ramosissima.
'D, Detritivore; H, Herbivore; P, Predator. Reference for all (Borror et al. 1981) except Apioceridae (Cole 1969) and Andrenidae, Formicidae, and Tettigoniidae (Essig 1926).
Salticidae, Habronattus sp.; Thomisidae, Misumenops sp.
'Araneidae, Hypsosinga sp.; Salticidae, Metaphidippus sp. & Habronattus sp.; Thomisidae, Misumenops sp.
'Adults and immatures.
7Immatures.







Wiesenborn: Nitrogen Contents in Riparian Arthropods


peroxide and heated samples 1 h at 400'C with a
block digestor (model 2040, Tecator, Herndon,
VA). After cooling, water was added to 60 mL.
The ammonia concentration formed in the clear,
digested samples was measured by colorimetry,
against standards prepared from dried ammo-
nium-sulfate, with a segmented flow analyzer
(model FS-4, OI Analytical, College Station, TX).
Salicylate, hypochlorite, and sodium nitroprus-
side were used as the indicator. I converted am-
monia concentration to mg N.
I adjusted estimates of mg N in arthropod
samples with chitin samples containing known N
masses. Chitin is a nitrogenous polysaccharide
(C8H,,NO,), abundant in arthropod exoskeleton,
or cuticle (Neville 1975), that typically comprises
25-40% of exoskeleton dry-mass in insects (Rich-
ards 1978). Various masses (2, 4, 8, 16, 32, 64 mg)
of powdered chitin (Tokyo Chemical Industry)
containing 6.89% N were weighed, placed in 20
mL water, digested, and measured for ammonia
within each batch (n = 4) of arthropod samples. I
increased estimates of mg N in arthropod sam-
ples in each batch to correct for the batch's mean
underestimate of %N (5.76, 6.23, 6.44, 6.08%) in
chitin samples. I calculated %N in arthropod
samples as 100(mg N/mg dry mass). Two arthro-
pod samples of Acinia and C'i ...., t.i with un-
usually low N concentrations (<0.9%) were ex-
cluded as outliers. Dry mass and mg N of each ar-
thropod sample were divided by the number of
specimens in the sample to estimate dry mass
and N mass per specimen.

Statistical Analysis

Body masses of arthropods, transformed
log(mg) to normalize residuals, were compared
among trophic levels with analysis of variance
(SYSTAT version 12, San Jose, CA). Nitrogen
masses in spiders and insects were analyzed se-
quentially. I first determined the relationship be-
tween N mass and body dry mass by regressing
log(mg N) against log(mg body mass) for each ar-
thropod sample. I verified that the relationship
was allometric (exponential) by testing with an
approximate t test the null hypothesis that the re-
gression coefficient b, = 1 (Neter et al. 1996).
Transformed N masses were adjusted for their al-
lometric relationship with transformed body
mass by adding the residuals from the regression
to the overall mean of transformed N mass (Sokal
& Rohlf 1981).
Adjusted, transformed N masses were com-
pared among arthropod orders with analysis of
variance. Hemiptera were split into suborders
Heteroptera and Homoptera, because the diges-
tive systems of most homopterans have filter
chambers that concentrate nitrogenous com-
pounds (Borror et al. 1981). I tested if classifying
arthropods by family instead of order or suborder


explained more variation in adjusted log(mg N)
with the general linear test approach (Neter et al.
1996). This approach tests if mean square error in
an analysis of variance decreases significantly
when the model becomes more complex. Samples
containing more than 1 family (3 samples of Ara-
neae, or spiders) were classified only to order.
Arthropod N-contents adjusted for body mass
were compared among trophic levels across and
within orders or suborders. I compared N masses
among trophic levels across orders or suborders
with analysis of variance. Separate analyses were
performed within Heteroptera, Diptera, and Hy-
menoptera, the 3 orders or suborders with 2 or
more trophic levels each containing more than 1
sample. Analyses within orders or suborders
weighted adjusted values of log(mg N) by 1/s2 in
each trophic level to correct for uneven variances
among trophic levels (Neter et al. 1996).

RESULTS

Collected Arthropods

I collected 121 samples of spiders and insects
containing 1,490 specimens in 9 orders or subor-
ders, 33 families, and 43 subfamilies or genera
(Table 1). All of the arthropods collected were
adults except for 8 samples in 3 taxa (families,
subfamilies, or genera) with adults and imma-
tures and 6 samples in 1 taxon with only imma-
tures. Body dry-masses of adult arthropods
ranged from 0.35 mg in Typhlocybinae leafhop-
pers (Cicadellidae) to 115 mg in the fork-tailed
bush katydid Scudderia furcata Brunner (Tet-
tigoniidae).
Two orders or suborders (Orthoptera and Ho-
moptera) of collected spiders and insects were
only herbivorous, 3 orders (Araneae, Odonata,
and Neuroptera) were only predaceous, and 4 or-
ders or suborders (Heteroptera, Coleoptera,
Diptera, and Hymenoptera) included both trophic
levels. All Coleoptera were predaceous except for
1 sample. The only detritivores collected were
flies (Diptera). Across orders or suborders, herbi-
vores included 42 samples in 22 taxa, predators
included 62 samples in 24 taxa, and 17 samples in
3 taxa were detritivores (Table 1). Trophic levels
contained arthropods with different body dry-
masses (F = 25.5; df = 2, 118; P < 0.001). Preda-
tors were largest (back-transformed mean = 6.37
mg) followed by herbivores (4.03 mg) and detriti-
vores (0.55 mg).
Allometric Nitrogen Contents

Nitrogen mass in riparian spiders and insects
was allometrically related to body dry mass
(Fig. 1). Transformed N mass per specimen in ar-
thropod samples was positively related (F 4, 066;
df = 1, 119; P < 0.001) to transformed body dry-
mass per specimen by:







76 Florida Entomologist 94(1) March 2011



I I 1 I I I I I 11
Or, Tettigoniidae


10.0

SD, Apioceridae
SAsilidae e Pentatomidae
Od, Libellulidae He, ntatomidae
Hy, Vespidae He, Largidae


Or Acrididae* Hy, Sphecidae
E D, Tabanidae
W N, Myrmeliontidae
1.0 Hy, Tiphiidae* He, Reduviidae
Z -Hy, Halictidae*/ D, Tachinidae
c Ho, Membracidae, Ho, Fl
ro Ho, Flatidae
SA, Salticidae o C, Coccinellidae
Araneae, Ho, Cicadellidae
A, Thomisidae
A, Philodromidae C, Bruchidae
D, Sarcophagidae**
N, Chrysopidae Hy, Andrenidae
Ho, Cixiidae 0* D, Lauxaniidae
0.1 Hy, Formicidae
0
S D, Tephritidae

S 0 He, Lygaeidae
SD, Dolichopodidae
I I 1''"1 I I 1 ''"'1

1 10 100
Mean body dry-mass (mg)
Fig. 1. Mean N mass us. mean body dry-mass in riparian arthropods from the lower Colorado River classified by
family. Abbreviations are orders or suborders (in Hemiptera): A, Araneae; C, Coleoptera, D, Diptera; He, Het-
eroptera; Ho, Homoptera; Hy, Hymenoptera; N, Neuroptera; Od, Odonata; Or, Orthoptera. Single point labeled Ara-
neae represents mixed samples of Araneidae, Salticidae, and Thomisidae. Axes are log scales. Line fit to
transformed data by linear regression weighted by sample size.


log mg N = -1.006 + 1.039(log mg dry mass) Nitrogen Content in Arthropod Orders
Back-transforming this equation produced:
Back-transforming this equation produced: Nitrogen mass adjusted for body mass in ripar-
mg N = 0.0986(mg dry mass)1039 ian arthropods (Fig. 2) differed (F = 3.64; df = 8,
112; P < 0.001) among orders or suborders. These
The exponent (1.039 0.016 SD) differs from taxonomic levels explained 20.7% of variation in
unity (t* = 2.43; df = 119; P = 0.008), verifying adjusted N mass. Orthoptera (mean 14.0% N),
that the relationship is exponential rather than Hymenoptera (12.4% N), Araneae (11.9% N), and
linear. This allometric relationship explained Odonata (12.3% N) contained the highest ad-
97.2% of variation in N mass. Percentage of N in justed N contents, and Coleoptera (8.2% N) con-
riparian arthropods (Table 1) increased as body trained the lowest adjusted N content. Orthoptera
mass increased. were mostly immature slant-faced grasshoppers







Wiesenborn: Nitrogen Contents in Riparian Arthropods


0.5,
-3 '

&0.4-
S)

E
z
S0.3.




0.2-


n4l-


I I I I I I


I I I


01




Fig. 2. Nitrogen mass allometrically adjusted for
body mass in riparian arthropods from the lower Colo-
rado River classified by order or suborder (in Hemi-
ptera). Letters are means ( SE) and trophic levels: D,
detritivores; H, herbivores; P, predators. Adjacent num-
bers are sample sizes. Y-axis is log scale.


(Acridinae) along with the sole katydid S. furcata.
Hymenoptera included ants (Formicidae), 2 fami-
lies of bees (Andrenidae and Halictidae), and 3
families of wasps (Sphecidae, Tiphiidae, and
Vespidae). Spider samples contained several fam-
ilies (Table 1). The only odonate collected was the
dragonfly Pachydiplax longipennis Burmeister.
Coleoptera included 1 sample of the herbivorous
seed beetle (Bruchidae) Algarobius prosopis Le-
Conte, collected from Prosopis spp., and 6 sam-
ples containing 2 species of predaceous ladybird
beetles (Coccinellidae), Chilocorus cacti L. and
the widespread Hippodamia convergens Guerin-
Meneville. Insects in other orders, including the 2
Hemiptera suborders, contained intermediate N
concentrations (Fig. 2).
Classifying arthropods by family instead of or-
der or suborder did not explain more variation in
N mass adjusted for body mass. Error variance of
adjusted N mass did not decrease (F = 1.45; df =
26, 86; P = 0.10) when arthropods were classified
by family compared with order or suborder. Clas-
sifying arthropods by family instead of order or
suborder explained 22.1%, a 1.4% improvement,
of variation in adjusted N mass.

Nitrogen Content in Trophic Levels

Differences in N content among the trophic
levels of herbivore, predator, and detritivore de-


H7
4 p H17, 3p
P10 H5


H
I I

?6 4
IH


I


pended on classification (Fig. 2). Across orders or
suborders, N mass did not vary (F = 0.62; df = 2,
118; P = 0.54) among trophic levels. Trophic levels
explained 1.0% of variation in N mass after ac-
counting for body mass. Back-transformed means
of adjusted N mass (and mean % N) were 0.413
mg (11.1% N) in herbivores, 0.397 mg (10.9% N)
in predators, and 0.380 mg (9.44% N) in detriti-
vores, the smallest arthropods collected. Within
orders or suborders, N mass varied among trophic
levels in Diptera (F = 4.60; df = 2, 35; P = 0.017)
but not in Heteroptera (F = 0.62; df = 1, 12; P =
0.45) or Hymenoptera (F = 0.13; df = 1, 11; P =
0.91). Adjusted N contents in flies (Fig. 2) were
lower in herbivores (mean 5.1% N) compared with
predators (10.9% N) or detritivores (9.4% N). All
phytophagous flies collected were 2 samples of
the fruit fly (Tephritidae) Acinia picturata
(Snow), swept from P. fremontii. Adjusted N con-
centrations in predaceous or parasitic flies (Apio-
ceridae, Asilidae, Sarcophagidae, Tabanidae, and
Tachinidae) and detritivorous flies (Dolichopo-
didae and Lauxaniidae) were similar.

DISCUSSION

Allometric Nitrogen Contents

The allometric relationship between N mass
and body mass in riparian arthropods resembles
a similar relationship between exoskeleton mass
and body mass in terrestrial arthropods. Ander-
son et al. (1979) dissected the exoskeletons from 3
species of immature and adult spiders, weighing
between 25 mg and 1.2 g, and determined exosk-
eleton dry-mass and body wet-mass were posi-
tively related by:

g exoskeleton = 0.078(g body mass)1135

Body mass in spiders explained 94.1% (their r-
value squared) of variation in exoskeleton mass.
Anderson et al. attributed this allometric rela-
tionship to scaling. The exoskeleton of terrestrial
arthropods must increase in thickness as body
weight increases to support the organism and
withstand the stresses of bending and twisting
(Prange 1977; Anderson et al. 1979).
Allometric relationships between N mass and
body mass, and between exoskeleton mass and
body mass, may be primarily due to exoskeleton
N. Trim (1941) estimated N concentrations of
11.8% in abdominal cuticles of 2 Orthoptera spe-
cies, approximating the mean concentration
(10.7%) in riparian arthropods. A large proportion
of N in terrestrial arthropods likely resides
within the exoskeleton due to its greater density
compared with internal tissues and hemolymph.
The allometric relationship between exoskeleton
mass and body mass may have produced the sim-
ilar relationship between N mass and body mass.







Florida Entomologist 94(1)


A linear increase in N mass in internal tissues as
body mass increases would dampen the exponen-
tial increase in cuticular N mass. The lower expo-
nent relating N mass to body mass (1.039) com-
pared with the exponent relating cuticle mass to
body mass (1.135) may reflect this dampening.

Nitrogen Contents in Orders or Suborders

Exoskeleton composition may have contrib-
uted to different N concentrations among orders
of spiders and insects (Fagan et al. 2002). Arthro-
pod cuticle is composed primarily of protein and
chitin (Neville 1975), and concentrations of N are
higher in the former. For example, I estimated
%N in arthropod cuticular protein from percent-
ages of amino acids in pronotal and abdominal cu-
ticles of adult Tenebrio beetles (Andersen et al.
1973; reported in Table 3.4 in Neville 1975) by as-
suming the amino acids were bonded into
polypeptides. The estimated N concentration of
cuticular protein (17.4%) exceeded that of chitin
(6.89%). Based on the maximum range of chitin
concentration (10-60% of dry mass) in insect cuti-
cle (Richards 1978; see also Table 1 in Hackman
1974), and assuming cuticle is entirely chitin and
protein, N concentrations in insect exoskeleton
may vary from 11.1% to 16.4%.
Greater concentrations of protein in arthro-
pod cuticle, producing higher N contents, have
been associated with concentrations of resilin
(Andersen 1979). Resilin is a flexible, elastic
protein that occurs in cuticle in near-pure con-
centrations or combined with other proteins
and chitin (Richards 1978). I estimated as
above that resilin contains 19.0% N from per-
centages of amino acids in resilin from Schisto-
cerca grasshoppers (Andersen 1966; reported in
Table 3.4 in Neville 1975). Various mechanical
structures in arthropods are elastic due to resi-
lin (Table 2.1 in Neville 1975). Resilin is espe-
cially prevalent in the wing tendons and hinges
of Odonata and Orthoptera (Andersen & Weis-
Fogh 1964), primitive orders with synchronous
flight muscles. Andersen and Weis-Fogh also de-
tected resilin in the abdominal sclerites of
Schistocerca grasshoppers, presumably allow-
ing the abdomen to stretch. Abundances of resi-
lin in riparian Odonata and Orthoptera may
have contributed to their high N contents. Al-
though resilin has not been found in spiders
(Andersen & Weis-Fogh 1964), the high degree
of abdominal stretching by spiders (Browning
1942) suggests their cuticles contain a similar
elastic protein. Cuticles of Coleoptera are likely
less elastic. A dominant feature of beetles is the
elytra, hardened front-wings that act only to
cover the folded hind-wings and abdomen. The
likely absence of resilin and resultant high con-
centrations of chitin, in elytra may have low-
ered %N in Coleoptera.


Nitrogen Contents in Trophic Levels

I did not detect an overall difference in N con-
centration among herbivorous, predaceous, and
detritivorous arthropods after accounting for the
allometric effects of body mass. Trophic level did
not appear to generally affect arthropod %N. This
contradicts the overall difference in N concentra-
tion between herbivorous and predaceous arthro-
pods detected by Fagan et al. (2002). Different re-
sults may have been due to statistical methodol-
ogy. Fagan et al. controlled for body length and
taxonomic group, to account for phylogeny,
whereas I controlled only for body mass. Control-
ling for phylogeny is difficult, because different
frequencies of herbivores compared with preda-
tors among taxonomic groups cause trophic level
and phylogeny to be confounded. Phylogeny and
trophic level cannot be statistically separated.
Similar N contents between trophic levels
agree with the concept that most insects satisfy
nutrient requirements by adjusting food intake
(Waldbauer 1968; reviewed in Simpson et al.
1995). An example in riparian arthropods may be
found in the 2 suborders of Hemiptera, insects
with piercing-sucking mouthparts. Phytophagous
Heteroptera, such as Lygus leaf bugs (Backus et
al. 2007), typically rupture, dissolve with saliva,
and ingest mesophyll from a variety of plant
structures. All Homoptera are herbivorous, and
many homopterans feed on phloem which is high
in water and carbohydrates but low in other nu-
trients including N. The Opsius stactogalus Fie-
ber leafhoppers collected here increase food in-
take, concentrate nutrients within their filter-
chamber digestive tracts (Wiesenborn 2004), and
void excess water and sugars. Concentrations of
N in Homoptera, phytophagous Heteroptera, and
predaceous Heteroptera were similar despite dif-
ferent diets and physiologies.
An exception was Diptera. Herbivorous flies,
all Tephritidae, contained lower N concentra-
tions than predaceous or detritivorous flies after
considering body mass. Fagan et al. (2002) com-
pared phylogenetic categories of herbivorous in-
sects and found lower N concentrations in
Diptera and Lepidoptera, combined as the recent
lineage Panorpida, after accounting for body
length. The database analyzed by Fagan et al. in-
cluded the herbivorous flies Bibionidae, Chlorop-
idae, and Drosophilidae, each in a different su-
perfamily separate from Tephritidae. The diver-
sity of phytophagous Diptera found to contain
low N concentrations suggests N contents in flies
generally vary by trophic level. Fagan et al.
(2002) suggested several explanations for lower
N contents in herbivores than in predators.
These included the direct effects of diet N, indi-
rect effects of trophic niche unrelated to diet, and
selection for low body N in response to low diet N.
The A. picturata tephritids that I collected de-


March 2011







Wiesenborn: Nitrogen Contents in Riparian Arthropods


velop as larvae in the flower heads of Pluchea
spp. (Foote et al. 1993), corresponding with the
flowering P. sericea at the study site. Infestations
byA. picturata reduce seed production (Alyokhin
et al. 2001), suggesting larvae eat ovaries or
seeds. The species does not appear to concentrate
N from food, because its N concentration (5.1%) is
within the range (1-7% of dry mass) reported for
seeds (Mattson 1980). The structural or biochem-
ical features correlated with low N concentration
in A. picturata and other plant-feeding flies are
unknown. Low exoskeleton mass in tropical, her-
bivorous beetles has been attributed to low diet
N, short larval-development time, and high fe-
cundity (Rees 1986). Equivalent N concentra-
tions in predaceous or parasitic flies and detritiv-
orous flies suggest their diets contain similar
amounts of N.

Arthropod Nitrogen as a Nutrient for Birds

Not all N in arthropods is digested by insectiv-
orous birds. Bird diets are frequently determined
by identifying undigested fragments of exoskele-
ton in fecal samples (e.g., Wiesenborn & Heydon
2007). Digestion of arthropod cuticle by verte-
brates likely depends on its sclerotization (Kara-
sov 1990). Sclerotized proteins are bonded to-
gether, frequently with chitin, forming an irre-
versibly-hardened cuticle that cannot be hydro-
lyzed into amino acids (Richards 1978).
Unsclerotized proteins, like resilin, can be hydro-
lyzed (Richards 1978). Relative proportions of
sclerotized and unsclerotized proteins vary
greatly among species (Richards 1978) producing
cuticles with different digestibilities. Arthropod
orders with high amounts of elastic protein, such
as Odonata and Orthoptera and probably Ara-
neae, may provide insectivorous birds with high
concentrations of digestible protein.
Riparian arthropods presented insectivorous
birds with prey containing a range (5.1-14.0%) of
N concentrations. Foraging by insectivorous birds
in relation to prey N concentration can be difficult
to discern, because birds frequently forage in re-
sponse to prey availability which is transitory
and hard to estimate. Selective foraging may be
inferred by comparing arthropods eaten by adults
with those concurrently captured by adults but
fed to nestlings. Insectivorous nestlings depend
on diet nutrients in addition to calories (Johnston
1993). Adult great tits (Parus major L.) and blue
tits (Parus caeruleus L.) in woodlands ate mostly
Lepidoptera larvae but provided 3-9 day-old nest-
lings with more spiders, earwigs (Dermaptera),
and flies (Cowie and Hinsley 1988). Including
other arthropods, especially spiders, as prey may
have augmented the low N content of Lepidoptera
(Fagan et al. 2002). Spiders also provide different
amino-acid compositions (Ramsay & Houston
2003).


The importance of prey N-concentration to in-
sectivorous birds that feed on more-diverse prey
is less clear. An example is the southwestern wil-
low flycatcher (Empidonax traillii (Audubon) ssp.
extimus Phillips), a migrant that winters in Cen-
tral America and breeds in southwestern U.S. ri-
parian habitats. Adult flycatchers ate mostly het-
eropterans, flies, and beetles but fed more odo-
nates and beetles to nestlings (Drost et al. 2003).
Diet N may be increased by including odonates,
especially dragonflies due to their large biomass.
Diets of nestling flycatchers in other localities
contained more Diptera than those of adults
(Durst et al. 2008) or prey compositions similar to
adults (Wiesenborn & Heydon 2007). The high-N
orders of Araneae, Odonata, and Hymenoptera,
taken together, were eaten with similar frequency
by flycatchers at different localities and habitats.
These orders comprised 21% of prey in California
(Drost et al. 2003), 31% of prey in Arizona (Durst
et al. 2008), and 21% of prey at 3 localities in Ar-
izona and Nevada (Wiesenborn & Heydon 2007).
In summary, N concentrations in riparian ar-
thropods are primarily dependent on body mass
and order and less dependent on trophic level.
Variation in prey N concentration may affect for-
aging by insectivorous birds and the qualities of
food they obtain.

ACKNOWLEDGMENTS

I am grateful to A. Stephenson, USBR Lower Colo-
rado Regional Laboratory, for measuring ammonia con-
centrations. I appreciate the help identifying insects
provided by S. L. Heydon, L. S. Kimsey, and T. J. Zavort-
ink at the Bohart Museum of Entomology, and C. A.
Tauber and P. S. Ward at the Entomology Department,
UC Davis. I am grateful to J. E. O'Hara at Agriculture
and Agrifood Canada for identifying tachinids and to D.
Ubick for identifying spiders. I thank J. Allen of the U.S.
Fish and Wildlife Service for the collection permit. This
work was funded by the Lower Colorado River Multi-
Species Conservation Program.

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Florida Entomologist 94(1)







Paraiso et al.: Egg Parasitoids of the Cactus Moth


EGG PARASITOIDS ATTACKING CACTOBLASTIS CACTORUM
(LEPIDOPTERA: PYRALIDAE) IN NORTH FLORIDA

OULIMATHE PARAISO1, STEPHEN D. HIGHT2, MOSES T. K. KAIRO1 AND STEPHANIE BLOEM3
'Center for Biological Control, College of Engineering Sciences, Technology and Agriculture,
Florida Agricultural & Mechanical University, Tallahassee, FL 32307

2USDA, ARS, CMAVE, Tallahassee, FL 32308

3USDA, APHIS, PPQ, CPHST, Plant Epidemiology and Risk Analysis Laboratory, Raleigh, NC 27606

ABSTRACT

Interest in the natural enemies of Cactoblastis cactorum (Berg) has increased since the moth
was found in Florida in 1989. Previous surveys for natural enemies in Argentina identified
egg parasitoids in the family Trichogrammatidae as potentially important control agents of
C. cactorum. A study was conducted in north Florida to identify and to assess occurrence of
egg parasitoids attacking this invasive moth in its new homeland. Surveys undertaken at 6
locations in north Florida from Jul 2008 to Dec 2009 revealed that eggsticks of C. cactorum
were attacked by egg parasitoids from the Trichogramma genus: T pretiosum Riley, T fuen-
tesi Torre, and an additional unidentified Trichogramma species belonging to the T pretio-
sum group. In order to assess the importance of these egg parasitoids, the fate of individual
C. cactorum eggsticks was determined during weekly visits to each site. This assessment
showed that the combined level of parasitism of C. cactorum eggsticks was very low with less
than 0.2% of host eggs attacked at any one of the 6 sites. While parasitoids attacked smaller
eggsticks, there was no correlation between the numbers of eggs in an eggstick attacked with
increasing number of eggs/eggstick. Comparing the mean number of eggs/eggstick, there
was no difference between the 3 flight periods of C. cactorum, but there was a difference be-
tween the 6 sites. Based on these results, the use of Trichogramma wasps as an inundative
biological control agent, complementary to the Sterile Insect Technique application, is dis-
cussed.

Key Words: Cactoblastis cactorum, cactus moth, Trichogramma, egg parasitoids, North Florida

RESUME

El interns en los enemigos naturales de Cactoblastis cactorum (Berg) ha aumentado desde
que esta especie fue encontrada en el estado de Florida en 1989. Busquedas de enemigos na-
turales de C. cactorum hechas en anos pasados en Argentina identificaron a parasitoides de
huevos de la familiar Trichogrammatidae como enemigos naturales de possible importancia
para esta especie. Llevamos a cabo un studio en seis localidades en el norte del estado de
Florida con el objetivo de identificar y evaluar la ocurrencia de parasitoides de huevos ata-
cando a esta especie en su nueva area de distribuci6n. La busqueda de parasitoides llevada
a cabo entire julio del 2008 y diciembre del 2009 indico que los bastoncitos de huevos de C.
cactorum son atacados por parasitoides del genero Trichogramma: T pretiosum Riley, T
fuentesi Torre, y una especie adicional no identificada perteneciente al grupo taxon6mico de
Trichogramma pretiosum. Para evaluar la importancia de estos parasitoides en el control de
C. cactorum, seguimos el destino de bastoncitos individuals a trav6s de visits semanales
a cada una de las localidades. Estas observaciones demostraron que el grado de parasitismo
en esos basoncitos es muy bajo, con menos de 0.2% de los huevos parasitados en cualquiera
de las seis areas. Mientras que observamos que los parasitoides atacaron bastoncitos de hue-
vos de tamaho pequeno, no hubo correlaci6n entire el nmmero de huevos parasitados por bas-
toncito y el tamano del mismo. Comparando el numero promedio de huevos por bastoncito,
no detectamos diferencia en los bastoncitos ovipositados en las tres generaciones anuales de
C. cactorum pero detectamos diferencias dependiendo del area. Basado en estos resultados,
discutiremos el uso de parasitoides del genero Trichogramma como agents inundativos de
control biol6gico, complementando la aplicaci6n de la T6cnica del Insecto Est6ril contra
C. cactorum.

Translation provided by the authors.


fect example of a successful weed biological con-
trol agent (Moran & Zimmermann 1984). In 1925,


The cactus moth, Cactoblastis cactorum (Berg)
(Lepidoptera: Pyralidae), is often cited as the per-







Florida Entomologist 94(1)


the cactus moth was introduced from its native
Argentina into Australia to control prickly pear
cactus, Opuntia spp., which had originally been
brought into Australia for commercial purposes
(Dodd 1940; Mann 1970). The cactus had become
invasive and made large tracts of rangeland unfit
for grazing cattle. Within a few years after the in-
troduction of C. cactorum into Australia, US $6
million worth of rangeland was restored, equiva-
lent to more than US $60 million in today's dol-
lars (Dodd 1940; Williamson 2009). Based on
these promising results, C. cactorum was im-
ported from Australia to South Africa, Mauritius,
and Hawaii to manage other non-native and inva-
sive Opuntia spp. (Moran & Zimmermann 1984).
In 1957, C. cactorum was introduced into several
Caribbean islands (Nevis, Montserrat, and Anti-
gua) to control non-native as well as native Opun-
tia spp. (Simmonds & Bennett 1966). Unfortu-
nately, the implementing agencies did not fully
consider the potentially injurious environmental
impacts ofC. cactorum if this insect were to move
to neighboring countries where some species of
Opuntia are important native species and some
are commercially important (Stiling et al. 2004).
The first record of C. cactorum in the U.S. was
from Bahia Honda Key, Florida, in Oct 1989
(Dickel 1991). It is uncertain how the moth ar-
rived in Florida, but several interceptions of Car-
ibbean ornamental Opuntia spp. infested with C.
cactorum were found at ports of entry in south
Florida during the 1980s and 1990s (Pemberton
1995; Zimmermann et al. 2001; Stiling 2002; Si-
monsen et al. 2008). Since its appearance in Flor-
ida, C. cactorum has become a threat to native
Opuntia spp. in North America. Current manage-
ment options include the use of Pherocon 1-C
Wing traps (Trece Incorporated, Salinas, CA)
baited with a 3-component synthetic sex lure (Su-
terra, LLC, Bend, OR) to identify the presence of
the moth, coupled with removal of infested plants
to reduce C. cactorum populations (Bloem et al.
2005; Hight & Carpenter 2009). Complementary
to the detection, monitoring, and removal efforts,
implementation of the Sterile Insect Technique
(SIT) is being used to slow the geographic expan-
sion of C. cactorum in the U.S. (Hight et al. 2002;
Bloem et al. 2005; Bloem et al. 2007). In Mexico,
localized invasions of C. cactorum on 2 islands
were eradicated in 2008 with a program of phero-
mone traps, host removal, and SIT (NAPPO 2006;
NAPPO 2008; NAPPO 2009).
Bennett & Habeck (1995) suggested biological
control as an additional control option that should
be considered for C. cactorum. Pemberton &
Cordo (2001) reported that several larval and pu-
pal parasitoids attacked the cactus moth in South
America, including species of Hymenoptera (Bra-
conidae, Chalcidae, and Ichneumonidae), and 1
Diptera (Tachinidae). They also reported on 2
chalcid species (Brachymeria ovata (Say) and B.


pedalis Cresson) and 1 unidentified egg parasi-
toid from the family Trichogrammatidae attack-
ing C. cactorum in Florida. Logarzo et al. (2008)
found the larval parasitoid Apanteles alexander
Brethes (Hymenoptera: Braconidae) and the egg
parasitoid Trichogramma pretiosum Riley (Hy-
menoptera: Trichogrammatidae) attacking C.
cactorum in Argentina.
Trichogrammatid egg parasitoids have been
used successfully for inundative biological control
against major lepidopteran pests such as corn
borers, sugarcane borers, and cotton bollworm
(Li-Ying 1994; van Lenteren 2000). Egg parasi-
toids are easy to rear in mass quantity in labora-
tory conditions and to release over wide areas. Bi-
ological control can be used to complement and
synergize the application of SIT (Gurr & Kve-
daras 2010). Recent studies showed that the com-
bination of both techniques was more efficient in
controlling pest population of the codling moth,
Cydia pomonella (L.) (Bloem et al. 1998). Syner-
gistic interactions between SIT and fruit fly bio-
logical control with parasitoids increased the sup-
pression of pest fruit flies, even leading to eradi-
cation (Sivinski 1996; Rendon et al. 2006). SIT
and biological control have been successfully com-
bined to combat several lepidopteran pests, in-
cluding C. pomonella (Bloem et al. 1998) and
painted apple moth, Orgyia anartoides (Walker)
(Suckling et al. 2007). Radiation doses for steril-
izing C. cactoblastis adults have been determined
to produce partially sterile but more fit males
which, when mated with wild females, generate
sterile offspring (Carpenter et al. 2005). The com-
bination of egg parasitoid releases and SIT has
the advantage that parasitoids manage high pest
densities, while SIT works best at low pest densi-
ties. In addition, release of sterile insects provides
an egg resource for egg parasitoids increasing the
ratio of natural enemies to adult hosts. Egg para-
sitoids and sterile insects have the characteristic
of being self dispersing and consequently are able
to cover wide areas (Sivinski 1996).
We conducted field surveys in order to identify
egg parasitoids already established in North Flor-
ida that attack C. cactorum. Cactoblastis cac-
torum adults have 3 annual flight periods in
north Florida (Apr-May, Jul-Aug, and Oct-Nov)
(Hight et al. 2005; Hight & Carpenter 2009). We
report on the distribution, seasonality, and para-
sitism parameters of the Trichogramma species
attacking C. cactorum in northern Florida. The
number of eggs/eggstick was compared between
different flight periods and sites to assess host
egg resource for egg parasitoids. The effect of C.
cactorum eggstick size on level of parasitism was
evaluated by comparing number of eggs from par-
asitized versus un-parasitized eggsticks. These
data will be beneficial in promoting discussions
on possible implementation of biological control
for the cactus moth and, in particular, assessing


March 2011







Paraiso et al.: Egg Parasitoids of the Cactus Moth


the potential of an inundative biological control
program against C. cactorum in North America.

MATERIALS AND METHODS

Field surveys were carried out at 6 locations
(Fig. 1) in north Florida from Jul 2008 to Dec
2009. The selection of study sites was based on ex-
isting records of infestations from the literature,
personal observations from preliminary surveys,
and information provided by experts. Female C.
cactorum place their eggs end to end to form a
chain that looks like a short "stick", and the egg
mass is referred to as an eggstick. Although no ex-
tensive field surveys were conducted from May to
Jul 2008 at St. Marks and St. George Island, egg-
sticks with eggs that appeared parasitized were
collected and held in laboratory conditions until
parasitoids emerged. At survey locations, 20 to 30
healthy Opuntia spp. plants were chosen with no
to minor feeding damage by cactus moth larvae
and an average of at least 50 pads per plant. Dur-
ing weekly visits throughout all 3 flight periods,
any new eggstick was identified by plant, pad,
and its general location on the plant so the egg-
stick could be found during subsequent checks. A
mark was made on the plant at the base of the
eggstick with a felt tip pen and a red tape "flag"
affixed to an insect pin placed near the eggstick to
aid in finding the eggstick. The flag was labeled
with a unique number to identify each eggstick.
The oviposition preferences of C. cactorum fe-
males on host plants were recorded by classifying
the attachment of the eggstick to either a glochid
at an areole, to a spine, or on the fruit. Observa-
tions on plant habitat and host eggstick distribu-
tion within the surveyed site and within the se-
lected plant were collected to provide additional
information on the host finding behavior of egg


Penmcola RBeach
N3 .33525: W87.48928 Okaloosa Island
N30.8674 W6 3.7807 St. Maks(NWR)
ImN30.07772: W84.18242


,\ %, \ N30o3,2:WK43935
Mcxico Bcach
N29.94133; W85.40636 \( 1- ,
St. George Island
N2m .506S W4.9:120 1


Fig. 1. Locations and their coordinates surveyed for egg
parasitoids of Cactoblastis cactorum in North Florida.


parasitoids. The number of eggs per eggstick was
determined either by a direct count or by a corre-
lated estimate of eggstick length to egg number
(2.62 0.013 eggs/mm). The ratio of eggstick
length to egg number was calculated in this study
by counting the number of eggs in a segment of
eggstick, replicated on 20 eggsticks. Eggstick
length was estimated in situ by placing a plastic
string next to the eggstick and cutting a piece of
equivalent length. The length of the piece of
string was then measured to the nearest 0.01 mm
with a metric micrometer. Measurements of egg-
sticks were obtained so that the number of eggs
per eggstick could be estimated if the eggstick
was lost before it could be collected and directly
counted. The fate of each eggstick was deter-
mined by making weekly visits to each site to
evaluate the status of previously tagged egg-
sticks. The fate of each eggstick was categorized
as follows: eggstick lost; predated (visible chew-
ing damage) eggs in the eggstick versus non pre-
dated eggs; or parasitized eggs in the eggstick
(black eggs formed before C. cactorum larvae suc-
cessfully developed). Eggsticks were collected if
they were damaged during evaluation or mea-
surement, eggs of the eggstick had hatched, or
eggs appeared predated or parasitized. Eggsticks
with viable eggs were collected and held in small
plastic cups (30 mL) under laboratory conditions
(25 1 C, 16:8 L:D and 40-60% RH) to record
hatch rate. Eggsticks with parasitized eggs were
collected and monitored in the laboratory to de-
termine the emergence rate, number of eggs/egg-
stick attacked by parasitoids, number of parasi-
toids emerging per parasitized egg, and to ascer-
tain the identity of the parasitoids. Parasitoid
specimens were submitted to R. Stouthamer, De-
partment of Entomology, University of California,
Riverside, for molecular identification. The se-
quencing of ribosomal DNA Internal Transcribed
Spacer 2 (ITS 2) was used to identify the different
species of egg parasitoids.

Data Analysis

The numbers of eggs/eggstick at different
flight periods for each surveyed location and the
average number of eggs/eggstick at each site were
log transformed before analyses to satisfy the as-
sumptions of the analysis of variance. One way
analysis of variance (PROC GLM) was applied to
the log transformed data and the separation of
means was made with the least significant differ-
ence (LSD) test. Comparison of number of eggs/
eggstick that was parasitized versus number of
eggs/eggstick not parasitized was also evaluated
with a one-way analysis of variance (PROC
GLM). Since only a few eggsticks with parasitized
eggs were collected in this study (see text below),
comparisons between eggsticks with parasitized
eggs were made against the same number of ran-







Florida Entomologist 94(1)


domly selected eggsticks with un-parasitized
eggs. Variation between the number of eggs for
parasitized eggsticks and the number of eggs for
the randomly selected un-parasitized eggsticks
was analyzed by a folded F test (Davis 2007). Be-
cause the variances in numbers of eggs for egg-
sticks with parasitism and number of eggs in egg-
sticks without parasitism were not significantly
different, means of these 2 groups were compared
with a two-sample t-test. A Pearson's Correlation
Coefficient (r) was calculated to determine
whether the numbers of eggs parasitized were de-
pendent on the number of eggs/eggstick. The SAS
Statistical Software Version 9.2 (SAS Institute,
Cary, North Carolina) was used to perform the
statistical analyses. Estimates of central tenden-
cies were reported as mean standard error of
mean.

RESULTS AND DISCUSSION

Although host plant species of Opuntia strict
(Haworth) Haworth, 0. humifusa (Rafinesque)
Rafinesque, and 0. ficus-indica (L.) P Miller var-
ied among the different geographic regions sur-
veyed, the oviposition preferences of C. cactorum
females was similar on the various species (Table
1). In this study, parasitized eggsticks of C. cac-
torum appeared mostly on the areole/glochid
structure of the pads (Table 1).
Altogether, 1,527 eggsticks with 91,013 C. cac-
torum eggs, not including 344 eggsticks missing
from the field or lost during collection, were
tagged on plants of Opuntia spp. (Table 2). Of all
the eggsticks checked, 62% were collected on Oka-
loosa Island and had a mean of 59 (+/- 1.83) eggs/
eggstick. The proportion of eggsticks examined in
the laboratory as percentage of all eggsticks sur-
veyed at the 6 field sites ranged from 53 to 100%,
except for summer 2008 at St. George Island and
St. Marks National Wildlife Refuge (NWR) in
which only 30% and 24%, respectively, of the mon-
itored eggsticks were examined (Table 2). The
majority of the eggsticks from these 2 locations
for this flight period were recorded as lost (Table
2). The cause for this high number of lost egg-
sticks is not clear. Several biotic and abiotic fac-
tors could have contributed to the high number of
lost eggsticks. During summer 2008, 23% of egg-
sticks examined from St. Marks had eggs that
were preyed upon compared with less than 3% in
other locations. Although not directly observed at
St. George Island or St. Marks, substantial preda-
tion ofC. cactorum eggs by ants has been recorded
in South Africa (Robertson 1984). Because the
plants surveyed at St. Marks were located within
100 m of the waters of the Gulf of Mexico, strong
winds characteristic of coastal regions could have
knocked eggsticks off the plants. All other study
sites were along the Gulf Coast; in none of them
were the plants as close to the water as at St.


Marks. In addition, heavy rainfall may have sep-
arated the eggsticks from plants, but we do not
have any data on the severity of the rain storms
at different study sites. Cactoblastis cactorum life
table studies in Argentina (Logarzo et al. 2009)
and South Africa (Robertson & Hoffmann 1989)
identified rain and wind as major factors contrib-
uting to mortality of eggs.
Surveyed sites and oviposition periods were
analyzed to evaluate their influence on number of
eggs/eggstick. Eggsticks were collected for multi-
ple oviposition periods at 3 sites (St. George Is-
land, St. Marks, and Okaloosa Island) (Table 2).
The numbers of eggs/eggsticks for the different
oviposition periods were not significantly differ-
ent for St. George Island (F = 1.84, df = 1, P =
0.18), St. Marks (F = 93.86, df = 3, P = 0.07), or
Okaloosa Island (F = 0.22, df = 3, P = 0.88). Be-
cause the numbers of eggs/eggstick for multiple
oviposition periods were not different, eggsticks
from all flight periods were pooled to calculate the
means for those sites (St. George Island (62 2.8),
St. Marks (53 2.8), and Okaloosa Island (59
1.8)). The pooled eggsticks were used to compare
the number of eggs/eggstick between all 6 sites
and significant differences were found (F = 11.44,
df = 5, P < 0.0001) (Table 2).
Female C. cactorum laid similar numbers of
eggs/eggstick for each of the 3 oviposition periods
but not at all 6 survey sites along the Florida pan-
handle. The longest eggsticks were observed at
St. George Island, Pensacola Beach, and Oka-
loosa Island (Table 2). Significantly smaller egg-
sticks were recorded at St. Marks and Mexico
Beach (Table 2). Panacea had significantly
smaller number of eggs/eggstick than all other
sites (Table 2). The cause of differences between
eggsticks at the various sites was unclear. Studies
in South Africa identified differences in total fe-
cundity of C. cactorum due to host plant species,
the flight period when eggs were laid, and the
temperature during oviposition (Robertson 1989).
We did not distinguish eggsticks collected from
different host plants (Table 1). While South Afri-
can female C. cactorum had significantly higher
fecundity during the summer flight (Robertson
1989), our study did not show any difference in
number eggs/eggstick between flight periods in
north Florida. Cactoblastis cactorum has a ten-
dency to oviposit on plants with high nitrogen
(Myers et al. 1981; Robertson 1987), but we have
no direct measurements of plant quality at our
sites.
Comparing the number of eggs/eggstick for
eggsticks that were parasitized (38 13.7) (Table
3) against un-parasitized eggsticks (61 13.1) re-
vealed a significant difference (pooled t test =
3.14, df= 12, P = 0.0085). Although the number of
eggs/eggstick was highly variable, the variation of
the number of eggs/eggstick for parasitized ver-
sus the randomly selected un-parasitized group


March 2011









TABLE 1. SITES SURVEYED IN NORTH FLORIDA FOR TRICHOGRAMMA EGG PARASITOIDS OF CACTOBLASTIS CACTORUM EGGSTICKS ON OPUNTIA SP. AND ADDITIONAL INFORMA-
TION ON MOTH OVIPOSITION PREFERENCE.

Percent Eggsticks at Attachment
Location'
Species of Number Host Total Number
GPS Dates Eggsticks Total Number Opuntia Host Plant Eggsticks Areole/
Site Coordinate Surveyed Surveys Plant Examined Evaluated Glochid Spine Fruit Missing2

Pensacola Beach N30.33525 Summer 2008 10 0. strict 20 120 50 34 16 0


St. George Island




St. Marks (NWR)


Mexico Beach


Panacea


Okaloosa Island


W87.48928 (Jul 10-Sep 10, 08)


N29.65068 Summer 2008
W84.9120 (Jull7-Sepl9, 08)
Fall 2008
(Sep 25, 08-Feb 25, 09)
N30.07772 Summer 2008
W84.18242 (Jul 15,-Sep 12, 08)
Fall 2008
(Oct 01, 08-Feb 25, 09)
Spring 2009
(Apr 17-Jul 15, 09)
Fall 2009
(Oct 07, 09-Jan 12, 10)
N29.94133 Fall 2009
W85.40636 (Oct 21-Nov12, 09)
N30.03127 Fall 2009
W84.39353 W84.39353

N30.08674 Fall 2008
W86.37807 (Oct 08, 08-Feb 27, 09)
Spring 2009
(Apr 08-Jul 08, 09)
Summer 2009
(Jul 01-Sep 25, 09)
Fall 2009
(Sep 18, 09-Jan 12, 10)


0. humifusa
0. ficus-indica
10 0. strict


18 0. stricta
0. humifusa


3 0. ficus-indica


3 0. stricta
0. ficus-indica

21 0. ficus-indica


105 63 30 7 0

28 89 11 0 0


45 80 13 7 0

9 78 0 22 0

47 88 2 0 10

151 n/a n/a n/a n/a


29 n/a n/a n/a n/a


65 n/a n/a n/a n/a


186 81 18.5 0.5 0


79 15 1 4

77 19 2 2


151 n/a n/a n/a n/a


Attachment locations of eggsticks that were not determined is indicated by "n/a".
Information about eggstick attachment failed to be recorded.


















TABLE 2. NUMBER OF CACTOBLASTIS CACTORUM EGGSTICKS COLLECTED, LOST IN THE FIELD, EXAMINED IN THE LABORATORY, AND MEAN NUMBER OF EGGS PER EGGSTICK
SE AT DIFFERENT SITES IN NORTH FLORIDA FOR DIFFERENT OVIPOSITION PERIODS.

Total Number Percent Total Number Mean Number Overall Mean Eggs/
Total Number (Percent) Eggsticks Moth Eggs Eggs/Eggstick Eggstick
Site Flight Period Eggsticks Tagged Eggsticks Lost Examined Examined + SE SE at Each Site'

Pensacola Beach Summer 2008 120 69 (58) 42 7,402 62+/-1.5 62 1.5 a
St. George Island Summer 2008 105 84 (70) 30 6,685 64+/-2.0 62 2.8 a
Fall 2008 28 13 (46) 53 1,614 58+/-3.6

St. Marks (NWR) Summer 2008 45 35 (77) 24 3,088 68+/-2.9 53 2.8 b
Fall 2008 9 4 (44) 55 513 57+/-3.3
Spring 2009 47 23 (46) 54 2,561 54+/-3.1
Fall 2009 151 0 (0) 100 6,917 46+/-1.4

Mexico Beach Fall 2009 29 0 (0) 100 1,522 52+/-3.0 52 3.0 b
Panacea Fall 2009 65 0 (0) 100 2,892 45+/-1.9 45 1.9 c
Okaloosa Island Fall 2008 186 61(29) 71 11,118 60+/-1.4 59 1.3 a
Spring 2009 308 3(1) 99 20,527 61+/-1.2
Summer 2009 280 21(8) 92 17,126 60+/-1.1
Fall 2009 151 1(0.6) 99 8,638 57+/-1.2

Means with different letter are significantly different (P < 0.05).





















TABLE 3. LOCATION AND DATE PARASITIZED CACTOBLASTIS CACTORUM EGGSTICK WAS COLLECTED, IDENTITY OF PARASITOID SPECIES, NUMBER OF EGGS PER EGGSTICK,
NUMBER OF PARASITIZED EGGS, NUMBER OF PARASITOIDS EMERGED, FEMALE RATIO, AND PARASITISM LEVEL OF EGG PARASITOIDS ATTACKING C. CACTORUM
IN NORTH FLORIDA.

Collection Flight Number Eggs/ Number Parasitized Number Parasitoids Percent Level of
Site Date Period Trichogramma sp. Eggstick Eggs/Eggstick # (%) Emerged Females Parasitism (%)

St. Marks 05/15/08 Spring 08 T pretiosum 73 5 (7) 8 75 n/a1
05/15/08 T pretiosum 78 17(22) 34 85

Pensacola Beach 04/22/08 Spring 08 unknown 88 19 (22) 18 77 n/a
08/06/08 Summer 08 T pretiosum 44 8(18) 5 40 0.2
08/13/08 T pretiosum 18 6(33) 10 70

Okaloosa Island 10/16/08 Fall 08 T fuentesi 20 2(10) 11 73 0.1
10/16/08 T pretiosum 42 10(24) 7 71
10/16/08 T fuentesi 56 6(11) 16 62
11/03/08 unknown 52 3 (6) 9 56
10/23/09 Fall 09 T fuentesi 25 13 (52) 63 89 0.1

'Indicates that the level of parasitism was not determined.







Florida Entomologist 94(1)


was similar (folded F test = 1.10, df= 6,P = 0.91),
suggesting that the difference found between the
two groups was not driven by unequal or extreme
variation. However, there was not a significant
correlation between the number of eggs/eggstick
and number of eggs parasitized by Trichogramma
spp. (n = 7, r = -0.16, P = 0.74). Therefore, while fe-
male Trichogramma spp. parasitized eggsticks
with fewer eggs, they did not parasitize more eggs
as the number of eggs in an eggstick increased.
The average number of eggs parasitized in an
eggstick was 9 (5.8).
Ten eggsticks were found parasitized at 3 of
the 6 sites surveyed (Pensacola Beach, St. Marks,
and Okaloosa Island). Five of the parasitized egg-
sticks were found at Okaloosa Island. Parasitized
eggsticks were found during all 3 oviposition pe-
riods of C. cactorum females: the spring flight (St.
Marks and Pensacola Beach), summer flight
(Pensacola Beach), and fall flight (Okaloosa Is-
land). Of the 496 eggs in the 10 parasitized egg-
sticks, a total of 89 eggs (or 18%) were parasit-
ized, resulting in the emergence of 181 adult par-
asitoids with a sex ratio of 70% (+14) females (Ta-
ble 3). The level of parasitism by Trichogramma
spp., relative to the total number of eggs exam-
ined during the different flight periods for each
site, was less than 0.2% of total C. cactorum eggs
collected (Table 3). We did not observe any para-
sitized eggsticks at St. George Island, Mexico
Beach, or Panacea.
Two species of Trichogramma were reared
from C. cactorum eggsticks in north Florida (Ta-
ble 3) and identified by differences in IST2 se-
quences. Trichogramma pretiosum was collected
at St. Marks, Pensacola Beach, and Okaloosa Is-
land, while T fuentesi Torre was recovered only
from Okaloosa Island. It was not possible to iden-
tify 2 collections of Trichogramma spp. from Oka-
loosa Island; one because a good molecular se-
quence could not be obtained and for the other the
sequence was not in the database and possibly
represents a new species in the T pretiosum
group (R. Stouthamer, UC-Riverside, personnel
communication).
More than 15 million ha of agriculture and
forestry worldwide are treated annually with
Trichogramma egg parasitoids (van Lenteren
2000). Trichogrammatid wasps have been used
successfully in inundative release programs
against lepidopteran pests in greenhouses and
crop production worldwide (Smith 1996). Inun-
dative releases of Trichogramma spp. have been
implemented in Florida to control major lepi-
dopteran pests of collards, cabbages, soybeans,
bell peppers, tomatoes, corn, and tobacco produc-
tion (Martin et al. 1976). Trichogramma pretio-
sum is commonly found in the Western hemi-
sphere. This Trichogramma species has been re-
leased commercially against major lepidopteran
pests such as cotton leafworm (Alabama argilla-


cea) (Hiibner), corn earworm (Helicoverpa zea)
(Boddie), tomato pinworm (Keiferia lycopersi-
cella) (Walshingham), sugarcane borers (Di-
atraea spp.), and cabbage looper (Trichoplusia
ni) (Hiibner) (Pinto et al. 1986; Hassan 1993; Li-
Ying 1994; Monje et al. 1999). Trichogramma fu-
entesi have been recorded in countries in South
America (Argentina, Columbia, Mexico, Peru,
and Venezuela) and in the U.S. (Alabama, Cali-
fornia, Florida, Louisiana, New Jersey, South
Carolina and Texas) (Fry 1989, Pinto 1999). Its
primary hosts are species from the Noctuidae
family such as H. zea and Heliothis virescens (F.)
and from the Pyralidae family such as Diatrea
saccharalis (F.), Ephestia kuehniella Zeller, and
Ostrinia nubilalis (Hiibner) (Fry 1989; Wilson &
Durant 1991; Pintureau et al. 1999; Querino &
Zucchi 2003). Trichogramma parasitoids also are
widely used for pest control in orchards
(Olkowski & Zang 1990). The observed low inci-
dence of the wasps in natural areas might be ex-
plained by unfavorable environmental factors or
natural plant chemicals (Smith 1996; Romeis et
al. 1997, 1999). However, contrary to other natu-
ral enemies, Trichogramma can be easily and
cheaply mass-reared for the implementation of
an inundative biological control program.
The potential for inundative releases of Tri-
chogramma spp. as a strategy against C. cac-
torum is currently being investigated with sus-
tainable laboratory colonies of T fuentesi origi-
nating from field collected insects reared from
parasitized C. cactorum eggsticks. Biological
characteristics (sex ratio, egg load, and longevity)
and different behavioral mechanisms (influence
of parasitoid age, density, and host age on parasit-
ism) involved in host finding of T fuentesi reared
on C. cactorum eggs are being evaluated. The in-
undative releases of Trichogramma wasps could
be integrated in the current pest management
system based on SIT applications during the 3
flight periods by building Trichogramma popula-
tions. This field survey was useful in identifying a
potential inundative biological control agent that
could be integrated within a pest management
strategy against C. cactorum.

ACKNOWLEDGMENTS

We thank Shalom Benton (FAMU) for field collection
and laboratory assistance and Chris Albanese, Michael
Getman, and John Mass (USDA-ARS-CMAVE, Talla-
hassee) for field assistance. We thank Stuart Reitz
(USDA-ARS-CMAVE, Tallahassee, FL) and Jim Nation
(University of Florida) for comments on earlier drafts of
this manuscript. This work is funded under the FAMU-
USDA APHIS Cooperative Agreement, 07-10-8100-
0755-CA. Mention of trade names or commercial prod-
ucts in this publication is solely for the purpose of pro-
viding specific information and does not imply
recommendation or endorsement by the U.S. Depart-
ment of Agriculture.


March 2011







Paraiso et al.: Egg Parasitoids of the Cactus Moth


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Florida Entomologist 94(1)







Rengifo et al.: Host Status of Purple Passionfruit for Ceratitis capitata


HOST STATUS OF PURPLE PASSIONFRUIT
FOR THE MEDITERRANEAN FRUIT FLY (DIPTERA: TEPHRITIDAE)


JOSE A. RENGIFO1, JAVIER G. GARCIA1, JOHN F. RODRIGUEZ1 AND KRIS A. G. WYCKHUYS2
1Colombian Institute for Agriculture and Livestock, ICA, Bogota, Colombia

2International Center for Tropical Agriculture CIAT, Cali, Colombia

ABSTRACT

The Mediterranean fruit fly Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) is a key
pest of a wide range of fruit crops and the focus of rigid quarantine restrictions and eradica-
tion measures in several countries. In Colombia, the susceptibility of purple passionfruit
(Pasiflora edulis f edulis Sims; Violales: Passifloraceae) to C. capitata is uncertain. Field col-
lections of fruit were made to evaluate natural infestation. Forced infestation studies were
conducted in the laboratory with punctured and intact fruit to determine the acceptability
of fruit at different stages of maturity and physiological suitability of fruit to development.
No C. capitata larvae were found and no adults emerged from a total of 976 hand-picked fruit
and 623 fallen fruit. In the meantime, trap data indicated that C. capitata is not present in
the principal passionfruit production regions. For intact fruit, C. capitata females oviposited
exclusively in fruit of maturity level zero, with 41.67% of fruit accepted for oviposition and
an average of 183.1 33.8 eggs per fruit. No oviposition was recorded in fruit of maturity lev-
els 2 and 4. For punctured fruit, C. capitata oviposited a total of 84,410 and 84,250 eggs into
fruit of maturity levels 0 and 2, respectively, but no C. capitata adults emerged from fruit at
either maturity level. Laboratory tests suggest that purple passionfruit is a non-host for C.
capitata.

Key Words: quarantine pest, Ceratitis capitata, host status, risk analysis, fruit fly

RESUME

La mosca del Mediterraneo Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) es una
plaga clave de una amplia gama de frutales y es el foco de estrictas restricciones cuarente-
narias y medidas de erradicaci6n en various paises. En Colombia, la susceptibilidad del ma-
racuyd morado (Pasiflora edulis f edulis Sims; Violales: Passifloraceae) a C. capitata es
incierta. Se hicieron colectas de frutos en campo para evaluar el nivel de infestaci6n. En el
laboratorio se desarrollaron studios de infestaci6n forzada con frutos perforados e intactos
para determinar la aceptabilidad del fruto en los diferentes estados de maduraci6n e idonei-
dad fisiol6gica del desarrollo de los frutos. No se encontraron larvas de C. capitata ni adults
emergidos en un total de 976 frutos recogidos manualmente y 623 frutos caidos. Mientras
tanto, los datos de capture indicaron que C. capitata no esta present en las principles re-
giones de producci6n del maracuya. Para frutos intactos, las hembras de C. capitata ovipo-
sitaron exclusivamente frutos de nivel de maduraci6n cero, con 41.67% de aceptaci6n de
frutos para oviposici6n y en un rango de 183.1 33.8 huevos por fruto. No se registry ovipo-
sici6n en frutos con niveles de maduraci6n 2 y 4. Para frutos perforados, C. capitata oviposit6
un total de 84,410 y 84,250 huevos dentro de frutos con nivel de maduraci6n 0 y 2 respecti-
vamente, pero no emergieron adults de C. capitata de los frutos en ningun nivel de madu-
raci6n. Las pruebas de laboratorio sugieren que el maracuyd morado no es hospedero para
C. capitata.

Translation provided by the authors.


Tephritid fruit flies are key pests of a wide va- (Wiedeman), a destructive pest of multiple fruit
riety of fruit species, affecting crop yield, quality crops worldwide (Liquido et al. 1991). In assess-
of harvested produce, and (international) market ing risk of C. capitata arrival in the U.S. and de-
access (e.g., Robinson & Hooper 1989; Aluja & veloping associated quarantine protocols, su-
Mangan 2008). Given the polyphagous nature of preme precaution is taken to avoid entry of poten-
many fruit fly species, quarantine restrictions are tial host fruits of this pest. Listings of the status
in place to avoid their introduction in certain of particular fruits as hosts of C. capitata are the
countries or geographical regions. A key quaran- cornerstone of quarantine restrictions (Liquido et
tine pest for the continental United States is the al. 1991). However, current restrictions include
Mediterranean fruit fly, Ceratitis capitata fruit species for which there is poor information







Florida Entomologist 94(1)


regarding C. capitata host status. Hence, re-
search is needed to revise and update C. capitata
host information and thereby improve quarantine
decision making (Aluja et al. 2004; Peia et al.
2006; Jenkins & Goenaga 2008; Staub et al. 2008;
De Graaf 2009; Follett et al. 2009).
Purple passionfruit (Passiflora edulis f. edulis
Sims) is one of several tropical fruits that is well-
positioned in local markets and gradually becom-
ing popular internationally (Ocampo 2007; Wyck-
huys et al. in press). In Colombia, purple passion-
fruit is mainly grown by small-scale, resource-
poor farmers on a total area of 100-400 ha. It is a
profitable crop and fresh fruit is increasingly be-
ing exported to northern Europe and Canada
(Wyckhuys, unpublished data). Entry of fresh
fruit into the continental U.S. is not permitted
currently, based upon its presumed suitability as
a host forAnastrepha spp. and C. capitata.
Liquido et al. (1991) list C. capitata as a poten-
tial pest of P edulis, but provide no evidence of
adult fly emergence from field-collected fruit.
Other reports indicate C. capitata is an occasional
pest of Passiflora sp., without specifying the exact
crop species, botanical form or variety (Thomas et
al. 2001). Yellow passionfruit (P edulis f. fla-
vicarpa Degener) is reported as a possible host of
C. capitata in Hawaii (Akamine et al. 1954), while
many tephritids attack certain Passiflora species
in Brazil (Aguiar-Menezes et al. 2002). In Colom-
bia, national pest survey records for C. capitata
maintained since 1986 have not detected this pest
in the principal production regions of purple pas-
sionfruit (ICA, 2009). As a note of caution, it is im-
portant to indicate that climate change could
cause altitudinal range shifts of pest tephritids
and may eventually bring C. capitata into those
production regions in the future (Hill et al. 2011).
Considering a lack of scientific information re-
garding purple passionfruit host status for C. cap-
itata and the importance of its production as
source of income for rural smallholders, we at-
tempted to determine the host suitability of Co-
lombia-grown purple passionfruit for C. capitata
using standard methods (Cowley et al. 1992). This
information can be used to re-evaluate the quar-
antine status of this fruit for market access to the
United States.

MATERIALS AND METHODS

All methodologies for host status screening
were adopted from Cowley et al. (1992), taking
into account parameters set by RSPM No. 30
(NAPPO 2008) and APPPC RSPM No. 4 (FAO
2005; Follett & Hennessey 2007).

Field Collections

Between Sep 2008 and May 2010, sampling
was done during 4 distinct events in the princi-


pal purple passionfruit production regions, lo-
cated in the departments of Boyaca, Cundi-
namarca, Tolima, and Huila (Colombia). During
each sampling event, 9-16 different purple pas-
sionfruit orchards were visited and fruit was col-
lected from each orchard. Fruit samples con-
sisted of hand-picked fruit of different maturity
levels (i.e., fruit harvested from vines) and fallen
fruit, collected from the ground. Fruit was sam-
pled in a random fashion, and the number of
fruit collected from each orchard depended upon
phenological stage of the crop. We collected a to-
tal of 405, 285, 183, and 113 hand-picked fruit
from Boyaca, Cundinamarca, Huila, and Tolima,
respectively. Respective numbers of fallen fruit
collected from each department were 345, 124,
96, and 58.
Fruit samples were counted, weighed and
taken to the Horticulture Research Center CIAA
(Chia, Colombia) in ventilated plastic containers
(70 x 50 x 50 cm) for further laboratory process-
ing. In the laboratory, fruit samples were kept at
22.0 + 2.0C, 65% RH and 12:12 L:D. Within 1
week following the collection, containers were
screened for presence of fruit fly puparia, and
fruit were dissected to assess presence of te-
phritid larvae. Larvae were subsequently trans-
ferred to ventilated plastic Petri dishes with
moistened vermiculite. Petri dishes were checked
daily for adult emergence. We recorded the num-
ber of tephritid larvae and C. capitata adults for
each sampling event and production region.
Simultaneous with field collections, McPhail
traps (baited with protein hydrolysate; Cebofrut,
AgroBiologicos SAFER, Medellin, Colombia) were
deployed in orchards in each production region
and visited bi-weekly to record the number of C.
capitata adults. A total of 6 traps were deployed
per orchard, of which 5 were placed within the or-
chard itself and a sixth trap was placed outside
the orchard in the dominant surrounding habitat
type. To check trap attractiveness, we recorded
captures of other tephritids.

Laboratory Experiments

Insect material was collected from coffee fruit
(Coffea arabica L.) in commercial orchards in Fre-
donia (Antioquia, Colombia), at 1,400 m altitude,
and Medellin (Antioquia), at an altitude of 1,493
m. Upon field collection, fruits were transferred to
the ICA Entomology Laboratory in Bello (Antio-
quia). Each fruit was dissected and any tephritid
larvae were allowed to pupariate in vermiculite.
Puparia of C. capitata were subsequently taken to
the Quarantine Treatment Laboratory of the Co-
lombian Institute for Agriculture and Lifestock
ICA in Mosquera (Cundinamarca) for further ex-
perimenting. Adults from field collected puparia
were exposed to mango (Mangifera indica L.), a
preferred host of C. capitata (NAPPO 2008).


March 2011







Rengifo et al.: Host Status of Purple Passionfruit for Ceratitis capitata


Adult flies were maintained within mesh cages
(25x25x25 cm), allowed ample access to water
and fed ad libitum with torula yeast and sugar.
All insect developmental stages were maintained
within climate-controlled rearing chambers at 25
+ 1C, 65 5% RH and 12:12 L:D. Second genera-
tion C. capitata adults were then used for host
status trials. Laboratory experiments were car-
ried out between Oct 2008 and May 2009.Voucher
specimens of study insects were kept at the ICA
laboratory.
All fruits used in the experiment were selected
and harvested in several purple passionfruit or-
chards in Venecia (Cundinamarca) or mango or-
chards in La Mesa (Cundinamarca). Fruit of dif-
ferent maturity levels were selected based on
commonly-used color tables for either mango or
purple passionfruit (ICONTEC 1999; Pinz6n et
al. 2007). Prior to use in experiments, fruit was
disinfected by immersion in a 0.05% sodium hy-
pochlorite solution for 10 min. Subsequently, each
fruit was dried and stored in plastic containers to
use in host status trials. Fruit was used for exper-
imenting within 72 h of harvest.

Oviposition Preference Assay

A total of 120 C. capitata pairs, aged 14 d, were
placed within a mesh cage (70 x 50 x 50 cm) (Vidal
et al. 2005) and allowed access to water and ad li-
bitum torula yeast and sugar. Within each cage,
we placed 8 purple passionfruit of each of 3 matu-
rity levels (i.e., maturity 0, 2, and 4; see Pinzon et
al. 2007). Purple passionfruit are approximately 5
cm in diameter. After 24 h, fruit was removed
from the cages and dissected to determine the to-
tal number of C. capitata eggs. Over the course of
3 d, fruit were placed within each cage and sub-
ject to the same ovipositing C. capitata females.
The experiment was carried out with 3 replicates,
thus screening 72 fruit per maturity level. The
number of eggs within fruit of differing maturity
level was compared by one-way analysis of vari-
ance (ANOVA). For all analyses, the statistical
package SAS was used.

Host Status Trials

Based upon results of the previous assay, fur-
ther trials were conducted to determine purple
passionfruit host status to C. capitata. To stimu-
late fly oviposition, fruit was punctured with
standard dissection pins (10 pinholes 1-2 mm into
the fruit) before placing them within experiment
cages (FAO 2005; NAPPO 2008). Purple passion-
fruit of maturity levels 0 and 2 were included in
trials, while mango fruit (maturity degree 2 or 3)
was used as a positive control. We placed 11 fruit
per cage (70 x 50 x 50 cm) with 120 C. capitata
pairs, aged 14-19 d and provided with water and
ad libitum torula yeast and sugar. There were 3


replicates of each fruit type and maturity level,
and simultaneous trials were conducted. In total,
990 purple passionfruit and 495 mango fruits
were subjected to an infestation pressure of 10.9
C. capitata females per fruit.
Over the course of 15 d, fruits within each cage
were replaced on a daily basis, and subsequently
kept within ventilated plastic containers. In a
random fashion, a subsample of 45 fruits of either
species or maturity degree was dissected upon re-
moval from experimental cages to assess the
number of C. capitata eggs. Remaining fruits
were kept at 25 1C, 65 + 5% RH and 12:12 L:D
and were checked daily for larval emergence, pu-
paria formation, or adult eclosion. After 15 d, all
fruits were dissected and C. capitata larvae (per
fruit) were counted and placed within vermiculite
to allow pupariation.

RESULTS

Field Collections

From 2008 up to 2010, a total of 976 purple
passionfruit were hand-picked and 623 fallen
fruit were collected. No C. capitata adults
emerged from any fruit. Diptera larvae were
found within (immature) fruit; all of which suc-
cessfully developed into lonchaeid adults. No C.
capitata adults were caught in McPhail traps de-
ployed in or near orchards in any of the produc-
tion regions. Trap effectiveness was confirmed
through capture of Lonchaeidae (Diptera: Tephri-
toidea) at all locations.

Laboratory Experiments: Oviposition Preference Assay

The number of C. capitata eggs significantly
differed between fruit of distinct maturity de-
grees (F = 18.84, df = 2, P < 0.0001). The highest
number of eggs per fruit (183.7 33.8; mean SE)
was oviposited in purple passionfruit of maturity
level 0, while no eggs were laid in maturity levels
2 and 4.

Host Status Trials

Ceratitis capitata successfully completed its
development on the preferred host mango, but no
adults emerged from punctured fruit of maturity
levels 0 and 2 (Table 1). Few C. capitata larvae
developed in passionfruit, with larval weights
ranging from 2.5 to 3.2 mg. In mango, the weight
of third instars ranged from 9.7 to 10.3 mg. Of the
194 C. capitata that were obtained from passion-
fruit (maturity 0), <10% successfully pupariated.
Puparial weights of individuals developing on
passionfruit ranged from 2.3 to 3.1 mg, compared
to C. capitata puparia from mango that weighed
between 9.3 and 10.2 mg. Also, most C. capitata
puparia that developed from passionfruit were







Florida Entomologist 94(1)


TABLE 1. OVIPOSITION AND SUBSEQUENT DEVELOPMENT OF C. CAPITATA ON MANGO AND PURPLE PASSIONFRUIT (PPF)
OF 2 MATURITY LEVELS UNDER LABORATORY CONDITIONS. DATA REPRESENT CONSOLIDATED NUMBER OF INDI-
VIDUALS WITHIN EACH C. CAPITATA DEVELOPMENT STAGE ON A TOTAL OF 990 PPF FRUIT OR 495 MANGO FRUIT.

C. capitata development stages

Tested commodity Eggs Larvae Puparia Adults

Mango 139,410* 64,990 53,854 46,920
PPF maturity degree 0 84,410 194 18 0
PPF maturity degree 2 84,250 0 0 0

*The total number of eggs was determined by counting the number of C. capitata eggs on 10% of (dissected) fruits, and extrap-
olating this for all tested fruits.


malformed. No adults closed from purple pas-
sionfruit puparia, whereas 46,920 adults
emerged from infested mangos.

DISCUSSION

Fruit fly host status determination lies at the
basis of trade and can help connect small-scale
fruit producers in the developing world to lucra-
tive export markets. To aid developing nations in
the process of assessing whether a given fruit is a
host to a particular fruit fly species, well-defined
protocols and experimental guidelines have been
defined (FAO 2005; Hennessey 2007; Aluja &
Mangan 2008; NAPPO 2008). Natural field infes-
tation trials and a set of screen-house or labora-
tory experiments all help determine whether a
given fruit crop is natural host, non-host or condi-
tional host (e.g., Jenkins & Goenaga 2008; De
Graaf 2009). These protocols have been adopted
for a wide range of fruit crops, such as mamey sa-
pote (Pouteria sapota (Jacq.)), litchi (Litchi chin-
ensis Sonn.), rambutan (Nephelium lappaceum
L.), avocado 'Hass' (Persea americana (Mill.)
'Hass'), highbush blueberry (Vaccinium corymbo-
sum L.), green mango (Mangifera indica L.
'Tommy Atkins' and 'Keitt'), and others.
Although data from natural field infestation
trials provide the most accurate assessment of
host status of a given fruit (NAPPO 2008), a key
limitation of these trials is that one cannot con-
trol variability in fruit fly abundance. In our ex-
periments, no C. capitata adults were reared from
field-collected passionfruit in the principal pro-
duction regions of Colombia. However, McPhail
trapping in orchards and surrounding habitats
also did not encounter any wild C. capitata popu-
lations in any of these zones. Purple passionfruit
crops are located at 2016.1 250.9 m (mean SD)
above sea level (Wyckhuys et al. in press), while
C. capitata has not been reported above 1,600 m
(ICA 2009). Thus, under the current altitudinal
and geographic distribution of C. capitata in Co-
lombia it is very unlikely that this species affects
purple passionfruit orchards. Climate change
could eventually bring C. capitata into actual


cropping regions and equally shift current pas-
sionfruit production zones to higher altitudes
(Hill et al. 2011). At present however, natural
field infestation data remain inconclusive with re-
spect to passionfruit host status.
Forced infestation trials under laboratory con-
ditions proved critical in delineating purple pas-
sionfruit host status to C. capitata. Even though
C. capitata females oviposited in intact fruit (ma-
turity degree 0) as in punctured fruit of different
maturity degrees, larval development was very
poor and no adults emerged. No adult emergence
from fruit under laboratory conditions is either
indicative of its character as non-host under ex-
perimental conditions (NAPPO 2008) or as non-
host overall (FAO 2005). Nevertheless, we need to
indicate that adult development from purple pas-
sionfruit could have been affected by dissecting
infested fruit 15 d after oviposition. On less suit-
able hosts, C. capitata likely develop slow and
take longer to complete larval development. How-
ever, fruit was dissected according to its deterio-
ration status (see FAO 2005; NAPPO 2008), while
taking into account an upper C. capitata egg-lar-
val development time of 15 d (EPPO 2010). In
conclusion, even though early dissection of purple
passionfruit may have affected pupation and
adult eclosion, the poor larval development and
lack of emergence of adults from 18 'C capitata
puparia clearly indicate the poor suitability of
this fruit.
For intact fruit, maturity level 0 was preferred,
while fruit of more advanced maturity were not
accepted for oviposition by C. capitata. Fruit ma-
turity state can greatly affect its acceptability as
an oviposition substrate by certain fly species
(Armstrong 2001; Willink & Villagran 2007). Cer-
tain physical stimuli determined by fruit matu-
rity level (e.g., color) influence C. capitata accep-
tance or rejection of fruit of particular maturity
levels (Prokopy et al. 1984; Suarez et al. 2007).
Also, fruit maturity level can affect physical resis-
tance to oviposition and interfere with successful
C. capitata oviposition (Gould & Hallman 2001).
To circumvent such, C. capitata tend to oviposit in
existing oviposition holes, bird pecks or crevices


March 2011







Rengifo et al.: Host Status of Purple Passionfruit for Ceratitis capitata


(Aluja & Mangan 2008). This could further ex-
plain high degrees of oviposition in punctured
fruits and low acceptability of intact fruit, more so
at advanced maturity degrees at which purple
passionfruit has an exceptionally firm epicarp.
Fruit fly oviposition in hosts that are inade-
quate for larval development is commonly ob-
served (Joachim-Bravo et al. 2001). Especially for
highly polyphagous species such as C. capitata,
behavioral adaptations cause oviposition in a
wide range of fruit crops (Aluja & Mangan 2008).
Additionally, under highly artificial conditions,
time-limited gravid females may accept a broad
range of substrates for oviposition (see Robacker
& Fraser 2002). A high level of acceptance for ovi-
position of intact and punctured fruit does not
necessarily imply suitability of the infested fruit
for further larval development or adult emer-
gence. Increased mortality, poor larval develop-
ment and reduced puparia size or weight all are
indicative of antibiosis and biochemical defenses
(Greany et al. 1983) that cannot be detected by
ovipositing females. Passiflora species are cyano-
genic and liberate hydrogen cyanide in fruits or
leaves when under (insect) attack (Spencer & Sei-
gler 1983). Possibly, these compounds disrupt lar-
val development in passionfruit.
As presence of low numbers of larvae in fruit is
not indicative that it is an acceptable host (Gould
& Hallman 2001; Jenkins & Goenaga 2007; Will-
ink & Villagran 2007), we can conclude that Co-
lombia-grown purple passionfruit is a non-host
under the experimental conditions used in these
tests and may be a non-host in the field. Since C.
capitata is currently not established in the princi-
pal growing areas in Colombia, it is very unlikely
that this pest will infest purple passionfruit un-
der natural conditions. There may therefore be
significant potential for the establishment of pest
free areas to allow exports to the United States or
a systems approach based upon low C. capitata
prevalence and poor host status.


ACKNOWLEDGMENTS

We are grateful to Catherine Varela, Maribel Hur-
tado, Jaime Abell6, Juan Camilo Rodriguez, Paola Ur6n,
Liliana Cardenas, Nubia Esmeralda Guerrero, Rub6n
Molina, Jose Rey, Gloria Palma, Oscar Menjura, Jorge
Enrique Arias, and Yaneth Bernal at the ICA Quaran-
tine Treatment Laboratory for help in fruit dissections
and fruit fly colony maintenance. We are grateful to Glo-
ria Marlene Vidal (USDA-APHIS) for help with data in-
terpretation and research conceptualization. This
research was financed by the Colombian Ministry of Ag-
riculture and Rural Development, with project grant
MADR 2008L6772-3445 to KW.

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Florida Entomologist 94(1)







Salas et al.: Population Dynamics of Greenidea on Guava and Ficus


POPULATION DYNAMICS OF TWO SPECIES OF GREENIDEA (HEMIPTERA:
APHIDIDAE) AND THEIR NATURAL ENEMIES ON PSIDIUM GUAJAVA
(MYRTACEAE) AND FICUS BENJAMIN (MORACEAE) IN CENTRAL MEXICO


MANUEL DARIO SALAS-ARAIZA1, ROBERT W. JONES2, ALEJANDRO PENA-VELASCO',
OSCAR ALEJANDRO MARTINEZ-JAIME1 AND EDUARDO SALAZAR-SOLIS1
Departamento de Agronomia, Divisi6n Ciencias de la Vida, Universidad de Guanajuato, Mexico
dariosalasaraiza@hotmail.co

Facultad de Ciencias Naturales, Universidad Aut6noma de Queretaro, Juriquilla, Queretaro, Mexico

ABSTRACT

Greenidea psiidi van der Goot and Greenidea ficicola Takahashi (Hemiptera: Aphid-
idae), are Asiatic species that feed on guava, Psidium guajava and Ficus spp.; both of
these aphids were reported as exotic pests in Florida in 2002 and in Mexico in 2003. The
present study characterized the population dynamics of both aphid species and their
natural enemies on guava and ornamental figs in the Bajio region of Central Mexico.
This report represents the first record of G. psiidi on Ficus sp. in Mexico and the first re-
port of the presence of both species in the state of Guanajuato. Greenidea psiidi and G.
ficicola were detected on guava in Mar 2007 and on fig trees during the same year in Apr
near Irapauto, Guanajuato. Populations of both alate and apterous forms of G. psiidi in
Apr were greater on guava than on fig trees (W = 119.0; P = 0.0122), which coincided with
new vegetative growth after leaf loss in winter on guava. In Apr populations of apterous
forms of both species were significantly greater than winged forms on both guava and
figs. No correlation was found between temperature changes and population densities of
aphids. The indigenous predators, Chrysoperla comanche Banks, Chrysoperla exotera
(Navas) and Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae), fed readily on
the aphids and were found on both guava and fig trees, although densities of all 3 species
were in greater numbers on Ficus. The combined population densities of the 3 predators
had a positive correlation with that of G. ficicola (r = 0.74), with a best fit found with a
quadratic model of simple regression: y (densities of Chrysoperla spp.) = 1.2479x2 -
4.3073x + 9.6493, and R = 0.703. Nine species of coccinelid beetles (Coleoptera: Coccinel-
lidae) were identified, the most common being of the genus Scymnus. Results suggest
that non-deciduous ornamental fig trees may serve as reservoirs of beneficial insects for
deciduous guava trees. Results from the present study provide basic biological data to
aid in management of these 2 exotic species of Greenidea on guava in central Mexico.

Key Words: guava, aphids, Greenidea, Psidium guajava, Ficus benjamin

RESUME

Greenidea psiidi van der Goot y Greenidea ficicola Takahashi (Hemiptera: Aphididae), son
de origen asidtico, en 2002 fueron reportados como plaga ex6tica en Florida y en M6xico en
2003. En el present studio, se describe la dinamica poblacional de estas dos species de afi-
dos y sus enemigos naturales en guayaba y Ficus en la region de El Bajio en el centro de
M6xico. Es el primer report de G. psiidi en Ficus spp. en M6xico y el primer report de am-
bas species en el estado de Guanajuato. Greenidea psiidi y G. ficicola fueron observadas en
guayaba desde marzo y en Ficus desde mediados de abril, en el area de studio. Las pobla-
ciones de afidos alados y apteros de G. psiidi fueron mas altas en abril en guayaba que en
Ficus (W = 119.0; P = 0.0122), esto coincide con los nuevos brotes despu6s de que el arbol de
guayaba pierde las hojas. Las poblaciones de las formas apteras de ambas species fueron
significativamente mayores que las formas aladas tanto en guayaba como en Ficus. No se en-
contr6 correlaci6n entire la temperature y las poblaciones de afidos. Los enemigos naturales
Chrysoperla comanche Banks, Chrysoperla exotera (Navas) y Chrysoperla carnea (Stephens)
(Neuroptera: Chrysopidae), se alimentan de estos afidos y fueron encontrados tanto en gua-
yaba como en Ficus, aunque la densidad de las tres species fueron mas altas en Ficus. Las
poblaciones combinadas de las tres species de depredadores presentaron una correlaci6n
positive con la especie G. ficicola (r = 0.74), lo cual se explica con la ecuaci6n cuadratica de
regresi6n simple: y (densidades de Chrysoperla spp.) = 1.2479x2- 4.3073x + 9.6493, donde el
coeficiente R2= 0.703. Se identificaron nueve species de Coccinellidae, siendo el mas comun
el g6nero Scymnus. Estos resultados sugieren que los arboles de Ficus cuyo follaje es pe-
renne, puede servir como refugio de insects ben6ficos para los arboles de guayaba, cuyo ha-







Florida Entomologist 94(1)


bito es caducifolio. Los resultados de esta investigaci6n aportaran conocimientos sobre la
biologia de estas species ex6ticas de afidos lo que ayudard en el manejo de la plaga en huer-
tas de guayaba en la region central de M6xico.

Translation provided by the authors.


Mexico contributes 25% of the world produc-
tion of guava (Psidium guajava L.; Myrtaceae), of
which the states of Michoacan, Aguascalientes,
and Zacatecas of the central Altiplano region are
the principal producers (Gonzalez-Gaona et al.
2002). In 2008, the exotic aphid pests, Greenidea
psidii van der Goot and Greenidea ficicola Taka-
hashi were observed on guava and ornamental fig
trees, Ficus benjamin (L.), in the central Altipl-
ano state of Guanajuato (Salas-Araiza, unpub-
lished data). There is concern among producers
and agricultural researchers of central Mexico
that these new pests may pose a threat to guava
production in Mexico.
The genus Greenidea Schouteden belongs to
the subfamily Greenideinae within the Aphididae
and includes approximately 45 species (Perez
Hidalgo et al. 2009). The natural distribution of
the genus is Asiatic and species are found to favor
young foliage of plants of the families, Fagaceae,
Betulaceae, Juglandaceae, Myrtaceae, Rosaceae,
and Rubiaceae (Blackman & Eastop 1994). The
presence of Greenidea in the New World may be
the result of importation of infested ornamental
fig trees which are widely commercialized
throughout the world, and they have been impli-
cated as vehicles for the introduction of exotic
pests (O'Donnell & Parrella 2005).
Greenidea psidii was first discovered in the
New World in1916 on guava and its relative, Psid-
ium cattleianum Sabine in Brazil (Noemberg-
Lazzari et al. 2006). Halbert (2004) reported that
G. ficicola was collected in Florida in 2002, and to-
gether with G. psidii were found to feed on guava
and ornamental fig trees (Ficus benjamin (L.)).
Perez Hidalgo et al. 2009 reported the presence of
G. psidii in Costa Rica in 2009. The first reports of
Greenidea in Mexico come from Pena-Martinez et
al. (2003), who recorded G. psidii feeding on
guava in the states of Hidalgo, Morelos, Guerrero,
and the Federal District, and G. ficicola feeding
on ornamental fig trees in the state of Guana-
juato, Mexico. Trejo-Loyo et al. (2004) reported G.
ficicola occurs on Myrtaceae in Cuernavaca, Mo-
relos. Although further data is lacking, it is prob-
able that both G. psidii and G. ficicola are widely
distributed in Mexico.
The biology of species of Greenidea on guava
and other host plants is poorly known (Halbert
2004; Sousa-Silva et al. 2005; Noemberg-Lazzari
et al. 2006). Northfield et al. (2008) noted that
many insect pests feed on alternative host plants
and that the understanding of the relationships of
pest populations on wild hosts with those on cul-


tivated hosts is crucial in the development of
management strategies. The objective of the
present study was to determine the population
dynamics of Greenidea psidii, G. ficicola and their
natural enemies on Psidium guajava and Ficus
benjamin in the state of Guanajuato, Mexico.
The data generated from this study will help eval-
uate the importance of these aphids within the
pest complex attacking guava in the Bajio Region,
as well as aid in the development of integrated
pest management strategies.

MATERIALS AND METHODS

The study site was conducted at the experi-
mental field station of the Division of Life Sci-
ences of the University of Guanajuato at the Ex-
Hacienda El Copal (10101'01"N, 2049'49"0) at
1,750 masl in the municipality of Irapuato, Gua-
najuato, Mexico. The region has a mean annual
precipitation of 750 mm, a mean temperature of
19C and mean relative humidity of 56% (INEGI
2009). Two orchards, 1 guava (P. guajava), and
the other ornamental figs, (F benjamin) were
chosen for the study sites.
All trees were 10 years old and the 2 orchards
were separated by approximately 1 km. Weekly
samples were made of aphids alatee and apterous
forms) and predators (adults of Coccinellidae and
larvae of Chrysopidae) from 10 trees of each host
species. For each tree, samples consisted of 20
beats of a 1-m wooden rod on the branches of the
trees, from which insects fell onto a 1-m2 beating
sheet. All insects on the sheet were collected and
placed in a labeled vial with 70% alcohol. The sam-
ple period was from 23 Mar to 22 Jun 2007 which
corresponded to the early spring growth period of
leaves of guava and to the reproductive activity of
aphids. Samples were not taken on subsequent
dates because aphids had ceased reproductive
growth and individuals were virtually undetect-
able. All material from each tree was preserved in
individual vials containing 70% alcohol, with a cor-
responding label and brought to the laboratory.
The collected specimens were identified in the
Entomology Laboratory of University of Guana-
juato, with a compound and stereo microscope.
Aphids were mounted for species determination
following techniques given by Peia-Martinez
(1995). The keys of Blackman & Eastop (1994,
2000) were used for species identification of
aphids. For the neuropteran and coccinellid pred-
ators, the keys of L6pez-Arroyo et al. (2008) and
Gordon (1985) were used, respectively. Meteoro-


March 2011







Salas et al.: Population Dynamics of Greenidea on Guava and Ficus


logical data was obtained from a weather station
maintained at the study site located 200 m from
the orchards. All identified specimens were depos-
ited in the Entomological Collection "Leopoldo Ti-
noco Corona" of the University of Guanajuato.
Data were analyzed with the statistical soft-
ware program SAS (SAS 1995). Abundance mea-
sures were calculated for G. psiidi and G. ficicola
and the various predator species on guava and
figs. Due to non-linearity of data determined by
the Shapiro-Wilks test, mean comparisons of
aphid and predator numbers between hosts and
among samples dates were made with Mann-
Whitney non-parametric procedures. In addition,
correlation analyses were conducted between
population numbers of aphids with those of the
various predators and also with climatic vari-
ables (temperature and precipitation).

RESULTS AND DISCUSSION

Species of Greenidea in Guava and Figs

Both G. psidii and G. ficicola were present on
guava and the ornamental fig, F benjamin (Figs.
1, A, B, C). This is the first report of G. psiidi on
the widely planted F benjamin from Mexico, and
for this species for the state of Guanajuato. Al-
though the habitual host for G. ficicola is Ficus
spp. (Noemberg-Lazzari et al. 2006), Halbert
(2004) reports that this aphid species also occurs
on guava, as confirmed in the present study. Al-
though the apparent habitual hosts of G. psidii is
guava, and that of G. ficicola is Ficus spp., it is un-
clear how populations on the habitual and other
infrequent host plants interact and which of the
infrequent hosts can maintain viable, reproduc-
tive populations in the absence of the habitual
host plants.
Both species of Greenidea were aggregated on
new shoots and leaves of their hosts, a feeding pref-
erence previously reported by Perez Hidalgo et al.
(2009). On guava, G psiidi and G ficicola were
present on new leaf buds and on either side of young
leaves, whereas on F benjamin they were found
principally on the underside of young leaves. The
preference by aphids for new plant growth is a com-
mon behavior in aphids. Gould et al. (2007) state
that certain stages of aphids have preferences for
specific tissues of host plants. For example, Chaito-
phorus populicola Thomas, preferably feeds on new
growth with diverse and high levels of amino acids.
These authors also note that high sugar levels and
low amino acid concentration in leaves increases
the production of winged individuals.

Population Dynamics of Greenidea psidii and Greenidea
ficicola

Both G. psiidi and G. ficicola were first re-
corded on guava trees in late Mar 2007, although


the abundance of G. psiidi was notably greater
during Mar and early Apr. This appearance and
population growth coincided with the emergence
of new shoots and leaves on guava, the timing of
which corresponds to that described by Damian-
Nava et al. (2004). Colonizing alate aphids are of-
ten attracted to specific volatiles of host plants
(Chapman et al. 1981; Nottingham et al. 1991;
Powell & Hardie 2001). Because G. psiidi and G.
ficicola are found initially on young shoots and
leaves, it is probable that initial alate colonizers
are attracted to volatiles associated with new
growth of guava plants.
It important to note that the 2 tree species
studied have marked differences in leafing pat-
terns and the appearance of new growth. Guava
is deciduous, with complete leaf loss occurring
generally in Nov with new growth beginning in
Mar in the study area. This is in contrast to F
benjamin which is a non-deciduous tree that
produces new growth apparently in response to
environmental factors. However, both G. psiidi
and G. ficicola appeared first on guava, and then
on F benjamin (Fig. 1, A, B, C), although foliage
was available on the latter host throughout win-
ter months. These data suggest that G. psiidi and
G. ficicola first colonize and establish on guava
and then later move to F benjamin. This behav-
ior was expected for G. psiidi for which guava is
considered a habitual host, but not for G. ficicola
which Ficus is considered the habitual host. Fur-
ther study is needed to establish the initial colo-
nization behavior of these 2 species and whether
the behaviors are the result of greater attractive-
ness of guava in comparison to F benjamin and/
or the leaf quality of F benjamin is inadequate
until late Apr.
The peak abundance of both species alatee and
apterous forms) occurred in mid and late Apr (7.5
and 6.0 aphids/sample, for G. psiidi and G. fici-
cola, respectively) (Fig. 1, A, B). The densities of
G. psiidi were greater on guava than on F ben-
jamina during the first 5 sample periods, and sig-
nificantly so at peak densities during the third
week of Apr (w = 119.0; P = 0.0122; Fig. 1, A).
By mid May, densities of G. psiidi and G. fici-
cola were barely detectable, and remained at very
low densities through late Jun. These low densi-
ties are presumably the result of the maturation
of leaves and the deterioration of the physical and
nutritional requirements for these species (Fig. 2,
A).
The appearance and abundance of alate forms
in relation to apterous forms of G. psiidi and G.
ficicola followed patterns expected from observa-
tions reported by Noemberg-Lazzari et al. (2006).
Alates were found in very low numbers initially
on both guava and F benjamin and differences
between the 2 forms were not significantly differ-
ent until the third week of Apr (Fig. 2, B, C). At
that time, the abundance of apterous forms in-







Florida Entomologist 94(1)


A Greendea psdi i apterous and aiate) on Psildum gua)ava
a Greendea r' dil i apterous andalates)on Fcu s bn amina


a

b
a aa


III I I I I I





B
SGreendea ficola apterous anialal s)on Psidiu
guapva
o Greendea fiowola (apterous andalas) on Fcus
a bernam ina




a

__J--


f F- I% P F h Fe
N I


00 0 0 0 0 0 0 0
Fig. 1. Population dynamics of apterous and alate forms of Greenidea on 14 samples dates in Irapuato, Guana-
juato, M6xico. Means with the same letter not significantly different based on Mann-Whitney non parametric test
(A: 04/20/07, w = 119.0, P =0.0122).


a mGreendea psdii(aptao sand alales) onP sdiimgLuaava
I IDGreenmfea fiCcolIa(aptel us and ala so n P &diLumguajava


1


! I l l I


March 2011
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