• TABLE OF CONTENTS
HIDE
 Genetic variation in neonate behavior...
 A new species of Anteon (Hymenoptera:...
 Female and male polymorphism in...
 Dacnusa cicerina (Hymenoptera:...
 Vitex agnus castus and Euphorbia...
 Selection of Atriplex lentiformis...
 Three new species of Phyllophaga...
 A new genus Chrysonasma (Lepidoptera,...
 A safe and effective propylene...
 Life history and damage of a new...
 Natural ocurrence of hymenopterous...
 Host status of litchi and rambutan...
 A new gall-inducing species of...
 Biology of the queen of Spain fritillary,...
 Temporal and spectral features...
 Performance of Bemisia tabaci (Hemiptera:...
 New species of American larginae...
 Identification of grape juice aroma...
 Efficacy of selected bait and residual...
 Five new species of the genus Microplitis...
 Molecular survey of endosymbionts...
 Phenology of Maconellicoccus hirsutus...
 Egg parasitoids of citrus weevils...
 Identification of mature larvae...
 First host plant record for Anastrepha...
 Herbivorous insect fauna of mile-a-minute...
 Aegopsis bolboceridus (Coleoptera:...
 Effects of the exotic crustacean,...
 The effect of season of fire on...
 A new exotic pest for Florida’s...
 Mass rearing of Pseudophilothrips...
 W. W. Yothers, a pioneer in citrus...
 Book reviews
 Back Matter














Group Title: Florida Entomologist
Title: The Florida entomologist
ALL VOLUMES CITATION THUMBNAILS PAGE IMAGE ZOOMABLE
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STANDARD VIEW MARC VIEW
Permanent Link: http://ufdc.ufl.edu/UF00098813/00356
 Material Information
Title: The Florida entomologist
Uniform Title: Florida entomologist (Online)
Abbreviated Title: Fla. entomol. (Online)
Physical Description: Serial
Language: English
Creator: Florida Entomological Society
Florida Center for Library Automation
Publisher: Florida Entomological Society
Place of Publication: Gainesville Fla
Gainesville, Fla
Publication Date: June 2008
Frequency: quarterly
regular
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Subject: Entomology -- Periodicals   ( lcsh )
Insects -- Periodicals -- Florida   ( lcsh )
Genre: review   ( marcgt )
periodical   ( marcgt )
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Additional Physical Form: Also issued in print.
System Details: Mode of access: World Wide Web.
Language: In English; summaries in Spanish.
Dates or Sequential Designation: Vol. 4, no. 1 (July 1920)-
Issuing Body: Official organ of the Florida Entomological Society; online publication a joint project of the Florida Entomological Society and the Florida Center for Library Automation.
General Note: Title from caption (JSTOR, viewed Sept. 13, 2006).
General Note: Place of publication varies.
General Note: Latest issue consulted: Vol. 87, no. 4 (Dec. 2004) (JSTOR, viewed Sept. 13, 2006).
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Bibliographic ID: UF00098813
Volume ID: VID00356
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 Related Items
Preceded by: Florida buggist (Online)

Table of Contents
    Genetic variation in neonate behavior of fall armyworm (Lepidoptera: Noctuidae)
        Page 151
        Page 152
        Page 153
        Page 154
        Page 155
        Page 156
        Page 157
        Page 158
    A new species of Anteon (Hymenoptera: Dryinidae) from Argentina
        Page 159
        Page 160
        Page 161
    Female and male polymorphism in two species of Melittobia parasitoid wasps (Hymenoptera: Eulophidae)
        Page 162
        Page 163
        Page 164
        Page 165
        Page 166
        Page 167
        Page 168
        Page 169
    Dacnusa cicerina (Hymenoptera: Braconidae: Alysiinae), a new species of endoparasitoid of Liriomyza cicerina (Diptera: Agromyzidae)
        Page 170
        Page 171
        Page 172
        Page 173
        Page 174
        Page 175
        Page 176
        Page 177
        Page 178
    Vitex agnus castus and Euphorbia characias ssp. wulfenii as reservoirs of aphid parasitoids (Hymenoptera: Braconidae: Aphidiinae)
        Page 179
        Page 180
        Page 181
        Page 182
        Page 183
        Page 184
        Page 185
        Page 186
        Page 187
        Page 188
        Page 189
        Page 190
        Page 191
    Selection of Atriplex lentiformis host plants by Hesperopsis gracielae (Lepidoptera: Hesperiidae)
        Page 192
        Page 193
        Page 194
        Page 195
        Page 196
        Page 197
    Three new species of Phyllophaga from Mexico (Coleoptera: Scarabaeidae: Melolonthinae)
        Page 198
        Page 199
        Page 200
        Page 201
        Page 202
        Page 203
        Page 204
    A new genus Chrysonasma (Lepidoptera, Gelechioidea, Lecithoceridae), with description of a new species from the Philippines
        Page 205
        Page 206
        Page 207
        Page 208
        Page 209
    A safe and effective propylene glycol based capture liquid for fruit fly (Diptera: Tephritidae) traps baited with synthetic lures
        Page 210
        Page 211
        Page 212
        Page 213
    Life history and damage of a new Baradinae weevil (Coleoptera: Curculionidae) on amaryllis
        Page 214
        Page 215
        Page 216
        Page 217
        Page 218
        Page 219
    Natural ocurrence of hymenopterous parasitoids associated with Anastrepha fraterculus (Diptera: Tephritidae) in Myrtaceae species in Entre Rios, Northeastern Argentina
        Page 220
        Page 221
        Page 222
        Page 223
        Page 224
        Page 225
        Page 226
        Page 227
    Host status of litchi and rambutan to the West Indian fruit fly (Diptera: Tephritidae)
        Page 228
        Page 229
        Page 230
        Page 231
    A new gall-inducing species of Holopothrips (Thysanoptera: Phlaeothripinae) from Tabebuia trumpet trees in the Caribbean region
        Page 232
        Page 233
        Page 234
        Page 235
        Page 236
        Page 236a
        Page 236b
        Page 236c
        Page 236d
    Biology of the queen of Spain fritillary, Issoria lathonia (Lepidoptera: Nymphalidae)
        Page 237
        Page 238
        Page 239
        Page 240
    Temporal and spectral features of sounds of wood-boring beetle larvae: identifiable patterns of activity enable improved discrimination from background noise
        Page 241
        Page 242
        Page 243
        Page 244
        Page 245
        Page 246
        Page 247
        Page 248
    Performance of Bemisia tabaci (Hemiptera: Aleyrodidae) on healthy and Cotton leaf curl virus infected cotton
        Page 249
        Page 250
        Page 251
        Page 252
        Page 253
        Page 254
        Page 255
    New species of American larginae (Heteroptera: Largidae) and keys to known species of Largulus and Theraneis
        Page 256
        Page 257
        Page 258
        Page 259
        Page 260
        Page 261
        Page 262
        Page 263
        Page 264
        Page 265
    Identification of grape juice aroma volatiles and attractiveness to the Mexican fruit fly (Diptera: Tephritidae)
        Page 266
        Page 267
        Page 268
        Page 269
        Page 270
        Page 271
        Page 272
        Page 273
        Page 274
        Page 275
        Page 276
    Efficacy of selected bait and residual toxicants for control of bigheaded ants, Pheidole megacephala (Hymenoptera: formicidae), in large field plots
        Page 277
        Page 278
        Page 279
        Page 280
        Page 281
        Page 282
    Five new species of the genus Microplitis (Hymenoptera: Braconidae: Microgastrinae) from China
        Page 283
        Page 284
        Page 285
        Page 286
        Page 287
        Page 288
        Page 289
        Page 290
        Page 291
        Page 292
        Page 293
    Molecular survey of endosymbionts in Florida populations of Diaphorina citri (Hemiptera: Psyllidae) and its parasitoids Tamarixia radiata (Hymenoptera: Eulophidae) and Diaphorencyrtus aligarhensis (Hymenoptera: Encyrtidae)
        Page 294
        Page 295
        Page 296
        Page 297
        Page 298
        Page 299
        Page 300
        Page 301
        Page 302
        Page 303
        Page 304
    Phenology of Maconellicoccus hirsutus (Hemiptera: Pseudococcidae) in Florida based on attraction of adult males to pheromone traps
        Page 305
        Page 306
        Page 307
        Page 308
        Page 309
        Page 310
    Egg parasitoids of citrus weevils in Guadeloupe
        Page 311
        Page 312
        Page 313
        Page 314
    Identification of mature larvae of Hydaticus cinctipennis and H. bimarginatus (Coleoptera: Dytistidae) based on morphology and breeding seasons
        Page 315
        Page 316
    First host plant record for Anastrepha elegans (Diptera: Tephritidae)
        Page 317
        Page 318
    Herbivorous insect fauna of mile-a-minute weed, Persicaria perfoliata (Polygonaceae), in Japan
        Page 319
        Page 320
        Page 321
        Page 322
        Page 323
    Aegopsis bolboceridus (Coleoptera: Melolonthidae): an important pest on vegetables and corn in Central Brazil
        Page 324
        Page 325
        Page 326
        Page 327
    Effects of the exotic crustacean, Armadillidium vulgare (Isopoda), and other macrofauna on organic matter dynamics in soil microcosms in a hardwood forest in central Florida
        Page 328
        Page 329
        Page 330
        Page 331
    The effect of season of fire on density of female garden orbweavers (Araneae: Araneidae: Argiope) in Florida scrub
        Page 332
        Page 333
        Page 334
        Page 334a
        Page 334b
        Page 334c
        Page 334d
        Page 334e
    A new exotic pest for Florida’s natural areas: Crypticerya genistae (Hemiptera: Monophlebidae)
        Page 335
        Page 336
        Page 337
    Mass rearing of Pseudophilothrips ichini (Thysanoptera: Phlaeothripidae), an approved biological control agent for Brazilian peppertree, Schinus terebinthifolius (Sapindales: Anacardiaceae)
        Page 338
        Page 339
        Page 340
    W. W. Yothers, a pioneer in citrus entomology
        Page 341
        Page 342
        Page 343
        Page 344
        Page 345
        Page 346
        Page 347
        Page 348
    Book reviews
        Page 349
        Page 350
        Page 351
    Back Matter
        Page 352
Full Text



Stuhl et al.: Fall Armyworm Larval Preferences


GENETIC VARIATION IN NEONATE BEHAVIOR
OF FALL ARMYWORM (LEPIDOPTERA: NOCTUIDAE)

CHARLES J. STUHL1, ROBERT L. MEAGHER AND ROD N. NAGOSHI
Center for Medical, Agricultural and Veterinary Entomology Agricultural Research Service,
U.S. Department of Agriculture, Gainesville, FL 32608

'University of Florida, Entomology and Nematology Department, Gainesville, FL 32611

ABSTRACT

Bioassays were developed to test plant selection of fall armyworm (Spodoptera frugiperda (J.
E. Smith)) host strains to corn (Zea mays L.) and stargrass, a forage grass closely related to
bermudagrass (Cynodon nlemfuensis Vanderyst var. nlemfuensis). Neonate larvae from 3
corn strain and 3 rice strain colonies preferentially selected corn over stargrass in petri dish
choice tests. However, bioassays of whole plants and whole plant volatiles showed that se-
lection of a particular host was not clear and there were no significant differences in plant
choice. Two additional bioassays were conducted to determine if larvae would continue to
disperse once they came in contact with a plant source. One colony was always biased to-
wards corn regardless of which plant was encountered first. For 4 colonies, the attraction to
corn was reduced such that when stargrass was first contacted, equal or greater numbers of
larvae stayed and did not migrate to corn. Finally, the attraction to corn was lowest in 1 col-
ony in which significantly more larvae moved away from corn even when it was presented
first. Results of our study, along with behavioral and feeding trials from other studies, sug-
gest that there is a consistent attraction of neonates to damaged corn regardless of "strain"
designation, but substantial variability in the strength of that attraction if they come in con-
tact with another plant host first. These differences among colonies most likely reflect ge-
netic and phenotypic variation in wild populations. More information at http://
www.ars.usda.gov/pandp/people/people.htm?personid=l 1809

Key Words: Spodoptera frugiperda, choice tests, larval behavioral traits, host plant selection

RESUME

Se desarrollaron bioensayos para probar la selecci6n de plants por cepas del gusano cogo-
llero (Spodoptera frugiperda (J. E. Smith)) segun su hospedero hacia el maiz (Zea mays L.),
el past estrella, y un past de foraje cercano a "bermudagrass" (Cynodon nlemfuensis Van-
deryst var. nlemfuensis). Las larvas reci6n nacidas de 3 colonies de la cepa de maiz y 3 colo-
nias de la cepa de arroz preferian secciones de maiz sobre las secciones del past estrella en
las pruebas de opciones realizadas en plates petri. Sin embargo, bioensayos de la plant en-
tera y volatiles de toda la plant mostraron que la selecci6n de un hospedero particular no
fue clara y no hubo una diferencia significativa en la selecci6n de plants. Se realizaron dos
bioensayos adicionales para determinar si las larvas continuaran su dispersion una vez que
est6n en contact con la plant. Una colonia siempre tuvo como preferencia la selecci6n del
maiz sin importar cual de las plants fueron encontradas inicialmente. Para 4 de las colo-
nias, la atracci6n al maiz fue reducida de tal manera que cuando fueron puestas en contact
con el past estrella, un numero igual o mayor de las larvas se quedaron y no emigraron al
maiz. Finalmente, la atracci6n al maiz fue la mas baja en 1 colonia donde un numero signi-
ficativamente mayor de las larvas se apartaron del maiz aun despu6s de ser presentado ini-
cialmente. Los resultados de nuestro studio, junto con las pruebas de comportamiento y
alimentaci6n hechas en otros studios, sugieren que hay una atracci6n consistent de las lar-
vas reci6n nacidas a maiz daiado sin importar la clase de la cepa, pero hay una variabilidad
substantial en el grado de la atracci6n de las larvas si se ponen en contact con otra plant
inicialmente. Estas diferencias entire las colonies a lo mejor reflejan la variaci6n gen6tica y
fenotipica en poblaciones de campo.


Fall armyworm, Spodoptera frugiperda (J. E. jury and economic damage to many crops, includ-
Smith) is a polyphagous insect that migrates each ing corn (Marenco et al. 1992) and pasture
season from overwintering areas in southern grasses (Martin et al. 1980). Fall armyworm is
Florida and southern Texas to the eastern and composed of two sympatric and morphologically
central U.S. (Luginbill 1928; Mitchell 1979; Pair identical strains that are defined by their host
et al. 1986; Pair et al. 1991; Westbrook & Sparks plant preferences (Nagoshi & Meagher 2004).
1986; Mitchell et al. 1991). This pest can cause in- One strain was identified from populations feed-







Florida Entomologist 91(2)


ing on corn (Zea mays L.) and sorghum (Sorghum
spp.) (corn strain) and the other strain was iden-
tified from populations feeding on rice (Oryza sa-
tiva L.) and forage grasses (Cynodon spp.) (rice
strain) (Pashley et al. 1985; Pashley 1986). The 2
strains can be distinguished by genetic markers
(Levy et al. 2002; Nagoshi & Meagher 2003a; Na-
goshi & Meagher 2003b; Nagoshi et al. 2006).
Based on capture of adult males in pheromone
traps, corn strain individuals were found primarily
in agricultural areas, whereas rice strain individu-
als were found in agricultural, natural, and urban
habitats (Meagher & Nagoshi 2004; Nagoshi &
Meagher 2004). However, rice strain larvae were
found in both corn and forage grasses but corn
strain larvae were only found in corn (Meagher &
Gallo-Meagher 2003; Nagoshi et al. 2006).
There have been several reports of behavioral
and physiological differences between strains
that could explain their asymmetrical distribu-
tion in the wild. For example, rice strain larvae
feeding on corn were observed to display a slower
rate of weight gain, longer developmental time,
lower pupal weight, and reduced survival than
when reared on bermudagrass (Pashley 1988; Pa-
shley et al. 1995; Veenstra et al. 1995). However,
in many cases these biological differences have
been difficult to reproduce. For example, while
Whitford et al. (1988) also reported reductions in
larval and pupal weights when rice strain larvae
were raised on corn, they did not observe differ-
ences in developmental time or survival. They
similarly showed that rearing corn strain larvae
on rice or bermudagrass had no consistent nega-
tive effect on larval development or fitness, a find-
ing that we have also observed (Meagher et al.
2004; RLM unpublished data).
The inability to reproduce strain-specific be-
haviors under controlled laboratory settings has
stymied attempts to dissect the mechanism of
strain divergence and limited our overall under-
standing of fall armyworm biology. A better de-
scription of why this occurs could allow for better
bioassays that would make possible the experi-
mental identification of strain-specific character-
istics. One explanation is that these observa-
tional disagreements are due to environmental
influences, in particular the effects of artificial
culturing practices on more complex behaviors
(Quisenberry & Whitford 1988; Jamjanya et al.
1990). An alternative explanation is that there is
substantial genetic variability even within
strains such that individual isolates can exhibit
substantial differences in physiology and behav-
ior. To examine the latter possibility, we took ad-
vantage of an observation by Pashley et al. (1995)
that =90% of neonate larvae of both strains pre-
ferred corn over bermudagrass in petri dish
choice-tests. Simple modifications of this bioassay
were made to test plant specificity and to compare
the behaviors of several independently isolated


colonies of both strains. The behavioral differ-
ences are described and their ramifications on our
understanding of strain biology discussed.

MATERIALS AND METHODS

Strain Isolation and Plant Growth

Fall armyworm egg masses and larvae were
collected during 2003 and 2005 from multiple
sites in Florida (field corn, University of Florida
Dairy Research Unit, Hague, Alachua Co.; forage
grasses, University of Florida Range Cattle Re-
search and Education Center, Ona, Hardee Co.;
and sweet corn, Miami-Dade Co.), and 1 location
in Mississippi (Washington Co. by J. Adamczyk
from bermudagrass). Single adult pair matings
were performed in small oviposition cages which
consisted of a cylindrical 473-mL plastic food con-
tainer (Solo Cup Co., Urbana, IL) lined with a 7-
cm x 7.6-cm coffee filter (Bunn, Springfield, IL).
Holes (5 mm) were placed in the bottom of the
containers to allow for airflow. Two holes (1.5 cm)
were placed in the lid (Solo, ML8) so that braided
cotton rolls (Richmond Dental, Charlotte, NC)
could be inserted. The cage was inverted and
placed over a 177-mL container (S306, Sweet-
heart Products Group, Owings Mills, MD), which
held 2 plastic souffle cups (Solo, P100), one with
deionized water and the other with 10% honey/
sugar solution. This system allowed for absorp-
tion of liquids for adult nourishment. Females
were allowed to freely deposit eggs on the inner
surface of the coffee filter. At least 20 pairs of F,
moths were used to establish the colonies.
Upon death, male and female moths were
analyzed separately for strain identification
with mitochondrial markers (Levy et al. 2002;
Meagher & Gallo-Meagher 2003). Eggs were
collected daily, and labeled according to pair
mating. Newly emerged larvae were reared on
pinto bean diet (Guy et al. 1985) until strain
identification was verified. Once strain associa-
tion was confirmed with the 2003 colonies, F2
larvae were placed either on a corn ('Truckers
Favorite') or stargrass (Cynodon nlemfuensis
Vanderyst var. nlemfuensis 'Florona') foliage
diet according to their host strain (CS-Hag03
and RS-Ona03, respectively). In 2005, two more
colonies of each strain were established accord-
ing to the procedures above. However, larvae
from these colonies (CS-Hag05, CS-JS05, RS-
MS05, and RS-Ona05) were continuously
reared on pinto bean diet.
Plants were grown in 550-mL pots in a green-
house at ambient temperature (22-40C) and
were fertilized weekly with Miracle-Gro (Marys-
ville, OH) 15-30-15 plant food. Plant age during
experimentation was approximately 3 weeks for
both corn and 'Florona' stargrass. 'Florona' star-
grass is a long-lived, persistent perennial grass,


June 2008







Stuhl et al.: Fall Armyworm Larval Preferences


similar to types of bermudagrass, that was ob-
served growing in Ona in 1973 (Mislevy et al.
1989; Mislevy et al. 1993). Previous research
showed this grass to be an excellent host for fall
armyworm (Meagher et al. 2007).

Choice Test Bioassays

Three separate bioassays were designed to
compare preferences of neonate larvae of both
host strains for either corn or stargrass. The first
experiments were conducted in 9-cm diameter
polystyrene petri dishes (Thomas Scientific,
Swedesboro, NJ) as a choice arena to present cut
leaf sections to neonates. New growth leaf sec-
tions were taken from each plant type, and
trimmed along the top and sides to achieve a uni-
form size (5 cm x 1.5 cm). Since stargrass leaves
are smaller than corn leaves, 2 leaf sections were
used to obtain the same area as the corn leaves.
One section of each plant host was placed on filter
paper discs (9 cm; Thomas Scientific) moistened
with 1 mL of deionized water. Sections were
placed 2 cm from the center, along the outer edge
of the petri dish. Twenty newly-hatched larvae (0-
24 h) were placed in the center of each dish, and
the lid put into place. Ten replicates were per-
formed for each colony. Petri dishes were placed in
an incubator at 23.9 2'C with a 14/10 day/night
cycle, 80% RH. The number of larvae on or under
each leaf section was counted 24 h after introduc-
tion. Three corn strain (CS-Hag03, CS-Hag05,
and CS-JS05) and 3 rice strain (RS-Ona03, RS-
Ona05, and RS-MS05) colonies were tested.
The second bioassay was conducted with a Y-
tube olfactometer made of 2.5 cm-diameter clear
Plexiglas tubing with a 58.0-cm body and 15.2-cm
arms to test neonate responses to odors alone.
Odor sources for the bioassay consisted of whole
plants in 550-mL pots housed in a 3.8-L glass jar.
Charcoal-filtered house air was passed over the
plants in the jar and through an arm at 0.2 L/min.
Airflow was pulled through the base of the Y-tube
by a vacuum at 0.4 L/min.
Larval attraction to the volatiles of 1 corn
plant or a small group of stargrass plants was
tested by placing an eggmass at the midpoint in
the body of the Y-tube. A black 9-cm filter paper
disk (Thomas Scientific) was placed encircling the
area outside of the tube above the eggmass to re-
duce light interference. Larvae were allowed free
movement within the olfactometer. The number
of larvae in each arm was counted after 24 h.
There were 7 replicates each of corn (CS-Hag03)
or rice strain (RS-Ona03) larvae; the position of
the host plant was alternated for each replicate.
The third bioassay was made with a clear
acrylic plastic cage (measuring 51 (L) x 25 (W) x
28 (H) cm with a testing area of 51 x 25 x 18 cm)
to test neonate responses to whole plants rather
than plant sections. Potted plants were placed in


rectangular receptacles (15 x 7.5 cm) that were
removable and allowed for the soil/plant interface
to be level with the floor surface. The corn recep-
tacle contained 4 plants and the stargrass recep-
tacle contained between 15 and 20 plants. During
testing, the cage was placed in an environmen-
tally controlled room at 23.9 2C with a 14/10
day/night photoperiod and 80% RH. Eggmasses
(CS-Hag03 and RS-Ona03) were placed in the
center, and the number of neonate larvae on ei-
ther the corn plants or stargrass plants was
counted after 24 h. Plant location was alternated
for each of the 5 replicates.

Passing-Over Tests

These tests were conducted to determine if
larvae would continue to disperse once they
came in contact with a plant source. Two bioas-
says were conducted. Sections of corn and star-
grass leaf material were placed on filter paper
discs (Thomas Scientific) moistened with ca. 1
mL deionized water and cut to fit the dimen-
sions of a 14-cm diameter polystyrene petri dish
(Thomas Scientific). The plant material first en-
countered (and potentially "passed-over") by
the neonates was cut to dimensions large
enough to span the diameter of the petri dish.
Another more distally-located plant section was
trimmed to a uniform size (5 cm x 1.5 cm) and
placed 30 mm from the center, and 20 mm along
the outer edge of the petri dish. As with the
choice test petri dish bioassays, extra sections
of stargrass were used to provide the same sur-
face area as the corn. Twenty neonate larvae
were placed in the dish opposite the leaf sec-
tion. Petri dishes were placed in the incubator
and the number of larvae on or under each leaf
section was counted after 24 h. Since bioassays
on larvae from the 2 initial host strain isolates
provided interesting results (CS-Hag03 and
RS-Ona-03), 2 more colonies of each strain were
examined (CS-Hag05, CS-JS05, RS-Ona05, and
RS-MS05). For each colony, there were 10 repli-
cates passing over corn to stargrass and 10 rep-
licates passing over stargrass to corn.
The second passing-over bioassay used the
choice cage as described above, except it was mod-
ified to allow inflow and outflow of air over the
odor sources. Charcoal-filtered house air was
passed sequentially over the distal plant material
and then over the proximal plant material before
being presented to the larvae. Air was vented to
the outside to prevent plant volatiles from re-en-
tering the cage. Plants were arranged in the cage
so that larvae would have to pass through one
plant host to reach the other plant. Four corn
plants and between 15-20 stargrass plants were
used. Newly emerged larvae from an eggmass
were placed in the downwind position from the
first plant. For each strain (CS-Hag03 and RS-







Florida Entomologist 91(2)


Ona03) examined, there were 5 replicates of lar-
vae passing through corn to stargrass and 5 rep-
licates of larvae passing through stargrass to
corn. The number of larvae on each plant was
counted after 24 h.

Statistics

Data were analyzed as binomial experiments
where the null hypothesis was that corn and star-
grass were chosen equally. Since larval numbers
differed among replicates (r), the proportion of
larvae (p = number of larvae selecting corn / total
number of larvae) selecting corn was converted to
z scores [z = (p 0.5) / I (p (1 p) / total]. For
each test, z scores for all r values were averaged
and t-tests (t = z scores mean / SEM; r 1 df) were
calculated. When p was either 0 or 1.0, a conver-
sion factor was used: for p = 0, p = 0.375 / (total
number + 0.75); forp = 1.0, p = (number selecting
corn + 0.375) / (total number + 0.75); (Ott & Long-
necker 2001).

RESULTS

CS-Hag03 and RS-Ona03 Colonies

Initial experiments were performed with 1
corn strain and 1 rice strain colony generated in
2003 from Florida populations. In the petri dish
bioassays, larvae were provided a choice between
2 equal-sized cut sections from corn or stargrass
leaves. Larvae of both strains showed a strong
preference for corn sections, with almost 80% of
corn strain and over 92% of rice strain neonates
selecting corn (Fig. 1). A different behavior was
observed in olfactometer studies where larvae
were exposed only to volatiles from whole plants.
No statistically significant differences were ob-
served (Fig. 1). Less than 1% of the larvae moved
opposite of the airflow.
To reduce the artificiality of the bioassay, a
choice test was designed where the larvae were ex-
posed to and could make contact with whole plants.
This required a larger assay chamber, but other-
wise the overall design was similar to the petri dish
experiment. Neonates of both strains rapidly
moved to the plant material, but showed no signif-
icant preference to corn or stargrass (Fig. 1).
The choice cage passing-over bioassay was de-
signed to introduce forced air flow thereby facili-
tating exposure to volatiles while also allowing
contact with whole plants. In addition, the assay
required that the larvae contact 1 plant type be-
fore reaching the second. This was to test whether
neonates simply chose the first suitable food
source encountered, or were able to detect and ac-
tively search for the preferred host. The results
presented the first indication of colony-specific
and perhaps strain-specific larval feeding behav-
ior. Corn strain neonates were found at statisti-


Petri dish

SCS-Haog03
S RS-Ona03


S=2.91,df 9
P= 0.0174 = .6, df=9

P < 0.0001


corn stargrass corn stargrass
Plant selected

Olfactometer
O CS-Hag03 RS-Ona03
-t = 0.47, df 6
P = 0.6519
S= 148. d(=6
P= 0.1890





corn stargrass corn stargrass
Plant selected



Choice cage

[I CS-Hag03 2 RS-Ona03
= 055. dt=4
- 1 22, d = 4 P= 06145
P=02907


.__


corn stargrass com
Plant selected


Fig. 1. Percentage of corn strain (CS-Hag03) or rice
strain (RS-Ona03) larvae that chose either corn or star-
grass in a petri dish, olfactometer, or choice cage bioassay.


cally equal proportions on corn and stargrass in-
dependent of which was encountered first (Fig. 2).
In contrast, there was a general tendency for rice
strain neonates to remain on the first plant con-
tacted whether it was corn (69.6%) or stargrass
(82.4%), although only the latter was statistically
significant (Fig. 2).
Because the strongest plant host preferences
were observed with leaf sections in petri dishes,


stargrass


June 2008








Stuhl et al.: Fall Armyworm Larval Preferences


Choice cage passing over CS-Hag03

con -, grass OiM] grass -r corn

==084.d= 4 .. 07s6.=4
P = 0 4502 P 0 4901







corn stargrass corn stargrass
Plant selected


corn stargrass corn stargrass
Plant selected


Fig. 2. Percentage of corn strain (CS-Hag03) or rice
strain (RS-Ona03) larvae that chose either corn or star-
grass in a choice cage passing-over bioassay. For each
host strain, larvae encountered either corn (corn ->
grass) or stargrass (grass -> corn) first and then dis-
persed to the second plant.



the passing-over tests were repeated with that
simplified experimental design. The results iden-
tified 2 distinct behavioral patterns characteristic
of the 2 colonies. Neonates from the CS-Hag03
colony tended to remain on the first plant mate-
rial contacted regardless of plant type (Fig. 3). In
contrast, rice strain neonates were influenced by
host plant. In the configuration where the corn
section was contacted before the stargrass mate-
rial, the RS-Ona03 larvae distributed themselves
equally on the 2 plant types. If stargrass was en-
countered first, over 90% of larvae remained on
that section (Fig. 3).

CS-JS05, CS-Hag05, RS-MS05, and RS-Ona05 Colonies

Subsequent studies were performed with 4 col-
onies established in 2005 from populations in
southern and central Florida and the Mississippi
delta. In the petri dish choice bioassay, neonates
from all 4 colonies displayed a significant bias to
the corn material, consistent with that observed
with the 2003 colonies (Fig. 4).


100


80


S 60


40


20


0


100


80


Petri dish passing over CS-Hag03




- I corn grass II] grass corn




t=103, s d7=9 474.9
P<00001P 0o0011
___ FR


corn stargrass corn
Plant selected


stargrass


corn stargrass corn stargrass
Plant selected


Fig. 3. Percentage of corn strain larvae (CS-Hag03)
or rice strain larvae (RS-Ona03) that chose either corn
or stargrass in a petri dish passing-over bioassay. Lar-
vae encountered either corn (corn -> grass) or stargrass
(grass -> corn) first and then dispersed to the second
plant.



The passing-over experiments with plant sec-
tions in petri dishes identified 2 distinct behav-
iors, both different from that observed with the


100
Petri dish

80 CS-Hagm hIM CS-JS05 S RS-MS06 RS-OnaOS


S 60
gI 12 5, =f 2 3
C. P.0 293 P=232 0V1Mol
40 01.9 a
4')U

20


Fg n 4. m n g m P000 1m n enn
Plant selected

Fig. 4. Percentage of corn strain (CS-Hag05 and CS-
JS05) or rice strain (RS-MS05 and RS-Ona05) larvae that
chose either corn or stargrass in a petri dish bioassay.







Florida Entomologist 91(2)


2003 colonies. Over 90% of the neonates from all
4 colonies remained on the corn section when it
was contacted first (Fig. 5). In the reciprocal con-
figuration, larvae from the CS-JSO5, RS-MSO5,


and RS-Ona05 colonies distributed themselves
equally on the 2 plant types (Fig. 5). In contrast,
larvae from the CS-Hag05 colony always pre-
ferred the corn section (Fig. 5).


80 -



60 -


corn stargrass corn
Plant selected


corn stargrass corn
Plant selected


stargrass


60 -



40 -


Petri dish -
passing over




S0co0m -> grass
[r1 grass com





t=8.4, df= 9

P< 0.0001



n-


CS-JS05





t=-0.14, df=9

P = 0.8940


corn stargrass corn stargrass
Plant selected


stargrass


corn stargrass corn stargrass
Plant selected


Fig. 5. Percentage of corn strain (CS-Hag05 and CS-JS05) or rice strain (RS-MS05 and RS-Ona05) larvae that
chose either corn or stargrass in a petri dish passing over bioassay. Larvae encountered either corn (corn -> grass)
or stargrass (grass -> corn) first and then dispersed to the second plant.


Petri dish CS-Hag05
passing over





[ corn --) grass
rM grass -- corn

t=4.6, df= 9
P = 0.0013
t=22.5, df= 9

P < 0.0001



Inrm


June 2008







Stuhl et al.: Fall Armyworm Larval Preferences


DISCUSSION

Our objective in this study was to assess vari-
ation among different independently-isolated fall
armyworm populations based upon simple behav-
ioral bioassays targeted at the youngest larval
stage. Since the colonies were treated identically
for several generations, the presumption is that
any observed differences will most likely be the
result of genetic variation.
The attraction of fall armyworm neonates to
corn plant sections was previously observed by
Pashley et al. (1995), who found that most neo-
nate larvae of both strains preferred corn over
bermudagrass. Our findings that this phenotype
can be reproduced consistently with 6 indepen-
dently-isolated colonies generated over a two-
year period strongly suggest that this behavior is
ubiquitous and relatively insensitive to the influ-
ences of artificial culturing and genetic inbreed-
ing. Why the rice strain should exhibit this bias
toward corn is not clear.
Interestingly, a simple change in the orienta-
tion of the plant sections in the petri dish, so that
physical contact with one must occur before the
other can be reached resulted in significant behav-
ioral differences among colonies. The first stage of
the experiment was designed to have neonates
first encounter corn. Based on the choice test re-
sults, it was expected that both strains would tend
to remain on the corn section. This was observed
for 5 of the 6 colonies tested. Larvae from the 1 ex-
ception, RS-Ona03, were equally distributed over
the 2 plant sections, suggesting this colony might
have lost its attraction to corn volatiles.
We anticipated several possible outcomes when
stargrass was encountered first, depending on the
relative strength of the attraction to corn volatiles
compared to the tactile or taste attractiveness of
the intervening stargrass sections. In particular,
since the rice strain is the predominant fall army-
worm population in stargrass, we anticipated that
a substantial proportion of the neonates would
recognize it as an acceptable host and remain on
the stargrass section. This was the case for 2 of the
3 rice strain colonies tested, where a statistically
equal proportion of larvae was found on both plant
materials. Larvae from the third colony, RS-
Ona03, showed the same tendency but to a much
greater degree, consistent with the proposition
that attraction to corn volatiles was lost in this
colony. Our initial expectation with the corn strain
colonies was that these would always show a pref-
erence to corn, the presumed preferred plant host.
This was not the case, as each of the 3 corn strain
colonies gave a different response when the star-
grass section was encountered first.
Overall, the passing-over experiments with 6
different colonies uncovered 4 distinct behavioral
patterns with no clear indication of strain-specific
behavior. One simple way of explaining these re-


sults is to assume variability in the attraction to
corn sections relative to the attraction to star-
grass sections. Larvae from CS-Hag05 were al-
ways biased to the corn section, regardless of
which plant type was encountered first. In the
next 3 colonies (CS-JS05, RS-MS05, RS-Ona05),
the attraction to corn appeared reduced such that
when stargrass was first contacted, a substantial
number of larvae remained rather than move to
the corn section. Further reduction in corn attrac-
tion was found with larvae from CS-Hag03, where
now a majority of the neonates remained on star-
grass when it was encountered first. Finally, at-
traction to corn was so low with RS-Ona03 larvae
that a substantial number moved onto stargrass
even when corn was first encountered.
We conclude from this study that while there is
a consistent attraction of neonates to damaged
corn, there is substantial variability in the rela-
tive strength of that attraction that can be uncov-
ered in the passing-over experiments. Since the 6
colonies were cultured under the same laboratory
conditions, we believe these differences most
likely reflect genetic and phenotypic variability in
wild populations in addition to that associated
with strain differences.

ACKNOWLEDGMENTS

We thank H. Burnside, C. Dillard, and N. Lowman
for technical assistance with the experiments and in
culturing the fall armyworm colonies. We thank Mark
Carroll (USDA-ARS) and Heather McAuslane (Univer-
sity of Florida) for critical review of an early manu-
script. The statistical assistance of M. C. Christman,
University of Florida, Department of Statistics, IFAS
Statistical Consulting Unit, is appreciated. The use of
trade, firm, or corporation names in this publication is
for the information and convenience of the reader. Such
use does not constitute an official endorsement or ap-
proval by the United States Department of Agriculture
or the Agricultural Research Service of any product or
service to the exclusion of others that may be suitable.

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R. N. STORY. 1990. Comparison of bermudagrass
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LEVY, H. C., A. GARCIA-MARUNIAK, AND J. E. MARU-
NIAK. 2002. Strain identification of Spodoptera fru-
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gene. Florida Entomol. 85: 186-190.
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MARENCO, R. J., R. E. FOSTER, AND C. A. SANCHEZ.
1992. Sweet corn response to fall armyworm (Lepi-







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doptera: Noctuidae) damage during vegetative
growth. J. Econ. Entomol. 85: 1285-1292.
MARTIN, P. B., B. R. WISEMAN, AND R. E. LYNCH. 1980.
Action thresholds for fall armyworm on grain sor-
ghum and coastal bermudagrass. Florida Entomol.
63: 375-405.
MEAGHER, JR., R. L., AND M. GALLO-MEAGHER 2003.
Identifying host-strains of fall armyworm (Lepi-
doptera: Noctuidae) in Florida using mitochondrial
markers. Florida Entomol. 86: 450-455.
MEAGHER, R. L., AND R. N. NAGOSHI. 2004. Population
dynamics and occurrence of Spodoptera frugiperda
host strains in southern Florida. Ecol. Entomol. 29:
614-620.
MEAGHER, R. L., P. MISLEVY, AND R. N. NAGOSHI. 2007.
Caterpillar (Lepidoptera: Noctuidae) feeding on pas-
ture grasses in central Florida. Florida Entomol. 90:
295-303.
MEAGHER, R. L., R. N. NAGOSHI, C. STUHL, AND E. R.
MITCHELL. 2004. Larval development of fall army-
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MISLEVY, P., W. F. BROWN, L. S. DUNAVIN, D. W. HALL,
R. S. KALMBACHER, A. J. OVERMAN, O. C. RUELKE,
R. M. SONODA, R. L. STANLEY, JR., AND M. J. WILL-
IAMS. 1989. 'Florona' Stargrass. Florida Agric. Exp.
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MISLEVY, P., W. F. BROWN, L. S. DUNAVIN, D. W. HALL,
R. S. KALMBACHER, A. J. OVERMAN, 0. C. RUELKE, R.
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June 2008







Olmi & Virla: New Species ofAnteon from Argentina


A NEW SPECIES OF ANTEON (HYMENOPTERA: DRYINIDAE)
FROM ARGENTINA

MASSIMO OLMI1 AND EDUARDO G. VIRLA2
'Department of Plant Protection, University of Tuscia, I-01100 Viterbo, Italy

2PROIMI-Biotechnology, Biological Control Division, Av. Belgrano y Pje. Caseros (T4001 MBV),
San Miguel de Tucuman, Argentina

ABSTRACT

A new species, Anteon tucumanense, is described from San Miguel de Tucuman (Tucuman
Province, Argentina). This new species can be distinguished from the related Neotropical
species A. molle Olmi andA. parkeri Olmi by differences in segment 5 of the fore tarsus (dis-
tal part large and straight inA. tucumanense, slender and curved in the other 2 species). A
key is provided for the determinations of the above species with an illustration of the female
chela of the new species.

Key Words: Taxonomy, Hymenoptera, Dryinidae, Anteon tucumanense, new species, Argentina

RESUME

Se describe la nueva especieAnteon tucumanense de San Miguel de Tucuman (Provincia de
Tucuman, Argentina). A. tucumanense puede ser diferenciada de las species neotropicales
relacionadas, A. molle Olmi y A. parkeri Olmi, por diferencias en la forma del quinto seg-
mento de los tarsos anteriores (con la parte distal ancha, robusta y derecha enA. tucuman-
ense; con la parte distal angosta, delicada y curva en las otras dos speciess. Se provee una
clave para la determinaci6n de las species antedichas y la ilustraci6n de la quela de la
nueva especie.


Translation provided by the authors.


Anteon Jurine, 1807 was last revised by Olmi
(1984). It contains about 326 described species
from all continents (Olmi 1999). However, since
then many new species have been described, an-
other revision is necessary.
The species of Anteon inhabiting Argentina
were studied mainly by Ogloblin (1938); Olmi
(1984, 1987, 1991, 1992); Olmi et al. (2000); Olmi
& Virla (2004); Virla (1998); and Virla & Olmi
(1998). In 2007 we have examined additional
specimens of Anteon from Argentina and have
found the new species described below.

MATERIALS AND METHODS

The measurements reported are relative, except
for the total length (head to abdominal tip, without
the antennae), which is expressed in millimeters.
The description uses the terminology of Olmi (1984,
1994, 1999), with the following additional abbrevia-
tions: POL is the distance between the inner edges
of the 2 lateral ocelli; OL is the distance between the
inner edges of a lateral ocellus and the median ocel-
lus; OOL is the distance from the outer edge of a lat-
eral ocellus to the compound eye; OPL is the dis-
tance from the posterior edge of a lateral ocellus to
the occipital carina; TL is the distance from the pos-
terior edge of an eye to the occipital carina.


The material studied in this paper is deposited
in the Hymenoptera collection of Instituto de Zoo-
logia, Fundaci6n Miguel Lillo (IMLA), San
Miguel de Tucuman, Argentina.

Anteon tucumanense sp. nov. (Fig. 1)

Description. Holotype Female. Fully winged.
Length 2.37 mm. Head black, except mandibles
testaceous; antennae testaceous, except segments
7-10 darkened; mesosoma black, except anterior
margin of pronotum testaceous-brown; gaster
brown; legs testaceous. Antennae clavate; anten-
nal segments in the following proportions:
12:5:5:4:4:5:5:5.5:5:7. Head granulated and with
many irregular slight areolae; frontal line com-
plete; occipital carina complete; POL = 7; OL = 4;
OOL = 5; OPL = 4; TL = 5; greatest diameter of
posterior ocelli: 1. Pronotum dull, granulated and
crossed by numerous transverse keels; posterior
surface of pronotum much shorter than scutum
(4:15), broader than long; pronotal tubercles
reaching the tegulae. Scutum dull, smooth, gran-
ulated. Notauli incomplete, reaching approxi-
mately 0.2x length of scutum. Scutellum and met-
anotum shiny, smooth, without sculpture. Propo-
deum reticulate rugose, with a strong transverse
keel between dorsal and posterior surface; poste-







Florida Entomologist 91(2)


Fig. 1.Anteon tucumanense: chela of holotype (scale bar = 0.3 mm).


rior surface without longitudinal keels. Forewing
hyaline, without dark transverse bands; distal
part of stigmal vein much shorter than proximal
part (2:6). Fore tarsal segments in the following
proportions: 4:2:2:4.5:12. Segment 5 of fore tarsus
(Fig. 1) with basal part slightly longer than distal
part (7:5), 2 rows of 17 lamellae without interrup-
tion to the distal apex, and enlarged claw with a
proximal prominence bearing a long bristle. Tib-
ial spurs 1,1,2.
Male: Unknown.
Holotype: Female, Argentina, Tucuman Prov.,
San Miguel de Tucuman, 8.i.2007, E. Virla reared


ex Xerophloea viridis (Fabricius) (Hemiptera: Ci-
cadellidae) (IMLA).
Etymology: The species is named tucuman-
ense = inhabiting Tucuman.
Remarks. For the pronotum not crossed by a
transverse raised carina (Fig. 34 in Olmi, 1998),
A. tucumanense is similar to A. molle Olmi, 1984
and A. parkeri Olmi, 1998. The main difference
among the above species concerns the shape of
the chela. Following the description of A. tucu-
manense, the key to the females of NeotropicaAn-
teon presented by Olmi (1998) can be modified by
replacing couplet 14 as follows:


14 Segment 5 of fore tarsus with distal part large and straight (Fig. 1)................. tucumanense sp. nov.
-Segment 5 of fore tarsus with distal part slender and curved (Fig. 311 in Olmi, 1984;
fig. 37 in Olmi, 1998) ................. ............................................. 14'
14' Posterior surface of pronotum shorter than half of scutum. ................................. molle Olmi
-Posterior surface of pronotum longer than half of scutum ............................... parkeri Olmi


ACKNOWLEDGMENTS

The research described in the present paper was
supported by the scientific and technological co-opera-
tion between Italy and Argentina (IT-PA05-AYE/XV/
040). The authors thank Dr. Susana Paradell (Facultad
de Ciencias Naturales y Museo, Universidad Nacional
de La Plata) for identification of the host ofAnteon tucu-
manense sp. nov.


REFERENCES CITED

OGLOBLIN, A. A. 1938. Descripciones de Bethylidae y
Dryinidae de las colecciones del Museo Argentino de
Ciencias Naturales. An. Mus. Argentino Cie. Nat.
Bernardino Rivadavia, Buenos Aires 40: 35-50.
OLMI, M. 1984. A Revision of the Dryinidae (Hy-
menoptera). Mem. American Entomol. Inst. 37: xii +
1913 pp.


June 2008







Olmi & Virla: New Species ofAnteon from Argentina


OLMI, M. 1987. New species of Dryinidae (Hy-
menoptera, Chrysidoidea). Fragmenta Entomol. 19:
371-456.
OLMI, M. 1991. Supplement to the revision of the world
Dryinidae (Hymenoptera Chrysidoidea). Frustula
Entomol. (1989), N.S. xii (xxv): 109-395.
OLMI, M. 1992. New species of Dryinidae (Hymenop-
tera). Acta Zool. Hungarica 38: 281-292.
OLMI, M. 1994. The Dryinidae and Embolemidae (Hy-
menoptera: Chrysidoidea) of Fennoscandia and Den-
mark. Fauna Entomologica Scandinavica 30. Brill,
Leiden. 100 pp.
OLMI, M. 1998. New Embolemidae and Dryinidae (Hy-
menoptera Chrysidoidea). Frustula Entomol. (1997),
N. S. xx (xxxiii): 30-118.


OLMI, M. 1999. Hymenoptera Dryinidae-Embolemidae.
Fauna d'Italia 37. Edizioni Calderini, Bologna: xvi +
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mol. Chilena 22: 19-35.







Florida Entomologist 91(2)


June 2008


FEMALE AND MALE POLYMORPHISM IN TWO SPECIES OF MELITTOBIA
PARASITOID WASPS (HYMENOPTERA: EULOPHIDAE)


JORGE M. GONZALEZ1 AND ROBERT W. MATTHEWS2
'Department of Entomology, Texas A & M University, College Station, TX 77843, USA

2Department of Entomology, University of Georgia, Athens, GA 30602,USA

ABSTRACT

Polymorphism appears to be fundamental in Melittobia wasps, but uncertainty exists as to
its extent and form. Most researchers recognize 2 basic female forms-a long-winged disper-
sive "type form" and an early-maturing, short-winged gravid "second form." However, some
investigators have recognized two macropterous forms, "jumpers" and "flyers." Many others
have suggested that males, which normally comprise only about 5% of the population, also
may exist in 2 forms in various Melittobia species. This study examined the role of matura-
tional factors in 2 widespread representatives of different species groups, M. australica (ha-
waiiensis group) and M. digitata (acasta group). Individuals of both sexes from different
points in the emergence curve were examined and measured immediately after eclosion and
5 d later. Both sets of measurements supported the existence of 2 clearly defined female mor-
phs. However, when newly closed macropterous females were randomly assigned to 5-d
placements in empty vials or with prepupal hosts (Trypoxylon politum Say), subsequent
tests in a flight arena demonstrated that "jumpers" and iIl.... 'were simple reflections of
macropterous female physiological state, with heavier, gravid females reluctant to do more
than a slow crawl, and lighter, more nutritionally stressed females being more inclined to fly.
Thus we conclude that there is no justification for recognizing a "jumper" morph. Male mor-
phometrics indicated continuous variability in size and pigmentation of simple eyes and
ocelli, but no morphologically distinctive male morphs at any point in the emergence curve.
However, reports of distinct male morphs in other Melittobia species cannot be dismissed.

Key Words: brachyptery, morphometrics, flight, hopping, locomotion, eclosion order

RESUME

El polimorfismo parece ser fundamental en avispas del g6nero Melittobia, aunque no existe
certeza de su forma y extension. Muchos investigadores reconocen dos formas femeninas ba-
sicas-una dispersiva, con alas largas, la formaa tipica", y una de maduraci6n temprana, las
gravidas formaa secundarias" de alas cortas. Sin embargo, algunos investigadores han reco-
nocido dos formas macr6pteras, "saltadoras" y "voladoras". Otros han sugerido que entire ma-
chos, los cuales conforman cerca del 5% de la poblaci6n, pueden existir tambi6n dos formas
en varias species de Melittobia. Este studio examine el papel que cumplen los factors de
maduraci6n en dos representantes ampliamente distribuidos de dos grupos, M. australica
(grupo hawaiiensis) y M. digitata (grupo acasta). Se examinaron individuos de ambos sexos
de diversos puntos en la curva de emergencia, midi6ndolos al eclosionar y luego de cinco 5
dias. Ambos juegos de medidas apoyan la existencia de dos formas femeninas. Sin embargo,
cuando a hembras macr6pteras recientemente eclosionadas se colocaron al azar a los 5 dias
en contenedores vacios o con prepupas de sus hospedadoras (Trypoxylon politum Say), prue-
bas subsecuentes en una arena de vuelo demostraron que las supuestas formas "saltadoras"
y "voladoras" son solo el reflejo del estado fisiol6gico de las hembras macr6pteras, en las cua-
les las gravidas, mas pesadas, son reacias a hacer algo diferente a caminar, mientras que las
mas ligeras, estresadas nutricionalmente, estan mas dispuestas a volar. Es asi como conclui-
mos que no existe justificaci6n para reconocer una forma "saltadora". La morfometria entire
machos indica que existe una variabilidad continue en tamano y pigmentaci6n de ojos y oce-
los, pero no necesariamente existen formas masculinas distintas en cualquier punto en la
curva de emergencia. Sin embargo, reports sobre formas distintas en machos de otra espe-
cie de Melittobia no pueden ser descartados.

Translation provided by the authors.


Polymorphism is extensive in the wasp genus other in color, wing size, eye structure, and gross
Melittobia (Hymenoptera: Eulophidae). Males morphology. In addition, researchers have repeat-
and females of these small, arrhenotokous, gre- edly noted that each sex also seems to exhibit
gariously developing parasitoids differ from each more than one morphological form (Assem & Ma-







Gonzalez & Matthews: Polymorphism in Melittobia Parasitoids


eta 1980; Dahms 1984; Freeman & Ittyeipe 1982;
Schmieder 1933).
The most visibly prominent element of female
Melittobia polymorphism is wing length. Wing
polymorphism is widespread in insects, with deter-
minants that can be genetic, environmental, or
both. Its significance relates to dispersal, and the
phenomenon usually involves tradeoffs between
flight capability and ovarian investment (see re-
view in Zera & Denno 1997). The first to describe
this in Melittobia was Schmieder (1933), who dis-
cerned 2 female forms in M. chalybii, labeling them
as the "type form" and "second form" based on wing
development. Some 40 years later, Freeman & It-
tyeipe (1976, 1982) identified 3 female morphs in
M. sp. (hawaiiensis group), which are now consid-
ered to be M. australica. Their brachypterous
"crawlers" were clearly equivalent to Schmeider's
"second form", but within the macropterous, posi-
tively phototactic group, they distinguished 2 forms
that differed in their propensity for flight. One
form, the "jumpers", had slightly swollen abdomens
and wing tips that did not extend beyond the abdo-
men tip; the other form, the "flyers," had more slen-
der abdomens, and their wings extended beyond
the tip of their abdomens. The 2 researchers sug-
gested that this female trimorphism was an adap-
tation to exploit 3 spatial levels of host distribution
(Freeman & Ittyeipe 1982).
Meanwhile, studies were suggesting that male
Melittobia also might be polymorphic. Early stud-
ies ofM. chalybii by Schmeider (1933, 1938, 1939)
and his collaborators (Schmieder & Whiting 1947;
Whiting 1947; Whiting & Blauch 1948) were
joined by observations of Freeman & Ittyeipe
(1982) on M. australica. In both of these species,
researchers noted that some males seemed to be
larger and had pigmented eyespots and ocelli,
whereas other males were smaller and did not
have pigmented eyespots and ocelli. There also
were tantalizing signs that male Melittobia of
various other species might exhibit 2 forms (As-
sem & Maeta 1980; Hartley & Matthews 2003;
Lapp 1994).
But how many of these polymorphic forms ac-
tually exist as distinct entities? Rearing various
species of Melittobia in our laboratory, we had
questions. While we could easily discern 2 female
forms-a long-winged, positively phototactic dis-
perser and a short-winged non-disperser-we
could not reliably distinguish between jumpers
and flyers. Moreover, males (which comprise only
5% of most populations) are pugilistic and even
cannibalistic toward one another, and most com-
monly are observed only after they have died; our
observations of dead males showed considerable
morphological variation, which might or might
not be attributable to such factors as male age or
desiccation. Furthermore, the majority of pub-
lished observations suggesting male polymor-
phism had been based on the catch-all group,


M. chalybii; knowing that this designation has
been applied to at least 4 different species
(Gonzalez & Matthews 2002), we thought it plau-
sible that males identified as different morphs
might even have been males of different species.
Therefore, the objectives of this study were to
quantify differences purported to distinguish
Melittobia morphs, and to clarify the role of vari-
ous life factors in the expression of behavior and
morphology within each sex of Melittobia. To do
so, we undertook morphometric and behavioral
examinations of individuals of 2 common North
American species, M. digitata Dahms and M. aus-
tralica (Girault), comparing females reared iden-
tically except for feeding regimen before being
placed in a flight test arena, and comparing iden-
tically reared males grouped by emergence se-
quence. Our hypotheses were that apparent male
polymorphism and apparent female macropter-
ous subgroup polymorphisms were artifacts of
natural male size variation and female nutri-
tional condition, respectively.

MATERIALS AND METHODS

Rearing Protocol

Each culture of M. australica or M. digitata
was initiated by placing 1 mated female upon a
naked prepupa of Trypoxylon politum Say (Hy-
menoptera: Crabronidae) within a shell vial
maintained at 25C under constant darkness. Af-
ter 18 d, males and females were separated as
late pupae, at which time the sexes can readily be
established. Eclosion occurs over a week or more
in these species. Each day, as new adults of both
sexes appeared, they were removed and slide-
mounted for morphology measurements. Females
from identically reared cultures were removed
and maintained for behavioral studies as outlined
below.

Morphological Measurements

Using an ocular micrometer, we measured
wing length, hind tibia length, and abdomen
length of each individual. Also for males we mea-
sured head width and recorded the occurrence of
pigmentation in their simple eyes eyespotss) and
ocelli. To assess whether male morphometrics or
pigmentation patterns were related to develop-
ment or emergence schedule, we partitioned male
data into 3 age categories-arly (d 1-3), middle (d
4-7), and late (d 8-11)-based on a previous study
chronicling emergence patterns (Adams 2002).

Behavioral Studies

Additional cultures of each species, reared
identically, were used to further assess mac-
ropterous female locomotory behavior and test







Florida Entomologist 91(2)


the hypothesis that macropterous "jump vs.
flight" propensity might be due to fluid/food inges-
tion rather than being a genetically determined
proclivity.
A simple flight arena was constructed by
standing a round wooden toothpick (25 mm long)
on a platform in the center of a white poster board
marked with concentric rings at 50, 100, and 150
mm. Pilot tests confirmed that a female wasp re-
leased at the base of the pick and prodded gently
with the bristles of a small camel's hair brush
would almost invariably respond by climbing to
the top of the pick. From here she had the choice
of flying off, jumping off, turning around and
climbing back down, or simply becoming a pole-
sitter. Pilot trials showed that the overwhelming
majority of newly closed females of both species
preferred the latter 2 options, but flight and
jumping both occurred regularly in these popula-
tions. The longest jumping distance recorded was
less than 100 mm.
Within 1 d of eclosion from our experimental
cultures, females were tested individually with
this system. Once on the pick, the wasp was al-
lowed up to 5 min to either launch herself into the
air, crawl down and off, or remain on the pick. If
genetically determined sub-morph differentiation
into "jumpers" and "flyers" were present within
this macropterous population, it should manifest
itself most clearly in the behavior of those individ-
uals that chose to launch themselves off the pole
at this early stage of their adult lives. Thus, indi-
viduals that crawled down and off the pick or
were still crawling on the pick after 5 min were
disregarded. Those that launched from the pick
were assigned to 4 distance-based groups until
the sample size of each group reached at least 20
individuals. As determined by landings within
the concentric rings on the flight arena floor,
launches were scored as less than 50 mm, 51-100
mm, 101-150 mm, and more than 150 mm.
Females landing less than 50 mm from the
pick were considered equivocal and were excluded
from further study; these wasps may have fallen
off the pick, jumped, or possibly flown in a tight


loop. At the other extreme, females that covered
distances of more than 150 mm in the arena
clearly had flown, and thus were considered to be
equivalent to the "flyers" of Freeman & Ittyeipe
(1982). Because the pilot study indicated that
jump distance never exceeded100 mm, those cov-
ering 101-150 mm most probably were all flyers,
but to reduce uncertainty we excluded them as
well. Only wasps that went 51-100 mm were con-
sidered equivalent to Freeman & Ittyeipe's
"jumpers."
Those "flyers" and "jumpers" meeting the above
criteria then were randomly assigned to one of 2
nutritional subgroups. Individuals in 1 subgroup
("fed") were provided with a T politum prepupa;
individuals of the other subgroup ("unfed") were
placed in an identical empty container. After 5 d,
all females were again individually tested in the
flight arena by the same protocol, and their abdo-
men lengths were again measured. Video record-
ings were made of selected individuals' perfor-
mances in the flight test arena to assist in discern-
ing specific behaviors related to flight and jumping.

RESULTS

Male Morphometrics

In all, the wing lengths, hind tibia lengths, and
head widths of 52 freshly closed males ofM. aus-
tralica and 133 freshly closed males of M. digi-
tata were measured, and ratios of hind tibia:wing
length and hind tibia: head width were calculated
(Table 1).
In both species, when measurements taken
upon the first and last group of males to emerge
were compared, clear differences in both wing
length and head width were apparent. However,
when the middle group was included, a continu-
ous size variation and loss of a bimodal size distri-
bution resulted (Table 1).
Ratios of the hind tibia length to wing length
or head width revealed no consistent differences
relating to emergence time. However, wing lengths
of the first few M. digitata males to eclose were


TABLE 1. VARIATION IN MALE SIZE AND FORM FOR M. AUSTRALICA AND M. DIGITATA BY ECLOSION SEQUENCE.

Eclosion interval Forewing length Hind tibia length Head width Hind tibia/head
(d) (mm) (mm) (mm) Hind tibia/wing width

M. australica (n = 15 per eclosion interval; 45 total)
1-3 0.49+ 0.005 0.29 0.005 0.39 0.01 0.60 0.007 0.75 0.03
4-7 0.43 0.02 0.28 0.01 0.34 0.02 0.62 0.06 0.83 0.05
8-11 0.39 0.005 0.29 0.005 0.30 0.01 0.67 0.04 0.89 0.08
M. digitata (n = 21 per eclosion interval; 63 total)
1-3 0.50 0.005 0.30 0.01 0.40 0.01 0.61 0.02 0.76 0.02
4-7 0.45 0.02 0.30 0.009 0.39 0.009 0.65 0.02 0.75 0.02
8-11 0.43 0.03 0.28 0.01 0.37 0.02 0.66 0.04 0.76 0.05


June 2008







Gonzalez & Matthews: Polymorphism in Melittobia Parasitoids


significantly different from their later-eclosing
brothers (Kruskal-Wallis Test, P < 0.05).
Males of Melittobia lack compound eyes, but
have undeveloped simple eyespots and ocelli, and
differences in their extent of pigmentation also
have been suggested to signal the existence of 2
male morphs. Our analysis (Table 2) showed that
overall pigmentation of both structures was the
commonest condition, and that while variation
did occur, there was no consistent pattern of pig-
ment presence or absence in relation to emer-
gence time. However, almost half (40%) of our
sample exhibited variation in eyespot and/or
ocelli presence and pigmentation. Unpigmented
eyespots were invariably correlated with unpig-
mented ocelli, but 13/42 (31%) of the males with
pigmented eyespots lacked pigmented ocelli.

Female Morphometrics

Samples of 100 females of M. australica and
M. digitata representing different developmental
stages of the cultures were measured and com-
pared (Table 3). Both species exhibited a clearly
bimodal distribution of measures assignable to 2
morphological groups most obviously separated
by wing length. In each species, the wing lengths
of the 2 morphs could be clearly distinguished and
were statistically significantly different from
each other. Likewise, the ratio of hind tibia length
to wing length clearly showed that females of
each species exhibit only 2 clear morphs.
Absolute body sizes varied, but females of M.
digitata were slightly larger than M. australica
overall. Wings of brachypterous M. digitata fe-
males were more uniform in size than those of
brachypterous M. australica. Variations in abdo-
men length (a highly variable character) and in
abdomen: hind tibia length ratio were both con-


tinuous, thus also providing no support for the hy-
pothesis of 2 distinct long-winged female morphs
in the 2 species we studied.

Behavioral Studies

Even with the naked eye, it is apparent that
macropterous females of both species mostly walk
about, but occasionally will hop and/or take short
flights as well as rarer long ones (Matthews et al.
1996). Videotape recordings of females crawling
about on toothpick towers revealed that the fe-
males often repeatedly raised the anterior part of
their body and rapidly flexed their wings 1 or
more times in a few seconds, inducing a contrac-
tion of the thorax. These behaviors typically pre-
ceded a launch from the pick and we consider
them to be intention movements. Whether fe-
males also simply jumped off or dropped from the
picks without accompanying wing flips was not
confirmed, but probably also occurred.
At 1 d of adult age, most newly emerged long-
winged females simply crawled and showed no
strong tendency to launch from the toothpick, al-
though flight >150 mm was observed.
All females provided with a host for 5 d became
physogastric due to host feeding and consequent
ovarian development. This state was easily observ-
able, as the tips of their swollen abdomens now ex-
tended beyond the wing tips (Fig. 1). In contrast,
long-winged females placed in empty containers
for 5 d became more slender as their internal re-
serves were depleted; in consequence, their abdo-
mens visibly shrunk so that their wing tips clearly
extended beyond the tip of abdomen (Fig. 1).
Confronted with a second time atop the pole in
the flight arena (Table 4), 5-d-old physogastric fe-
males displayed an extremely low propensity to
launch themselves in any manner. (For example,


TABLE 2. EYESPOT AND OCELLAR PIGMENTATION IN MALES OF MELITTOBIA AUSTRALICA AND M. DIGITATA IN RELATION
TO ECLOSION ORDER.

Eyespots fully pigmented

All ocelli fully No ocellar Mixed ocellar No pigmented
Eclosion interval pigmented pigmentation pigmentation eyespot or ocelli'

M. australica (n = 45)
Early (d 1-3) 8 5 1 1
Middle (d 4-7) 10 3 1 1
Late (d 8-11) 9 5 0 1
All males 27 (60%) 13 (29%) 2 (4.5%) 3 (6.5%)
M. digitata (n = 64)
Early 12 4 2 3
Middle 10 7 2 2
Late 12 8 1 1
All males 34 (53%) 19 (30%) 5 (8%) 6 (9%)

In both species, no individuals that lacked eyespot pigmentation were observed to have ocellar pigmentation.







Florida Entomologist 91(2)


TABLE 3. FEMALE MORPH DIFFERENCES IN MELITTOBIA AUSTRALICA AND M. DIGITATA.

Forewing length Hind tibia length Abdomen length Hind tibia/wing Hind tibia/
Morph (mm) (mm) (mm) length abdomen length

M. australica (n = 100)
Long winged 0.61 0.08 0.29 0.004 1.14 + 0.13 0.49 0.07 0.26 0.02
Short winged 0.84 0.09 0.30 0.004 1.04 0.06 0.35 0.03 0.28 0.01
M. digitata (n = 100)
Long winged 0.65 0.10 0.30 0.01 1.03 0.05 0.47 0.06 0.29 0.01
Short winged 1.09 0.06 0.34 0.01 1.18 0.06 0.31+ 0.01 0.28 0.01


in the earlier pilot study, only 1 of 12 M. austral-
ica and 0 of 18 M. digitata left the pole.) When
they did launch themselves, few became air-
borne-only 15% of fed M. australica launchers
flew, whereas 60% of the unfed launchers flew. For
M. digitata, the differences were even more dra-
matic; none of the fed launchers flew, but 86% of
the unfed launchers did.

DISCUSSION

How Many Female Morphs Does Melittobia Have?

Our data confirmed the existence of only 2
clearly defined morphological forms, brachypter-
ous and macropterous, in both M. australica and
M. digitata. These results concur with previous
findings in other species (e.g., Schmieder 1933;
Lith 1955; Gonzalez 1994; Gonzalez et al.1996;
Lapp 1994). Recent work confirms that these mor-


phs are nutritionally determined (C6nsoli & Vin-
son 2002). The first several offspring on a singly
parasitized large host become brachypterous fe-
males that develop several days faster than their
macropterous siblings, apparently due to better
food quality, an assumption made originally by
Schmieder (1933) and supported by Freeman & It-
tyeipe (1982). However, if 2 or more Melittobia fe-
males superparasitize a host, few or often no brac-
hypterous females are produced (unpublished ob-
servations), and only macropterous females re-
sult. In any case, the number of brachypterous
females rarely exceeds 40 individuals, and is usu-
ally closer to 20 (Freeman & Ittyeipe 1982).
Although most behavioral studies ofMelittobia
have concentrated solely upon the long-winged
form, it is clear that once set in motion, develop-
mental and morphological differences play out in
different behaviors throughout the lives of the 2
morphs. For example, the 2 morphs ofM. digitata


Fig. 1. Grossly apparent differences in Melittobia digitata after 5 d of feeding upon a natural Trypoxylon politum
prepupal host. (Left) The abdomen of fed females swells with ovarian development so that it extends beyond the
wing tips. (Right) Unfed 5-d-old females have shrunken abdomens, such that the tips of the forewings extend be-
yond the abdomen tip.


June 2008







Gonzalez & Matthews: Polymorphism in Melittobia Parasitoids


TABLE 4. LAUNCHING BEHAVIOR OF MACROPTEROUS 5-D-OLD MELITTOBIA FEMALES IN RELATION TO HOST-FEEDING
STATUS.

Behavior inferred through landing distance from pick

Nutritional status "Jump" (51-100 mm) "Flight" (>150 mm)

M. australica
Fed (n = 20) 17 (85%) 3 (15%)
Unfed (n = 37) 15 (40%) 22 (60%)
M. digitata
Fed (n =10) 10(100%) 0
Unfed (n = 22) 3 (14%) 19 (86%)


vary in courtship details (Gonzalez & Matthews
2005). Ability to become airborne appears to be
simply one more difference, in this case appar-
ently reflecting body weight. As gravid, fed fe-
males of M. australica and M. digitata become
"heavier", they show a reduced propensity to fly.
On the other hand, in the absence of feeding, nu-
trient reserves diminish and the wasp becomes
noticeably more slender, displaying a correspond-
ingly greater tendency to fly. Thus, Freeman & It-
tyeipe's (1976, 1982) findings probably simply re-
flected the confounding influence of food intake.
Freeman & Ittyeipe (1982) indicate that their
jumper morph comprised only about 20-40 indi-
viduals intermediate in morphology to crawlers
and flyers. However, they acknowledged that
their 3 morphs overlapped in their morphologies,
and recognized them primarily as "functionally
distinct". In our experience, females use crawling
as their primary form of locomotion, and undis-
turbed females rarely hop or jump spontaneously.
However, jumping can be readily elicited if one
"threatens" a female, e.g. with the tip of a pencil
or paint brush (Matthews et al. 1996). Jumping
thus appears to be primarily a predator avoidance
response and, contrary to Freeman & Ittyeipe's
assertion, seems unlikely to be used as a principal
dispersal mechanism, The lack of unique morpho-
logical attributes further weakens the case for
recognizing a distinct jumper morph.
Because host feeding induces ovarian develop-
ment, a cascade of physiological and behavioral
changes inevitably follows this act. However, ova-
rian development normally occurs only after suc-
cessful dispersal and consequent host location;
because gravid females do not usually leave their
host, they would not be likely to engage in further
extensive locomotion.
Whether these changes in locomotion should
be attributed to feedingper se or to ovarian devel-
opment is still an open question. It is worth re-
membering that host feeding is not the only
source of food available to Melittobia; females
readily ingest carbohydrates in the laboratory
(unpubl. observe ) and may obtain honeydew and


possibly floral nectar during dispersal. Ageing no
doubt also affects behavior and physiology. Fur-
ther work will be required to tease apart the roles
of such factors as they affect dispersal and loco-
motion behavior.
What adaptive significance might there be in
this little parasite's female polymorphism? As sug-
gested for other examples of polymorphism in in-
sects, the brachypterous morph may provide a
trade-off of rapid fecundity against being flightless
(Harrison 1980; Roff 1986; Tanaka 1993; Wheeler
1995). According to C6nsoli & Vinson (2002a) brac-
hypterous M. digitata females emerge with a load
of about 30 developed eggs, and following mating,
start immediately laying them on their natal host.
Although Freeman & Ittyeipe (1982) suggest that
brachypterous females may disperse short dis-
tances, we have seen no evidence that they ever
leave their natal host. Rather, they stay to further
exploit their partially consumed natal host and
thus fully realize their fecundity.
Simultaneously, it is obvious that total investi-
ture in a non-dispersing morph would be a dead-
end route, both in the short term and evolution-
arily. The long-winged, powerfully jawed, and
strongly phototactic females of the macropterous
form are true dispersal machines. To compensate
for their very small chances of successfully locat-
ing a new host, estimated by Freeman & Ittyeipe
(1982) as a probability of 1 in 485, these females
have much higher potential fecundity. We have
recorded over 800 offspring on a single host for
some Melittobia species (unpubl. observ.). To-
gether, the 2 female morphs provide Melittobia
with a strategy in which the whole could truly be
said to be greater than the sum of its parts.

How Many Male Morphs Are There?

Though some authors have postulated the ex-
istence of 2 male morphs in various species of
Melittobia, data from our study fail to support the
existence of 2 clear morphs in males inM. austral-
ica and M. digitata, and instead suggest that a
continuum of morphological variation exists. For







Florida Entomologist 91(2)


M. australica, this supports data in Gonzalez et
al. (1996). Behavior-based male morphs inM. aus-
tralica also seem unlikely. Whereas Freeman & It-
tyeipe (1982) postulated 2 morphological forms of
males in M. sp. (hawaiiensis complex) (=M. aus-
tralica), they acknowledged that they "saw no dif-
ference in the behavior of male morphs".
The early work of Schmieder (1933, 1938,
1939) and collaborators (Schmieder & Whiting
1947; Whiting 1947; Whiting & Blauch 1948) with
M. chalybii has formed the basis for much of the
speculation about male morphs in Melittobia, but
is problematical because we now know that those
researchers were actually working with up to 4
different species (Gonzalez & Matthews 2002).
Thus, the possibility cannot be discounted that
males of 2 different species were considered as 2
different morphs of a single species.
Freeman & Ittyeipe (1982) reported that M.
australica males in large broods (>40 adults) were
more fully pigmented overall (including fully pig-
mented eyespots and ocelli) in comparison to more
lightly pigmented "second form" males with "un-
derdeveloped ocelli" that appeared only in small
broods (up to 30 adults per standard host). Inter-
estingly, Schmeider (1933) recognized a type form
male with pigmented eyespots and ocelli but ap-
parently his second "totally blind" form was darker,
not lighter, than his type form. Our data indicated
a great deal of variation in eyespots and ocelli, and
failed to show an association between this varia-
tion and eclosion sequence. It is also worth noting
that the M. australica males in our study were
reared in very large broods (ca. 400 adults) that
should have led solely to "type form" development
by Freeman & Ittyeipe's (1982) criterion. Instead,
we found pigmentation variants representing both
supposed morphs and a range between.
Melittobia digitata is perhaps the most thor-
oughly studied of all the species in this genus.
Most of the published research has centered upon
the long-winged females. However, important
studies by C6nsoli and his collaborators (C6nsoli
& Vinson 2002a, b; 2004; C6nsoli et al. 2004) have
confirmed that the brachypterous and macropter-
ous female morphs are real, and are determined
by host quality and quantity. The question as to
whether this species also produces distinct male
morphs by similar or analogous means has yet to
be addressed. Certainly the present study found
only continuous variation over time, as males be-
came progressively smaller in absolute size but
kept their basic same body proportions. However,
it is worth remembering that our study addressed
a male population all produced under one stan-
dard scenario; different rearing regimens might
yield different results.
We also cannot discount the possibility of male
morphs in some other Melittobia species. Assem &
Maeta (1980) reported 2 distinct morphs in males
of a Melittobia species from Japan (=M. sosui) and


suggested that this dimorphism was not related to
food supply. Working with M. femorata Dahms,
Lapp (1994) distinguished 2 possible male morphs
based upon their eclosion before and after the ex-
tended prepupal diapause that is apparently
unique to this species (see Matthews et al. 2005).
In a forthcoming study (Matthews & Gonzalez, un-
publ. data), we present evidence that the first few
M. femorata males from the pre-diapause clutch do
consistently differ morphologically from those in
the post-diapause, late-developing group.

REFERENCES CITED
ADAMS, R. F. 2002. Dynamics of daily progeny produc-
tion in Melittobia digitata Dahms: Baseline studies.
Undergrad. Sci. Bull. 3:17-21.
ASSEM, J. VAN DEN, AND Y. MAETA. 1980. On a fourth
species ofMelittobia from Japan. Kontyu 48: 477-481.
CONSOLI, F. L., AND S. B. VINSON. 2002a. Clutch size,
development and wing morph differentiation of
Melittobia digitata Dahms (Hymenoptera: Eu-
lophidae). Entomol. Exp. Appl. 102: 135-143.
CONSOLI, F. L., AND S. B. VINSON. 2002b. Larval devel-
opment and feeding behavior of the wing dimorphics
of Melittobia digitata Dahms (Hymenoptera: Eu-
lophidae). J. Hymenop. Res. 11(2): 188-196.
CONSOLI, F. L., AND S. B. VINSON. 2004.Wing morph de-
velopment and reproduction of the ectoparasitoid
Melittobia digitata: Nutritional and hormonal ef-
fects. Entomol. Exp. Appl. 112: 47-55.
CONSOLI, F. L., H. S. TIAN, S. B. VINSON, AND C. J.
COATES. 2004. Differential gene expression during
wing morph differentiation of the ectoparasitoid Melit-
tobia digitata (Hym. Eulophidae). Comp. Biochem.
Physiol. (A): Molec. Integr. Physiol. 138(2): 229-239.
DAHMS, E. 1984. A review of the biology of species in the
genus Melittobia (Hymenoptera: Eulophidae) with
interpretations and additions using observations on
Melittobia australica. Mem. Queensland Mus. 21(2):
337-360.
FREEMAN, B. E., AND K. ITTYEIPE. 1976. Field studies on
the cumulative response ofMelittobia sp. (hawaiien-
sis complex) (Eulophidae) to varying host densities.
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FREEMAN, B. E., AND K. ITTYEIPE. 1982. Morph determi-
nation in Melittobia, a eulophid wasp. Ecol. Entomol.
7: 355-363.
GONZALEZ, J. M. 1994. Taxonomia, Biologia y Compor-
tamiento de Avispas Prasiticas del Genero Melit-
tobia Westwood (Hymenoptera: Eulophidae) en
Venezuela. Ph.D. Thesis, Universidad Central de
Venezuela, Maracay. 118 pp.
GONZALEZ, J. M., AND R. W. MATTHEWS. 2002. Develop-
ment and sex ratio of Melittobia australica and M.
digitata (Hymenoptera: Eulophidae) on Megachile
rotundata (Hymenoptera: Megachilidae) and Try-
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GONZALEZ, J. M., AND R. W. MATTHEWS. 2005. Court-
ship of the two female morphs of Melittobia digitata
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(3): 258-267.
GONZALEZ, J. M., R. W. MATTHEWS, AND J. B. TERAN.
1996. Parasitoides del g6nero Melittobia Westwood
(Hymenoptera: Eulophidae) en Venezuela. Dis-


June 2008







Gonzalez & Matthews: Polymorphism in Melittobia Parasitoids


tribuci6n y hospederos. Bol. Entomol. Venezuela
(N.S.) 11(2): 139-147.
HARRISON, R. G. 1980. Dispersal polymorphisms in in-
sects. Annu. Rev. Ecol. Syst. 11: 95-118.
HARTLEY, C. S., AND R. W. MATTHEWS. 2003. The effect
of body size on male-male combat in the parasitoid
wasp Melittobia digitata Dahms (Hymenoptera: Eu-
lophidae). J. Hymenop. Res. 12: 272-277.
LAPP, S. 1994. The Mating System of Melittobia Para-
sitic Wasps: Fluctuating Asymmetry, Sexual Selec-
tion, and Comparative Courtship Behaviour. M.Sc.
Thesis, University of Georgia, Athens. 63 pp.
LITH, J. P. VAN. 1955. Biologie van Melittobia acasta
Walker (Hymenoptera, Chalcididae). Tijdschr. Ento-
mol. 98: 29-42.
MATTHEWS, R. W., T. R. KOBALLA, JR., L. R. FLAGE, AND
E. J. PYLE. 1996. WOWBugs: New Life for Life Sci-
ence. Riverview Press, Athens, GA. 318 pp.
MATTHEWS, R. W., L. D. DEYRUP, AND J. M. GONZALEZ.
2005. Increased male sex ratio among brachypterous
progeny in Melittobia femorata, a sib-mating parasi-
toid wasp (Hymenoptera: Eulophidae). Insect Sci.
12: 443-448.
ROFF, D. A. 1986. The evolution of wing dimorphism in
insects. Evolution 40: 1009-1020.
SCHMIEDER, R. G. 1933. The polymorphic forms of Melit-
tobia chalybii Ashmead and the determinant factors


involved in their production (Hymenoptera: Chalci-
doidea, Eulophidae). Biol. Bull. 74: 135-163.
SCHMIEDER, R. G. 1938. The sex ratio in Melittobia cha-
lybii Ashmead, gametogenesis and cleavage in fe-
males and in haploid males (Hymenoptera:
Chalcidoidea). Biol. Bull. 74: 256-266.
SCHMIEDER, R. G. 1939. The significance of the two
types of larvae in Sphecophaga burra (Cresson) and
the factors conditioning them (Hymenoptera: Ich-
neumonidae). Entomol. News 50: 125-131.
SCHMIEDER, R. G., AND P. W. WHITING. 1947. Reproduc-
tive economy in the Chalcidoid wasp Melittobia. Ge-
netics 32: 29-37.
TANAKA, S. 1993. Allocation of resources to egg produc-
tion and flight muscle development in a wing dimor-
phic cricket, Modicogryllus confirmatus. J. Insect
Physiol. 39: 493-498.
WHEELER, D. 1995. The role of nourishment in oogene-
sis. Annu. Rev. Entomol. 41: 407-431.
WHITING, P. W. 1947. Some experiments with Melit-
tobia and other wasps. J. Hered. 38: 11-20.
WHITING, P. W., AND B. M. BLAUCH. 1948. The genetic
block to free oviposition in the Chalcidoid wasp
Melittobia sp. Biol. Bull. 95: 243-244.
ZERA, A. J., AND R. F. DENNO. 1997. Physiology and
ecology of dispersal polymorphism in insects. Annu.
Rev. Entomol. 42: 207-231.







Florida Entomologist 91(2)


June 2008


DACNUSA CICERINA (HYMENOPTERA: BRACONIDAE: ALYSIINAE),
A NEW SPECIES OF ENDOPARASITOID OF LIRIOMYZA CICERINA
(DIPTERA: AGROMYZIDAE)

JOSEP TORMOS1, XAVIER PARDO2, JOSEP DANIEL ASiS1 AND SEVERIANO F. GAYUBO1
'Unidad de Zoologia, Facultad de Biologia, Universidad de Salamanca, 37071-Salamanca, Spain
E-mail: tormos@usal.es

2Universitat de Valencia. Institute Cavanilles de Biodiversitat i Biologia Evolutiva.
Apartat Oficial 2085-46071 Valencia, Spain

ABSTRACT

The larvae, pupa, adults, and venom apparatus of Dacnusa cicerina sp. n., an endoparasi-
toid of Liriomyza cicerina (Rondani), found on Cicer arietinum Linnaeus in Spain, are de-
scribed, illustrated, and compared with those of allied species. The mature larva of
Eurytoma sp., possibly a hyperparasitoid of D. cicerina, also is described, illustrated, and
compared with those of allied species. Keys to discriminate adults are provided and morpho-
logical structures of phylogenetic value are discussed. The adults of D. cicerina are similar
to those of Dacnusa rodriguezi Docavo & Tormos (1997). The immature larvae are similar to
those of Dacnusa areolaris (Nees) and Dacnusa dryas (Nixon), and the mature larva is very
similar to that ofD. dryas, from which it differs in having scale-like sensilla setaeae") on the
thorax and abdomen. The cast skin of the final instar, like those described for Dacnusa Ha-
liday, has a pleurostoma with well differentiated mandible processes and a long stipital
sclerite. The venom apparatus of this species is very similar to that of Dacnusa flavicoxa Th-
omson, differing from it in length of the reservoir and the number of gland filaments. The
mature larva of Eurytoma Illiger described here, despite its endoparasitoid nature, has well
differentiated pleural and ventral setae.

Key Words: Hymenoptera, Braconidae, new species, host, immature stages, venom appara-
tus, hyperparasitoid, Spain

RESUME

Se described, ilustran, y comparan con las species mas pr6ximas, las larvas, pupa, adults
y el aparato del veneno de Dacnusa cicerina n. sp., endoparasitoide obtenido en Espana a
partir de Liriomyza cicerina (Rondani) sobre Cicer arietinum Linnaeus. Adicionalmente, se
describe, ilustra y compare con las species mas afines, la larva madura de Eurytoma sp., un
probable hiperparasitoide de D. cicerina. Se proporcionan claves dicot6micas para la separa-
ci6n de los images, a la vez que se discuten estructuras morfol6gicas con valor filogen6tico.
Los images de D. cicerina son similares a los de Dacnusa rodriguezi Docavo & Tormos (1997).
Las larvas inmaduras son similares a las de Dacnusa areolaris (Nees) y Dacnusa dryas
(Nixon). La larva madura es muy similar a la de D. dryas, de la que se separa por presentar
sensilas con forma de escama "setas" sobre el t6rax y abdomen. La exuvia del ultimo estado
larvario, al igual que las descritas de Dacnusa Haliday, present un pleurostoma con proce-
sos mandibulares bien diferenciados y un esclerito estipital largo. El aparato del veneno de
esta especie es muy similar al de Dacnusa flavicoxa Thomson, del que se separa por la lon-
gitud del reservorio y el numero de filamentos glandulares. La larva madura de Eurytoma
Illiger descrita, a pesar de su naturaleza endoparasitoide, present setas pleurales y ventra-
les bien diferenciadas.

Translation provided by the authors.



From a study undertaken in 1987-1992 on Dac- the Mediterranean area it mainly feeds on chick-
nusini (Hymenoptera: Braconidae: Alysiinae) and peas, for which it may be a serious pest.
their agromyzid (Diptera: Agromyzidae) hosts in In the study cited above, Dacnusa cicerina sp.
the Iberian Peninsula, data were gathered on the n. was obtained from L. cicerina feeding on chick-
parasitoids of Liriomyza cicerina (Rondani), a pea plants, and Eurytoma sp., a possible hyper-
species injurious to Cicer arietinum Linnaeus parasitoid of D. cicerina, also was obtained. An-
(chickpea) cultures. Although this species also other species of Braconidae, which has been ob-
feeds on Ononis and Hymenocarpus elsewhere, in served parasitizing L. cicerina on chickpeas in







Tormos et al.: Larvae and Venom Apparatus of D. cicerina sp. n.


Spain, Opius monilicornis Fischer (Garrido et al.
1992) also was found. Detailed information on the
economic importance and the biology of L. cice-
rina has been given by Spencer (1973, 1976,
1990).
The genus Dacnusa Haliday belongs to the
subfamily Alysiinae and to the tribe Dacnusini,
most of whose members attack agromyzid flies.
The classification and biology of this genus, which
has approximately 77 European species (Achter-
berg 2004), have been studied by Griffiths (1964,
1966, 1968, 1984); Tobias (1995); and Belokobyl-
skij et al. (1998). The structures that allow char-
acterization of the final instar have been de-
scribed in depth only in Dacnusa rodriguezi
Docavo & Tormos (Pardo et al. 2000). Detailed
studies addressing the variation in gland and res-
ervoir morphology of the venom apparatus in spe-
cies of Dacnusini recently have been conducted by
Quicke et al. (1997) and Tormos et al. (2003).
The genus Eurytoma Illiger is the largest of
the family Eurytomidae (Noyes 2001), and in-
cludes species displaying diverse larval feeding
habits. This study addresses the larval morphol-
ogy of an undetermined species of this genus,
probably an hyperparasitoid of the new species of
Dacnusa described here. A recent study of the ma-
ture larvae of Eurytoma has been carried out by
Tormos et al. (2004).

MATERIALS AND METHODS

Adults and exuviae of the final instar ofD. cic-
erina, and images of Eurytoma sp. were obtained
in Aug 1988 from pupae ofL. cicerina whose lar-
vae were mining leaves of C. arietinum in Ayora
(Valencia, Spain). To collect specimens in both
cases, we picked structures from plants that were
infested with larvae of the agromyzids and placed
them in plastic bottles of suitable dimensions
whose openings were covered with gauze held in
place with a rubber band. These receptacles were
kept under environmental conditions of tempera-
ture, relative humidity (RH), and photoperiod.
The methodology used for opening the puparium
and preparing the cast skins was that proposed
by Tormos et al. (2003).
To study the different larval stages ofD. cice-
rina, in Jun 1990, we collected leaves ofC. arieti-
num that were mined by L. cicerina, and took
them to the laboratory, where they were placed in
the same kind of receptacle as above and kept un-
der environmental conditions. The parasitoids
that emerged from the host puparia were fed with
honey impregnated on strips of blotting paper or
with sugar and water. Females aged between 24
and 72 h were placed individually in Petri dishes
(9 x 1.5 cm) and provided daily with leaves of C.
arietinum infested with larvae of different stages
of L. cicerina. To study the larval development of
the parasitoid, parasitized material was kept in a


chamber at 21-23C, 60-80 RH, and a photoperiod
of 16L:8D, and the hosts were periodically dis-
sected. Where possible, these observations were
complemented with the dissection of specimens of
hosts parasitized in the field. These dissections
allowed us to study the mature larva of an unde-
termined species of Eurytoma. All dissections
were performed in 0.9% saline solution. To study
the development of the larvae ofD. cicerina, the
methodology of Tormos et al. (2003) was used. For
the microscopic preparation of the larval stages of
both the braconid and the eurytomid, the meth-
ods of Tormos et al. (2003, 2004) were employed.
The venom apparatus was prepared and
drawn according to the method described by
Quicke et al. (1992, 1997) with clorazol black
staining for dry museum specimens. The venom
apparatus was treated with a hydroxide solution,
after which the soft tissue could be removed. It
was then possible to observe the characteristics of
the remaining chitinous gland intima, which are
not apparent from examination of an intact gland
and reservoir. The material examined (adults, im-
mature stages, and venom apparatus) is depos-
ited at the "Torres Sala" Entomological Founda-
tion (Valencia, Spain).
The terminology for the body morphology, bio-
metric data, and wing venation of the adults of
Dacnusa follows Wharton et al. (1997). The termi-
nology used in the description of the different
structures of the immature stages of the braconid
and of the eurytomid is that used by Tormos et al.
(2003, 2004). The terminology used for character-
istics of the gland and reservoir parts of the
venom apparatus follows Tormos et al. (2003).

RESULTS AND CONCLUSIONS

Dacnusa cicerina sp. n. (Fig. la-d)

Type Material. SPAIN: Valencia: Ayora
(30SXJ6825), 25-VI-1988 (date of host capture)/8-
11-VIII-1988 (emergence date of the parasitoids):
Holotype: 2, from puparium ofL. cicerina (host)/
on C. arietinum (host's food plant). Paratypes: 1 Y,
1 6, ditto.
Holotype, length of body 1.7 mm.
Head-Width of head 2.0 times its length, 1.9
times distance between eyes. Height of head 1.3-
1.5 times its length. Antenna with 23 antenno-
meres; maxillary palpi moderately long; length of
eye in dorsal view 0.8 times temple; fairly smooth
medially and finely setose towards sides and at
center of its foremost part; clypeus width 0.70
times distance between eyes; mandible three-
toothed, expanded apically, 0.5 times length of
head, with middle tooth long and pointed. Meso-
soma-Length of mesosoma 1.3 times its height,
1.9 times its width; pronotum with a median pit;
sternaulus smooth, weak, short; metapleuron se-
tose towards the posterior coxae; notauli weakly







Florida Entomologist 91(2)


-11
~1


/4




UI la


0.5 mm


Fig. 1. Dacnusa cicerina sp. n. (Y): a-head in lateral view; c-detail of mesopleuron showing the sternaulus;
d-petiole in dorsal view. Dacnusa cicerina sp. n. (6): b-left mandible.


impressed; mesoscutum with dorsal pit, largely
smooth, shiny, setose, with setae longer in its
middle-posterior part, covering all its surface;
scutellar sulcus simple; surface of propodeum
wrinkled and finely setose. Wings-Pterostigma
moderately wide and dark, 2.0 times longer than
R1; 3Rsb sinuate; m-cu antefurcal. Legs-Hind
tarsus shorter than hind tibia. Metasoma-Peti-
ole glabrous, as long as wide apically, with large
dorsope; third tergite smooth, without setae on its
base; ovipositor sheath not extending beyond api-
cal tergite in retracted position. Color-Head and
mesosoma black; face black; clypeus dark brown;
labrum and palpi yellow; antennae dark brown,
centre of mandibles orange-yellow; legs pale yel-


low, with slightly darkened tarsi; wings hyaline,
with dark pterostigma; metasoma brownish, be-
coming darker apically.
Allotype-Similar to 9, but pterostigma wider
and dark.
Differential Diagnosis. This new species is sim-
ilar to D. rodriguezi, from which it is distinguished
by the following character states: (a) mandibles
expanded, with middle tooth pointed; (b) sternau-
lus smooth; and (c) first metasomal tergite brown-
ish, glabrous, as long as wide apically.
Etymology: The specific name of this species
refers to Liriomyza cicerina, of which it is a para-
sitoid.


This species can be identified with the keys provided by Docavo & Tormos (1997, page 387) modified
as follows:

Males
159 (144) Antennae 22-23 segmented. Mandibles three-toothed, not expanded. First metasomal tergite dark brown-
ish red, 1.7 times longer than wide apically. Pterostigma narrower than in Dacnusa melicerta (Nixon) (Fig.


June 2008


0.5 mm
II


\r.


pi


r1 '
"'O'.


la, lb ,


1c 0.5 mm
I I







Tormos et al.: Larvae and Venom Apparatus of D. cicerina sp. n.


4). Sternaulus absent. Body 1.3 mm. Parasitoid of Liriomyza dracunculi Hering, L. artemisicola de Meijere.
Center, Central Ural; East Germany; Austria .................. .............. D. austriaca Fischer
159' (215) Antennae 20-23 segmented. Mandibles three-toothed, expanded. First metasomal tergite 1.3 times, or
less, longer than wide apically. Pterostigma much longer than the R1 (Fig. 4b, see Docavo & Tormos 1997).
Sternaulus present ................................................................... 159"
159" (144) Antennae 20-22 segmented. Mandibles weakly expanded, with middle tooth blunt (Fig. 2). First metaso-
mal tergite black, 1.3 times longer than wide apically. Sternaulus weakly crenulate. Body 1.5 mm. Parasi-
toid of Chromatomyia horticola (Goureau). Spain ..................... D. rodriguezi Docavo & Tormos
159" (144) Antennae 23 segmented. Mandibles expanded, with middle tooth long and pointed (Fig. Ib). First meta-
somal tergite brownish, glabrous, as long as wide apically. Sternaulus smooth. Body 1.7 mm. Parasitoid of
Liriomyza cicerina (Rondani). Spain ...................................... ... .D. cicerina sp. n.

Females
214 (215) Antennae 21-24 segmented. Mandibles three-toothed, not expanded. First metasomal tergite reddish
dark brown, slightly pubescent, 1.7 times longer than wide apically. Pterostigma yellowish dark brown,
parallel-sided, few longer than R1. Sternaulus absent. Body 1.3 mm .............. D. austriaca Fischer
214' (215) Antennae 20-23 segmented. Mandibles three-toothed, expanded. First metasomal tergite 1.3 times, or
less, longer than wide apically. Pterostigma much longer than the R1 (Fig. 4a, to see Docavo & Tormos
1997). Sternaulus present .............................................................. 214"
214" (215) Antennae 20-22 segmented. Mandibles weakly expanded, with middle tooth blunt. First metasomal terg-
ite (petiole) black, fairly glabrous, 1.3 times longer than wide apically (Fig. 3). Pterostigma dark brown,
much longer than the R1. Sternaulus crenulated (Fig. 2, see Docavo & Tormos 1997). Body 1.5 mm
..................................... ........................ D. rodriguezi Docavo & Torm os
214" (215) Antennae 23 segmented. Mandibles expanded, with middle tooth long and pointed (Fig. la). First meta-
somal tergite (petiole) brownish, glabrous, as long as wide apically (Fig. Id). Pterostigma dark brown, much
longer than the R1. Sternaulus smooth (Fig. Ic). Body 1.7 mm ..................... .D. cicerina sp. n.

Females and males ofD. cicerina can be also distinguished from those ofD. basirufa Tobias, in Be-
lokobylskij et al. (1998: 339, 340, 353) after the following modifications:

Males
56 (55) 1 -3"d metasomal tergite brownish-yellow .................................................... 57
-First tergite black. Antennal segments more than 24 ........................................... 58
57 (56) Radial cell almost not shortened. Pterostigma long, narrow, yellowish. 1t-3'd flagellar segments yellow. An-
tenna 25-segmented; median segments about twice as long as wide. 1.7 mm (Fig. 128, 5, see Belokobylskij
et al. (1998)). Primorskiy Territory. ......................................... D. (P.) basirufa Tobias
-Radial cell shortened. Pterostigma wider, brown. Antenna 20-23-segmented, dark; median segments 2-2.5
times as long as wide................................................................. 57'
57'(57) Antennae 22-23 segmented. Mandibles three-toothed, not expanded. First metasomal tergite dark brownish
red, 1.7 times longer than wide apically. Pterostigma narrower than in Dacnusa melicerta (Nixon) (Fig. 4).
Sternaulus absent. Body 1.1-1.5 mm. Magadan Province, Primorskiy Kray...... D. (P.) austriaca Fischer
-Antennae 20-23 segmented. Mandibles three-toothed, expanded. First metasomal tergite 1.3 times, or less,
longer than wide apically. Pterostigma much longer than the R1 (Fig. 4b, see Docavo & Tormos 1997). Ster-
naulus present ...................................................................... 57"
57" (57') Antennae 20-22 segmented. Mandibles weakly expanded, with middle tooth blunt (Fig. 2). First metasomal
tergite black, 1.3 times longer than wide apically. Sternaulus weakly crenulate. Body 1.5 mm. Parasitoid
of Chromatomyia horticola (Goureau). Spain ........................ D. rodriguezi Docavo & Tormos
-Antennae 23 segmented. Mandibles expanded, with middle tooth long and pointed (Fig. Ib). First metasomal
tergite brownish, glabrous, as long as wide apically. Sternaulus smooth. Body 1.7 mm. Parasitoid of Liri-
omyza cicerina (Rondani). Spain ..............................................D. cicerina sp. n.

Females
114 (106) M esosoma entirely dark colored ........................................................ 115







Florida Entomologist 91(2)


-First metasomal tergite yellowish brown ................................................... 117
117 (114) Sternauli absent ................ ............................................. 117'
-Sternauli present ................ ................................................ 117"
(117') (114) Median segments of antenna 2.5 times as long as wide; basal segments of antenna dark. Pterostigma
weakly wedge-shaped. Antenna 21-24-segmented. Only first metasomal tergite pale colored, but infuscate
medially. 1.3-1.4 mm ................ ............................... D. (P.) austriaca Fischer
-Median segments of antenna 1.5-1.8 times as long as wide; four basal segments of antenna yellow.
Pterostigma lineal (in male elongate wedge-shaped and as in female pale colored). Antenna 21-22-seg-
mented. First-third metasomal tergite pale yellowish brown. 1.1-1.4 mm ........ .D. (P.) basirufa Tobias
117"(114) Antennae 20-22 segmented. Mandibles weakly expanded, with middle tooth blunt. First metasomal terg-
ite (petiole) black, fairly glabrous, 1.3 times longer than wide apically (Fig. 3). Stigma dark brown, much
longer than the R1. Sternaulus crenulated (Fig. 2, see Docavo & Tormos 1997). Body 1.5 mm
.................................... ...................... D. rodriguezi Docavo & Tormos
-Antennae 23 segmented. Mandibles expanded, with middle tooth long and pointed (Fig. la). First metasomal
tergite (petiole) brownish, glabrous, as long as wide apically (Fig. Id). Stigma dark brown, much longer
than the R1. Sternaulus smooth (Fig. Ic). Body 1.7 mm. ........................ D. cicerina sp. n.


Immature Stages ofD. cicerina sp. n. (Figs. 5a-d, 6a, b)

The first instar was found in different larval
stages of the host; second and third instars were
only found in host pupae.
Larva. 1" instar. General Aspect (Fig. 5a).
Body [length (1) and width (w) (at the level of the
mesothoracic segment): 0.5 x 0.15 mm] with head
well defined and 13 body segments, caudate, ver-
miform, transparent, curved to the ventral side.
Last abdominal segment slightly modified into a
short blunt, rounded organ in the form of a tail (1
= 80-85 pm, number of specimens = 3), with 25 se-
tae (1 = 50-70 pm) distributed in a fan around the


2 0.5 mm


3 0.5 mm


Figs. 2-4. 2-Dacnusa rodriguezi (6): mandible in
lateral view. 3-D. rodriguezi (9): a-petiole in dorsal
view; 4-Dacnusa austriaca ( ): anterior wing accord-
ing Tobias (1995).


anus. Segments 2-12 with a row of short setae (1 =
7-12 pm) on their posterodorsal part, the numbers
corresponding to 8 (mesothorax), 12 (metatho-
rax), and between 17 and 35 (abdominal seg-
ments). Cranium (Fig. 5b) (length and width: 160
x 170 pm) with sclerites strongly sclerotized with
the exception of the epistoma (weakly sclero-
tized). Mouthparts: Mandibles well defined, with
an oblong molar lobe and one blade (1 = 32 pm)
sharp, curved, and well sclerotized.
2nd instar. General Aspect (5c). Body [1 = 1 mm;
w (at the level of the mesothoracic segment) =
0.20 mm] cylindrical, long with respect to meso-
thoracic width, slightly spindle-shaped at ends.
Integument bare. Without cephalic sclerites or
mouthparts.
3rd instar. General Aspect (5d). Typical Hy-
menopteriform (1 = 1.70, w = 0.60 mm), with head,
thoracic and abdominal segments well defined;
yellowish. Integument with scale-like sensilla
setaeae") (1 = 3 pm) covering the thoracic and ab-
dominal segments, except the intersegmental
zones and around the spiracles and anus. Nine
pairs of spiracles (diameter (di) = 8 pm), with the
atrium and closing apparatus well differentiated,
one pair on the prothorax and another on the an-
terior edge of each of the first eight abdominal seg-
ments.
Pupa. Exarate. Without cocoon.
Exuvia of Final Instar. Of the 2 exuviae avail-
able for study only 1 was measured. General as-
pect. Tegument weakly sclerotized, except spira-
cles and scale like sensilla setaee) [1 = 3 pm]. Spi-
racles (Fig. 6a) situated on prothorax and first 8
abdominal segments; atrium (atr) [di = 6 pm]
sparingly developed, round, unarmed, separated
from the closing apparatus (ca) [1 = 11 pm, w = 9
pm] by a section of the trachea (t) [1 = 52 pm, a =
5-7 pm]. Cranium (Fig. 6b) [w (maximum) = 0.50
mm, h (taken from the base of the mandibles) =
0.20 mm] reduced; weakly sclerotized; with sen-


June 2008







Tormos et al.: Larvae and Venom Apparatus of D. cicerina sp. n.


G


md
PPP
5b 0.1 mm


,sp
OL


5c 0.5 mm

Fig. 5. Dacnusa cicerina sp. n.: 5-Larval phase: a-1l
c-2nd instar (lateral view); d-3'd instar (general aspect).


silla (se) [di = 3 pm] and setae (st) [1= 3 pm]; or-
bital antennal circular (a) [di = 0.08 mm], weakly
protuberant; pleurostoma (plst), superior (app)
and inferior (ppp) mandible processes, hypostoma
(h) and stipital sclerite (sl) well differentiated and
sclerotized; the latter joined to the labial sclerite
(ls), which is weakly sclerotized. Mouthparts.
Mandibles (md) [1 = 0.06 mm] with broad base
and relatively long blade, curved, thin, unarmed
(smooth) unidentate, sclerotized; maxillary (mp)
and labial (lp) palpi circular, slightly protuberant,
with a highly developed sensilla [di = 6 pm] in the
case of the labial palpi, and with two sensilla, one
of them highly developed [di = 5 pm] and the
other minute [di = 2 pm] in the case of the maxil-
lary palpi; salivary orifice well defined (so) (1 = 12
pm).

Description and Comments of Venom Apparatus (Fig. 7)

The venom apparatus exhibits the characters
specified by Quicke et al. (1997) for Dacnusa: (a)
an undivided reservoir; (b) a reservoir neck region
without narrowing; (c) a reservoir more than six
times longer than maximally wide; (d) a second-
ary venom duct absent; (e) an extensively
branched venom gland; (f) a venom gland inserted
at the extreme posterior end of the reservoir; and


5d 0.5 mm sc

Sinstar (lateral view); b-mandibles and head sclerites;



(g) a secondary venom duct that is not narrow.
The venom apparatus ofDacnusa cicerina is very
similar to that of D. flavicoxa Thomson (both spe-
cies are included in the subgenus Pachysema
Foerster), the morphological differences being
that the reservoir length is more than 12 times
longer than maximally wide in D. cicerina (less
than 12 times in D. flavicoxa) and that the num-
ber of gland filaments is 6 in D. cicerina, while D.
flavicoxa has 8 filaments (Fig. 7).

Notes on the Hyperparasitoid Eurytoma sp. (Fig. 8a, b)

A mature larva of this genus was collected, to-
gether with a second larval instar of D. cicerina,
from a puparium ofL. cicerina at Ayora (Valencia,
Spain) on 10-VI-1989. This appears to indicate
that Eurytoma sp. probably oviposits into the
phytophagous host, representing then a hyper-
parasitoid of D. cicerina (Sullivan 1999). An adult
male Eurytoma sp. was obtained on 15-VIII-1988,
from the same locality, from a puparium ofL. cic-
erina that was originally collected on 6-VI-1988.
General aspect (Fig. 8a). Body (1 = 2.2 mm, w =
0.61 mm), shape varying between barrel-shaped
and cylindrical, anterodorsal protuberances
present on thoracic segment 3 (th3) and first nine
abdominal segments (al-a9), with three thoracic


5a 1.5 mm


Sepst
app
S -plst






Florida Entomologist 91(2)


dg

(~- c


O a

,Z p1st


- at



t




ca


6a 25 um

Figs. 6-7. Dacnusa cicerina sp. n.: 6-Larval phase: exuvia of final instar: a-spiracle; b-cephalic structures 7.
Venom apparatus of D. cicerina sp. n. showing the terminology used for the venom gland and reservoir parts. Let-
tering: antennal orbit (a), anterior pleurostomal process (app), atrium (at), closing apparatus (ca), venom gland
(dg), secondary venom duct (f), gland filament (sack) (e), epistoma (epst), hypostoma (h), labial palpi (lp), labial
sclerite (ls), mandible (md), maxillary palpi (mp), pleurostoma (p1st), posterior pleurostomal process (ppp), reser-
voir with spiral culture (r), mesothoracic, metathoracic and abdominal setae (s), anal setae (sa), scale like setae
setaeae") (sc), sensilla (se), stipital sclerite (sl), salivary orifice (so), spiracle (sp), setae (st), trachea (t).


and ten abdominal segments. Color yellowish.
Weakly sclerotized, except for mandibles (md),
spiracles (sp) and setae. Anus small, subterminal,
transverse. Pleural lobes scarcely developed. Teg-
ument setose, with: (a) dorsal setae (1 = 62-120
pm): two pairs of setae on (thl-a7), a pair on the
(a8) and (a9); (b) dorsal terminal setae (1 = 30 pm):
one pair; (c) pleural setae (1 = 55-110 pm): four
pairs on (thl-a2); two pairs on (a3-a9); (d) ventral
setae (1 = 150-420 pm): one pair on (thl-a9), (sp)
on (th2), (th3), and on (al-a7); atrium of spiracle (1
= 20 pm, d maximum = 10 pm) funnel-shaped,
with approximately twelve chambers; closing ap-
paratus of spiracle (1 = 7 pm; w = 3 pm) adjacent
to atrium. Cranium (Fig. 8b). Wider than high (w
= 210 pm, height (from apex of cranium to base of
(md)) = 105 pm), narrower than (thl), weakly
sclerotized, with two pairs of setae (1 = 4-5 pm):


superior frontal setae (1 = 4 pm), hypostomal se-
tae (1 = 5 pm). antenna approximately 2.5 times
as long as broad, located below middle of cranium,
with two small sensilla on apex. Clypeus and la-
brum without setae or sensilla; epipharynx with a
pair of small sensilla (a). Tentorium with the
pleurostoma and its anterior and posterior pleu-
rostomal processes sclerotized and differentiated.
Epistoma almost indistinct, and very weakly scle-
rotized. Mouthparts. (md) (1 = 5 pm, w = 3 pm)
sclerotized, more heavily sclerotized at their
blade, unidentate, with a wide base; maxillae
(mx) and labium (lum) completely fused: (mx)
with a pair of maxillary setae (1 = 4 pm) and a pro-
tuberant maxillary papilla (4 x 2 pm); (lum) with-
out setae, with a pair of small prelabial sensilla.


O


- r




7 0.4 mm
II


6b 0.2 mm
I I


June 2008







Tormos et al.: Larvae and Venom Apparatus of D. cicerina sp. n.


thl-3


8a 2 mm


epx(a)


bl md U alu 1
8b 0.1 mm

Fig. 8. Eurytoma sp.: a-mature larva in lateral
view; b-cranium. Lettering: abdominal segments (al-
9), antenna (an), anal segment (as), anterodorsal protu-
berances (adp), base of mandibles (ba), blade of mandi-
ble (bl), clypeus (clp), dorsal setae (d), dorsal terminal
seta (dt), sensilla of the epipharynx (epx): a), superior
frontal setae (fs), hypostomal setae (hy), labrum (lm),
labium (lum), mandibles (md), maxillae (mx), pleural
setae (p), pleural lobes (pl), spiracles (sp), thoracic seg-
ments (thl-3), ventral setae (v).



DISCUSSION

The first instar ofD. cicerina can be considered
of the caudate-mandibulate type, according to the
classifications of Clausen (1962) and Hagen
(1964). Its larva is similar to the first instar of
Dacnusa areolaris (Nees) (Haviland, 1922) and
Dacnusa dryas (Nixon) (Guppy & Meloche 1987).
Like D. areolaris, it has a semicircle of stouter se-
tae arranged fanwise round the anus. Like D.
dryas, it has rows of setae on the posterodorsal
part of several body segments. The strongly scle-
rotized mandibles of this first instar could serve
to break the chorion; alternatively, they could
contribute to preventing super- or multiparasi-
toidism, as has been indicated for the caudate-
mandibulate larvae of Alysiinae (Tormos et al.
2003).
The second instar is fairly similar to the ma-
ture larva, having lost the tail, tegumental differ-
entiations, cephalic sclerites and mouthparts; it is


very similar to those of D. dryas (Guppy & Me-
loche 1987) and Dacnusa sibirica Telenga (Croft
& Copland 1994).
The mature larva is very similar to that de-
scribed for D. dryas (Guppy & Meloche 1987),
from which it differs by having scale-like sensilla
setaeae") on the thorax and abdomen. Differences
in the cephalic sclerites are addressed in the dis-
cussion of the exuviae. The description of D. are-
olaris (Haviland 1922) does not allow compara-
tive studies to be carried out.
The cast skin of the final instar of D. cicerina,
like those described for Dacnusini, displays sim-
ple, unarmed mandibles and a reduction in the la-
bial sclerite (Capek 1970, 1973). It shares a pleu-
rostoma with well-differentiated mandible pro-
cesses and a long stipital sclerite with those of the
genus Dacnusa. The only appreciable differences
from D. rodriguezi and D. dryas (species whose
mature larvae have been adequately described)
lie in the presence/absence, type, number and ar-
rangements of tegumental differentiations:
spinules, setae, and sensilla. Unlike D. rodrigu-
ezi, D. cicerina does not have spinules on the teg-
ument; the tegumental sensilla are scale-like (not
bluntly pointed) and the labial palpi only have 1
sensillum. Unlike those ofD. dryas, the tegumen-
tal, thoracic, and abdominal sensilla are scale-
like (not bluntly pointed) and the maxillary palpi
have 2 sensilla.
The mature larva of Eurytoma sp. shares the
following character states with other known Eu-
rytoma spp.: (a) body mainly barrel-shaped,
broader in mid-region; (b) head hemispherical,
without pronounced clypeus, with hypostomal se-
tae longer or about as long as half the width of the
labrum, and with inconspicuous and unpig-
mented craneal sclerites; (c) integument with se-
tae arranged in distinct rows along all body seg-
ments, and with ventral setae arranged in paired
rows; (d) atrium of spiracle long. Like E. nodu-
laris Boheman, this larva has the antennae lo-
cated below the middle of the cranium and more
than 2 dorsal setae present on abdominal seg-
ments A6-7; the cranium, without FI setae, is
similar to that ofE. heriadi Zerova.
This mature larva can be characterized and
distinguished from the similar mature larvae of
Eurytoma: E. nodularis and E. heriadi, by the
combination of the following character states: (a)
tegument with two pairs of setae on (thl-a7); (b)
cranium without inferior frontal setae or setae on
genae; and (c) epipharynx with a pair of small
sensilla. Additionally, it has an atrium with 12
chambers, an intermediate number between
those ofE. nodularis (14) and E. heriadi (10). The
smaller size of this larva may represent a mor-
pho-functional adaptation to its possible nature
as hyperparasitoid.







Florida Entomologist 91(2)


ACKNOWLEDGMENTS

We are indebted to G. C. D. Griffiths (University of
Alberta, Canada) and I. Docavo (Universidad de Valen-
cia, Spain) for corroboration of the determinations of the
agromyzid host and braconid parasitoid, respectively.
Maria Jesus Verdu (I.V.I.A., Spain) confirmed the deter-
mination of the Eurytoma. Thanks to Sergey Alexan-
drovich Belokobylskij (Zoological Institute of Russia)
and Charles Godfray (University of Oxford, UK) for ob-
servations on and critical reading of the manuscript.
This study was carried out in the laboratories of the De-
partamento de Protecci6n Vegetal, I.V.I.A. (Valencia,
Spain). Financial support for this paper was provided
from the Junta de Castilla y Le6n, project SA012A05,
and Fundaci6n Entomol6gica "Torres-Sala"


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June 2008







Kavallieratos et al.: Vitex and Euphorbia spp.


VITEXAGNUS CASTUS AND EUPHORBIA CHARACIAS SSP.
WULFENII AS RESERVOIRS OF APHID PARASITOIDS
(HYMENOPTERA: BRACONIDAE: APHIDIINAE)

NICKOLAS G. KAVALLIERATOS1, ZELJKO TOMANOVIC2, PETR STAR3 AND NICKOLAS E. EMMANOUEL4
'Laboratory of Agricultural Entomology, Department of Entomology and Agricultural Zoology,
Benaki Phytopathological Institute, 8 Stefanou Delta str., Kifissia, Attica, Greece

2Institute of Zoology, Faculty of Biology, University of Belgrade, Studentski trg 16, 11000, Belgrade, Serbia

'Institute of Entomology, Academy of Sciences of the Czech Republic,
Branigovska 31, 37005 Cesk6 Budejovic6, Czech Republic

4Laboratory of Agricultural Zoology and Entomology, Agricultural University of Athens,
75 Iera Odos, 11855 Athens, Attica, Greece

ABSTRACT

The role of the self-sown shrubs Vitex agnus castus and Euphorbia characias ssp. wulfenii as
reservoirs of aphid parasitoids was investigated. In field studies, V. a. castus grew adjacent
to an orchard of Citrus sinensis and E. characias ssp. wulfenii adjacent to an orchard of Pru-
nus dulcis. The relative abundance of the parasitoids ofAphis viticis Ferrari on V a. castus,
Toxoptera aurantii (Boyer de Fonscolombe) on C. sinensis,Aphis euphorbiae Kaltenbach on
E. characias ssp. wulfenii and Brachycaudus amygdalinus (Schouteden) on P. dulcis in var-
ious parts of Greece was assessed during 1996-2005. Aphidius colemani Viereck predomi-
nated on A. viticis and T aurantii in all sampling cases. In contrast, Ephedrus persicae
Froggatt was the dominant species parasitizingA. euphorbiae on E. characias ssp. wulfenii
and B. amygdalinus on P. dulcis. Furthermore, to illustrate the agro-ecosystem relation-
ships through the reservoirs, we present the distribution and host range patterns of parasi-
toids associated with aphids on V. a. castus and Euphorbia spp. in southeastern Europe. A
key for the identification of aphid parasitoids on V a. castus and Euphorbia spp. is provided.

Key Words: aphids, parasitoids, reservoirs, farmland management, SE Europe

RESUME

Se investigo el papel de los arbutos autosembrados Vitex agnus castus y Euphorbia characias
ssp. wulfenii como reservorios de parasitoides de afidos. En studios de campo, V a. castus
crecio adyacente a un huerto de Citrus sinensis y E. characias ssp. wulfenii creci6 adyacente
a un huerto de Prunus dulcis. Se evalu6 la abundancia relative de los parasitoides deAphis
viticis Ferrari en V a. castus, Toxoptera aurantii (Boyer de Fonscolombe) en C. sinensis,
Aphis euphorbiae Kaltenbach en E. characias ssp. wulfenii y Brachycaudus amygdalinus
(Schouteden) en P. dulcis en varias parties de Grecia durante los anfos 1996-2005.Aphidius
colemani Viereck fue predominante enA. viticis y T aurantii en todos los muestreos. Al con-
trario, Ephedrus persicae Froggatt fue la especie de parasitoide mas dominant en A. eu-
phorbiae on E. characias ssp. wulfenii y B. amygdalinus en P. dulcis. Ademas, para ilustrar
las relaciones del ecosistema agricola a travez de los reservorios, presentamos los patrons
de la distribuci6n y el rango de hospederos de los parasitoides asociados con afidos en V a.
castus y Euphorbia spp. en el sureste de Europe. Se provee una clave para la identificaci6n
de los parasitoides de afidos en V a. castus y Euphorbia spp.


Many studies have considered the importance
of various plants as reservoirs of aphid parasi-
toids due to their possible useful role in aphid bi-
ological control. The present account elucidates
the role of self-sown shrubs Vitex agnus castus L.
Verbenaceae and Euphorbia characias L. ssp.
wulfenii (Hoppe ex Koch) A. R. Sm. Euphorbi-
aceae as reservoirs of aphid parasitoids when
they grow in the vicinity of Citrus sinensis (L.) Os-
beck Rutaceae and Prunus dulcis (Miller) D.A.


Webb Rosaceae crops, respectively. The present
study is an attempt to establish some guidelines
required for IPM programs. Furthermore, to illus-
trate the agro-ecosystem relationships through
the reservoirs, we present the distribution and
host range patterns of parasitoids associated with
aphids on V a. castus and Euphorbia spp. in
southeastern Europe. A key for the identification
of aphid parasitoids on V a. castus and Euphorbia
spp. is provided.







Florida Entomologist 91(2)


MATERIALS AND METHODS

In the years 1996-2005, during spring and sum-
mer, plant samples bearing mummified aphids
were collected randomly from fields of C. sinensis
neighboring plants of V a. castus in Central and
Southern Greece. Similarly, plant samples were
collected from fields of P dulcis in the vicinity of E.
characias ssp. wulfenii in Central Greece. Each
sample was placed separately in a plastic con-
tainer covered with nylon mesh and the containers
were brought to the laboratory where each sample
of aphids was separated, preserved in a 2:1 ratio of
90% ethyl alcohol and 75% lactic acid and identi-
fied later (Eastop & van Emden 1972). Mummies,
attached on a small leaf piece each, were placed
separately in small plastic boxes, which were put
inside a growth cabinet. On the lid of each box
there was a circular opening covered with muslin
for ventilation in order to maintain conditions in-
side the boxes similar to those existing in the
growth cabinet (22.5C, 65% RH, 16L:8D) (Kaval-
lieratos et al. 2001, 2005a, b, 2006).
In spring and summer 2003, samples were
taken every 10 d from 2 untreated fields culti-
vated with C. sinensis and P dulcis located in
Southern (Sykaminon, Attica) and Central (Ri-
zomylos, Magnissia) Greece, respectively.
The C. sinensis plantation covered an area of
10000 m2. At each sampling date, 10 shoots, 20 cm
long, were randomly collected from 10 trees (1
stem per plant). The area adjacent to this planta-
tion was uncultivated and bore abundant and ran-
domly dispersed plants of V a. castus. At each sam-
pling date, 10 shoots, 20 cm long, were collected
from different plants of V a. castus (1 shoot per
plant) and treated as above. The aphid samples
were maintained in air-conditioned rooms (22.5C,
65% RH, 16L:8D) and checked daily for emerging
parasitoid adults. Emerged parasitoids were
stored in 70% ethanol and identified to species.
The P dulcis plantation covered an area of
15000 m2.At each sampling date, 10 shoots, 20 cm
long, were randomly collected from 10 trees (1
stem per plant). Along 1 side of the P dulcis area
there was a rural road along which plants of E.
characias ssp. wulfenii were growing densely, cre-
ating a natural field bank. At each sampling date,
10 shoots, 20 cm long, were collected from differ-
ent plants of E. characias ssp. wulfenii (1 shoot
per plant) and treated as above, also monitoring
the species composition, abundance, and seasonal
occurrence of aphids and parasitoids.
These plantations and nearby habitats were
chosen for our study because previous samplings
had shown that (a) Toxoptera aurantii (Boyer de
Fonscolombe) (Hemiptera: Aphididae) and
Brachycaudus amygdalinus (Schouteden) (Hemi-
ptera: Aphididae) were the major C. sinensis and
P dulcis pests, respectively, at these locations,
and (b) apart from Aphidiinae the observed num-


bers of other predators or parasitoids (such as
Coccinellidae or Aphelinidae) were limited. In
contrast, V a. castus and E. characias ssp. wulfe-
nii are associated with a single aphid species,
Aphis viticis Ferrari (Hemiptera: Aphididae) and
Aphis euphorbiae Kaltenbach (Hemiptera: Aphid-
idae), respectively, in Greece (Kavallieratos et al.
2001, 2004, 2007). These aphid species are not re-
ported to use Citrus and Prunus spp. as hosts.
Sampling started with the first infections of V
a. castus and E. characias ssp. wulfenii by aphids
and continued until the aphid colonies had col-
lapsed on C. sinensis and P dulcis.
Data were analyzed by one-way ANOVA with
the statistical package JMP (Sall et al. 2001).
ANOVA was used to test whether there were dif-
ferences in the total number of species of aphidi-
ines which emerged from each aphid species (per
shoot) on V a. castus, E. characias ssp. wulfenii,
C. sinensis and P dulcis during the period of the
study. Separation of means was done with the
Tukey-Kramer (HSD) test (at a = 0.05).
Samples from V a. castus and Euphorbia spp.
consisting of both live and mummified aphids
were taken from many localities in several coun-
tries of southeastern Europe (Greece, Serbia,
Montenegro) during 1996-2005. When necessary,
plants were preserved as herbarium specimens
for identification. The samples were treated as
above. Aphid parasitoid nomenclature in the key
follows Kavallieratos et al. (2001).

RESULTS

Parasitoid Complexes, Abundance and Associations

The species of parasitoids associated with A.
viticis on V a. castus, T aurantii on C. sinensis,A.
euphorbiae on E. characias ssp. wulfenii and B.
amygdalinus on P dulcis as well as their relative
abundance during 1996-2005 are shown in Tables
1 and 2.Aphidius colemani Viereck predominated
on A. viticis and T aurantii on C. sinensis in all
experimental areas (Table 1). In contrast, Ephe-
drus persicae Froggatt was the only species para-
sitizing A. euphorbiae on E. characias ssp. wulfe-
nii and the dominant one on B. amygdalinus on P.
dulcis in all experimental areas (Table 2).
All 4 species of aphids were found to be para-
sitized in the experiments conducted in the year
2003. Toxoptera aurantii was parasitized by A.
colemani, Aphidius matricariae Haliday, Diaere-
tiella rapae (M'Intosh), Praon volucre (Haliday)
and E. persicae. Aphis viticis was parasitized by
A. colemani, A. matricariae, P. volucre, Binodoxys
acalephae (Marshall) and Binodoxys angelicae
(Haliday). Brachycaudus amygdalinus was para-
sitized by E. persicae,A. matricariae andD. rapae
whereas A. euphorbiae by E. persicae only. The
relative abundances of aphidiines on T aurantii,
A. viticis, B. amygdalinus and A. euphorbiae are


June 2008







Kavallieratos et al.: Vitex and Euphorbia spp.


TABLE 1. RELATIVE ABUNDANCE (%) OF APHIS VITICIS AND TOXOPTERA AURANTII APHIDIINE PARASITOIDS FOUND ON
VITEX AGNUS CASTUS AND CITRUS SINENSIS, RESPECTIVELY (1996-2005) (V.a.c-VITEX AGNUS CASTUS,
C.s-CITRUS SINENSIS, A.v-APHIS VITICIS, T.a-TOXOPTERA AURANTII, A.c-APHIDIUS COLEMANI, A.m-
APHIDIUS MATRICARIAE, D.r-DIAERETIELLA RAPAE, P.v-PRAON VOLUCRE, B.ac-BINODOXYS ACALEPHAE,
B.an-BINODOXYS ANGELICA, E.p-EPHEDRUS PERSICAE).

Parasitoids Total
number of
Regions Plants Aphids A.c A.m D.r P.v B.ac B.an E.p specimens

Attica V.a.c A.v 82.4 3.5 14.1 711
(Central Greece)
C.s T.a 74.1 22.0 3.7 0.2 820
Argolis V.a.c A.v 90.8 0.4 0.3 2.1 6.4 796
(Southern Greece)
C.s T.a 81.9 10.2 4.8 0.6 2.6 502
Korinthia V.a.c A.v 97.1 2.9 612
(Southern Greece)
C.s T.a 75.3 10.5 14.2 683


shown in Tables 3 and 4, respectively. ANOVA
showed significant differences among the species
of aphidiines parasitizing T aurantii on Citrus
sinensis (F = 38.0, df = 4, 535; P < 0.0001), B.
amygdalinus on P. dulcis (F = 53.3, df = 2, 297; P
< 0.0001) andA. viticis on V a. castus (F = 57.7, df
= 3, 428; P < 0.0001). In the case of C. sinensis,
there were significantly more A. colemani (x =
3.8) than A. matricariae (x = 1.0), D. rapae (x =
0.1), E. persicae (x = 0.01) and P. volucre (Y =
0.2). In the case of V a. castus there were signifi-
cantly more A. colemani (x = 8.1) than B. acale-
phae (x = 0.3), B. angelicae (x = 0.8) and P volu-
cre (x = 0.1). In the case of P dulcis there were
significantly more E. persicae (x = 6.4) than A.
matricariae (x = 0.3) and D. rapae (x = 0.1).

Seasonal Occurrence and Co-incidence of Aphids and
Parasitoids

A comparison of both situations documents
also the seasonal occurrence and interactions of


both aphid-key parasitoid species participants.
The dominant A. colemani occurs on both aphid
participants of the interactionA. viticis (V a. cas-
tus) and T aurantii (C. sinensis) throughout the
season (Table 3). Similarly, the dominant E. per-
sicae occurs on both aphid participants of the in-
teraction A. euphorbiae (E. characias ssp. wulfe-
nii) and B. amygdalinus (P dulcis) throughout
the season (Table 4).

Parasitoid Transfer Trials

Some of the host associations of each parasi-
toid species were additionally verified by transfer
trials in the laboratory, aiming to prove the capa-
bility of each parasitoid to alternate between dif-
ferent host species and populations. The following
laboratory transfers were successful: A. colemani
fromA. uiticis-V a. castus to T aurantii-C. sin-
ensis and E. persicae from A. euphorbiae-E.
characias ssp. wulfenii to B. amygdalinus-P.
dulcis and vice versa.


TABLE 2. RELATIVE ABUNDANCE (%) OF APHIS EUPHORBIAE KAI BRACHYCAUDUS AMYGDALINUS APHIDIINE PARASI-
TOIDS FOUND ON EUPHORBIA CHARACIAS SSP. WULFENII AND PRUNUS DULCIS, RESPECTIVELY (1996-2005)
(E.c-EUPHORBIA CHARACIAS SSP. WULFENII, P.d-PRUNUS DULCIS, A.e-APHIS EUPHORBIAE, B.a-
BRACHYCAUDUS AMYGDALINUS, E.p-EPHEDRUS PERSICAE, A.m-APHIDIUS MATRICARIAE, D.r-DIAERE-
TIELLA RAPAE).

Parasitoids
Total number
Regions Plants Aphids E.p A.m D.r of specimens

Voiotia (Central Greece) E.c A.e 100.0 -467
P.d B.a 86.0 14.0 -321
Thessaly (Central Greece) E.c A.e 100.0 1092
P.d B.a 95.4 3.1 1.5 936







Florida Entomologist 91(2)


June 2008


TABLE 3. RELATIVE ABUNDANCE (%) OF APHIS VITICIS AND T. AURANTII APHIDIINE PARASITOIDS FOUND ON VITEXAG-
NUS CASTUS AND CITRUS SINENSIS IN SYKAMINON (2003) (V.a.c-VITEX AGNUS CASTUS, C.s-CITRUS SIN-
ENSIS, A.v-APHIS VITICIS, T.a-TOXOPTERA AURANTII, A.c-APHIDIUS COLEMANI, A.M-APHIDIUS
MATRICARIAE, D.r-DIAERETIELLA RAPAE, P.v-PRAON VOLUCRE, B.ac-BINODOXYS ACALEPHAE, B.an-
BINODOXYS ANGELICA, E.p-EPHEDRUS PERSICAE).

Parasitoids Total
number of
Dates Plants Aphids A.c A.m D.r P.v B.ac B.an E.p specimens

13/4 V.a.c A.v -
13/4 C.s T.a -
20/4 V.a.c A.v -
20/4 C.s T.a -
27/4 V.a.c A.v -
27/4 C.s T.a 88.6 11.4 44
4/5 V.a.c A.v 79.5 2.9 2.9 14.7 34
4/5 C.s T.a 62.5 35.4 2.1 48
11/5 V.a.c A.v 66.3 3.6 7.2 22.9 83
11/5 C.s T.a 76.0 21.4 1.3 1.3 75
18/5 V.a.c A.v 83.3 1.9 6.5 8.3 108
18/5 C.s T.a 68.5 27.4 2.7 1.4 73
25/5 V.a.c A.v 87.3 3.8 8.9 212
25/5 C.s T.a 74.2 24.0 1.2 0.6 175
1/6 V.a.c A.v 93.0 1.1 5.9 187
1/6 C.s T.a 90.7 9.3 75
6/6 V.a.c A.v 90.1 1.4 2.8 5.7 141
6/6 C.s T.a 100.0 26
13/6 V.a.c A.v 94.8 0.7 4.5 134
13/6 C.s T.a 100.0 12
20/6 V.a.c A.v 93.8 1.0 1.0 4.2 96
20/6 C.s T.a -


Parasitoid-aphid Associations on Vitex agnus castus and
Euphorbia spp.

The country abbreviations are: GRE-Greece,
SER-Serbia, MNG-Montenegro.

Aphidius colemani Viereck.
Aphis viticis Ferrari: on Vitex agnus castus (GRE).
Aphidius matricariae Haliday.
Aphis viticis Ferrari: on Vitex agnus castus (GRE).
Aphidius urticae Haliday.
Macrosiphum euphorbiae (Thomas): on Euphorbia salic-
ifolia (SER), Macrosiphum sp.: on Euphorbia
amygdaloides (MNG).
Binodoxys acalephae (Marshall).


Aphis euphorbiae Kaltenbach: on Euphorbia cyparissias
(SER), Aphis viticis Ferrari: on Vitex agnus-castus
(GRE).
Binodoxys angelicae (Halliday).
Aphis viticis Ferrari: on Vitex agnus-castus (GRE, SER).
Diaeretiella rapae (M'Intosh).
Aphis viticis Ferrari: on Vitex agnus castus (GRE).
Ephedrus persicae Froggatt.
Aphis euphorbiae Kaltenbach: on Euphorbia characias
ssp. wulfenii (GRE); Aphis sp.: on Vitex agnus-castus
(MNG).
Praon volucre (Haliday).
Macrosiphum euphorbiae (Thomas): on Euphorbia
amygdaloides (MNG), Aphis viticis Ferrari: on Vitex
agnus-castus (GRE).


Diagnostic Characters used in the Key

The following characters were used in the key: number of antennal segments, length of first
flagellomere (= F,), width of F, number of segments of maxillary palps, number of segments of labial
palps, existence of m-cu vein, degree of development of SR1 vein, existence of Rs+M vein, degree of pig-
mentation of Rs+M vein, existence of r-m vein, existence of M vein, existence of M+m-cu vein, length
of petiole, width of petiole at level of spiracles, length between spiracular and secondary tubercles,
sculpture of anterolateral area of petiole, color of last tergites, existence of prongs of the last sternite,
type of pupation.
The following key for the identification of aphid parasitoid species associated with Vitex agnus cas-
tus and Euphorbia spp. in southeastern Europe is based on females.







Kavallieratos et al.: Vitex and Euphorbia spp.


1. SR1 vein reaching forewing margin (Fig. 10). Mummy black .......................... Ephedrus persicae

-SR1 vein not reaching forewing margin (Figs. 4-9, 11). Mummy yellow or brown or whitish ............. 2

2. Forewing Rs+M vein present and pigmented at its basal part (Fig. 11). Pupation under aphid's
empty skin .................................................................. Praon volucre

-Forewing Rs+M vein absent (Figs. 4-9). Pupation inside aphid mummy ............................. 3

3. Forewing r-m vein present. Forewing M and m-cu veins united forming M+m-cu vein, developed throughout
(F igs. 4 -6) .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. 4

-Forewing r-m, M, m-cu veins absent (Figs. 7-9) ................................................. 6

4. Anterolateral area of petiole costate (Fig. 12). ..................................... Aphidius colemani

-Anterolateral area of petiole costulate (Figs. 13-14) .............................................. 5

5. Antennae 18-19-segmented. Flagellomere 1 3.3-3.8 times as long as wide. Maxillary palp 4-segmented. Labial
palp 3-segmented. Petiole 3.4-3.9 times as long as wide at level of spiracles (Fig. 17) .... Aphidius urticae

-Antennae 14-15-segmented. Flagellomere 1 2.6-3.0 times as long as wide. Maxillary palp 3-4-segmented
(Figs. 2, 3); when 3-segmented, last segment may bear a trace of 2 segments (Fig. 3). Labial palp 2-seg-
mented (Fig. 2, 3); sometimes one palp 1-segmented (Fig. 2). Petiole 2.5-3.2 times as long as wide at level
of spiracles (Fig. 16). ................................................... Aphidius matricariae

6. Last metasomal sternite with prongs (Figs. 23-24) ................................................ 7

-Last metasomal sternite without prongs (Fig. 22) .................. .............. Diaeretiella rapae

7. Distance between primary and secondary tubercles on petiole shorter than width at spiracles with enlarged
parallel sides (Fig. 18). Petiole and last tergites dark brown ..................... Binodoxys acalephae

-Distance between primary and secondary tubercles on petiole longer than width at spiracles; sides of petiole
between primary and secondary tubercles are neither enlarged nor parallel (Fig. 19). Petiole dark brown to
yellow. Last tergites yellow ................................................ Binodoxys angelica


TABLE 4. RELATIVE ABUNDANCE (%) OF APHIS EUPHORBIAE AND BRACHYCAUDUS AMYGDALINUS APHIDIINE PARASO-
TOIDS FOUND ON EUPHORBIA CHARACIAS SSP. WULFENII AND PRUNUS DULCIS IN RIZOMYLOS (2003) (E.c-
EUPHORBIA CHARACIAS SSP. WULFENII, Pd-PRUNUS DULCIS, A.e-APHIS EUPHORBIAE, B.a-BRACHYCAU-
DUS AMYGDALINUS, E.p-EPHEDRUS PERSICAE, A.m-APHIDIUS MATRICARIAE, D.r-DIAERETIELLA RAPAE).

Parasitoids
Total number
Dates Plants Aphids E.p A.m D.r of specimens

12/4 E.c A.e. 100.0 7
12/4 Pd B.a -
19/4 E.c A.e 100.0 18
19/4 P.d B.a 76.0 24.0 25
26/4 E.c A.e 100.0 39
26/4 P.d B.a 89.4 10.6 66
3/5 E.c A.e 100.0 89
3/5 P.d B.a 98.0 1.0 1.0 102
10/5 E.c A.e 100.0 143
10/5 P.d B.a 92.3 4.3 3.4 117
17/5 E.c A.e 100.0 151
17/5 P.d B.a 95.5 3.0 1.5 133
24/5 E.c A.e 100.0 187
24/5 P.d B.a 98.9 1.1 183
31/5 E.c A.e 100.0 150
31/5 P.d B.a 100.0 45
7/6 E.c A.e 100.0 116
7/6 Pd B.a 2
14/6 E.c A.e 100.0 102
14/6 P.d B.a -







Florida Entomologist 91(2)


Ni


0.5 mm


0.5 mm 0.5 mm


Fig. 1. Maxillary and labial palps of female Aphidius colemani Viereck. Fig. 2. Maxillary and labial palps of fe-
male Aphidius matricariae Haliday. Fig. 3. Maxillary and labial palps of female Aphidius matricariae Haliday. Fig.
4. Forewing of female Aphidius colemani Viereck. Fig. 5. Forewing of female Aphidius matricariae Haliday. Fig. 6.
Forewing of female Aphidius urticae Haliday.


DISCUSSIoN topical research items in many countries. Archi-
tecture and species composition of stabilizing ele-
Biodiversity in the Cultivated Landscape ments ecotoness, biocorridors, refugia etc.) are the
core of efforts to increase biodiversity. Diversifica-
Enhancement and conservation of biodiversity tion inside the crops and landscape in general re-
in the landscape, including the agroecosystems as lated to the phytophages and their antagonists
well as their effect on neighboring ecosystems, are (including parasitoids) was dealt with by a num-


June 2008


" ~ij







Kavallieratos et al.: Vitex and Euphorbia spp.


0.3 mm 0.5 mm


0.5 mm 0.5 mm


0.5 mm


0.3 mm


Fig. 7. Forewing of female Binodoxys acalephae (Marshall). Fig. 8. Forewing of female Binodoxys angelicae (Ha-
liday). Fig. 9. Forewing of female Diaeretiella rapae (M'Intosh). Fig. 10. Forewing of female Ephedrus persicae Frog-
gatt. Fig. 11. Forewing of female Praon volucre (Haliday). Fig. 12. Anterolateral aspect of petiole of femaleAphidius
colemani Viereck.


ber of authors (Altieri et al. 1993; Thies & Ts- which respective biodiversity studies can be real-
charntke 1999; Tscharntke et al. 2005). Aphids ized. The knowledge of plant-aphid-associations
and their associated parasitoids were shown to together with specific biological phenomena of
represent one of the model group associations in both the aphids and parasitoids allow the identi-







Florida Entomologist 91(2)



14


June 2008


0.3 mm


0.3 mm


0.15 mm


17








0 0











0.3 mm

Fig. 13. Anterolateral aspect of petiole of female Aphidius matricariae Haliday. Fig. 14. Anterolateral aspect of
petiole of female Aphidius urticae Haliday. Fig. 15. Dorsal aspect of petiole of female Aphidius colemani Viereck.
Fig. 16. Dorsal aspect of petiole of female Aphidius matricariae Haliday. Fig. 17. Dorsal aspect of petiole of female
Aphidius urticae Haliday. Fig. 18. Dorsal aspect of petiole of female Binodoxys acalephae (Marshall).







Kavallieratos et al.: Vitex and Euphorbia spp.


0.15 mm


0.15 mm


0.15 mm 0. mm


Fig. 19. Dorsal aspect of petiole of female Binodoxys angelicae (Haliday). Fig. 20. Lateral aspect of ovipositor
sheath ofAphidius colemani Viereck. Fig. 21. Lateral aspect of ovipositor sheath ofAphidius matricariae Haliday. Fig.
22. Lateral aspect of ovipositor sheath of Diaeretiella rapae (M'Intosh). Fig. 23. Lateral aspect of last sternite prong
of Binodoxys acalephae (Marshall). Fig. 24. Lateral aspect of last sternite prong of Binodoxys angelicae (Haliday).

fiction of the food webs, their seasonal changes, fruit crops which play a positive role as sources of
peculiarities and interactions. The present contri- alternative hosts of the parasitoid antagonists at-
bution deals with the interactions between some tacking also aphid pests in the nearby orchards







Florida Entomologist 91(2)


and habitats. The occurrence of the V a. castus
and E. characias ssp. wulfenii groves is a typical
example of an interaction of both the ecosystems
where the presence of the antagonists (parasi-
toids) is positive for aphid pest control, but where
there is no interaction through the aphid species.
Thus, biodiversification of the nearby orchard
community tends to enhance the over-all species
diversity and contributes also to the pest control
by the antagonists which interfere between both
the systems.

Reservoirs

Reservoirs of aphid parasitoids may be the
crops themselves (for example: Stary 1964, 1978;
Eikenbary & Rogers 1974; Pons & Stary 2003) as
well as non-crop plants in areas such as field
banks, roadsides, ruderal areas, uncultivated
places, abandoned or fallow grounds, hedges, or-
chards, parks, in and around residential areas,
near ruins of old buildings, meadows, orchard un-
dergrowth (Stary 1964; Stary & Lyon 1980;
Kavallieratos et al. 2002; Tomanovic et al. 2006;
Levie et al. 2001; Frere & Hance 2001). A complex
landscape with a significant presence of unculti-
vated and perennial habitats may lead to the en-
hancement of natural enemies, which can provide
a more successful natural biological control in an-
nual crops (Thies et al. 2005). In the case of aphid
parasitoids, however, that are characterized by
very specific trophic interactions, investigation in
a specific way is required (Thies & Tscharntke
1999). In non-crop areas, parasitoids may parasit-
ize economically unimportant aphid species from
where they may disperse to the neighboring crops
and parasitize target aphid pests there
(Stary 1962, 1964; Stary & Lyon 1980; Kavallier-
atos et al. 2002; Tomanovic et al. 2006). It should
also be emphasized that the respective relation-
ships among the agro-ecosystems can range from
identical to basically different if the host ranges
(food web associations) of aphids and their antag-
onists (parasitoids) are considered (Stary 1972,
1978).

Interactions of Parasitoid-aphid Associations

The predominance of A. colemani on A. viticis
and T aurantii could be attributed to the dis-
persal ofA. colemani from V a. castus to C. sinen-
sis and vice versa. A similar dispersal could be as-
sumed in the case of E. persicae that predomi-
nates onA. euphorbiae and B. amygdalinus. How-
ever, it is necessary to compare such situations
both from the seasonal and the abundance as-
pects: on the one hand, the dominant parasitoid
species occurs simultaneously on the 2 different
hosts (plants) in nearby habitats. Thus, every
habitat can be classified as reservoir of a target
parasitoid which may occur there independently


throughout the season. This phenomenon is im-
portant for parasitoid conservation in the land-
scape. On the other hand, interactions of both
parasitoid populations can be presumed which is
important both for parasitoid population conser-
vation and aphid pest control (Citrus).
Other researchers have recorded similar obser-
vations concerning the seasonal exchange of the
parasitoid populations between different crops as
well as between crops and uncultivated plants in-
fested by different aphid species. Stary (1978)
and Stary & Lyon (1980) stated that Aphidius
ervi Haliday can be dispersed by Acyrthosiphon
pisum (Harris) on lucerne to Metopolophium
dirhodum (Walker) on barley and vice versa,
Aphidius avenue (Haliday) byA. pisum on lucerne
to M. dirhodum on barley and vice versa and
Aphidius eadyi Stary, Gonzalez & Hall by
Acyrthosiphon pisum ononis (Koch) on Ononis
spp. to A. pisum on leguminous crops and vice
versa. Eikenbary & Rogers (1974) found that par-
asitization of Schizaphis graminum (Rondani) by
Lysiphlebus testaceipes (Cresson) on sorghum in-
creased when Helianthus annuus L. Compositae
(Aphis helianthi Monell) were grown nearby. Pons
and Stary (2003) determined similar seasonal
shifts between wheat and maize. Langer (2001)
studied hedges as reservoirs of parasitoids of ce-
real aphids in organic agriculture, including the
seasonal host alternation on different crops and
habitats. Frere & Hance (2001) analyzed the role
of grassy strips related to the period of activity of
parasitoids on nearby cereal crops. Similarly,
Kavallieratos et al. (2002) stated that A. matri-
cariae can be dispersed by Capitophorus inulae
(Passerini) on Dittrichia viscosa to Rhopalosi-
phum padi (L.) on durum wheat and barley and
Lysiphlebus fabarum (Marshall) and Lysiphlebus
confusus Tremblay & Eady, by Aphis ruborum
(Bdrner) on Rubus ulmifolius to Aphis gossypii
Glover on cotton and vice versa. Tomanovic et al.
(2006) presumed that Salix spp. represent reser-
voirs of L. confusus and L. fabarum, in agroeco-
systems, through Aphis farinosa Gmelin, an eco-
nomically unimportant aphid on willows.

Transfer Trials

Our successful laboratory transfers of A. cole-
mani and E. persicae support the evidence de-
rived from field samples of different aphid spe-
cies. The ability of parasitoids to switch from 1
species of aphid to another on different plants has
also been confirmed in the laboratory in numer-
ous cases. Stary (1986) confirmed that laboratory
transfers ofL. fabarum originating fromAphis fa-
bae cirsiiacanthoidis Scopoli on Cirsium arvense
in the field to Aphis fabae Scopoli on Faba vul-
garis were positive. Stary & N6mec (1986) re-
ported that the field populations of Praon abjec-
tum (Haliday) and B. angelicae emerging from


June 2008







Kavallieratos et al.: Vitex and Euphorbia spp.


Aphis sambuci L. on Sambucus nigra were suc-
cessfully transferred to A. fabae on F vulgaris in
the laboratory. Similarly, Stary & Gonzalez (1991)
tested successfully all transfer combinations of
D. rapae populations emerging from Brevicoryne
brassicae (L.) on Brassica napus, to Hayhurstia
atriplicis (L.) on Chenopodium album and Myzus
persicae (Sulzer) on F vulgaris. Furthermore,
Ephedrus nacheri Quilis reared from Cryptosi-
phum artemisiae Buckton on Artemisia vulgaris
and H. atriplicis on C. album were transferred
successfully to M. persicae on beans. Stary (2006)
brings still a set of transfers cases; sometimes,
however, what is achieved in the laboratory, may
not occur in the field; the latter cases may be due
to actually laboratory situations (for example,
Aphidius transcaspicus Telenga), or due to un-
solved taxonomic problems (indications of new spe-
cies-for example, the history ofA. ervi vs. Aphid-
ius microlophii Pennacchio & Tremblay).
The alternation of 2 or more host aphid species
by a parasitoid may have a higher or lower effect
on the intra-population composition. The transfer
information or trials are commonly explained as
host preference and they are illustrated or ex-
plained by such phenomena such as the olfactory
clues, the effect of original host species etc. Trans-
fer trials need to possess population genetic anal-
ysis targeting the identification of different lines,
their respective eventual abundance and/or oc-
currence changes as one of the key points. Some
up-dated studies have determined that at least in
broadly oligophagous parasitoids such lineages
did not become diversified due to host species al-
ternation (Antolin et al. 2006; Baer et al. 2004).
These results tend to confirm the earlier hypoth-
esis (Nemec & Stary 1984, 1985) on the role of
host species alternation as a more or less selective
net affecting the (lineage) composition of respec-
tive filial populations.

Reservoirs and Their Interactions in SE Europe

Ephedrus persicae, a species native to south-
eastern Europe, is now distributed worldwide. It
is common in most of the habitats/ecosystems in
southeastern Europe, including agroecosystems
in both continental and broader coastal areas
(Kavallieratos et al. 2004). Aphidius colemani is
an exotic species, and in southeastern Europe is
restricted to the Mediterranean (mainly coastal)
part. Originally an oriental species, now pantrop-
ical and subtropical in distribution (Stary 1975;
Takada 1998), it was most probably accidentally
introduced into southeastern Europe. Ephedrus
persicae and A. colemani have a relatively broad
host spectrum in southeastern Europe, with 23
and 25 aphid hosts respectively (Kavallieratos et
al. 2004, 2006).
It should be noted thatA. viticis is monoecious
holocyclic on V a. castus (Blackman & Eastop


1994) andA. euphorbiae is monoecious on several
Euphorbia species (Stroyan 1984). However, nei-
ther of these species is a crop pest. Furthermore,
the plant V a. castus is very common in sandy,
costal and riverside areas whereas E. characias
ssp. wulfenii is common in dry, uncultivated areas
and neither of them grow inside the orchards or
cultivated fields (Kavadas 1956; Sarlis 1999).
Generally, Euphorbia spp. grow in dry and stony
places or in desert areas while some species are
weeds (Kavadas 1956).
Maintenance of the existing plants V a. castus
and E. characias ssp. wulfenii close to crop fields
should be recommended to serve as reservoirs of
parasitoids of aphids that infest C. sinensis and P.
dulcis. The presence of these common plants near
the crops is very important, because of the rela-
tively weaker dispersion ability of parasitoids in
comparison with aphids (Thies et al. 2005). Simi-
lar recommendations for the usefulness of various
plants as reservoirs of aphid parasitoids as well
as of other natural enemies of aphids have been
proposed for other plants such as numerous
honey plant species (Stary 1962), as Galium spp.
(Stary 1974), Fraxinus excelsior (Stary 1982), Ur-
tica dioica (Stary 1983), C. arvense (Stary 1986),
Philadelphus coronarius (Stary 1991), Ononis
spp. (Stary & Lyon 1980), S. nigra (Stary &
Nemec 1986), C. album (Stary & Gonzalez 1991),
D. viscosa, R. ulmifolius (Kavallieratos et al.
2002) and Salix spp. (Tomanovic et al. 2006).
The plants V a. castus and E. characias ssp.
wulfenii could be considered as possible reser-
voirs of aphid parasitoids for other crops as well
since the species of parasitoids identified in the
present study on the aphids A. viticis (A. cole-
mani, A. matricariae, D. rapae, B. acalephae, B.
angelicae, P volucre) and A. euphorbiae (E. persi-
cae) are important parasitoids of a number of
aphid pests in several cultivated plants (Kavalli-
eratos et al. 2004, 2005b, 2006).

ACKNOWLEDGMENTS

This study was supported by the State Scholarships
Foundation of the Hellenic Republic, The Ministry of
Science and Environment Protection of the Republic of
Serbia (143006B) and the Entomology Institute Project
Z5000508 (Academy of Sciences of the Czech Republic).

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Florida Entomologist 91(2)


June 2008


SELECTION OF ATRIPLEX LENTIFORMIS HOST PLANTS
BY HESPEROPSIS GRACIELAE (LEPIDOPTERA: HESPERIIDAE)

WILLIAM D. WIESENBORN1 AND GORDON F. PRATT2
'U.S. Bureau of Reclamation, Lower Colorado Regional Office, P.O. Box 61470, Boulder City, NV 89006

2Department of Entomology, University of California, Riverside, CA 92521

ABSTRACT

We counted Hesperopsis gracielae (MacNeill) (Lepidoptera: Hesperiidae) eggs on Atriplex
lentiformis T. ..1 .:. S. Watson (Chenopodiaceae) plants on 3 dates during 2006-2007 next to
the lower Colorado River in Cibola Valley, Arizona. The skipper has received conservation in-
terest due to its restricted geographic range and apparent rarity. Atriplex lentiformis, the
skipper's only known host species, is a large shrub capable of C, photosynthesis and N2 fix-
ation. We measured the size (canopy radius) and percent water of plants and the percent ni-
trogen of leaves. Percentages of water and nitrogen were partially correlated, whereas plant
size was not partially correlated with the percentage of water or nitrogen. Skipper eggs were
more likely to be present on shrubs with greater canopy radius, water content, or nitrogen
concentration. Likelihood of egg presence also increased with plant size when percent nitro-
gen was controlled and increased with percent nitrogen when plant size was controlled.
Numbers of eggs, adjusted for canopy radius, on shrubs with at least 1 egg were not related
to the percentage of water or nitrogen. Ovipositing H. gracielae appear to select host plants
when thresholds of plant size and water or nitrogen content are exceeded. These plant char-
acteristics should be considered when surveying or restoring the skipper's habitat.

Key Words: Insecta, Chenopodiaceae, oviposition, canopy size, plant water, leaf nitrogen

RESUME

Los huevos de Hesperopsis gracielae (MacNeill) (Lepidoptera: Hesperiidae) sobre plants de
Atriplex lentiformis T. ..1 .. S. Watson (Chenopodiaceae) en 3 fechas durante los anos 2006-
2007 cercanas a la parte baja del Rio Colorado en Cibola Valley, Arizona fueron contados.
Esta mariposa ha recibido interns en su conservaci6n debido a su rango geografico limitado
y aparente rareza.Atriplex lentiformis, el unico hospedero conocido de la mariposa, es un ar-
busto grande capaz de realizar la fotosintesis de C, y la fijaci6n de N,. Nosotros medimos el
tamano del radio de la copa del arbusto, el porcentaje de agua de las plants y el porcentaje
de nitr6geno en las hojas. El porcentaje de agua y el nitr6geno estaban correlacionados par-
cialmente, mientras que el tamano de la plant no estaba correlacionado con el porcentaje
del agua y nitr6geno. Era mas probable que los huevos de la mariposa estuvieran presents
en arbustos que tienen un mayor radio de la copa, contenido de agua y concentraci6n de ni-
tr6geno. La probabilidad de la presencia de huevos aumento en plants mas grandes cuando
el porcentaje de nitr6geno fue controlado y aumento con el porcentaje de nitr6geno cuando
el tamaio de la plant fue controlado. El numero de los huevos, ajustado por el radio de la
copa, en arbustos con al menos 1 huevo no fue relacionado con el porcentaje de agua o nitr6-
geno. La mariposa, H. gracielae parece seleccionar la plant hospedera para ovipositar
cuando se excede el umbral del tamano de la plant y el contenido de agua o nitr6geno. Las
caracteristicas de esta plant deben de ser consideradas cuando se realize un muestreo o
trabajo en la restauraci6n del habitat de la mariposa.


MacNeill's sootywing, Hesperopsis gracielae
(MacNeill) (= Pholisora gracielae MacNeill), is a
small (wingspread 18-24 mm, MacNeill 1970)
dark-brown skipper (Lepidoptera: Hesperiidae). It
is found along the Colorado River downstream of
the Grand Canyon and along the river's tributar-
ies in Utah, Nevada, California, and Arizona (Aus-
tin & Austin 1980; Stanford 1980; Nelson &
Andersen 1999). The skipper also occurs in Impe-
rial County in southeast California (MacNeill
1970). Hesperopsis gracielae was described in
1970 from specimens collected in California south


of Parker Dam (MacNeill 1970). Two flights of the
sootywing occur during Apr and Jul to Oct (Emmel
& Emmel 1973). Due to the skipper's rarity, it was
granted the U.S. state conservation ranks (Master
1991) of S1 (critically imperiled) in Nevada, S1 or
S3 (rare or uncommon but not imperiled) in Cali-
fornia, and S? (not yet ranked) in Arizona.
Larvae ofH. gracielae eat onlyAtriplex lentifor-
mis (Torrey) S. Watson (Chenopodiaceae) leaves
(MacNeill 1970).Atriplex lentiformis is a large (<3
m high) blue- or grey-green, dome-shaped shrub
that is wind pollinated, generally dioecious, and







Wiesenborn & Pratt: Host Selection by Hesperopsis gracielae


frequently found on floodplains where soils are sa-
line and groundwater is available to roots (Turner
et al. 1995). The plant assimilates carbon by the C4
photosynthetic pathway (Laetsch 1968), permit-
ting reduced rates of transpiration and water up-
take and increasing adaptation to its hot, arid en-
vironment. Atriplex lentiformis also is capable of
relatively-high rates of N, fixation by root-symbi-
otic bacteria (Malik et al. 1991).
Restoration of habitat and surveys for the
sootywing would be improved by a clearer under-
standing of the skipper's oviposition and repro-
ductive rate on different qualities ofA. lentiformis
plants. Two host-plant qualities that influence re-
productive rates of insects are concentrations of
water and nitrogen. Greater concentrations of
these nutrients in plants generally increase the
growth rate of lepidopteran larvae (Scriber 1984).
Although insects are expected to distribute eggs
in relation to plant quality, few studies have ex-
amined the influence of intraspecific host-plant
variation on oviposition behavior (Bernays &
Chapman 1994). Our objective was to examine
the selection of A. lentiformis by ovipositing H.
gracielae in relation to shrub size and water con-
tent and leaf nitrogen concentration. We exam-
ined 3 questions. (1) Are sizes and water and ni-
trogen contents of A. lentiformis shrubs interre-
lated? (2) Do these plant characteristics influence
host-plant selection by sootywings? (3) Does in-
traspecific variation in host plants affect the pres-
ence of sootywing eggs on plants or numbers of
eggs on selected plants?

MATERIALS AND METHODS

The study site is located (3317'N, 11443'W; el-
evation 62 m) on the Colorado River floodplain in
Cibola Valley, La Paz County, Arizona, 37 km
south-southwest of Blythe, California. The site,
within Cibola National Wildlife Refuge, contains
irrigated farm fields converted to wildlife habitat
and is bounded to the west by a remnant river ox-
bow and to the east by an excavated river channel.
The surrounding floodplain is farmed or dominated
by naturalized tamarisk, Tamarix ramosissima
Ledebour or Tamarix chinensis Loureiro (Tamari-
caceae), and bordered by Sonoran desert. Climate
at Blythe is summarized as maximum tempera-
tures averaging 42.5C during Jul, minimum tem-
peratures averaging 3.5C during Dec, and rainfall
averaging 97 mm yearly and occurring mostly dur-
ing Dec-Mar and Aug-Sep (NOAA 2007).
Atriplex lentiformis shrubs along both sides of
a dirt road were sampled for H. gracielae eggs.
Percent covers of plant species bordering the road
and measured with a tape were 47% for A. lenti-
formis, 28% for Prosopis glandulosa (L. Benson)
M. Johnston (Fabaceae), 19% for Pluchea sericea
(Nuttall) Coville (Asteraceae), 4% for T ramosis-
sima, 2% for Baccharis sarothroides A. Gray x


Baccharis emoryi A. Gray (Asteraceae), and <1%
for Sesuvium uerrucosum Rafinesque (Aizoaceae)
and Sida rhombifolia L. (Malvaceae). We recog-
nized sootywing eggs by their spherical shape and
heavily sculptured, ridged chorion (Fig. 71 Em-
mel & Emmel 1973) and reddish-brown color.
Vouchers of adult H. gracielae were deposited at
the Entomology Research Museum, University of
California, Riverside.
We used a retrospective design (Agresti 1990)
to ensure that approximate numbers ofA. lenti-
formis plants with and without sootywing eggs
were sampled. Separate plants >1 m tall were ar-
bitrarily selected and examined for unhatched
eggs by 2 persons for 10 min, by 1 person for 20
min, or until the entire shrub was searched. We
counted eggs on each shrub. If an egg was not
found, the nearest shrub that we previously ob-
served being visited by sootywings was selected
next for sampling. If at least 1 egg was found, we
next sampled the nearest shrub that we had not
observed being visited by sootywings. Two per-
sons sampled 20 plants on 24 May 2006 and 12
plants on 29 Jun 2006, and 1 person sampled 7
plants on 5 Jun 2007. We measured ( 0.1 m) the
height and minimum and maximum diameters of
each sampled plant and calculated its average
canopy-radius based on a hemisphere. We
snipped eight 20 cm-long cuttings from the ends
of branches around the circumference of each
plant. Cuttings from each plant were combined
and immediately weighed ( 0.5 g) with a 60-g ca-
pacity spring scale.
We reweighed branch cuttings from eachA. len-
tiformis after drying 24 h at 100C to calculate per-
cent water. We estimated percent nitrogen of dried
leaves from each plant by Kjeldahl digestion (Isaac
& Johnson 1976). A 25-mg subsample of ground
and sieved leaves was heated with a block digestor
1 h at 400C in 7 mL of concentrated sulfuric acid,
containing 4.2% selenous acid, and 3 mL of 30%
hydrogen peroxide. Water was added to 50 mL, and
the supernatant was diluted 1/10. We measured
the ammonia concentration of the supernatant
against standards by colorimetry with a seg-
mented flow analyzer (OI Analytical, College Sta-
tion, TX). We repeated the procedure on a second
subsample of leaf tissue and averaged percentages
of nitrogen between subsamples within plants.
Partial correlations between canopy radius,
percent water, and leaf percent nitrogen (trans-
formed 2 arcsin [X/100]1) ofA. lentiformis shrubs
were calculated (Neter et al. 1996, SYSTAT ver-
sion 10.2, Richmond, CA). We tested relationships
between these 3 plant measurements and pres-
ence and absence ofH. gracielae eggs with multi-
ple logistic regression (Agresti 1990; Neter et al.
1996). Sampling date was included in the regres-
sions as 2 indicator variables to account for differ-
ences in egg sampling and abundance. We evalu-
ated regressions predicting egg presence based on







Florida Entomologist 91(2)


significance (P < 0.05) of predictor variables and
values of D, a logistic-regression analog of R2
(Agresti 1990).
Dependences of egg abundance on the radius,
percent water, and percent nitrogen ofA. lentifor-
mis shrubs supporting at least 1 sootywing egg
were determined with sequential regressions
(Graham 2003). Plants sampled on 24 May 2006
were analyzed separately, because they had
higher water contents. We regressed numbers of
eggs, transformed (Y + 0.5)12, against canopy ra-
dius and calculated adjusted numbers of eggs by
adding the residuals and mean. Numbers of eggs
adjusted for canopy radius were regressed sepa-
rately against plant percent water and leaf per-
cent nitrogen. Regression of plants with eggs on
25 Jun 2006 and 5 Jun 2007 included an indicator
variable for date.

RESULTS

We found different associations between can-
opy radius (1.6, 0.9-2.5 m [mean, range]), water
content (63, 50-73%), and leaf nitrogen concentra-
tion (3.0 [back transformed], 1.5-4.7%) ofA. lenti-
formis shrubs (n = 39) sampled for H. gracielae
eggs. Percentages of water and nitrogen were pos-
itively correlated (partial r = 0.44; t = 2.96; df =
36; P = 0.003) when plant radius was controlled.
This correlation is not due to confounding, be-
cause we estimated percent nitrogen in dried
leaves. Shrubs high in water content usually con-
tained high leaf nitrogen concentrations. We de-
tected weak and non-significant correlations be-
tween plant radius and percent water (partial r =
0.22; t = 1.36; df = 36; P = 0.091), controlling for
percent nitrogen, and between plant radius and
leaf percent nitrogen (partial r = 0.26; t = 1.59; df


= 36; P = 0.061), controlling for percent water.
Neither water content nor nitrogen concentration
was appreciably associated with sizes of shrubs.
We found H. gracielae eggs on 23 of the 39 A.
lentiformis shrubs sampled. We counted 35 eggs
on 11 plants on 24 May 2006, 31 eggs on eight
plants on 29 Jun 2006, and 16 eggs on four plants
on 5 Jun 2007. Plant radius, plant percent water,
and leaf percent nitrogen each were related to egg
presence when separately considered (Table 1).
The presence of eggs also was related simulta-
neously to plant radius and percent nitrogen.
Shrubs were more likely to support at least 1 egg
as canopy radius increased and nitrogen concen-
tration was held constant and as nitrogen concen-
tration increased and canopy radius was held con-
stant (Fig. 1). For example, the odds of a shrub
supporting at least 1 egg (= probability of eggs
present/probability of eggs absent, see Neter et al.
1996) increased 104% (95% CI = 14-265%) as
plant radius increased 0.2 m with percent nitro-
gen held constant. Similarly, the odds of a plant
supporting at least 1 egg increased 104% (95% CI
= 5.8-291%) as leaf nitrogen increased from 3.0 to
3.5% with plant radius held constant. The likeli-
hood of egg presence also was related to plant wa-
ter content when simultaneously considered with
plant radius, whereas the latter was a weak and
nonsignificant predictor (Table 1; Fig. 2). The
odds of a shrub supporting at least 1 egg increased
83% (95% CI = 9.1-207%) with a two-percentage-
point increase in percent water and shrub radius
held constant. Variable likelihood of egg pres-
ence are indicated by these large confidence inter-
vals. Percent water also was related to egg pres-
ence when simultaneously considered with per-
cent nitrogen. Nitrogen concentration likely did
not significantly predict egg presence when con-


TABLE 1. MULTIPLE LOGISTIC REGRESSIONS OF PRESENCE OR ABSENCE OF HESPEROPSIS GRACIELAE EGGS ON ATRI-
PLEX LENTIFORMIS PLANTS AGAINST PLANT MEASUREMENTS.

No. of Plant Partial
predictors measurement' b SE t df P D2

1 canopy radius 3.95 1.41 2.81 35 0.005 0.25
1 % water 36.8 12.5 2.97 35 0.003 0.27
1 % nitrogen3 27.3 10.3 2.64 35 0.008 0.21
2 canopy radius 2.95 1.51 1.95 34 0.051 0.38
% water 30.2 13.2 2.29 34 0.022
2 canopy radius 3.56 1.49 2.40 34 0.017 0.36
% nitrogen 25.6 12.0 2.13 34 0.033
2 % water 32.8 14.7 2.24 34 0.025 0.33
% nitrogen 17.3 13.1 1.32 34 0.19
3 canopy radius 3.08 1.63 1.89 33 0.059 0.42
% water 23.2 14.3 1.62 33 0.11
% nitrogen 18.0 14.2 1.27 33 0.20

'Regressions include 2 indicator variables for sampling date.
Analog of R.
Measured in leaves, transformed 2 arcsin (X/100)"2.


June 2008







Wiesenborn & Pratt: Host Selection by Hesperopsis gracielae


2.3


2.0


1 2
% n

Fig. 1. Presence (solid syn
symbols) of Hesperopsis gracie
formis shrubs at Cibola Valley


0



a
o o
*

o
a
a
a


opy radius vs. leal perceCnI niiirogein. Pilanis saiiipledu on
24 May 2006 (circles), 29 Jun 2006 (triangles), and 5
Jun 2007 (squares). Some points slightly shifted to
avoid overlap.


sidered with water content due to intercorrelation
between the 2 plant measurements. Percentages
of water and nitrogen also explained less varia-


DISCUSSION

Atriplex lentiformis shrubs sampled for sooty-
wing eggs contained variable concentrations of
leaf nitrogen despite their ability to fix atmo-
spheric N2. Performance of nitrogen fixing bacte-
ria generally corresponds with conditions for


I I I I I


0





a E


A 0
U A


a.
U.
A



0. *


a 0 o
A


a


50 55 60 65 70


% water

Fig. 2. Presence (solid symbols) and absence (open
symbols) of Hesperopsis gracielae eggs onAtriplex lenti-
formis shrubs at Cibola Valley, Arizona, plotted as can-
opy radius vs. plant percent water. Shrubs sampled on
24 May 2006 (circles), 29 Jun 2006 (triangles), and 5
Jun 2007 (squares). Some points slightly shifted to
avoid overlap.


% nitrogen

Fig. 3. Numbers of Hesperopsis gracielae eggs, trans-
formed (Y + 0.5)"2, adjusted for canopy radius, and back
transformed, plotted against leaf percent nitrogen of
Atriplex lentiformis shrubs with >1 egg at Cibola Valley,
Arizona. Plants sampled on 24 May 2006 (circles), 29
Jun 2006 (triangles), and 5 Jun 2007 (squares).


tion in egg presence than other regressions with 2
1 predictors (Table 1), reinforcing the importance of
plant size as a predictor of egg occurrence. Corre-
lation between percentages of water and nitrogen,
and weak associations between these concentra-
Stions and plant radius, likely prevented these
measurements from significantly predicting egg
AO a on e presence when considered simultaneously.
Numbers of sootywing eggs (3.3 [back trans-
a formed], 1-8 eggs) onA. lentiformis shrubs with at
a least 1 egg were not dependent on plant size, wa-
*e ter content, or nitrogen concentration. Egg abun-
* o dance was not related to canopy radius on 24 May
2006 (b = 0.66 + 0.37 [SE];t = 1.79;df= 9;P = 0.11)
or on 29 Jun 2006 and 5 Jun 2007 (b = 0.82 + 0.41;
o t = 1.99; df = 9; P = 0.078). Egg numbers adjusted
o for canopy radius were not dependent on leaf per-
I cent nitrogen (Fig. 3) on 24 May 2006 (b = 3.8 +
3 4 5 3.7; t = 1.03; df = 9; P = 0.33) or on the later dates
nitrogen (b = 5.0 + 3.2; t = 1.58; df = 10; P = 0.15). Neither
was plant percent water related to adjusted num-
ibols) and absence (open bers of eggs (Fig. 4) on 24 May 2006 (b = 7.9 + 6.3;
lae eggs onAtriplex lenti- t = 1.26; df = 9; P = 0.24) or on 29 Jun 2006 and 5
Arizona, plotted as can- Jun 2007 (b = 5.3 + 3.0; t = 1.79; df= 10;P = 0.10).


2.3


2.0-


1.4


1.1 -


0.8
45


a


* m






* U


2

- 1
1
75 n
75 n


I I I







Florida Entomologist 91(2)


I I I
A
7

6

5 A

4 A
6
Z A
3 A

2-
0
1 -

0 I L I
50 55 60 65
% water
Fig. 4. Numbers of Hesperopsis graciel
formed (Y + 0.5)"2, adjusted for canopy rac
transformed, plotted against percent wa
lentiformis shrubs with >1 egg at Cibola V
Plants sampled on 24 May 2006 (circles)
(triangles), and 5 Jun 2007 (squares).


plant growth (Mengel & Kirkby 2001
ture is especially important, as wat
duces N2 fixation by root symbionts ar
tion of fixed nitrogen, as amino acid
to leaves. Association between conc
water and leaf nitrogen in A. lentil
was due to increased nitrogen fixation
location produced by increased so
Patchy soil moisture at our study sit
supported shrubs that varied in conc
water and nitrogen.
Presence or absence and number
cielae eggs onA. lentiformis plants pa
from the oviposition behaviors of hos
ing and acceptance. Ovipositing fe
randomly would be more likely t
larger plants. Larger shrubs also
greater visual or olfactory cues if fer
tracted to host plants. Hesperopsis gr
dency to fly within shrubs (MacNeil
increase the likelihood of ovipositior
lentiformis. MacNeill's sootywings to
sunlight less than the smaller, sympa
Brephidium exilis (Boisduval)
1999). Female sootywings may only
shrubs large enough to provide sha
limit body temperature.
Selection by H. gracielae of host
greater water and nitrogen contents
experimental evidence that develop
lepidopteran larvae generally increa
with higher concentrations of the!


(Scriber 1984) and the concept that phytophagous
insects place eggs on plants that maximize off-
spring growth, survival, and fecundity (Jaenike
1978; Thompson & Pellmyr 1991). Water is espe-
cially critical to sootywing larvae due to their arid
environment, and greater nitrogen contents in in-
sects (7-14% of dry mass) than in plants (0.03-
7.0%) require insect herbivores to concentrate
this element (Mattson 1980). An example of a but-
terfly ovipositing on plants in relation to host ni-
trogen content and suitability for larvae is pro-
vided by the nymphalid Heliconius erato (F.) (Ker-
** pel et al. 2006). Confined H. erato females placed
more eggs on shoots from Passiflora suberosa
(Passifloraceae) grown in nitrogen-rich soil that
increased leaf nitrogen content (from 2.4 to 3.9%)
and shortened larval development time (from 21
I to 19 d). More rapid development increases larval
70 7& survival by reducing exposure to predators and
parasites (Rhoades 1983).
Water and nitrogen contents of A. lentiformis
ae eggs, trans- appeared to dichotomously affect oviposition by H.
dius, and back gracielae. Shrubs low in water or nitrogen were not
ter of Atriplex found or accepted as hosts, whereas shrubs high in
alley, Arizona. these nutrients received variable numbers of eggs.
,29 Jun 2006 This observation suggests that concentrations of
plant nutrients exceed a threshold before accep-
tance by female sootywings. Ovipositing insects
are hypothesized to accept host plants when plant
1). Soil mois- stimuli exceed thresholds that are influenced by
er stress re- each insect's physiological state (Miller & Strickler
nd transloca- 1984). For example, females of the nymphalid but-
s, from roots terfly Euphydras editha Boisduval placed on vari-
entrations of ous plant species accepted less-preferred hosts as
Formis likely more time elapsed since the previous oviposition
n and trans- (Singer 1982). In nature, the rarity of suitable
il moisture. hosts would influence the frequency of oviposition
;e expectedly and the threshold of plant acceptance (Jaenike
entrations of 1978). Oviposition by a population of sootywings,
each with a different physiological state, would
rs of H. gra- tend to obscure the water and nitrogen concentra-
rtly resulted tions stimulating host-plant acceptance.
st plant find- Female H. gracielae may use visual and chem-
males flying ical cues to find and discriminate A. lentiformis.
o encounter Chlorophyll concentrations in leaves are associ-
may present ated with leaf nitrogen contents and affect plant
nales are at- color. Colors of plant models influenced landing
acielae's ten- and oviposition by Eurema hecabe (L.) (Pieridae)
11 1970) may butterflies (Hirota & Kato 2001). Nutrient con-
n on large A. centrations in leaves also may affect plant vola-
lerate direct tiles that influence host finding by olfactory cues
tric lycaenid and leaf-surface compounds that influence host
(Wiesenborn acceptance by contact chemoreception. Oviposi-
oviposit on tion by the moth Cochlyis hospes Walsingham
de needed to (Cochylidae) on green floral foam was stimulated
by moisture and by volatiles and contact-chemi-
plants with cals extracted from leaves and bracts of its host
agrees with plant, sunflower (Barker 1997).
nent rates of Preserving or creating habitat to increase pop-
se on plants ulations of H. gracielae should provide large A.
se nutrients lentiformis with high contents of water and leaf


June 2008







Wiesenborn & Pratt: Host Selection by Hesperopsis gracielae


nitrogen. Female sootywings oviposited on all
shrubs in our sampling that exceeded 1.6 m in
mean radius, 64% in water content, and 3.2% in
leaf nitrogen. Moisture appears to be the most im-
portant soil nutrient because it affects concentra-
tions of plant water and leaf nitrogen. Measuring
plant water content also provides an easy method
for estimating levels of leaf nitrogen inA. lentifor-
mis. Rates of N2 fixation and levels of leaf nitrogen
may be influenced by other soil nutrients, espe-
cially phosphate and potassium (Mengel &
Kirkby 2001). Plant water and nitrogen concen-
trations partly explain the apparent rarity of H.
gracielae compared with A. lentiformis through-
out the skipper's range. Soil moisture levels ade-
quate for shrub growth may be inadequate, or
asynchronous with sootywing oviposition due to
intermittent rainfall or changing groundwater
depth, for host plant finding and acceptance.

ACKNOWLEDGMENT

We thank Cissy Pratt for helping survey eggs, Amy
Stephenson (Reclamation Regional Lab, Boulder City,
NV) for measuring ammonia concentrations, and An-
drew Sanders (UCR Herbarium) for identifying Bac-
charis. We appreciate the study permit granted by
Cibola National Wildlife Refuge. This work was funded
by the Lower Colorado River Multi-Species Conserva-
tion Program.

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Florida Entomologist 91(2)


THREE NEW SPECIES OF PHYLLOPHAGA FROM MEXICO
(COLEOPTERA: SCARABAEIDAE: MELOLONTHINAE)

MIGUEL-ANGEL MORON1 AND ROBERT E. WOODRUFF2
1Departamento de Biologia de Suelos, Instituto de Ecologia, A.C. Apdo. Postal 63 Xalapa, Veracruz 91000 Mexico
E-mail: miguel.moron@inecol.edu.mx

2Florida State Collection of Arthropods. Florida Department of Agriculture and Consumer Services,
P.O. Box 147100, Gainesville, FL 32614-7100
E-mail: BobsGems@aol.com

ABSTRACT

Three new species of Phyllophaga from Mexico are described: P. (Phytalus) balli, new spe-
cies collected in pine and oak forests toward the northwest of Oaxaca valley; P. (Phylloph-
aga) navidad, new species obtained in tropical deciduous forests in the Pacific coast of
Jalisco; and P. (Phyllophaga) potosisalta, new species collected in tropical subdeciduous
forest of eastern San Luis Potosi. Illustrations of diagnostic characters are included.

Key Words: May beetles, taxonomy, tropical forests, oak forests, Mexico

RESUME

Se described tres species nuevas de Phyllophaga de M6xico: P. (Phytalus) balli nueva es-
pecie colectada en bosques de pinos y encinos situados al noroeste del valle de Oaxaca; P.
(Phyllophaga) navidad nueva especie capturada en bosques tropicales caducifolios locali-
zados en la costa de Jalisco; y P. (Phyllophaga) potosisalta nueva especie encontrada en
bosque tropical subcaducifolio ubicado en la Huasteca Potosina. Se incluyen ilustraciones de
algunos caracteres diagn6sticos.


Translation provided by the authors.


Curatorial work of Mexican May beetles depos-
ited in the Florida State Collection of Arthropods,
provided specimens of a number of undescribed
species. Many of these possess sets of external mor-
phological characters and male genital capsules
that are different from the groups of species pro-
posed by Mor6n (1986, 2003). Some of these may
support new groups of species, but others remain
isolated, or with uncertain position in the subge-
nus Phyllophaga (sensu stricto. In this paper we
describe 1 new species of Phyllophaga (Phytalus)
and 2 new species of Phyllophaga (s.str.).

MATERIALS AND METHODS

The characters and terms used in the descrip-
tions are those of Sanderson (1958), Saylor
(1942), Mor6n (1986), and Woodruff & Beck
(1989). Illustrations were made with JEOL:JSM-
5510LV Scanning Electron Microscope and with
the Auto-Montage Pro@ located at FSCA. Mea-
surements were obtained with an ocular mi-
crometer on a Zeiss stereomicroscope or with dig-
ital calipers. Specimens are deposited in the fol-
lowing collections: Florida State Collection of Ar-
thropods (FSCA); Instituto de Ecologia, Xalapa,
Mexico (IEXA); M. A. Mor6n, Xalapa (MXAL);
Universidad Aut6noma del Estado de Hidalgo,


Pachuca (UAEH); and United States National
Museum, Washington, D.C. (USNM).

Phyllophaga (Phytalus) balli, new species (Figs. 1-7)

Description

Holotype male. Body and legs yellowish brown,
shiny. Clypeus with numerous slender, erect, long
setae, 3.2 times wider than long, borders moder-
ately elevated, anterior margin briefly sinuate at
middle, disk surface widely convex, with many
uniformly distributed, round punctures. Fronto-
clypeal suture briefly sinuate and clearly im-
pressed. Frons 1.6 times wider than long, convex,
punctate rugose, with many slender, erect, long
setae. Frons 4.3 times wider than dorsal diameter
of eye. Eye canthi long and narrow, with 13-15
setae. Antenna 10-segmented, with 3-segmented
club, lamellae 1.3 times longer than length of pre-
ceding 6 segments combined; segments 3 or 4
shorter than 5; segments 6 and 7 wider than long,
with short flattened prominences on anterior
sides. Labrum bilobed, deeply notched, with
many slender, long setae along the borders. Men-
tum widely concave, impunctate, with scarce,
slender setae at sides, anterior border broadly
notched.


June 2008







Mor6n & Woodruff: New species of Phyllophaga from Mexico


Fig. 1-3. Phyllophaga balli, new species. 1. Male,
lateral view. 2. Male tarsal claws. 3. Male metatibial
spurs.


Pronotum 1.8 times wider than long and 2.1
times wider than frons. Pronotal disk slightly
shiny, with numerous slender, erect, long setae
(Fig. 1) and shallow, round punctures regularly
separated by 0.5-1 diameters; lateral borders
strongly angulate, lateral marginal bead crenu-
late, with long, curved setae; basal bead indicated
by punctures on most of it extension; anterior an-
gles obtuse, slightly prominent; posterior angles
obtuse, clearly prominent. Scutellum 1.3 times
wider than long, with 17 punctures and anterior
border widely sinuate. Elytron 2.3 times longer
than wide, shiny, densely and irregularly punc-
tate, with slender, erect, long setae near the base
and scutellum (Fig. 1), and some scattered, short
setae all along disc; epipleural border progres-
sively narrowed toward the apex, with slender,
long setae; humeral calla rounded, prominent;
apical calla rounded. Metathoracic wings com-
pletely developed. Pterosterna with long, dense,
yellowish setae. Visible abdominal sternites 2 to
4 slightly depressed at middle; 5th sternite
widely convex, shiny, with scattered setiferous
punctures; anal plate long, weakly concave, with
a central, moderate prominence covered by seti-
gerous granules (Fig. 4). Propygidium shiny, with
numerous punctures. Pygidium shiny, widely con-
vex, densely rugose, with scattered slender, long
setae mainly at sides, and lateral deep fovea; api-
cal margin with 20 long, slender setae; basal mar-
gin weakly defined at middle.


Fig. 4-5. Phyllophaga balli, new species. 4. Male
anal plate, ventral view. 5. Female pigidium and anal
plate, distal view.



Protibia slightly shorter than protarsus
(0.8:1), with 2 large teeth and a basal small tooth
on external border, preapical spur acute, straight,
1.8 times longer than 2nd protarsomerus. Meso-
tibia with one oblique, well marked, setiferous ca-
rina on external side, and curved, short spines
along dorsal border; upper apical spur slender,
with acute apex, 0.2 times longer than lower slen-
der spur, with acute apex. Metatibia slightly
shorter than metatarsus (0.9:1), with one oblique
setiferous carina on external side, and curved,
short spines along dorsal border; upper apical
spur articulated (Fig. 3), slightly curved, round
pointed, 1.3 times longer than basal metatarsom-
ere, and 1.1 times longer than lower spur; lower
apical spur articulated, apex rounded. Tarsomeres
semicylindrical, elongate, with enlarged apex,
some setae apically, and one line of short setae
along ventral side, especially on metatarsomeres.
Tarsal claws similar in all legs, deeply cleft, upper
tooth slightly longer than lower tooth (Fig. 2).


':i~"~b~'
.i*I- ~ Y
-".







Florida Entomologist 91(2)


Fig. 6-7. Phyllophaga balli, new species. 6. Male
genital capsule, lateral view. 7. Male genital capsule,
dorsal view.


Genital capsule with long, strongly curved
parameres not fused along ventral surface, dor-
sally fused at phallobase, apex rounded, slightly
flattened, with scattered short setae (Figs. 6-7).
Tectum with narrow, shallow depression on mid-
line. Aedeagus with sclerotized support, provided
with preapical, long sinuose projections at each
side. Length of genital capsule from apex of
parameres to border of basal piece: 4.2 mm. Total
body length: 14.1 mm. Humeral width: 6.0 mm.
Allotype Female. Similar to the male except as
follows: antennal club as long as preceding 4 seg-
ments combined. Visible abdominal sternites 20
to 5 convex, with scattered, setiferous punctures;
anal plate longer than in male, strongly convex,
punctate, with scattered long setae near the basal
border and row of 8 slender setae on apical border.
Pygidium densely rugose, with scattered long se-
tae on disc and preapical depression (Fig. 5). Both
apical spurs of metatibia widened, curved, with
rounded apex. Tarsal claws widely cleft, with
lower tooth slightly shorter than upper tooth.
Ventral genital plates moderately sclerotized,
nearly symmetrical, ovate, with few short setae
near distal border; dorsal genital plates with
rounded process directed mesially, each with a
tuft of setae at the apex. Total body length: 13.2
mm. Humeral width: 5.7 mm.


Variation
Total body length: 14.3-13.5 mm, humeral
width: 6.0-5.7 mm. Metatibial spurs of male from
Zaachila-San Miguel Peras with acute apex.

Type Materials
Described from 3 males and 5 females. Holo-
type male; MEXICO: Oaxaca, Rte. 190, 33.0 mi
NW Oaxaca, oak forest, 4/5-V-1967, G. Ball, T. L.
Erwin, R. E. Leech (FSCA). Allotype and 2 female
paratypes with same data as holotype (FSCA,
MXAL). Two males and two females paratypes:
MEXICO: Oaxaca, km 22 Zaachila-San Miguel
Peras, bosq. Pino-encino, alt. 2580 m, 28-VIII-
1991, P. Rojas (MXAL, IEXA).

Type Locality
Mazaltepec environs, 33.0 mi NW Oaxaca city,
state of Oaxaca, M6xico (approx. 1725'-1034'N;
96055'W).

Biological Data
This species inhabits dry pine-oak forests lo-
cated at 2500-2600 m altitude. Specimens were
collected during May (4) and Aug (4). Other spe-
cies flying at the same time and place were P.
(Phyllophaga) nisuens Saylor and (Phytalus) so-
lavegana Mor6n.


Taxonomic Remarks


Phyllophaga (Phytalus) ball belongs to the
species group "senicula" (sensu Mor6n 1986). It is
similar to P. (Phytalus) bolacoides (Bates), a rare
species known only from 2 localities in the south
of the state of Guerrero, Mexico, but the body
shape in both sexes of P balli is more elongate,
the male antennal club is larger and the dorsal
vestiture in both sexes is much longer and denser
than in the new species, the male anal plate of P.
bolacoides is nearly convex, and the parameres of
P balli are longer with downward curved apex.

Etymology
This species is dedicated to Dr. George E. Ball,
well known master professor of coleopterists, ded-
icated specialist on the Mexican Carabidae, and
who collected part of the type series of this inter-
esting new species.

Phyllophaga (Phyllophaga) navidad, new species
(Figs. 8-11)

Description

Holotype Male. All body surfaces shiny; head
dark brown; pronotum reddish brown; elytra,
sternites, pygidium and legs yellowish brown.


June 2008


IX59 500







Mor6n & Woodruff: New species of Phyllophaga from Mexico


... .. i : ,: ..:... . ................ .
.:...... .... .



... .. ,:.,.:.:.....


.


Fig. 8-9. Phyllophaga navidad, new species. 8.
Male, lateral view. 9. Male tarsal claws.

Clypeus 3.2 times wider than long, borders notice-
ably elevated, anterior margin briefly sinuate,
disk surface convex, with deep, irregular, round
punctures, and scarce, erect setae. Frontoclypeal
suture nearly straight, weakly impressed. Frons
2.3 times wider than long, widely convex, coarsely
rugo-punctate with scarce, erect setae. Frons 3.6
times wider than dorsal diameter of eye. Eye
canthi long and narrow, with 9 setae. Antenna 10-
segmented, with 3-segmented club, lamellae as
long as proceeding 6 segments combined; segment
4 as long as 3; segments 5 and 6 progressively
wider than long, with rounded prominences on
anterior sides; segment 7 much wider than long,
with flattened, acute prominence on anterior side.
Labrum bilobed, deeply notched, with scattered
slender setae along the borders. Mentum widely
concave, impunctate, with scarce, slender setae at
sides, anterior border briefly notched.
Pronotum 2.1 times wider than long and 2.3
times wider than frons. Pronotal disk with scat-
tered setae of different sizes (Fig. 8) and with
deep, round punctures irregularly separated by 1-
5 diameters; lateral borders strongly angulate,
lateral marginal bead crenulate, with slender,
long setae; basal bead indicated by punctures on
middle third; anterior angles acute, prominent;
posterior angles obtuse, rounded. Scutellum 1.5
times wider than long, with 4 punctures and an-
terior border widely sinuate. Elytron 2.5 times


Fig. 10-11. Phyllophaga navidad, new species. 10.
Male anal plate, ventral view. 11. Male genital capsule,
lateral view.


longer than wide, with scattered setae of different
size on all the surface (Fig. 8), densely and regu-
larly punctate; epipleural border progressively
narrowed toward the apex, with long setae; hu-
meral calla rounded, prominent; apical calla
rounded. Metathoracic wings completely devel-
oped. Pterosternum with moderate number of yel-
lowish, long setae. Visible abdominal sternites 2
to 40 convex at middle, with many short setae on
complete surface; 5th sternite widely convex,
shiny, with numerous short setae; anal plate
large, weakly furrowed at middle, shiny, granu-
lose-punctate with mixture of long and short se-
tae (Fig. 10). Propygidium moderately shiny,
densely granulose-punctate, with abundant short
setae. Pygidium moderately shiny, widely convex,
densely granulose-punctate, with abundant short
setae; apical margin with 14 medium sized, slen-
der setae; basal margin effaced at middle.
Protibia slightly shorter than protarsus
(0.9:1), with 2 large teeth and one basal small
tooth on external border, preapical spur acute,
straight, shorter than 2nd protarsomere. Me-
sotibia with an oblique, well-marked, setiferous
carina on external side; upper apical spur with


10


~1Cr~ll~ll~







Florida Entomologist 91(2)


acute apex, shorter than lower spur (0.7:1) lower
spur with round apex. Metatibia nearly as long as
metatarsus, with an oblique, weakly marked, se-
tiferous carina on external side; upper apical spur
articulated, curved, round pointed, slightly
longer than basal metatarsomere (1.2:1), and 1.3
times longer than lower spur; lower apical spur
articulated, wider than upper spur, curved, with
round apex. Basal 4 protarsomeres slightly short-
ened, with subapical tufts of setae. Meso- and
metatarsomeres semicylindrical, elongate, with
enlarged setose apex, and with a line of stout se-
tae along ventral side, best developed in metatar-
sus. All tarsal claws widely cleft, lower tooth wide
and slightly shorter than upper tooth (Fig. 9).
Genital capsule with long, narrow parameres dor-
sally fused with phallobase, ventrally convergent
and distally projected as strongly curved, acute
knives (Fig. 11). Aedeagus with strongly sclero-
tized tube-like support with lateral, preapical,
asymmetrical long spines (Fig. 11). Tectum widely
convex. Length of genital capsule from apex of
parameres to border of basal piece: 3.2 mm. Total
body length: 15.1 mm. Humeral width: 6.5 mm.
Female. Unknown.

Variation
Male Paratype. Pronotal and elytral setae less
abundant. Total body length: 14.9 mm, humeral
width: 6.3 mm.

Type Series
Described from 2 males. Holotype male; MEX-
ICO: Jalisco, 25 mi NW Barra de Navidad, 22-VII-
1974, blacklight, P. D. Perkins (FSCA). Paratype;
MEXICO: Jalisco, Est. de Biologia Chamela, 9-XI-
1985, M. Sanchez (MXAL).

Type Locality
Barra de Navidad, Melaque municipality, state
of Jalisco, Mexico (approx. 1918'N; 10449'W).

Biological Data
This species inhabits the tropical deciduous
forest located between 30 to 180 m of altitude in
the central part of the Pacific slopes of the state of
Jalisco. Specimens were collected during Jul (1)
and Nov (1). Another species flying at the same
time and place was P. (Phyllophaga) multipora
(Bates).

Taxonomic Remarks
Phyllophaga navidad does not belong to any
species group defined by Mor6n (1986; 2003). Body
surface, tarsal claws and general structure of
parameres are similar to P pilula (Moser) known
from the northern mountains of Chiapas (Moron
2001), but the shape of phallobase, distal part of


the parameres and details of the apex of tube-like
structure of aedeagus are clearly different. The
form of the parameres and phallobase of P
navidad is similar to P nisuens Saylor and P tsa-
jumiana Mor6n from the wet forests in northern
Oaxaca mountains (Mor6n 2001), but the body sur-
face and shape of tarsal claws are much different.

Etymology
Specific epithet derived from the name of type
locality, Barra de Navidad.

Phyllophaga (Phyllophaga) potosisalta, new species
(Figs. 12-15)

Description
Holotype Male. All body surfaces shiny; head
and pronotum reddish brown; elytra, sternites,
pygidium and legs yellowish brown. Clypeus 2.3
times wider than long, borders noticeably ele-
vated, anterior margin deeply notched, disk sur-
face progressively convex towards midline, with
deep, irregular, round punctures, and scarce,
erect, long setae. Frontoclypeal suture nearly
straight, weakly impressed. Frons 2.9 times
wider than long, widely convex, coarsely rugo-
punctate with sparse, erect long setae. Frons 6
times wider than dorsal diameter of eye. Eye
canthi long and narrow, with 12 setae. Antenna









S .. .....
W .o ..k 7. .



12


Fig. 12-14. Phyllophaga potosisalta, new species.
12. Male, lateral view. 13. Male tarsal claw. 14. Male
genital capsule, dorsal view.


June 2008







Mor6n & Woodruff: New species of Phyllophaga from Mexico


Fig. 15. Phyllophaga potosisalta, new species. Male
genital capsule, lateral view.


10-segmented, with 3-segmented club, lamellae
nearly as long as proceeding 5 segments com-
bined; segment 4 as long as 3; segments 5 and 6
progressively wider than long, with rounded
prominences on anterior sides; segment 7 much
wider than long, with large, rounded prominence
on anterior side. Labrum deeply bilobed, with
scattered slender setae along borders. Mentum
widely concave, impunctate, with scarce, slender
setae at sides, anterior border briefly notched.
Pronotum 1.6 times wider than long and 1.9
times wider than frons. Pronotal disk with scat-
tered erect setae of different sizes (Fig. 12) and
with deep, round punctures irregularly separated
by 1-3 diameters; lateral borders moderately an-
gulate, lateral marginal bead strongly crenulate-
dentate, with slender, long setae; basal bead re-
placed by an irregular line of round punctures
from side to side; anterior angles strongly acute,
prominent; posterior angles obtuse, prominent.
Scutellum 1.5 times wider than long, with 4
punctures irregularly placed and anterior border
widely sinuate. Elytron 2.4 times longer than
wide, surface regularly punctate with abundant
setae of different sizes on all the surface (Fig. 12),
erect long setae less abundant than short decum-
bent setae; epipleural border progressively nar-
rowed toward apex, with long setae; humeral
calla rounded, prominent; apical calla rounded.
Metathoracic wings completely developed.
Pterosternum with numerous yellowish, long se-
tae. Visible abdominal sternites 2 to 4 convex at
middle, with many short setae on all surface; 5th
sternite widely convex, shiny, with numerous
short setae; anal plate short, weakly excavated
with basal transverse ridge and scattered erect
setae. Propygidium shiny, densely with abundant
short setae. Pygidium shiny, widely convex,
densely punctate, with abundant decumbent
short setae and numerous erect, long setae; apical
margin with 12 medium sized, slender setae;
basal margin nearly effaced from side to side.


Protibia nearly as long as protarsus, with 2
large teeth and a basal small tooth on external
border, preapical spur acute, straight, as long as
2nd protarsomere. Mesotibia with an oblique,
well marked, setiferous carina near the middle of
external side and other short setiferous carina on
basal third of external side; both apical spurs
with acute apex and similar length. Metatibia
slightly shorter than metatarsus (0.9:1), with an
oblique, strong, setiferous carina near the middle
of external side, and 2 spines on basal third of
dorso-external side; upper apical spur articu-
lated, slightly curved, round pointed, longer than
basal metatarsomere (1.3:1), and 1.3 times longer
than lower spur; lower apical spur articulated,
curved, with round apex. Basal 4 protarsomeres
progressively shortened, with subapical tufts of
setae. Mesotarsomeres semicylindrical, elongate,
with enlarged setose apex. Metatarsomeres semi-
cylindrical, elongate, with enlarged setose apex
and with a line of stout setae along ventral side.
All tarsal claws dentate, lower tooth narrowed
and acute, placed near their bases (Fig. 13). Gen-
ital capsule with short, narrowed parameres dor-
sally fused with phallobase, ventrally closely par-
allel with rounded apex (Figs. 14-15). Aedeagus
with sclerotized, curved, symmetrical support
(Fig. 15). Tectum strongly projected over phallo-
base, wide and deeply bifurcate (Fig. 14). Length
of genital capsule from apex of parameres to bor-
der of basal piece: 4.5 mm. Total body length: 16.4
mm. Humeral width: 6.6 mm.
Allotype Female. Similar to the male except as
follows: anal plate slightly longer, convex, punc-
tate, with transverse row of long setae across mid-
dle and row of 10 slender setae on apical border.
Pygidium more convex, densely punctate, with
numerous short setae on all disk and scattered
very long, erect setae on distal half of disk. Both
apical spurs of metatibia widened, curved, with
rounded apex. Ventral genital plates moderately
sclerotized, nearly symmetrical, ovate, with
round process on the inner apical margin and long
process on the outer apical margin angled toward
midline; dorsal genital plates small, weakly scle-
rotized, partially covered by ventral plates. Total
body length: 19.3 mm. Humeral width: 7.1 mm.

Variation
Body color dark brown. Pronotum, elytra and
pygidium with more long setae. Lateral marginal
bead of pronotum deeply dentate. Total body
length: 16.4-19.3 mm, humeral width: 6.6-7.9 mm.

Type Series
Described from 4 males and 3 females. Holo-
type male; MEXICO: San Luis Potosi, El Salto
falls, 14-VI-1963, blacklight trap, R. E. Woodruff
(FSCA). Allotype and male paratype with same
data as holotype (FSCA, MXAL). One female







Florida Entomologist 91(2)


paratype: MEXICO: San Luis Potosi, Salto de
Agua, 1-V-1971, Blacklight trap, A. Newton
(FSCA). One female paratype: MEXICO: San
Luis Potosi, El Naranjo, 29-VI-1965, P. J. Span-
gler (USNM). Two male paratypes: MEXICO: San
Luis Potosi, El Naranjo, Cascada El Salto, 407 m,
17-VIII-2004, trampa-luz, J. Marquez, J. Asiain,
J. Canales (UAEH, MXAL).

Type Locality

El Salto, El Naranjo municipality, state of San
Luis Potosi, Mexico (2235'0.9.9"N; 99022'54.4"W).

Biological Data

This species inhabits the tropical subdecidu-
ous forest located at 350 to 470 m of altitude in
the eastern part of the state of San Luis Potosi.
Specimens were collected during May (1), Jun (4)
and Aug (4). Other species flying at the same time
and place were P (Phyllophaga) temora (Saylor),
P (P.) setifera (Burmeister), P. (Phytalus) tri-
chodes (Bates) and P (Chlaenobia) vexata (Horn).

Taxonomic Remarks

Phyllophaga potosisalta does not belong to any
species group defined by Mor6n (1986, 2003). Tar-
sal claws are similar to the species in the group
"scissa" (Mor6n 2003b) and the structure of the
phallobase is similar to some species in the group
"schizorhina" (e.g., P necaxa Saylor, P saylori
Sanderson) known from the mountains of Nuevo
Le6n, Tamaulipas, Hidalgo and Puebla (Moron
2003a), but the shape of clypeus and pronotum,
the abundant setae on all the body parts and legs,
and details of the tube-like structure of aedeagus
are clearly different.

Etymology

The specific epithet is formed with an anagram
of the name of the type locality, El Salto, San Luis
Potosi.


ACKNOWLEDGMENTS

Curatorial work ofMAM at the Florida State Collec-
tion ofArthropods (FSCA) was possible by a grant from
the Center for Systematic Entomology, Inc. (2004, 2006)
and the Instituto de Ecologia, A.C. (account 902-08-
011). We thank Dr. Paul E. Skelley for assistance with
the scanning electron microscope and automontage sys-
tem in FSCA. This publication was supported by the
project "Coleoptera Lamellicornia de Am6rica Latina,
Institute de Ecologia, A.C. (account 2000910011).

REFERENCES CITED

MORON, M. A. 1986. El g6nero Phyllophaga en M6xico.
Morfologia, distribuci6n y sistematica supraespeci-
fica (Insecta: Coleoptera). Institute de Ecologia, A. C.
Mexico. 341 pp.
MORON, M. A. 2001. New and rare species of Phylloph-
aga (s.str.) from Mexico (Coleoptera: Melolonthidae:
Melolonthinae). The Pan-Pacific Entomol. 77 (3):
168-189.
MORON, M. A. 2003a. Revision of the Phyllophaga s. s.
schizorhina species group (Coleoptera:
Melolonthidae: Melolonthinae). The Canadian Ento-
mol. 135: 213-302.
MORON, M. A. 2003b. Diversidad, distribuci6n e impor-
tancia de las species de Phyllophaga Harris en
M6xico (Coleoptera: Melolonthidae), pp. 1-27 In A.
Arag6n, M. A. Mor6n, y A. Marin [eds.], Estudios so-
bre Cole6pteros del Suelo en Am6rica. Publicaci6n
especial de la Benem6rita Universidad Aut6noma de
Puebla, M6xico. 359 pp.
SANDERSON, M. W. 1958. Faunal affinities of Arizona
Phyllophaga, with notes and descriptions of new
species. J. Kansas Entomol. Soc. 31: 158-173
SAYLOR, L. W. 1942. Notes on beetles related to Phyllo-
phaga Harris, with descriptions of new genera and
subgenera. Proc. United States Natl. Mus. 92 (3145):
157-165.
WOODRUFF, R. E., AND B. M. BECK. 1989. The Scarab
Beetles of Florida (Coleoptera: Scarabaeidae) Part II
The May or June Beetles (genus Phyllophaga). Ar-
thropods of Florida and Neighboring Land Areas.
Volume 13: 1-225. Florida Department of Agricul-
ture and Consumer Services, Division of Plant In-
dustry, Gainesville.


June 2008







Park & Byun: New Genus Chrysonasma and New Species


A NEW GENUS CHRYSONASMA (LEPIDOPTERA, GELECHIOIDEA,
LECITHOCERIDAE), WITH DESCRIPTION OF A NEW SPECIES
FROM THE PHILIPPINES

KYU-TEK PARK1 AND BONG-KYU BYUN2'3
'McGuire Center for Lepidoptera and Biodiversity, Florida Museum of Natural History,
University of Florida, Gainesville, FL 32611 USA
e-mail: keitpark@hanmail.net

2Division of Forest Biodiversity, Korea National Arboretum, Pocheon, 487-821 Korea

3Corresponding author; e-mail: bkbyun@korea.kr

ABSTRACT
A new genus, Chrysonasma Park, of Lecithoceridae (Gelechioidea) is described based on C.
cassiterota (Meyrick) and a new species, C. caliginosa, from the Philippines. The genus is
separable from its allies by the colorful wing-pattern with leaden metallic longitudinal
streaks. A key to the 2 species of the genus is given. Illustrations of the images, forewing ve-
nation, and male and female genitalia are provided.

Key Words: taxonomy, Lepidoptera, Lecithoceridae, Chrysonasma, new genus, new species

RESUME
Se describe un nuevo g6nero, Chrysonasma Park, de la familiar Lecithoceridae (Gelechioidea)
basado en C. cassiterota (Meyrick) y una nueva especie, C. caliginosa, de las islas Filipinas.
Se puede separar este g6nero de sus grupos cercanos por el patron colorido del ala con rayos
longitudinales metalicos de color plomo. Se provee una clave de las 2 species en el g6nero y
se ilustran los images, la nervadura del ala anterior, y las genitalias del macho y la hembra.


Diakonoff(1967) was the first to review Lecitho-
ceridae (referred to as Timyridae) from the Philip-
pines, and he reported 27 species belonging to 7
genera. Diakonoff reported C. cassiterota (Mey-
rick) under the genus Lecithocera Herrich-
Schaffer, with illustrations of the male genitalia
for the first time. Reviews of Tisis Walker (Park
2003) and Homaloxestis (Park & Byun 2007) of the
Philippines resulted in descriptions of 4 and 2 new
species, respectively. However, the fauna of Leci-
thoceridae in the Philippines is very poorly known
and incomplete. This paper is the result of a review
of the Lecithoceridae in the Philippines that were
or will be treated in a series of papers. The new ge-
nus C' .., .... .. ....i belongs to the family Lecitho-
ceridae (Gelechioidea), which includes small
moths occurring primarily in the Oriental Region.

MATERIALS AND METHODS

Examined material was borrowed from the
Zoological Museum, Copenhagen, collected in the
Philippines by O. Karsholt and his colleagues in
1961, and from the Museum fir Naturkunde,
Berlin, collected by W. Mey and his colleagues in
2000. The color standard for the description of
adults was Kornerup & Wanscher (1978). The ge-
nus is described by the first author and the spe-
cies is described by both authors.


RESULTS

Genus Chrysonasma Park, gen. nov.

Type species: C'i., .......' cassiterota Mey-
rick, 1923, Exot. Microlep. 3: 40 (Lecithocera).
The genus C'i, ........ .. is related to Torodora
Meyrick, but it is differentiated from the latter by
the following characters: forewing elongate, with
metallic blue longitudinal streak on the forewing;
termen strongly oblique, concave medially, with
black scales along margin; forewing venation with
R, absent, CuA1 and CuA2 coincident. The male
genitalia of C(l. .....1...1.. are separable from
those of the latter by the heavily sclerotized, beak-
like gnathos, which is similar to that of Epharmo-
nia Meyrick. However, the latter has different ve-
nation from this new genus by the presence of R5
in the forewing and absence of M2 in the hindwing.
Adults: Head with appressed scales, shining
metallic blue with golden yellow erect scales lat-
erally. Basal segment of antenna grayish brown
dorsally; flagellum pale grayish orange with pale
brown annulations. Second segment of labial pal-
pus thickened, light orange on outer and inner
surfaces, black ventrally; 3rd segment as long as
2nd. Hind tibia with long tufts dorsally Forewing
light orange to golden yellow, sparsely speckled
with dark brown scales; basal third of wing with
3 metallic blue, longitudinal streaks: first streak







Florida Entomologist 91(2)


Figs. 1-2. Adults of Chrysonasma species. (1) C. cas-
siterota (Meyrick); (2) C. caliginosa sp. nov.


runs between costa and R vein, rounded apically;
2nd median, dilated, truncate apically; 3rd nar-
rower, between cell and dorsum; median zone
broad, trapezoidal, gently dilated towards dor-
sum, uniform metallic blue or partly; distal 3rd of


wing golden yellow or pale brownish orange,
speckled with dark brown scales, with 2 metallic
blue, longitudinal streaks: a long one below costa,
terminated before apical patch, and shorter one
beyond cell, not so much extended; a large, oval,
metallic blue patch at apex and larger one at tor-
nus. Abdominal tergites II-VII with broad spinu-
lose zones.
Male Genitalia (Figs. 5, 6): Uncus slender, rel-
atively straight beyond curved base. Gnathos
heavily sclerotized, beak-like; acute apically.
Valva short, extremely broad at base and strongly
narrowed towards apex; costa with a long, digi-
tate process beyond middle (in the type species) or
without (in this new species); ventral margin
slightly concave medially. Juxta gently concave
on caudal margin, with slender, acute, latero-cau-
dal lobes. Aedeagus stout, cylindrical, longer than
valva; cornutus a long sinuate blade, serrated on
ventral margin posteriorly, with pointed apex.
Female Genitalia (Figs. 7, 8): Eighth sternite
deeply or slightly incised at middle, sometimes
with long sclerotized plates latero-ventrally. Os-
tium bursae with nearly straight caudal margin;
antrum cup-shaped, heavily sclerotized or not.
Ductus bursae narrowed beyond antrum, then ex-
panded; ductus seminalis arising before middle.
Corpus bursae, ovate; sigum a round plate with
short conical spines on surface.
Distribution. Oriental Region (the Philippines)
Etymology. The genus name is derived from
Greek, chrysos (= gold) + nasmos (= stream).


KEY TO SPECIES OF THE GENUS CHRYSONASMA PARK

1. Forewing ground color golden yellow; fringe pale orange, but distal part metallic blue; hind tibia with long hair-
tufts dorsally. Valva of male genitalia with digitate process on costa beyond middle... cassiterota Meyrick
2. Forewing ground color light orange to grayish orange; fringe pale orange, but distal part shining orange gray;
hind tibia without long hair-tufts dorsally. Valva of male genitalia without digitate process on costa
............... .................... .................................. caliginosa sp. nov.


Chrysonasma cassiterota (Meyrick, 1923), comb. nov.
(Figs. 1, 3, 5, 5a, 7)

Lecithocera cassiterota Meyrick, 1923. Exot.
Microlep. 3: 40; 1925: 240; Meyrick, 1925: 240;
Clarke, 1955: 74; 1965: 115pl. 57, figs 4-4d; Dia-
konoff, 1967: 135, Figs. 176, 200, 201, 606. TL: Lu-
zon, Philippines. [BMNH].
Diagnosis. Wingspan, 16.0-17.0 mm. The
forewing venation differs from species ofTorodora
by the absence of Rs, and the coincidence of CuA1
and CuA2, and the termen that is strongly con-
cave medially (Fig. 3). This species can be distin-
guished from the following new species by the
broad valva of the male genitalia with a digitate
process at the middle of the costa.
Material Examined. Three males & 1 female,
Philippines, Luzon, Mt. Makiling 400 m, 14-16 III
2000, LF (W. Mey & K. Ebert), gen. prep. no. CIS-


5023/Park, -5042/Park (female); 1 male, Santa Fe,
Bald Mts 1150 m, 11-13 XI 1997 (W. Mey & K.
Ebert) 7; 3 males, Luzon, Quezon, NP, LF, 20 III
2000 LF(W. Mey & K. Ebert). All specimens de-
posited in Museum fiir Naturkunde, Berlin.
Distribution. The Philippines (Luzon).
Remarks. The species was originally described
as a species ofLecithocera, based on a female from
Luzon, Philippines. The male genitalia subse-
quently illustrated by Diakonoff (1967). Meyrick
(1922) noted that he had an undescribed species
from the Philippines that was closely allied to
lamprodesma Meyrick. Diakonoff (1967) consid-
ered that the undescribed species mentioned by
Meyrick (1922) to be same as what Meyrick de-
scribed in 1923 as cassiterota. Diakonoff further
noted that cassiterota was allied with lam-
prodesma and that he knew of "two closely allied,
undescribed species" from Java and Borneo, but it


June 2008






Park & Byun: New Genus Chrysonasma and New Species


3


Vt


5a


Figs. 3-6. Forewing venation and male genitalia of Chrysonasma species. (3) C. cassiterota (Meyrick), forewing
venation; (4) C. caliginosa sp. nov., forewing venation; (5) C. cassiterota, male genitalia, (5a) aedeagus; (6) C. calig-
inosa sp. nov., male genitalia; (6a) aedeagus. Scale bar for the male genitalia: 1 mm.


is unclear if the 2 undescribed species by Dia-
konoff's notes are members of the new genus. We
had no chance to examine lamprodesma Meyrick
and cannot define its generic status in this paper.
However, we can expect additional species of this
new genus because of the presently limited col-
lecting in the Oriental Region.

Chrysonasma caliginosa Park and Byun, new species
(Figs. 2, 4, 6, 6a, 8)
Diagnosis. The new species is superficially sim-
ilar to C. cassiterota (Meyrick) in color pattern and
markings, with metallic, longitudinal streaks at
the basal part of the forewing. C' ., .... ....... calig-
inosa has a light orange to grayish orange ground
color of the forewing, a fringe that is shining or-
ange gray at distal half, and the male genitalia
with valva lacking a costal process, whereas C. cas-
siterota has a golden yellow color of the forewing, a
fringe that is metallic colored at distal half, and
male genitalia with a costal process of the valva.
Description. Male and Females. Wingspan,
15.0-16.0 mm. Head dark brown dorsally, with
orange gray erect scales laterally. Tegula dark


brown. Second segment of labial palpus moder-
ately thickened, pale grayish orange suffused
with dark scales on basal half on outer surface,
paler on inner surface; 3rd segment shorter than
2nd, dark brown ventrally. Antenna with basal
joint brown dorsally; flagellum pale grayish or-
ange with dark brown annulations. Hind tibia
dark brown scales, without long hair-tufs dor-
sally; mid spur very long, outer one nearly twice
length of the inner one. Forewing elongate;
ground color light orange to grayish orange; costa
nearly straight; apex more or less acute; basal 2/
5 of wing light orange to grayish orange, with
three shining metallic blue, longitudinal streaks;
first streak along subcosta, narrowed to apex; 2nd
median, dilated apically; 3rd shorter, runs between
cell and dorsum; median line almost vertical,
light orange, narrow, with blackish scales along
proximal side; median zone trapezoidal between
median and postmedian line with oblique outer
margin followed by a crescent blackish mark be-
yond upper corner of cell; a narrow leaden longi-
tudinal streak between costa and R3 vein and the
other broad similar streak beyond cell in distal 3rd
of wing; a large triangular metallic blue patch at


4


e"I







Florida Entomologist 91(2)


Figs. 7-8. Female genitalia of Chrysonasma species. (7) C. cassiterota (Meyrick); (8) C. caliginosa sp. nov. Scale
bar: 1 mm.
bar: 1 mm.


apex, edged by black scales along outer margin,
and the other larger one at tornus. Venation with
R2 closer to R3 than R, at base; R3 and R4 stalked
about 2/5 of R4; R4 reaching to costa before apex;
R5 absent; CuA1 and CuA, coincident; CuA1+2 aris-
ing from near lower corner of cell; apex acute (Fig.
4); termen strongly concave medially, slightly sin-
uate, with dense black scales along margin; fringe
pale orange at basal half, shining orange gray at
distal half. Hindwing with costa slightly ex-
panded before termination of Sc vein; Rs and M,
stalked well beyond end of cell; M2 present; M,
and CuA, shortly stalked; cell opened.


Male Genitalia (Figs. 6, 6a): Gnathos similar to
that of Epharmonia ardua (Meyrick), which was
described from N. India, but valva somewhat sim-
ilar to that of Hygroplasta lygaea Meyrick. Uncus
slender, gently bent downward. Gnathos beak-
like, heavily sclerotized, strongly bent at basal 1/
4. Valva elongate; costa slightly concave medially,
with dense long setae along ventral margin be-
yond middle, and with a thin row of short spines
near along ventral margin, reaching to middle of
apical margins; apex right angled dorso-apically;
ventral margin slightly emarginated near middle.
Juxta with digitate lateral lobes, about 1/3 length


June 2008







Park & Byun: New Genus Chrysonasma and New Species


of uncus. Aedeagus longer than valva; cornuti
consist of a pair of needle-like spines, about 3/4
length of aedeagus.
Female Genitalia (Fig. 8): Eighth sternite
slightly incised medially. Antrum short, weakly
sclerotized, about same width as posterior part of
ductus bursae. Ductus bursae with broad expan-
sion medially. Corpus bursae as long as ductus
bursae; signum elliptical, with dense conical
spines on surface.
Holotype: male, Palawan, Mantalingajan,
Pingisan 600 m, 17 IX 1961, Noona Dan Expedi-
tion. 61-62, gen. prep. No. CIS-5408/Park.
Paratype: 2 males, same locality, 16 IX 1961,
Noona Dan Exp. 61-62, gen. prep. no. CIS-5409/
Park; 3-, same locality, 13 IX 1961; 1 male, same
localiry, 19 IX 1961; 4 males and females, 23 & 24
IX 1961. gen. prep. no. CIS-5498/Park (female).
The holotype and paratypes are deposited in the
Zoological Museum, Copenhagen.
Distribution. The Philippines (Palawan).

ACKNOWLEDGMENTS

We are grateful to Mr. O. Karsholt, Zoological Mu-
seum, Copenhagen, and Dr. W. Mey, Museum fuir
Naturkunde, Berlin, for the loan of their valuable spec-


imens for this study. We thank Dr. D. Matthews Lott,
McGuire Center for Lepidoptera Research and Insect
Conservation, Florida Museum of Natural History, for
her reading of the manuscript with helpful comments.

REFERENCES CITED

CLARKE, J. F. G. 1965. Catalogue of the Type Specimens
of Microlepidoptera in the British Museum (Natural
History) Described by Edward Meyrick. Vol. 5. Lon-
don. 581 pp.
DIAKONOFF, A. 1967. Microlepidoptera of Philippine Is-
lands. United State National Museum Bulletin 257:
125-147. Smithsonian Institution Press, Washing-
ton, D.C.
KORNERUP, A., AND J. H. WANSCHER, 1978. Methuen
Handbook of Colour, 3'd ed. Methuen, London. 252 pp.
MEYRICK, E. 1922. New Microlepidoptera. Zoologische
Mededelingen, 7: 80-89.
MEYRICK, E. 1923, Exotic Microlepidoptera 3: 40.
MEYRICK, E. 1925. Lepidoptera Heterocera. Family
Gelechiidae. Genera Insectorum 184. Bruxelles. 290
pp.
PARK, K. T. 2003. Genus Tisis Walker in Philippines,
with description of four new species (Lepidoptera,
Lecithoceridae). Entomological Sciences, 6: 315-321.
PARK, K. T., AND B. K. BYUN. 2007. Review of Homalox-
estis Meyrick of the Philippines Islands, with de-
scription of two new species. Zootaxa 1449: 57-64.







Florida Entomologist 91(2)


June 2008


A SAFE AND EFFECTIVE PROPYLENE GLYCOL BASED CAPTURE
LIQUID FOR FRUIT FLY (DIPTERA: TEPHRITIDAE) TRAPS
BAITED WITH SYNTHETIC LURES

DONALD B. THOMAS
USDA-ARS Kika de la Garza, Subtropical Agriculture Research Center, 2413 East Hwy 83, Weslaco, TX 78596

ABSTRACT

Antifreeze is often used as the capture liquid in insect traps for its preservative and evapo-
rative attributes. In tests reported herein, fruit fly traps using non-toxic recreational vehicle
(RV) propylene glycol based antifreeze captured significantly more Anastrepha ludens
(Loew) than did traps with the automotive antifreeze. Automotive antifreeze has a charac-
teristic odor due to the additive tolytriazole. The odor may have been mildly repellent.
Whether better or equal in efficacy, fruit fly trapping programs should consider using the
non-toxic formulation as an environmentally friendly alternative over the automotive anti-
freeze, which contains a number of hazardous compounds.

Key Words: fruit fly traps, antifreeze, propylene glycol, Anastrepha

RESUME

La antecongelante esta frecuentamente usada para el liquid en trampas de insects para
sus caracteristicas preservativa y evaporativa. En pruebas reportada aqui, las trampas para
moscas de la fruta usando antecongelante basado en glicol de propilena tipo RV, usada en
sistemas de agua potable, ha capturada mas Anastrepha ludens (Loew) que las trampas
usando antecongelante tipo automovil. La antecongelante tipo automovil tiene un olor ca-
racteristica por parte del aditivo tolytriazola. La ausencia de este olor es posiblemente el fac-
tor responsible para el aumento en captures. Si es mejor o iqual en eficacia, las programs de
trampeo para moscas de la fruta necesitan considerar el uso de las formulaciones menos
toxicos para proteger el medioambiente en lugar de la antecongelante tipo automovil, que
contiene ingredients peligrosos.


Translation provided by the author.


Ethylene glycol based automotive antifreeze is
frequently used as a capture liquid in insect trap-
ping programs using flight-intercept, pitfalls or
pan-traps because of the preservative and evapo-
rative advantages. Antifreeze is also readily
available and less expensive than the technical
grade material ethylene glycol, although there
have been reports of effects on capture efficiency
due to either repellency or attraction (Koivula et
al. 2003; Schmidt et al. 2006) depending on the
targeted species.
Surveillance and detection programs aimed at
invasive species ofAnastrepha fruit flies (Diptera:
Tephritidae) have long relied on the McPhail trap
(McPhail 1939; Steyskal 1977), a bell-shaped, in-
verted glass jar baited with a liquid attractant.
The original design and its modern equivalents
are essentially bottle traps in which the entering
flies become immersed and die in the liquid bait,
typically an aqueous slurry of protein or yeast
(Lopez-Davila & Spishakoff 1963). Volatile com-
pounds released by the yeast or by the bacterial
degradation of the proteins are attractive to fruit
flies, especially females (Martinez et al. 1994; Lee
et al. 1995; Robacker & Bartelt 1997). Further re-


search identified the key volatiles responsible for
the attraction as acetic acid, ammonia, and
amines, especially putrescine and its derivative 1-
pyrroline (Keiser et al. 1976; Bateman & Morton
1981; Robacker & Warfield 1993; Robacker 2001).
Extensive testing of these synthetic lures dem-
onstrated their efficacy in the field (Heath et al.
1997; Heath et al. 2004; Robacker & Thomas
2007). The deployment of synthetic lures how-
ever, requires trap designs that, unlike the solid
McPhail trap, can be opened for access. Typically
the newer traps are two-piece plastic cylinders
that hold the lure in the upper part and the cap-
ture liquid in the lower part. Importantly, the de-
velopment of synthetic lures allowed the use of
capture liquids, such as the commercial polyalco-
hols, with better preservative properties than the
live baits. In selecting a capture liquid for deploy-
ment in mass trapping programs, cost and safety
are major considerations as well as the potential
influences on attraction and capture of the target
and non-target species.
Thomas et al. (2001) reported that captures of
fruit flies in synthetically baited traps containing
10% propylene glycol based automotive antifreeze







Thomas: Propylene glycol in Fruit Fly Traps


were significantly greater than when water and
surfactant was used as the capture liquid. Those
tests were conducted against Anastrepha ludens
(Loew) in Mexico and A. suspense (Loew) in Flor-
ida. The antifreeze effect was confirmed by Ro-
backer & Czokajlo (2006) against A. ludens in
Texas using both 2-component and 3-component
synthetic lures. Those researchers and Hall et al.
(2005) conducted tests for attractancy by automo-
tive antifreeze alone. Hall et al. (2005) captured
no A. suspense in traps baited with antifreeze
without the lures, and Robacker & Czokajlo
(2006) found no significant difference in captures
between traps with antifreeze alone and those
with water and surfactant alone. The latter au-
thors thus concluded that the increase in attrac-
tion with antifreeze was due to a synergism be-
tween the antifreeze and the synthetic lures.
Commercial automotive antifreeze comes in a
variety of formulations including those that are
clearly inappropriate for insect trapping pro-
grams. The use of ethylene glycol formulations
should be discouraged for insect trapping because
of its high mammalian toxicity (Hall 1991). Pro-
pylene glycol, on the other hand, is a GRAS mate-
rial, "generally regarded as safe" by the U.S. EPA
and FDA. It is a common food additive and ingre-
dient in cosmetics and medicines. Although the
acute oral toxicity of the parent compounds is not
very different, the metabolites are crucially dif-
ferent. Propylene glycol metabolizes in the blood-
stream to lactic acid and pyruvic acid, chemicals
produced in the body during normal glycolysis,
whereas ethylene glycol is metabolized to oxalic
acid. The oxalic acid precipitates in the kidneys as
calcium oxalate crystals resulting in renal failure
and rapid death, usually within 24 h (Barceloux
et al. 1999).
Weeks & McIntyre (1997) found no significant
difference in captures of arthropods in pitfall
traps where ethylene glycol or propylene glycol
based antifreezes were the preservative liquid.
Nevertheless, all automotive antifreeze formula-
tions, including the newer organic acid (OAT an-
tifreeze) formulations, are environmentally haz-
ardous because of the blend of additives (around
5%), including lubricants, buffers and corrosion
inhibitors. Therefore, tests were conducted with
recreational vehicle (RV) antifreeze; i.e., formula-
tions containing USP (food-grade) propylene gly-
col, such as those used to winterize swimming
pools and for drinking water systems in cabins
and mobile homes. These non-toxic formulations
should not be confused with "plumbers" anti-
freeze, which contains 20% methanol which is
toxic to humans. The primary consideration in
this study was whether the synergistic effect seen
with the automotive formulation was due to the
propylene glycol itself, or to one or more of the ad-
ditives, and its effects on attraction and trap cap-
tures.


MATERIALS AND METHODS

The automotive formulation of propylene gly-
col used in all of the aforementioned experiments
was Low Tox antifreeze/coolant (Prestone, Dan-
bury CT). For this experiment we compared this
same automotive formulation against a house-
hold formulation, Splash RV & Marine antifreeze
(Superclean Brands, Inc., St. Paul, MN). The lat-
ter formulation is an aqueous solution of 27.5%
propylene glycol, while the former is 95% propy-
lene glycol by weight, thus the industrial strength
products were diluted with water by 1:1 and 7:1
respectively, to achieve approximately equivalent
strength, i.e. about 13%, for the test comparison.
The traps used for this experiment were the
Multilure (Better World Manufacturing, Fresno
CA). The lure deployed in the trap was Biolure
(Suterra LLC, Inc., Bend OR), a two-component
lure consisting of ammonium acetate and pu-
trescine in dispensers suspended from the inside
top of the trap. Altogether, 25 traps were de-
ployed. Ten of the traps contained dilute Low Tox
as the capture liquid, 10 contained dilute Splash
as the capture liquid and 5 contained water with
3 drops of Triton X-100 (Dow Chemical, Midland,
MI) surfactant added. In all traps the amount of
capture liquid was 300 mL.
The experiment was conducted from mid-May
to late Aug 2007 (16 weeks, the Biolures were re-
newed at 8 weeks). The traps were serviced
weekly by filtering the capture liquid through a
screen mesh to remove the insects. The water/tri-
ton was replaced weekly. However, the antifreeze
liquids were recycled, only replacing that ab-
sorbed by the catch with an amount sufficient to
maintain levels at 300 mL. Propylene glycol is an
extremely stable compound, biodegradation oc-
curs at about half the rate of ethylene glycol, and
thus reuse is not only feasible but minimizes
waste disposal concerns, as well as being cost ef-
fective. Used antifreeze is not listed by the EPA as
a hazardous waste under 40 CFR 261, but under
Executive Order 13148 federal agencies are re-
quired to follow EPA recommendations for han-
dling solid waste. Those recommendations in-
clude injunctions against dumping automotive
antifreeze on the ground or discharging it into
sewage waste water systems (US-EPA, 2006). The
design of this experiment anticipated that the liq-
uid would be recycled in this manner if utilized in
an area wide trapping program.
Traps were deployed in a fruit orchard consist-
ing of alternate rows of orange and pear trees lo-
cated at Allende, Nuevo Leon, Mexico (10057'N;
2518'W; elev. 500 m) where wild flies were known
to be abundant. The traps were suspended at 2 m
in every other orange tree in the trap-row and
were rotated at each service interval to the suc-
ceeding trap-tree to minimize position effects
within the orchard.







Florida Entomologist 91(2)


For statistical analysis the means were com-
pared by a t-test and the resulting probabilities
calculated with the NCSS calculator (NCSS Sta-
tistical Software, Kaysville, UT).

RESULTS AND DISCUSSION

Weekly capture data are shown in Table 1. In
accordance with prior experience, both antifreeze
formulations captured far more flies than those
with water/surfactant as the capture liquid. The
traps with the household antifreeze formulation
captured significantly more flies than did the au-
tomotive formulation. The weekly mean of cap-
tures in the Splash traps was 89.5 flies versus
39.2 flies in the Low Tox traps (t = 3.08, df= 30, P
= 0.002). Moreover, while both trap-lure combina-
tions were strongly female biased, which is often
the case with food-based lures (Thomas et al.
2001; Conway & Forrester 2007), the Splash traps
caught significantly more males (as well as fe-
males) than did the Low Tox traps. The weekly
mean of male captures was 19.0 in the splash
traps versus 11.0 in the Low Tox traps (t = 2.16, df
= 30, P = 0.02).
Because the additives in antifreeze are propri-
etary the material safety data sheets provided by
the manufacturer list only those compounds
which are considered to be significant safety haz-
ards when used in accordance with the manufac-
turer's recommendations. The MSDS for the


Splash formulation cites a single additive, 0.2%
dipotassium phosphate, a water softener that is a
common ingredient in laundry detergent and cer-
tain dairy products. Although dipottassium phos-
phate is a surfactant, the concentration is so low
that addition of a drop of household detergent per
trap is recommended. This antifreeze formulation
is pink in color due to the addition of 0.002%
rhodamine B dye, commonly used in the hydro-
logic industry as a tracer. There are no other ad-
ditives in the Splash formulation though some
other brands of RV antifreeze contain methyl sal-
icylate or even corn syrup.
The additives in Low Tox antifreeze, according
to the MSDS, comprise approx. 5% of the formu-
lation. The only 1 of these materials listed in the
MSDS is the carcinogen tolytriazole, a corrosion
inhibitor. Based on industry-wide practices, the
other additives include, but are not limited to: so-
dium silicate, disodium phosphate, sodium mo-
lybdate, sodium borate, dextrin (hydroxyethyl
starch), and a green dye, disodium fluorescein
(dyes are added to antifreeze to help trace the
source of leaks, and as an identifier because the
different formulations are incompatible).
Because of its low volatility propylene glycol by
itself is odorless. The characteristic odor in the
automotive antifreeze formulation is caused by
the tolytriazole. According to Vogt (2005) the un-
pleasant odor in industrial use tolytriazole comes
from impurities in the product that are formed


TABLE 1. NUMBERS OF MALE AND FEMALE MEXICAN FRUIT FLIES CAPTURED WEEKLY IN TRAPS (n = 10) BAITED WITH
BIOLURE COMPARING SPLASH VS. LOW TOX PROPYLENE GLYCOL ANTIFREEZE AS THE CAPTURE LIQUID. CON-
TROL TRAPS (n = 5) HAD WATER AS THE CAPTURE LIQUID.

Splash (10 traps) Low Tox (10 traps) Water (5 traps)

Trap-Week S 2 Total S 2 Total S 2 Total

01 (May) 5 5 10 8 10 18 0 0 0
02 (May) 6 21 27 9 13 22 2 0 2
03 (May) 10 31 41 25 24 49 3 11 14
04 (Jun) 37 59 96 22 44 66 7 9 16
05 (Jun) 11 22 33 16 24 40 4 5 9
06(Jun) 3 8 11 3 12 15 1 2 3
07 (Jun) 11 40 51 15 44 59 1 0 1
08(Jul) 17 42 59 3 20 23 8 6 14
09(Jul) 19 75 94 10 22 32 2 5 7
10 (Jul) 31 86 117 8 21 29 0 0 0
11(Jul) 49 161 201 11 32 43 2 3 5
12(Aug) 16 77 93 15 40 55 2 3 5
13(Aug) 35 112 147 7 61 68 4 8 12
14(Aug) 23 142 165 4 12 16 1 2 3
15(Aug) 11 100 111 10 19 29 2 3 5
16(Aug) 20 156 176 12 51 63 2 6 8
Totals 304 1137 1432 176 449 627 41 63 104
Mean 19.0 71.1 89.5 11.0 28.1 39.2 2.6 3.9 6.5
SD 13.0 51.6 60.3 6.3 15.5 18.7 2.2 3.4 5.2


June 2008







Thomas: Propylene glycol in Fruit Fly Traps


from the toluidine isomers (ortho-, meta- and
para-toluidine) and meta-diamino toluene which
are side-products in the manufacture of tolytriaz-
ole. These side-products are highly reactive and
produce volatile aromatic amines which are re-
sponsible for the unpleasant odor.
While the present results are in accord with
the previously observed attractant synergy be-
tween the Biolures and the propylene glycol, it
may be that the odor in the automotive formula-
tion detracts from this effect. The latter hypothe-
sis needs to be confirmed with further study, as
does the result indicating the improved perfor-
mance of the antifreeze without the automotive
components. In either case, even if the non-toxic
formulation is no better or worse in terms of at-
tractancy, there is no reason to accept the envi-
ronmental hazard and waste disposal problems
associated with use of the automotive formula-
tions with the safer more economical, household
product readily available.

ACKNOWLEDGMENTS

Ronay Riley and Paco Daniel were diligent in the op-
eration and servicing of the trap grids in Mexico. Celes-
tino Cervantes provided technical assistance. I thank
Jim Daitner, production manager at Superclean, for
product information. David Robacker and Hugh Con-
way provided critical reviews of the manuscript. Men-
tion of trade names or commercial products in this
article is solely for the purpose of providing specific in-
formation and does not imply recommendation or en-
dorsement by the U.S. Department of Agriculture.

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Florida Entomologist 91(2)


June 2008


LIFE HISTORY AND DAMAGE OF A NEW BARADINAE WEEVIL
(COLEOPTERA: CURCULIONIDAE) ON AMARYLLIS

NANCY D. EPSKY, THOMAS J. WEISSLING1, ALISON WALKER2, ALAN W. MEEROW AND ROBERT R. HEATH
Subtropical Horticultural Research Station, United States Department of Agriculture, Agricultural Research
Service, 13601 Old Cutler Road, Miami, FL 33158 U.S.A.

'Current address: Dept. of Entomology, University of Nebraska, Lincoln, NE

2Current address: Fairchild Tropical Botanical Garden, 10901 Old Cutler Rd., Miami, FL


ABSTRACT

A small Baradinae weevil that feeds on amaryllis plants has been known in Florida for over
15 years. It is yet to be named taxonomically and its life history has not been studied previ-
ously. Observations on weevil damage were made on containerized amaryllis (Hippeastrum
hybrids) plants naturally infested in a greenhouse or used for colony rearing. Laboratory
studies were conducted at ambient room temperature (75 C) with excised leaves to obtain
information on weevil life history. Adults lived about 3 months, and fed on basal versus api-
cal leaf tissue. Females inserted eggs near the thickened leaf base, and eggs were 0.65 0.02
mm long by 0.40 0.01 mm wide. Females laid >400 eggs over their lifetime, with egg pro-
duction increasing over the first 7 weeks and then tending to decline. Eclosion ranged from
51% for eggs removed from host tissue within 24 h to 84% for eggs removed from host tissue
after 24 h of oviposition. In tests with excised leaf tissue, eggs hatched after 7.1 d and larval
development was complete after 28.8 d, of which 9.9 d were spent as prepupae. In no-choice
tests, survival was lower and pupal developmental time period was longer when larvae were
reared on excised bulb versus excised leaf tissue. Although larval development was poorer
on bulbs versus leaves in the laboratory studies, in intact plants larvae tunnel through leaf
tissue towards the bulb where they feed and complete development. In severe infestations,
larvae hollow out the inside of the bulb and may cause plant death. Adult damage is prima-
rily to the foliage through feeding and oviposition. This is the first report to quantify the life
history of this weevil.

Key Words: Amaryllidaceae, oviposition, fertility, damage


RESUME

En Florida por mas de 15 aios se ha conocido un pequeno picudo (gorgojo) de la subfamilia
Baradinae que se alimenta sobre las plants de amarilis. Todavia no se le ha dado un nombre
taxon6mico y su ciclo de vida no ha sido estudiado anteriormente. Se hizo observaciones so-
bre el dano causado por el picudo en plants de amaryllis (Hippeastrum hybrids) en recipien-
tes infestadas naturalmente en un invernadero o plants usadas para criar la colonia. Se
realizaron studios de laboratorio a la temperature ambiental del cuarto (75 C) con hojas
cortadas para obtener informaci6n sobre el ciclo de la vida del picudo. Los adults vivieron
aproximadamente 3 meses, y se alimentaron sobre el tejido basal versus el tejido apical de
la hoja. Las hembras insertaron los huevos cerca de la base engruesada de la hoja, y los hue-
vos fueron 0.65 0.02 mm de largo por 0.40 0.01 mm de ancho. Las hembras pusieron >
400 huevos por su ciclo de vida, con un aumento en la producci6n de huevos en las primeras
7 semanas y luego tendiendo a bajar. El rango de eclosi6n fue desde el 51% para los huevos
quitados del tejido del hospedero en el rango de 24 horas, hasta 84% para los huevos quita-
dos del tejido del hospedero 24 horas despu6s de la oviposici6n. En pruebas con tejido de ho-
jas cortadas, los huevos se eclosionaron despu6s de 7.1 dias y el desarrollo de larva fue
complete despu6s de 28.8 dias, de la cual 9.9 dias pasaron como prepupas. En pruebas de no-
opci6n, la supervivencia fue mas baja y el period del tiempo del desarrollo de la pupa fue
mas largo cuando las larvas fueron criadas en bulbos cortados versus tejidos de una hoja cor-
tada. Aunque el desarrollo de larvas fue pobre en bulbos versus en las hojas en los studios
del laboratorio, en plants intactas las larvas hacen tuneles por el tejido de la hoja hacia el
bulbo donde se alimentan y completan su desarrollo. En infestaciones several, las larvas ha-
cen un hueco adentro del bulbo y pueden causar la muerte de la plant. El dano hecho por
el adulto es principalmente al follaje por su alimentaci6n y oviposici6n. Este es el primer in-
forme para cuantificar la historic de vida de este picudo.







Epsky et al.: Baradinae Weevil on Amaryllis


Weevils in the subfamily Baridinae have been
described as difficult to characterize taxonomi-
cally and little is known about many of the spe-
cies outside of their original descriptions (Ander-
son 2002). Of the species that have been studied,
larvae tend to bore in flowers, petioles, stems, and
roots of herbaceous dicots, and some infest palm
fruits, grasses, and other monocots (Marvaldi
2003). Several Baridinae weevils have been iden-
tified as pests or potential pest. For example,
Palmelampius heinrichi O'Brien, is a pest of fruit
of the palm Bactris gasipaes H.B.K. in South
America (O'Brien & Kovarik 2000). Madarellus
undulatus (Say),Ampeloglypter ater (Riley),A. se-
sostris (LeConte) and Desmoglyptus crenatus (Le-
Conte) are weevil species known to feed on vines
of the genus Vitis (Vitaceae) in North America
(Bouchard et al. 2005). Stethobaris ovata (Le-
Conte) is a pest of native orchids (Orchidaceae) in
North America (Dunford et al. 2006). Others are
considered beneficial insects as palm pollinators
(Barfod & Uhl 2001) or as weed biological control
agents (Horner 2003).
In the early 1990s, a 5-mm long, solid black
weevil was observed feeding on and occasionally
killing amaryllis (Amaryllidaceae) plants in Flor-
ida, and it was determined to be an unknown ge-
nus and species in the subfamily Baridinae (Tho-
mas 2005). Reported host plants include amaryl-
lis Hippeastrum Herb. spp., spider lily Hymeno-
callis Salisb. spp., swamp lily Crinum L. spp., and
Amazon lily Eucharis x grandiflora Planch. &
Linden (Thomas 2005). In Sep 2005, we sent out
surveys to 30 amaryllis growers/distributors in
the southeast US, including 26 in Florida, 2 in
Georgia, and 1 each in Virginia and West Vir-
ginia. The survey included background informa-
tion, photos of adult weevils and damage to ama-
ryllis plants, and a questionnaire that included
the following questions on the new weevil: Have
they seen it, heard of it, or observed similar dam-
age? What are their growing conditions and pest
control practices? One Florida grower reported
damage but no insects, 1 Florida grower reported
weevils and damage on Hymenocallis spp., and 1
Florida distributor indicated that they had been
contacted by customers about insect damage to
Hippeastrum. Since Nov 2005, extension person-
nel in Florida and in Georgia have been contacted
by a landscaper and a homeowner, respectively,
about insect damage to Hymenocallis spp. and
Hippeastrum spp., respectively, presumably due
to this weevil (N.D.E., unpublished data).
The USDA/ARS, Subtropical Horticulture Re-
search Station (SHRS) located in Miami, FL, has
a Hippeastrum hybridization program, and plants
at the station have been subject to attack by this
weevil. Because little was known about this in-
sect, studies were initiated to quantify aspects of
weevil life history including developmental time,
adult longevity, oviposition, and fecundity.


MATERIALS AND METHODS

Insects

The insects used in this study were obtained
from a colony initiated from naturally infested
containerized amaryllis (Hippeastrum hybrids)
grown in a greenhouse at the USDA-ARS station
in Miami, FL. The colony was maintained on con-
tainerized amaryllis plants held in 3 screened en-
closures (1.5 m wide by 1.2 m deep by 2.3 m tall).
The screened enclosures were attached to a build-
ing on one side and had a roof that gave some pro-
tection to rain and direct sunlight, but the enclo-
sures were exposed to naturally fluctuating tem-
perature and relative humidity. To initiate the
colony and to augment the colony with wild stock
periodically, adults (Fig. 1A) that had been col-
lected by hand with a manual aspirator and in-
fested amaryllis plants (Fig. 1B, C) from the
greenhouse were added to the enclosures. Un-in-
fested amaryllis plants in 3.8- and 7.6-L pots were
added as needed to maintain active infestations.
When needed for experiments, adults were col-
lected by hand with a manual aspirator. To obtain
adults of known age, soil within plant culture con-
tainers was sifted and pupae were collected. Pu-
pae were placed individually in glass vials (10
mm diam. by 55 mm) and vials were filled half
way with moistened sand. Vials were checked
daily and adult emergence date was recorded. Ex-
periments on weevil life history were all con-
ducted under laboratory conditions under ambi-
ent temperature (75C) and relative humidity.
Experiments were conducted in rooms that had
windows to provide natural lighting and that
were supplemented with room lights set to a pho-
toperiod of 12:12 (L:D) h.

Feeding Location

Preferred feeding location was determined
from choice tests. Three-cm long basal and apical
pieces of leaf tissue were added to a large Petri
dish (100 x 15 mm) lined with water-moistened
filter paper. Two mated adults were added to each
arena, for a total of 10 replicates. Feeding damage
and frass production were determined after 24 hr.
Damage was reported as percent of total feeding
on either the basal or apical piece in each arena.

Oviposition

Recently emerged adults (<7 d old) were set up
in Petri dishes with moist filter paper and excised
leaf tissue. Mixed sex adults were held together
for 48 h to provide sufficient time for mating. Af-
ter 48 h, individual weevils were placed in small
Petri dishes (60 x 15 mm) lined with moist filter
paper and were provided with a piece (2.54 cm) of
basal amaryllis leaf. Adults that did not produce







Florida Entomologist 91(2)


Fig. 1. (A) Adult weevil finding harborage between leaf bases on an amaryllis plant. Dark brown streaks on ad-
jacent leaves are typically of damage due to adult feeding and oviposition activities. Amaryllis plants showing signs
of (B) light, (C) moderate and (D) heavy foliar damage due to adult weevil feeding and oviposition activity.


eggs within 2 weeks were discarded from the
study. Every 2-5 d, the leaf piece was removed and
carefully checked for eggs, and another basal leaf
piece was added. Sampling was continued until
the female died. Leaves were examined under a
stereomicroscope, the numbers of egg clutches
and the total numbers of eggs were recorded. Egg
production data were collected from 12 females.
In a separate experiment, 2 or 3 leaves from
potted plants in cages containing weevils were
collected and dissected. The distance eggs were
laid from the bulb was recorded, and egg length
and width were measured under a stereomicro-
scope. There were 7 replicate collections.

Egg Viability

Eggs that had been oviposited within a 48-h
time period were collected by dissecting leaf tis-
sue under a stereomicroscope and egg viability
was determined from percentage hatch. Eggs
were placed either on the surface of an excised
piece of leaf tissue or on moist filter paper in a


small Petri dish. There were 12 replicates of sets
of 10 eggs per dish.

Developmental Time Period

Weevil developmental time period was deter-
mined for individuals reared on amaryllis foliage
in experiment one. In experiment 2, developmen-
tal time period for individuals reared on amaryllis
foliage was compared to that of individuals reared
on bulb tissue (2.54 cm3). Eggs of known age were
obtained by removing leaf pieces that had been
placed with mixed sex adults for 24 h. Eggs were
dissected from the leaf tissue and placed individ-
ually in plastic cups (12 mL) on either moistened
filter paper or on foliage (experiment 1), or on ei-
ther foliage or excised bulb (experiment 2). For
experiment 1, eggs were dissected from the leaf
tissue within 24 h of oviposition. Additional neo-
nates were obtained from eggs of unknown age
that were placed in small Petri dishes lined with
moistened filter paper. Because of lower percent-
age hatch and potential damage in dissecting


June 2008







Epsky et al.: Baradinae Weevil on Amaryllis


eggs too soon after oviposition (see Results) in ex-
periment 1, eggs were dissected from leaf tissue at
least 24 h after oviposition for experiment 2. After
hatch, neonates (with leaf or bulb) were moved in-
dividually into plastic cups with a layer of vermic-
ulite (0.5-1 cm). Neonates from dishes with moist-
ened filter paper were transferred to an excised
piece of leaf tissue. Cups were checked daily, leaf
and bulb tissue replaced as needed, and date of
death, hatch, prepupal appearance, pupation, and
adult emergence were recorded. Descriptive sta-
tistics are presented as means and standard devi-
ations. Two sample t-tests from Proc TTEST (SAS
Institute 2000) were used for comparison of devel-
opmental time periods on leaf versus bulb tissue
in experiment 2.

RESULTS AND DISCUSSION

Feeding Location

In the field, adults were observed spending
considerable time seeking harborage and appar-
ently feeding on the leaf bases (Fig. 1, T.J.W. per-
sonal observation). Adults were observed tunnel-
ing into the leaf base, and abandoned galleries
were common in older leaves. Less frequently,
adults were found feeding on the surface of bulbs
just below the soil level. Results of the laboratory
choice test indicate a strong preference for adult
feeding on basal versus apical leaf tissue (97.0% +
3.0 and 3.0% + 3.0, respectively). Feeding was
confirmed by visual observation and the presence
of frass in the arenas. Preference for basal leaf tis-
sue could be due to several factors including dif-
ferences in nutrition and/or tissue quality. In ad-
dition, basal sections of amaryllis leaves are thick
(2.7 0.8 mm, n = 20) while apical pieces are thin
(0.6 0.1 mm, n = 20). Thicker tissue provides
more opportunity for harborage and weevils
readily bore into the thicker leaf bases.

Oviposition

After pairing recently emerged males and fe-
males, it took as few as 5 d for females to begin
laying eggs. Females inserted eggs into the tissue


o FemaEAs 12
S- *5 ..... ./\ A -o
o . .. '
E 4

12 a2
I i * *> \




1 3 5 7 9e 1 13 15 17 10 21 23
Weeks after mating
Fig. 2. Mean number of eggs per day per female
(black squares, solid line) in excised amaryllis leaf tis-
sue and survival (open diamond, dotted line) of female
weevils over time (weeks).


near the thickened leaf base. Average diameter of
oviposition holes measured on the leaf surface
was 0.2 mm and eggs measured 0.65 0.02 mm
long by 0.40 0.01 mm wide. Females lived 89.0
38.8 d (range 21-160 d) and laid 441.1 241.6
eggs (range 127-821 eggs). Eggs were laid 27.4
4.9 mm (range 5-65 mm) from the leaf base.
Mating during the 48 h that females were held
with males resulted in the transfer of adequate
quantities of sperm to fertilize eggs without sub-
sequent mating. The number of eggs laid per day
increased steadily through the first 7 wk (Fig. 2).
In general, egg production per female and female
survival steadily declined after the first 7 wk,
with only three females remaining alive by 18 wk
and all females dead by 23 wk (Fig. 2). Eggs were
laid in clutches, averaging 2.7 0.7 eggs/clutch
(range 1-5) and 2.7 1.6 clutches/day (range 0.3-
7). Percentage hatch averaged 83.6 67.3%
(range 0-97.2).

Developmental Time Period

Data on developmental time period and sur-
vival were obtained from 213 eggs of known age
and 83 eggs of unknown age, for a total of 296 eggs
evaluated (Table 1). The lower percentage hatch
(51.2% survival) obtained from these eggs com-


TABLE 1. WITHIN STAGE SURVIVAL AND DEVELOPMENTAL TIME (DAYS) OF A BARIDINAE WEEVIL REARED IN THE LABO-
RATORY ON EXCISED AMARYLLIS LEAF TISSUE.

Days per stage
Within stage
Stage Survival (%) n* Mean Std Dev Min Max

Egg** 51.2 104 7.1 1.19 5 12
Larvae 74.0 43 28.8 2.75 20 35
Pupa 94.3 17 14.2 1.47 12 17

*Number of individuals from which developmental time data was obtained.
**Eggs were dissected from amaryllis tissue within 24 h of oviposition.







Florida Entomologist 91(2)


Fig. 3. Weevil larvae and heavy damage to an ama-
ryllis bulb due to a high level of larval infestation.


pared with that obtained in the oviposition study
(above) may be because eggs were dissected from
the plant tissue within 24 h of oviposition (exper-
iment 1) versus between 24 and 48 h of oviposition
(oviposition study), respectively. Total time period
from oviposition to adult eclosion averaged 47.4 +
3.7 d. Larvae stayed within the leaf tissue so num-
ber of instars was not determined. Late instars
exited the tissue, moved into the vermiculite and
became non-feeding prepupae. Of the larval de-
velopmental time, 9.9 + 2.9 d (range 3-16 d) were
spent as prepupae. Newly closed, general adults
were light brown in color and did not feed. It took
an additional 3.8 1.1 d (range 1-6 d) for adults to
become solid black and begin feeding.
When neonates hatched from eggs placed di-
rectly on the cut edge of leaf tissue, they immedi-
ately burrowed into and fed within the leaf paran-
chyma tissue. As the old tissue was consumed,
new leaf tissue was added to the cups and larvae
readily moved into the new leaf. Except for move-
ment to new leaves, larvae remained in the leaf
until exiting and becoming prepupae. In our labo-
ratory colony, however, larvae are often recovered
from bulbs of containerized amaryllis plants used
for rearing. Experiment 2 compared developmen-
tal time periods for larvae on bulb tissue versus
foliage. There was no difference between larval
developmental time period for larvae reared on


foliage versus bulb tissue (32.8 2.8 d versus 31.3
+ 2.2 d, respectively; t = 1.28, df = 20, P = 0.2139).
However, more larvae survived to the pupal stage
on foliage versus bulb tissue, 17 of 31 (55%) ver-
sus 5 of 26 (19%), respectively. Pupal developmen-
tal time period was shorter for foliage-reared ver-
sus bulb-reared larvae (17.0 1.7 versus 22.3
3.1, respectively; t = 4.31, df = 15, P = 0.0005).
There was 82% eclosion (14 of 17) for pupae from
foliage-reared larvae versus 60% eclosion (3 of 5)
for pupae from bulb-reared larvae.
Adults primarily damage amaryllis foliage
through feeding and oviposition activities (Fig.
1B). However, they will tunnel through leaves
and on occasion feed on the outside of bulbs (Fig.
1C, D). Eggs are laid in the leaf tissue, early in-
star larvae tunnel through leaf tissue towards the
bulb where they feed and develop. If infestation
level is high enough, as we have observed in some
greenhouse-grown amaryllis or in plants used for
rearing weevils for this study, larvae can severely
damage the bulb (Fig. 3). Upon reaching maturity,
larvae exit the bulb and enter the soil to pupate
(Fig. 4). Under laboratory no-choice conditions,
larvae completed their development on excised
leaf tissue and on bulb tissue, but were more suc-
cessful on leaf tissue. Presumably larvae feeding
on intact plants could choose among leaf and/or
bulb tissue, which may increase survival and de-
crease developmental time period obtained in our
studies.
Due to their cryptic nature, infestations of am-
aryllis bulbs by weevils are difficult to determine
until host injury is expressed. Based on develop-
mental times, several generations a year are pos-
sible in south Florida. The host range is unknown
at this time but subsequent studies on host plant
preference by the weevil will result in better
choices for plant culture in regions with pest infes-
tations. Some varieties appear to be attacked
more often or are more susceptible to weevil infes-
tation (A.W.M., unpublished data). The emphasis
of commercial breeding for improved Hippeastrum
hybrids has been on large flower size and other fa-
vorable properties such as long-lasting flowers
with an unusual color range (Meerow 2000). Iden-
tification of weevil resistant varieties would be an
important tool for integrated pest management
and control of this new amaryllis pest.

ACKNOWLEDGMENTS

We thank Micah Gill, Pauline Andersen, Wayne
Montgomery, Carol Lee, Ingris Filpo, Karen Regas, and
Jeffrey Tefel (USDA/ARS, Miami, FL) for technical as-
sistance; Adrian Hunsberger (Univ. of FL, Miami-Dade
County Extension, Homestead), Lisa Ames (Univ. of
Georgia, Homeowner Insect & Weed Diag. Lab., Griffin),
for discussion of calls regarding insect damage to Ama-
ryllidaceae and, in addition Russell F. Mizell, III (Univ.
of FL, Quincy), and Robert Wright (Univ. of Nebraska-
Lincoln), for comments on an earlier version of this


June 2008







Epsky et al.: Baradinae Weevil on Amaryllis


Fig. 4. Weevil pupae that exited the amaryllis bulb as larvae and pupated in the soil.


manuscript. Voucher specimen have been placed with
Dr. C. O'Brien (FL A & M Univ., Tallahassee) and are
held at the USDA/ARS, Miami, FL. This study was par-
tially supported by a grant from The Fred C. Gloeckner
Foundation, Inc. (T.J.W. & A.W.M.). This article reports
the results of research only. Mention of a proprietary
product does not constitute an endorsement or recom-
mendation by the USDA for its use.

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Florida Entomologist 91(2)


June 2008


NATURAL OCURRENCE OF HYMENOPTEROUS PARASITOIDS ASSOCIATED
WITH ANASTREPHA FRATERCULUS (DIPTERA: TEPHRITIDAE) IN
MYRTACEAE SPECIES IN ENTIRE RIOS, NORTHEASTERN ARGENTINA


SERGIO M. OVRUSKI, PABLO SCHLISERMAN, LUIS E. ORONO, SEGUNDO R. NUNIZ-CAMPERO,
PATRICIA ALBORNOZ-MEDINA, LAURA P. BEZDJIAN AND GUIDO A. VAN NIEUWENHOVE
PROIMI Biotecnologia CONICET, Divisi6n Control Biol6gico de Plagas, Laboratorio Moscas de La Fruta.
Avda. Belgrano y Pje. Caseros s/n (T4001MVB) San Miguel de Tucuman, Argentina

ABSTRACT

Parasitoids ofAnastrepha fraterculus (Wiedemann) were monitored on ripe fruit of 3 native and
1 exotic, wild Myrtaceae species in the Province of Entre Rios, NE Argentina, between Jan and
Mar 1993 and 1994 with the aim of identifying indigenous parasitoid species and determining
natural parasitization rates and fruit infestation levels. The fruit species surveyed were Psidium
guajava L. (common guava), Feijoa sellowiana (0. Berg) O. Berg (feijoa), Eugenia uniflora L.
(Surinam cherry), and Myrcianthes pungens (Berg) Legrand (mato). Altogether 2,186 tephritid
puparia were obtained, 95% of which were A. fraterculus and 5% of which were Ceratitis capi-
tata (Wiedemann). Of 1,667 adult insects that emerged from these puparia, 1,378 wereA. frater-
culus, 89 C. capitata, and 200 larval-pupal parasitoids, representing 4 species of 2 Hymenoptera
families: Doryctobracon areolatus (Sz6pligeti), D. brasiliensis (Sz6pligeti), Utetes anastrephae
(Viereck) (all Braconidae, Opiinae), and Aganaspis pelleranoi (Brethes) (Figitidae, Eucoilinae).
All these parasitoid species are new reports for Entre Rios. Moreover, these records represent the
southernmost natural distribution range in the Americas for these species. Doryctobracon are-
olatus and A. pelleranoi were recovered from all of the Myrtaceae species sampled, and they
were the most abundant parasitoid species. Infestation patterns byA. fraterculus in mato, Suri-
nam cherry, guava, and feijoa varied from 15.2 to 41.8, 21.3 to 49.4, 34.1 to 109.2, and 78.9 to
140.6 larvae per kg of fruit, respectively. Highest levels of parasitism were recorded in P. gua-
java, whereas M. pungens had the lowest parasitization rates. However, overall mean parasit-
ism levels (i.e., considering all parasitoid species) did not appear to have great differences when
comparing Myrtaceae species, collection sites, and years. The relative abundance and parasiti-
zation rates data of the recovered parasitoids in the 4 Myrtaceae species suggest some degree of
host plant preference by U anastrephae and D. brasiliensis.

Key Words: fruit flies, parasitoids, Braconidae, Figitidae, Argentina, Myrtaceae

RESUME

Con el prop6sito de identificar species de parasitoides asociadas con Anastrepha fraterculus
(Wiedemann) y determinar niveles de parasitismo natural y de infestaci6n en fruta en la provin-
cia de Entre Rios, Noreste de Argentina, se colectaron tres species nativas y una ex6tica de Myr-
taceae entire Enero y Marzo de 1993 y de 1994. Las species frutales colectadas fueron Psidium
guajava L. (guayaba comun), Feijoa sellowiana (0. Berg) 0. Berg (feijoa), Eugenia uniflora L.
(arrayan), y Myrcianthes pungens (Berg) Legrand (mato). Se obtuvo un total de 2.186 puparios,
de los cuales el 95% correspondi6 aA. fraterculus y el 5% restante a Ceratitis capitata (Wiede-
mann). De los 1.667 insects adults que emergieron de estos puparios, 1.378 fueron A. frater-
culus, 89 C. capitata y 200 fueron parasitoides larvo-pupales, representando cuatro species de
dos families de Hymenoptera: Doryctobracon areolatus (Sz6pligeti), D. brasiliensis (Sz6pligeti),
Utetes anastrephae (Viereck) (todos Braconidae, Opiinae), yAganaspis pelleranoi (Brethes) (Fi-
gitidae, Eucoilinae). Todas estas species de parasitoides son nuevas citas para la provincia de
Entre Rios. Ademas, estos registros representan el rango natural de distribuci6n mas austral en
el continent americano para estas cuatro species de parasitoides. Doryctobracon areolatus yA.
pelleranoi fueron recuperados de todas las species de Myrtaceae colectadas, y fueron las espe-
cies de parasitoides mas abundantes. Los niveles de infestaci6n porA. fraterculus en mato, arra-
yan, guayaba, y feijoa variaron de 15.2 a 41.8, 21.3 49.4, 34.1 109.2, y 78.9 140.6 larvas por
kg de fruta, respectivamente. Sin embargo, los valores medios totales de parasitismo (incluyendo
todas las species de parasitoides) no presentaron grandes diferencias cuando se consideraron
las distintas species de Myrtaceae, lugares de colecta, y afios de muestreo. Los datos sobre
abundancia relative de parasitoides y las tasas de parasitismo en las cuatro species de Myrta-
ceae sugieren cierto grado de preferencia por la plant hospedera por parte de los parasitoides
U anastrephae y D. brasiliensis.


Translation provided by the authors.







Ovruski et al.: Parasitoids ofAnastrepha fraterculus in Entre Rios


The South American fruit fly, Anastrepha
fraterculus (Wiedemann), and the Mediterranean
fruit fly, Ceratitis capitata (Wiedemann), are
among the most significant insect pests of edible
fruit in Argentina (Spinetta 2004) and in the re-
maining South American countries (Malavasi et
al. 2000; Aluja et al. 2003a). The native A. frater-
culus is basically restricted to NE and NW Argen-
tina, where the climate mainly is warm and hu-
mid, whereas the exotic C. capitata is currently
distributed throughout all fruit-growing regions
of the country (Ovruski et al. 2003; Segura et al.
2006). Although A. fraterculus represents a cryp-
tic species ensemble (Steck 1991; Aluja et al.
2003a; Hernandez-Ortiz et al. 2004; Vera et al.
2006) distributed throughout continental Amer-
ica from Mexico to Argentina (Aluja 1999; Norr-
bom 2004), the Argentine populations of the com-
plex belong to a single biological species (Alberti
et al. 2002).
In the northeastern province of Entre Rios,
where A. fraterculus and C. capitata coexist in
large citrus crop areas (Segura et al. 2006), com-
mon guavas grown in thin patches of wild vegeta-
tion adjacent to commercial orchards support
larger local A. fraterculus populations (Turica &
Mallo 1961; Putruele 1996). Though A. fratercu-
lus has one of the broadest host ranges of all
known Anastrepha species (Norrbom 2004), fruit
in the Myrtaceae family are among its favored
host plant (Aluja 1999; Raga et al. 2005).
Poisoned bait sprays for A. fraterculus and C.
capitata control were tried with some success as
early as 1930 in Entre Rios (Ovruski & Fidalgo
1994). Some attempts to develop biological con-
trol programs were made in the 1940s and the
1960s. Thus, 3 exotic larval parasitoid species
(Tetrastichus giffardianus Silvestri, Acerato-
neuromyia indica (Silvestri), and Diachasmimor-
pha longicaudata (Ashmead)) were introduced
and released in limited numbers in citrus crop ar-
eas of Entre Rios (Ovruski et al. 1999). Tetrasti-
chus giffardianus and A. indica are 2 gregarious
eulophid parasitoids originally collected in West
Africa and Southeast Asia, respectively, whereas
D. longicaudata is a solitary braconid parasitoid
originally from the Malaysia-Philippine region
(Ovruski et al. 2000). OnlyA. indica and D. longi-
caudata were recovered (Turica 1968).
Studies on native parasitoids attacking te-
phritid pests in northeastern Argentina have been
largely neglected. Examples of fruit fly parasitoid
surveys are only those developed in Misiones
(Ogloblin 1937) and Corrientes (Turica & Mallo
1961; Ovruski & Schliserman 2003a). These stud-
ies were focused largely on commercial fruit in cit-
rus growing areas. Some parasitoid species also
were reported from fruit samples sporadically col-
lected in Entre Rios (Blanchard 1947) and Corri-
entes (Brethes 1924; Blanchard 1966). Most of the
available new information on native fruit fly par-


asitoids of Argentina is based on the surveys car-
ried out in NW Argentina (Turica & Mallo 1961;
Nasca 1973; Fernandez de Araoz & Nasca 1984;
Ovruski 1995; Wharton et al. 1998; Ovruski &
Schliserman, 2003b; Schliserman et al. 2004;
Schliserman & Ovruski, 2004; Ovruski et al. 2004,
2005, 2006; Orofo et al. 2005). As many as 11 na-
tive parasitoid species associated withA. fratercu-
lus on native and exotic host fruit species have
been so far recorded in Argentina (Ovruski et al.
2005). Most of them also have been found parasit-
izing diverse Anastrepha species in several Latin
American countries, including Brazil (Leonel et al.
1995; Aguiar-Menezes & Menezes 1997; Canal &
Zucchi 2000; Guimaraes et al. 2000; Carvalho
2001; Aguiar-Menezes et al. 2001; Uch6a-
Fernandes et al. 2003), Venezuela (Katiyar et al.
1995), Colombia (Yepes and Velez 1989; Carrejo &
Gonzalez 1999), Costa Rica (Jir6n & Mexzon 1989;
Wharton et al. 1981), Guatemala (Eskafi 1990),
and Mexico (Aluja et al. 1998, 2003b; L6pez et al.
1999; Sivinski et al. 1997, 2000, 2001; Guillen et
al. 2002; Hernandez-Ortiz et al. 1994, 2006).
The specific aims of this study were to survey
selected wild Myrtaceae species commonly in-
fested by A. fraterculus in order to provide infor-
mation on infestation levels in the fruit sampled,
degree of larval parasitization, and the parasitoid
fauna attacking this fruit fly pest in Entre Rios,
NE Argentina, as well as to document natural dis-
tribution ranges of these parasitoids along a lati-
tudinal gradient. This article complements our
previous study on the diversity and distribution
of native fruit fly parasitoid in the extremely en-
dangered subtropical rainforests in north Argen-
tina (Ovruski et al. 2004, 2005).

MATERIALS AND METHODS

Collections were made during Jan-Mar 1993
and 1994 in the following localities of Entre Rios,
NE Argentina: Parand (3164'S, 6032'W, eleva-
tion 90 m), La Paz (3044'S, 59038'W, 75 m), and
Concordia (3124'S, 5801'W, 48 m). The climate is
characterized as temperate-humid with long and
warm summers, the temperature of the hottest
month being >22C and the temperature of the
coldest month being <180C; it rains all year long,
but the proportion of winter rainfall is <5% of the
total, the rainfall being between 933 mm (Cakf
(w)) (Anonymous 1992).
Fruit samples included 3 native and 1 exotic,
wild species of Myrtaceae: Psidium guajava L.
(common guava) (exotic species), Feijoa sellowi-
ana (0. Berg) O. Berg (feijoa), Eugenia uniflora L.
(Surinam cherry), and Myrcianthes pungens
(Berg) Legrand (mato) (all native species). Fruit
were collected in backyard gardens in suburban
areas and patches covered with wild native vege-
tation adjacent to small citrus orchards (sour and
sweet oranges, tangerines, lime, sweet lemon, and








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grapefruit). All these fruit sampling sites were lo-
cated in a linear corridor along the banks of
Parand River (West sector of the NE Argentina re-
gion), except for the study site in Concordia, which
was located close to the banks of Uruguay River
(East sector of the NE Argentina region). The orig-
inal native vegetation was subtropical rainforest
locally known as "Selva en Galerfa", which contin-
ued along the banks of southern and northern
Parand and Uruguay River (Cabrera 1976).
Based on fruit size (measured as individual
fruit weight) of each plant species, approximately
10 fruit were randomly taken from each guava
and feijoa tree, and 35 fruit from each Surinam
cherry and mato tree. Guava and feijoa plants had
medium size fruit (mean SD: 45.8 4.9 g and
39.2 5.8 g, n = 100, respectively) while Surinam
cherry and mato plants had small size fruit (mean
SD: 6.8 2.3 g, and 7.2 5.8 g, n = 100, respec-
tively). Fruit was picked both from the tree and
from the ground under the tree canopy. Therefore,
each sample included fruit collected from tree plus
fruit of the same tree that had fallen on the
ground. Only ripe fruit that was about to fall from
the tree was harvested. This allowed fruit fly lar-
vae to complete development, and gave fruit fly
parasitoids the opportunity to parasitize larvae
throughout their development (Lopez et al. 1999).
Fruit samples were placed in styrofoam boxes
with sand in the bottom as pupation medium for
larvae, and they were taken to the laboratory. All
material collected in the field was processed in the
laboratory of the CIRPON institute (Centro de In-
vestigaciones para la Regulaci6n de Poblaciones
de Organismos Nocivos) in San Miguel de Tu-
cuman (2650'S, 6513'W, elevation 426 m), Tu-
cuman, Northwestern Argentina. Each styrofoam
box contained only one fruit sample and all cages
were kept inside a room at 27 2C and 60 10%
relative humidity. Fruit fly puparia were recov-
ered weekly during 1 month and then transferred
to plastic trays containing sterilized humid sand,
which was re-moistened every 3 d until all sam-
ples had been processed. Each tray was then
placed inside a sealed wooden box. All wooden
cages were kept inside a rearing room at 25 1C
and 75 5% relative humidity for 4 months. Pu-
paria of C. capitata and Anastrepha Schiner were
separated based on pupal characters (White & El-
son-Harris 1992). Emerged flies and/or parasi-
toids were identified, counted and sexed by sam-
ple and summarized for each locality. Afterward,
unemerged puparia also were counted. Additional
samples of parasitoid specimens collected in El
Palmar (3216'S, 5828'W, 44 m) and Concordia lo-
calities were received from colleagues.
Parasitoid specimens were identified to species
by S. Ovruski using Wharton and Marsh's (1978)
key and the taxonomic descriptions by Wharton et
al. (1998). Fruit flies were identified by S. Ovruski
using Zucchi's (2000) taxonomic key. Nomencla-


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Ovruski et al.: Parasitoids ofAnastrepha fraterculus in Entre Rios


ture for the Opiinae follows Wharton (1997) and
for the Eucoilinae Wharton et al. (1998). Parasi-
toid and fly specimens were placed in the entomo-
logical collection of the Fundaci6n Miguel Lillo
(FML) (San Miguel de Tucuman, Argentina). All
plant samples were compared to herbarium spec-
imens at the FML, and identified by Alejandra
Roldan (Facultad de Ciencias Naturales e Insti-
tuto Miguel Lillo, Universidad Nacional de Tu-
cuman). Nomenclature employed for plant identi-
fication was based on Morales et al. (1995).
All parasitism values and fruit infestation lev-
els reported here are based on the number of
emerging adult flies and parasitoids (Lopez et al.
1999), and on the number of fruit fly larvae per kg
of fruit (Aluja et al. 2000), respectively. Where ap-
propriate, means and standard deviations (mean
+ SD) were calculated as summary statistics for
the parasitism percentage, fruit size (weight),
and ovipositor length of opiine parasitoid species.

RESULTS

A total of 1,532 fruit, representing 33.1 kg was
processed during this study. Out of these, 420 (3.1
kg) fruit were E. uniflora, 310 (3.2 kg) M. pun-
gens, 615 (23.8 kg) P. guajava, and 107 (3.0 kg) F
sellowiana. In total, 2,186 tephritid puparia were
obtained from these fruit, 95% of which were A.
fraterculus and 5% of which were C. capitata. Of
1,667 adult insects that emerged from these pu-
paria, 1,378 were A. fraterculus, 89 C. capitata,
and 200 parasitoids, representing 4 species of 2
Hymenoptera families. No parasitoids were re-
covered from C. capitata puparia.
Only E. uniflora and P guajava were infested
by C. capitata, and infestation rates ranged from
0.1 to 1.9 and from 4.3 to 15.2 larvae/kg of fruit, re-
spectively. Approximately 17% and 33% of all
Surinam cherry and guava samples were simulta-
neously infested by both C. capitata and A. frater-
culus. Infestation patterns byA. fraterculus in M.
pungens, E. uniflora, P guajava, and F sellowiana
varied from 15.2 to 41.8, 21.3 to 49.4, 34.1 to 109.2,
and 78.9 to 140.6 larvae/kg of fruit, respectively.
Table 1 summarizesA. fraterculus and parasi-
toid species abundance, and parasitization rates
based on fruit species collected per study site and
year. Three species of opiine Braconidae parasi-
toids and 1 species of eucoiline Figitidae parasi-
toid were identified: Doryctobracon areolatus
(Szepligeti), D. brasiliensis (Szepligeti), Utetes
anastrephae (Viereck) (Braconidae), and Aganas-
pis pelleranoi (Brethes) (Figitidae). These species
constituted 45.4, 14.0, 12.5, and 28.1% of the total
recovered parasitoids, respectively. Doryctobra-
con areolatus was the most abundant parasitoid
species in 71% of total fruit samples and it was re-
covered from all of the Myrtaceae species sam-
pled.Aganaspis pelleranoi was also found in asso-
ciation with A. fraterculus in E. uniflora, F sell-


owiana, M. pungens and P. guajava. This eucoil-
ine parasitoid was recovered from 52% of the total
fruit samples. Doryctobracon brasiliensis was re-
covered only from 33 and 50% of the total guava
and feijoa samples, respectively. Approximately
76% of all U. anastrephae specimens was recov-
ered from both M. pungens and E. uniflora. This
opiine species was obtained from 8, 20, 67, and
100% of all guava, feijoa, mato, and Surinam
cherry samples, respectively.
Highest levels of parasitism were recorded in
P guajava, whereas M. pungens had the lowest
parasitization rates (Table 1). However, overall
mean parasitism levels (i.e., considering all para-
sitoid species) did not appear to have great differ-
ences when comparing Myrtaceae species, collec-
tion sites, and year (Table 1). Parasitization byA.
pelleranoi has accounted for only <10% of the to-
tal parasitization ofA. fraterculus on F sellowi-
ana, M. pungens and E. uniflora. In contrast, in P.
guajava the degree of larval parasitization by eu-
coiline species ranged from 24 to 73% of the total
parasitization.
From the additional parasitoid specimens col-
lected by colleagues in Entre Rios, 9A. pelleranoi
and 8 D. areolatus were identified. Six D. areola-
tus were recovered from guavas collected in El
Palmar, whereas the remaining 2 D. areolatus
and 9 A. pelleranoi were obtained from guavas in
Concordia.

DISCUSSION

New fruit fly parasitoid species records for En-
tre Rios are D. areolatus, D. brasiliensis, U. anas-
trephae andA. pelleranoi, all native of the Neotro-
pical region. Prior to this report, only the eucoil-
ine Rhoptromeris haywardi (Blanchard) had been
recorded from both C. capitata and A. fraterculus
in Entre Rios (Blanchard 1947). All of these spe-
cies belong to the fruit fly parasitoid guild num-
ber 2 described by Ovruski et al. (2000), which is
characterized by solitary, koinobiont larval-pupal
endoparasitoids ofAnastrepha spp. The braconids
D. areolatus and U. anastrephae, and the figitidA.
pelleranoi are widely distributed in Latin Ameri-
can, while D. brasiliensis is known to occur in
southern Brazil and northern Argentina (Ovruski
et al. 2005). The discovery of these 4 native A.
fraterculus-parasitoid species in Entre Rios rep-
resents their southernmost natural distribution
range in the Americas. During this study, D. bra-
siliensis and U. anastrephae were found at
3164'S latitude (Parand), whereas both D. are-
olatus and A. pelleranoi were also collected at
3216'S latitude (El Palmar). Previously, these
parasitoid species had been recorded from A.
fraterculus in the provinces of Corrientes (27015'-
3043'S and 59041'-5612'W) (Ovruski & Schliser-
man 2003a), Misiones (2530'-2810'S and 5338'-
56003'W) (Ogloblin 1937; Turica & Mallo 1961),







Florida Entomologist 91(2)


Catamarca (2509'-2916'S and 6525'-69006'W)
(Ovruski & Schliserman 2003b), Tucuman
(2605'-2801'S and 6428'-6613'W) (Nasca 1973;
Fernandez de Araoz & Nasca 1984; Ovruski 1995;
Schliserman et al. 2004; Ovruski et al. 2004), and
Salta (2202'-2621'S and 6221'-6834'W)
(Ovruski et al. 2005; Oroio et al. 2005). Pupal
parasitoids were not found because all of the A.
fraterculus and C. capitata were collected as lar-
vae, so they would not have been obtained even if
present at the collection site.
Rhoptromeris haywardi was not found during
this study. However, as discussed by Wharton et al.
(1998), published records for R. haywardi attack-
ing tephritids (Blanchard 1947; Turica & Mallo
1961; Nasca et al. 1980) are questionable and need
verification. All records of this eucoiline species
come from bulk samples of fruit, from which para-
sitoid species of both Drosophilidae and Tephriti-
dae could emerge. Without isolation of tephritid pu-
paria, correct host fly-parasitoid species associa-
tion cannot be made (Wharton et al. 1998).
Neither of the 2 introduced and released exotic
fruit fly parasitoid species were recovered from
fruit samples collected during this study, even
though both D. longicaudata and A. indica were
reported by Turica (1968) as established onAnas-
trepha spp. As noted by Ovruski et al. (1999), both
exotic parasitoid species were recovered immedi-
ately following release in Concordia in the 1960s,
and their establishment was not verified later. In-
terestingly, D. longicaudata was recently recov-
ered in the northeastern province of Misiones
(Schliserman et al. 2003) and in the northwestern
province of Salta (Oroio & Ovruski 2007) approx-
imately 40 years after its first release. Similarly,
recent fruit fly parasitoid surveys made in the
provinces of Jujuy (northwestern Argentina),
C6rdoba (Central Argentina), and Misiones
(northeastern Argentina) recorded the presence
ofA. indica (Ovruski et al. 2006).
Although no C. capitata parasitoids were recov-
ered in this study, some species, like A. pelleranoi,
were obtained from C. capitata pupae in Tucuman.
For example, in a recent fruit fly parasitoid survey
in Citrus crop areas of Tucuman,A. pelleranoi has
been recovered from C. capitata pupae in Citrus au-
rantium L. (Rutaceae), an exotic fruit mainly in-
fested by Medfly (Schliserman & Ovruski 2004).
According to Ovruski et al. (2004), C. capitata is not
heavily parasitized by neotropicalAnastrepha par-
asitoid species. With the exception of the figitidsA.
pelleranoi, A. nordlanderi Wharton, and Odon-
tosema anastrephae Borgmeier (Wharton et al.
1998), braconid parasitoids appear to adapt poorly
to the exotic C. capitata (Ovruski et al. 2004).
Even though the number and size of fruit sam-
ples collected during this study were relatively
small, the results seemingly suggest that U anas-
trephae and D. brasiliensis would have a certain
degree of fruit preference when searching host


larvae. For example, U anastrephae with the
shortest ovipositor of any of the opiine species ob-
tained (0.7 0.1 times as long as the metasoma,
n = 12), was most commonly found attacking A.
fraterculus larvae in M. pungens and E. uniflora,
the smallest fruit species sampled (7 and 8 times
smaller than the guava or the feijoa, respec-
tively). This relationship between smaller host
fruit species and U. anastrephae has been previ-
ously reported by Hernandez-Ortiz et al. (1994),
Lopez et al. (1999), Sivinski et al. (1997, 2000,
2001), and Aluja et al. (2003b) collecting fruit spe-
cies of Spondias (Anacardiaceae) in Mexico, and
by Aguiar Menezes et al. (2001) collecting Euge-
nia species (Myrtaceae) in Brazil. On the other
hand, D. brasiliensis, which has the longest ovi-
positor of the parasitoid species collected (3.3
0.2 times longer than the metasoma, n = 12), was
observed only in association withA. fraterculus in
F sellowiana and P guajaua, the largest fruit spe-
cies sampled. This peculiarity has also been noted
by Aguiar-Menezes & Menezes (1997) in Brazil,
when collecting A. fraterculus larvae parasitized
by D. brasiliensis exclusively from Prunus persica
(L.) Batsch (peach), and by Ovruski et al. (2004)
in NW Argentina, where D. brasiliensis was
mainly recovered from both peach and guava. In-
terestingly, wild fruits of Prunus persica, P gua-
java, and F sellowiana have a similar size
(Ovruski et al 2004; Ovruski & Schliserman
2003a). Doryctobracon areolatus, which has a rel-
atively long ovipositor (2.3 0.2 times longer than
the metasoma, n = 12) but 1.4 times shorter than
D. brasiliensis' ovipositor, may be occupying an
intermediate position between U. anastrephae
and D. brasiliensis in host fruit preference. As re-
ported previously Sivinski et al. (1997), Lopez et
al. (1999), and Aguiar Menezes et al. (2001),D. ar-
eolatus might not have any host fruit preference.
This opiine species has the broadest host-fly and
host-fruit range of any of the Mexican (Sivinski
1997, 2000, 2001; Aluja et al. 2003b) and Brazil-
ian (Canal & Zucchi 2000) fruit fly parasitoid spe-
cies. In NW Argentina, D. areolatus has been re-
ported foraging on a wide range ofA. fraterculus
host plant species (Ovruski et al. 2004). Appar-
ently, a short ovipositor would allow U. anas-
trephae to have access to fly larvae mainly in
small fruit, whereas a long ovipositor would allow
D. areolatus (or D. brasiliensis) to reach host lar-
vae in both large and small fruit. While studying
a MexicanAnastrepha-parasitoid guild of 5 opiine
species, including U. anastrephae and D. areola-
tus, Lopez et al. (1999) and Sivinski et al. (1997,
2001) found a strong correlation between oviposi-
tor length of parasitoids and the size of host-in-
fested fruits. This relationship would imply that
the ovipositor length is an important limitation
on foraging (Sivinski et al. 1997). Furthermore,
Sivinski et al. (2001) and Sivinski & Aluja (2003)
pointed out that the ovipositors in the opiine spe-


June 2008







Ovruski et al.: Parasitoids ofAnastrepha fraterculus in Entre Rios


cies attacking Mexican fruit fly would have origi-
nally diverged due to the action of environmental
factors such as temperature, humidity, seasonal-
ity, and/or host-fruit abundance and diversity.
Although data provided here are preliminary,
parasitization rates appear to be too low to con-
sider parasitoids as a significant natural mortal-
ity factor ofA. fraterculus in the study area. How-
ever, reliable data on the impact of the parasitoids
upon natural A. fraterculus populations can be
obtained only by closely monitoring the host and
parasitoid population fluctuations for many gen-
erations in exotic and native host plant species.
Thus, further studies on the ecology and behavior
ofA. fraterculus and their natural enemies should
be carried out in Entre Rios for implementing a
feasible, but cautious, biological control strategy
against this fruit fly pest. As suggested by Aluja
(1996, 1999), native host plants could be managed
to naturally augment parasitoid numbers and to
sustain parasitoid populations in wild vegetation
areas. Therefore, conservation of native A. frater-
culus-parasitoids would be an attractive alterna-
tive to the indiscriminate introduction of exotic
parasitoids (Sivinski & Aluja 2003).

ACKNOWLEDGMENTS

We express our gratefulness to Alejandra Roldan for
the identification of all plant samples and to Eduardo
Frias, Carolina Colin, and Jos6 Sa6z for technical sup-
port. Special thanks to Teresa Vera, Fanny Manso, Di-
ego Segura, Jorge Cladera, Ricardo Tomassi, Natalia
Petit, Norma Vaccaro, and Graciela Putruelle for send-
ing additional samples of parasitoid collected in Entre
Rios. We thank Martin Aluja (INECOL, A.C., Xalapa,
Veracruz, M6xico) for sharing with us his vast experi-
ence on ecology, biology, and ethology of fruit fly parasi-
toids. Financial support was provided by the Consejo
Nacional de Investigaciones Cientificas y T6cnicas de la
Republica Argentina (CONICET) (Grant PIP Nos. 4973/
97, 0702/98, and 5129/05).

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Florida Entomologist 91(2)


June 2008


HOST STATUS OF LITCHI AND RAMBUTAN TO THE
WEST INDIAN FRUIT FLY (DIPTERA: TEPHRITIDAE)

DAVID A. JENKINS AND RICARDO GOENAGA
USDA-ARS Tropical Agriculture Research Station, 2200 Pedro Albizu Campos Ave., Suite 201,
Mayaguez, Puerto Rico 00680-5470

ABSTRACT

Fruit of litchi, Litchi chinensis, and rambutan, Nephelium lappaceum, were collected from the
field in 2006 and 2007 and monitored for the emergence of West Indian fruit flies,Anastrepha
obliqua. Fruit clusters of rambutan and litchi, with a piece of the peel removed to allow access
to ovipositing females, were also placed in cages and exposed to 12-d-old post-eclosion male
and female West Indian fruit flies for 48 h. These exposed fruit were then monitored for the
emergence ofA. obliqua. Mango fruit were simultaneously exposed to male and female A. obli-
qua in separate cages and monitored for the emergence ofA. obliqua. Fruit fly traps baited
with putrescine and ammonium acetate were placed in orchards of litchi and rambutan, as
well as an adjacent orchard of carambola, Auerrhoa carambola, to demonstrate the presence
of fruit flies while litchi and rambutan were fruiting. Although we collected 3732 ripe litchi
fruit (40.34 kg) and 5534 ripe rambutan fruit (166.60 kg), none of these yielded tephritid lar-
vae. Litchi and rambutan fruit exposed to adult fruit flies in cages did not yield tephritid lar-
vae, though similarly exposed mangoes did. We conclude that litchi and rambutan have an
undetectably low probability of being infested byA. obliqua in Puerto Rico.

Key Words:Anastrepha obliqua, Litchi chinensis, Nephelium lappaceum, quarantine

RESUME

Frutas de litchi, Litchi chinensis, y rambutan, Nephelium lappaceum, fueron colectadas en
el campo en 2006 y 2007 y se monitorearon para detectar la presencia de la mosca de la fruta
de la Indias Occidentales,Anastrepha obliqua. Frutas de rambutan y litchi con un pedazo de
la cascara removida para permitir oviposici6n, fueron colocadas enjaulas y expuestas por 48
horas a machos y hembras de esta mosca. Las frutas fueron monitoreadas para detectar la
emergencia deA. obliqua. Trampas para la mosca de la fruta con carnada a base de putre-
cina y acetato de amonio fueron colocadas en huertos de litchi y rambutan y en huertos cer-
canos de carambola, Averrhoa carambola, para demostrar la presencia de la mosca de la
fruta mientras los arboles experimentales estaban en fruto. Se colectaron 3732 (40.34kg)
frutas de litchi maduras y 5534 (166.60 kg) de rambutan pero en ninguna se observe la pre-
sencia de larvas Tephritidae. Frutas de litchi y rambutan expuestas en una jaula a adults
de la mosca de la fruta tampoco mostraron emergencia de larva Tephritidae aunque frutas
de mango si. Se concluye, que frutas de litchi y rambutan tienen una baja probabilidad de
ser infectadas porA. obliqua en Puerto Rico.


Translation provided by the authors.


Litchi, Litchi chinensis Sonn., and rambutan,
Nephelium lappaceum L., both in the Sapindaceae
family, are valuable fruits native to southern
China and Malaysia, respectively (Morton 1987).
Although currently cultivated on a small scale on
the island of Puerto Rico, growers have expressed
an interest in expanding their market to include
the North American mainland. Currently, Puerto
Rico is home to 2 economically important tephritid
fruit flies, Anastrepha obliqua Marquart and A.
suspense (Loew) (Diptera: Tephritidae) (Martorell
1976).Anastrepha obliqua is only occasionally re-
ported from California and Texas, but otherwise is
not found within the continental U.S. (Epsky et al.
2003). As a result, there are understandable fears
that the importation of some fruit, such as litchi
and rambutan, from Puerto Rico to mainland


North America may facilitate the establishment of
A. obliqua there. A search of a host plant database
for Tephritidae (Norrbom 2004) found no reports
ofL. chinensis or N. lappaceum as hosts ofA. obli-
qua as of Aug 2007. In light of the difficulties reg-
ulatory agencies face in determining the relative
threat of importing exotic pests, particularly fruit
flies in the family Tephritidae, guidelines have
been published that outline methods thought to be
sufficient in establishing that threat for a particu-
lar fruit species (Cowley et al. 1992).
Our objective was to observe the incidence of
infestation in field-collected fruits, and in fruit ex-
posed to A. obliqua females in no-choice labora-
tory tests. In addition, adult fly populations were
monitored by trapping in orchards of litchi and
rambutan, and in adjacent orchards of other spe-







Jenkins & Goenaga: Host Status of Litchi and Rambutan


cies of fruit trees. We used the principles outlined
in Cowley et al. (1992) as guidelines for our inves-
tigation. Identical methods have been used to
demonstrate the non-host status of litchi and lon-
gan (Dimocarpus longan (Lour.): Sapindaceae)
and mamey sapote (Pouteria sapota (Jacq.) H.E.
Moore & Steam: Sapotaceae) toA. suspense and
A. obliqua, respectively (Gould et al. 1999; Gould
& Hallman 2001; Jenkins & Goenaga 2007).

MATERIALS AND METHODS

Between May 2006 and Aug 2007 mature litchi
fruits (Brewster, Bosworth-3, Groff, Mauritius,
Kaimana, and Salathiel varieties) were harvested
from an orchard in Adjuntas, PR, and mature
rambutan fruits (Benjai, Gulu Batu, Jitlee, R-134,
R-156, R-162, R-167, and Rongren varieties) were
harvested from an orchard in Corozal, PR. Har-
vested fruit were counted, weighed, placed on a
wire mesh over vermiculite in a screen-covered
plastic bin, and were stored at 25-27C in an envi-
ronment of approximately 60% RH (never less
than 50% RH). The vermiculite was monitored
weekly for fruit fly larvae or pupae. These were
collected and placed in a plastic Petri-dish with a
small amount of moistened vermiculite and stored
at 25C in an environmental chamber (12:12 D:L)
(White & Elson-Harris 1992). The Petri-dishes
were monitored daily for the emergence of adults.
In the summer of 2006, collapsible nylon cages
(60 x 60 x 60 cm) (Bioquip, Rancho Dominguez,
CA) were filled with clusters of ripe litchi with one
half of the peel removed from each fruit to expose
the fleshy pulp. Twenty female and 20 maleAnas-
trepha obliqua individuals, reared from mango,
Mangifera indica L. and/or fruit of Spondias mom-
bin L. (Anacardiaceae), were placed into each
cage 12-d post-eclosion. As simultaneous positive


controls, nylon cages were filled with mango fruit
that had been covered with brown paper bags
(Lawson pollination bags, No. 400) 2 weeks prior
to prevent field infestation byA. obliqua. Twenty
female and 20 male A. obliqua adults, 12 d old,
were placed into each cage. All cages were held in
a greenhouse (mean temperature 24.8C, RH
74%) and contained a single seedling of Ma-
nilkara zapota van Royen (Sapotaceae) to provide
a suitable microclimate for the flies. Prior to
placement in cages, flies were given slivers of car-
ambola, Averrhoa carambola L. (Oxalidaceae), to
provide carbohydrates and water. Mangoes that
had been bagged but not subsequently exposed to
ovipositing A. obliqua were monitored for the
emergence of fruit flies to ensure that the paper
bags prevented infestation. After 48 h of expo-
sure, all fruit in all cages were harvested and
monitored as described above for the emergence
of adult A. obliqua. All fruit were monitored for 3
weeks post-harvest and then discarded. Each lab-
oratory exposure was replicated 3 times for each
fruit variety and for the control exposures with
mangoes. This experiment was repeated with
fruits of rambutan in the summer of 2007.
In addition, 5 plastic Multilure traps@ (A Bet-
ter World, Inc., Fresno, CA) baited with ammonia
acetate and putrescine (Suterra, Bend, OR) were
placed in each litchi and rambutan orchard and
monitored weekly for fruit flies. Five traps also
were placed in a carambola orchard near the ram-
butan orchard and monitored weekly.

RESULTS

A total of 3732 litchi fruit weighing 40.34 kg
were collected from Adjuntas, PR, none of which
yielded tephritid pupae (Table 1). Similarly, 5534
rambutan fruits were collected, weighing a total


TABLE 1. COLLECTIONS OF LITCHI BY DATE AND VARIETY.

Variety Dates collected Number of fruit g of fruit Total fruit per variety Total g per variety
Brewster 27-Jun-2006 534 4986 1064 12110
13-Jun-2006 234 2567
31-May-2006 216 3510
29-May-2007 80 1047
Bosworth 3 23-May-2006 150 2120 705 6761
26-May-2006 305 2828
29-May-2007 250 1813
Groff 20-Jun-2006 171 1853 722 7534
1-Aug-2006 251 3818
21-Jun-2007 300 1863
Mauritius 31-May-2006 333 4008 583 7493
29-May-2007 250 3485
Kaimana 18-Jul-2006 147 1520 347 3785
29-May-2007 200 2265
Salathiel 11-Jul-2006 111 1015 311 2660
29-May-2007 200 1645
Total 3732 40343







Florida Entomologist 91(2)


June 2008


TABLE 2. COLLECTIONS OF RAMBUTAN BY DATE AND VARIETY.

Variety Dates collected Number of fruit g of fruit Total fruit per variety Total g per variety

Benjai 19-Jul-2006 127 3831 654 19830
22-Aug-2007 367 11103
29-Aug-2007 160 4896
Gulu Batu 28-Jun-2006 235 7134 692 20899
19-Jul-2006 275 8289
29-Aug-2007 182 5476
Jitlee 4-May-2006 284 8507 906 27149
13-Sep-2006 286 8453
22-Aug-2007 336 10189
R-134 17-May-2006 183 5324 706 20961
22-Aug-2007 308 9228
29-Aug-2007 215 6409
R-156 4-May-2006 249 7419 688 20633
22-Aug-2007 240 7252
29-Aug-2007 199 5962
R-162 17-May-2006 130 3906 682 20451
22-Aug-2007 245 7329
29-Aug-2007 307 9216
R-167 19-Jul-2006 219 6591 540 16273
22-Aug-2007 132 3961
29-Aug-2007 189 5721
Rongren 19-Jul-2006 267 8003 666 20401
9-Aug-2006 296 9433
29-Aug-2007 103 2965
Total 5534 166597


of 166.60 kg, none of which yielded tephritid pu-
pae (Table 2). Litchi and rambutan fruit exposed
to A. obliqua adults in laboratory studies yielded
no tephritid larvae, while mango fruit similarly
exposed yielded pupae (Tables 3 and 4, respec-
tively). Mangoes that had been bagged prior to
use but that were not exposed to A. obliqua fe-
males did not yield any tephritid pupae.


Multilure traps baited with putrescine and am-
monium acetate yieldedA. obliqua andA suspense
females simultaneous to fruit collection, indicating
that these flies were active in the area when fruit
were harvested. The number ofA. obliqua trapped
in the rambutan orchard was always 0, whereas a
nearby carambola orchard the number fluctuated
between 0 and 25 A. obliqua adults per trap per d.


TABLE 3 FLY PUPAE RECOVERED FROM LITCHI FRUIT EXPOSED TO 20 MALE AND 20 FEMALE ANASTREPHA OBLIQUA (12-
D POST-ECLOSION) FOR 48 H.

Tephritid pupae recovered

Date Variety Fruit/rep Rep 1 Rep 2 Rep 3 Mean SEM

13-Jun-2006 Brewster 200 0 0 0 0.0 0.0
13-Jun-2006 Mango 10 16 0 2 6.0 6.2
26-May-2006 Bosworth 3 189 0 0 0 0.0 0.0
26-May-2006 Mango 10 18 11 10 13.0 3.1
1-Aug-2006 Groff 234 0 0 0 0.0 0.0
1-Aug-2006 Mango 10 0 15 19 11.3 7.1
31-May-2006 Mauritius 200 0 0 0 0.0 0.0
31-May-2006 Mango 10 16 14 21 3.6 2.5
18-Jul-2006 Kaimana 212 0 0 0 0.0 0.0
18-Jul-2006 Mango 10 11 8 5 8.0 2.1
11-Jul-2006 Salathiel 125 0 0 0 0.0 0.0
11-Jul-2006 Mango 10 12 9 7 9.3 1.8







Jenkins & Goenaga: Host Status of Litchi and Rambutan


TABLE 4. FLY PUPAE RECOVERED FROM RAMBUTAN FRUIT EXPOSED TO 20 MALE AND 20 FEMALE ANASTREPHA OBLI-
QUA (12-D POST EMERGENCE) FOR 48 H.

Tephritid pupae recovered

Date Variety Fruit/rep Rep 1 Rep 2 Rep 3 Mean + SEM

29-Aug-2007 Benjai 50 0 0 0 0
Mango 5 8 3 6 5.66+ 1.8
29-Aug-2007 Gulu Batu 50 0 0 0 0
Mango 5 2 6 4 4.00+ 1.4
22-Aug-2007 Jitlee 50 0 0 0 0
Mango 5 5 8 3 5.33+ 1.8
22-Aug-2007 R-134 50 0 0 0 0
Mango 5 5 6 8 6.33+ 1.1
22-Aug-2007 R-156 50 0 0 0 0
Mango 5 4 4 3 3.67 +0.4
22-Aug-2007 R-162 50 0 0 0 0
Mango 5 5 7 7 6.33 +0.8
22-Aug-2007 R-167 50 0 0 0 0
Mango 5 4 8 3 5.00+ 1.9
29-Aug-2007 Rongren 50 0 0 0 0
Mango 5 2 4 3 3.00+ 0.7


The number ofA obliqua adults captured in the lit-
chi orchard ranged from 0 to 4.3 flies per trap per d
while these fruits were being harvested.

DISCUSSION

It is impossible to prove that a fruit is never a
host to a species of insect, only that a fruit is used
by a species of insect. However, our data show
that the likelihood of infestation of the varieties of
litchi or rambutan that we assayed byA. obliqua
is very small. We conclude that these fruit variet-
ies are extremely unlikely to contain A. obliqua
and therefore represent minimal threat of trans-
porting this pest when exported.

ACKNOWLEDGMENTS

Mention of trade names or commercial products in
this article is solely for the purpose of providing specific
information and does not imply recommendation or en-
dorsement by the U.S. Department of Agriculture. We
thank Elkin Vargas for his excellent field and labwork.

REFERENCES CITED

COWLEY, J. M., R. T. BAKER, AND D. S. HARTE. 1992.
Definition and determination of host status for mul-


tivoltine fruit fly (Diptera: Tephritidae) species.
J. Econ. Entomol. 85: 312-317.
EPSKY, N. D., P. E. KENDRA, AND R. L. HEATH. 2003. De-
velopment of lures for the detection and delimitation
of invasive Anastrepha fruit flies. Proc. Caribbean
Food Crops Society. 39: 84-89.
GOULD, W. P., M. K. HENNESSEY, J. PENA, A. CASTINEI-
RAS, R. NGUYEN, AND J. CRANE. 1999. Nonhost status
of lychess and longans to Caribbean fruit fly (Diptera:
Tephritidae). J. Econ. Entomol. 92: 1212-1216.
GOULD, W. P., AND G. HALLMAN. 2001. Host status of
mamey sapote to Caribbean fruit fly (Diptera: Te-
phritidae). Florida Entomol. 84: 730-375.
JENKINS, D. A., AND R. GOENAGA. 2007. Host status of
mamey sapote, Pouteria sapota (Sapotaceae), to the
West Indian fruit fly, Anastrepha obliqua (Diptera: Te-
phritidae) in Puerto Rico. Florida Entomol. 90:384-388.
MARTORELL, L. F. 1976. Annotated Food Plant Catalog
of the Insects of Puerto Rico. Agric. Exp. Stn., Univ.
Puerto Rico, Department of Entomology. 303 pp.
MORTON, J. F. 1987. Fruits of Warm Climates. Media In-
corporated, Greensboro, NC. 506 pp.
NORRBOM, A. L. 2004. Host plant database for Anas-
trepha and Toxotrypana (Diptera: Tephritidae: Tox-
otrypanini). Diptera Data Dissemination Disk (CD-
ROM) http://www.sel.barc.usda.gov:591/diptera/Te-
phritidae/TephHosts/search.html.
WHITE, I. M. AND M. M. ELSON-HARRIS. 1992. Fruit Flies
of Economic Significance: Their Identification and Bio-
nomics. CAB International, Wallingford. xii + 601 pp.







Florida Entomologist 91(2)


A NEW GALL-INDUCING SPECIES OF HOLOPOTHRIPS
(THYSANOPTERA: PHLAEOTHRIPINAE) FROM TABEBUIA
TRUMPET TREES IN THE CARIBBEAN REGION

IRMA CABRERA1 AND ALEJANDRO SEGARRA1
'Crop Protection Department, College of Agricultural Science, University of Puerto Rico, Mayaguez 00681-9030

ABSTRACT

In this research we describe Holopothrips tabebuia new species based on specimens col-
lected from Puerto Rico, Florida, and Dominican Republic. Holopothrips tabebuia differs
from the closely allied H. inquilinus (Bournier) in shape and reticulation patterns of the
metanotum, and in the number of epimeral setae. Most specimens have been collected from
Bignonaceae, particularly from host plants in the genus Tabebuia. Thus, we have chosen the
epithet tabebuia to denote its relationship with these plants.

Supplementary material (color pictures) on line at http://www.fcla.edu/FlaEnt/fe912.htm

Key Words: Thysanoptera, Phlaeothripinae, Holopothrips, Gall, Bignonaceae, Tabebuia Car-
ibbean Region

RESUME

En esta publicaci6n describimos a Holopothrips tabebuia nueva especie basado en especi-
menes colectados en Puerto Rico, Florida y Republica Dominicana. H. tabebuia difiere de su
aliado cercano, H. inquilinus (Bournier) en la forma y los patrons de las reticulaciones del
metanoto, y el numero de la seta epimeral. La mayoria de los especimenes fueron colectados
de las Bignonaceas, particularmente de las plants hospederas del g6nero Tabebuia. Se se-
leccion6 el epiteto tabebuia para denotar su relaci6n con estas plants.


Translation provided by the authors.


Holopothrips Hood is a Neotropical genus of 30
described species (Mound 2007). Several species
are known to induce leaf deformation, or even
galls, on the leaves of their host plants (Cavalleri
& Kaminski 2007). Mound & Marullo (1996) pro-
vided an identification key to more than 30 spe-
cies. Described species show a considerable range
of structural differences, and high morphological
variability within groups has resulted in five de-
scribed genera being synonymized (Mound &
Marullo 1996).
This paper describes a new species of Holopo-
thrips, first recorded from Puerto Rico in 2006
(Cabrera et al., in press). This thrips is possibly
widespread in the northern Caribbean, having
been taken from at least 2 species of Tabebuia (Bi-
gnoniaceae), in Puerto Rico, Florida, and Hispan-
iola. Adults and larvae feed on very young foliage,
inducing obvious gall-like deformations (Figs. 1
and 2), which become more noticeable as infested
leaves mature. All life stages of the new Holopo-
thrips species coexist within galled leaves. Anec-
dotally, attack severity seems more prevalent in
humid districts in Puerto Rico. Also, Tabebuia
heterophylla (D.C.) Britton shows the highest in-
festation when compared with other surveyed
trumpet tree species (Cabrera et al., in press).
These authors also found Montandoniola mora-


guezi Puton (Hemiptera: Anthocoridae) predating
on Holopothrips tabebuia n.sp. Similar predation
of Holopothrips sp. by M. moraquezi was observed
by Dobbs & Boyd (2006).

Holopothrips tabebuia,
new species

Female Macroptera. Body bicolored, mainly
yellow to brownish yellow with head and
pterothorax light brown, and abdominal seg-
ments VIII-X dark brown (Fig. 3); antennal seg-
ments I II light brown, III-VIII yellow (Fig. 4);
legs yellow; forewings uniformly pale; major setae
pale except dark anal setae.
Head slightly longer than wide; 1 pair of major
capitate postocular setae, arising behind inner
margin of eyes; maxillary stylets retracted to pos-
tocular setae (Fig. 5), about one third of head
width apart medially; mouth cone rounded, not
extending beyond fore coxae. Antennae 8-seg-
mented, segments III and IV with 3 sensoria, V
and VI with 2 sensoria; VIII slender. Pronotum
transverse, epimeral sutures incomplete; only 1
pair of epimeral setae, all 5 pairs of major setae
capitate. (Fig. 6). Fore tarsus without a tooth. Met-
anotum with longitudinal narrow reticulation, al-


June 2008







Cabrera & Segarra: A New Gall-Inducing Species of Holopothrips


Fig. 1-2. Damage caused by Holopothrips tabe-
buia, new species. 1, Tabebuia heterophylla; 2, Tabe
buia aurea.


most striate; 1 or more pairs of small setae present
anterior to median major setae (Fig.7). Forewing
parallel sided, with 10 duplicated cilia present
posteriorly on distal end of forewing; 3 capitate
sub-basal setae arising in a straight line. Pelta re-
ticulate, 1 pair of campaniform sensilla close to
posterior margin (Fig. 8). Segment IX with a swol-
len S-shaped spermatheca (Fig.9). Tergites II-VII
each with an additional almost straight seta ante-
rior to paired sigmoid wing-retaining setae (Fig
10). Tergite IX setae S1 and S2 blunt to weakly
capitate; S1 about as long as tube, S2 longer; S3
finely acute; tube much shorter than head.
Measurements holotypee female in microns).
Body length 2425. Head, length 275; width 255;
major postocular setae 50. Pronotum, length 180;
median width 360; major setae, anteromarginal
(am) seta 20, anteroangular (aa) seta 35, ml 50,
epimeral (epim) 76, posteroangular seta (pa) 55.
Forewing, length 800; sub-basal setae 35-40. Ab-


dominal tergite IX setae S1 (155) S2 (165), S3
(152.5); tube length 170. Antennal segments I-
VIII length 35, 40, 60, 35, 50, 45, 45, 30.
Male Macroptera. Similar to female but
slightly smaller; without fore tarsal tooth. Ab-
dominal sternites VII-VIII each with 3 glandular
areas, 1 close to posterior margin and 2 anterolat-
eral to row of discal setae (Fig. 11); on segment
VIII the posterior glandular area is prolonged lat-
erally onto the tergite and extends to tergal mar-
ginal setae S1 (Fig. 12); tergite IX setae S2
slightly longer than S1.
Measurements (paratype male in microns).
Body length 2100. Head, length 225; width 215;
major postocular setae 35. Pronotum, length 163;
median width 285, major setae, am 20, aa 27.5, ml
40, epim 55, pa 40. Forewing length 750; sub-
basal setae 30-35. Abdominal tergite IX setae S1
(142.5), S2 (147.5); tube length 160. Antennal seg-
ments I-VIII length same as holotype.

MATERIAL STUDIED

Holotype female, Puerto Rico, Toa Alta, from dis-
torted leaf of T heterophylla, 12.ix.2006 (S. Cruz),
deposited in Museum of Entomology and Tropical
Biodiversity (METB) Agricultural Experiment Sta-
tion, University of Puerto Rico, Rio Piedras, Puerto
Rico (PR. Acc. No. 07-2007). Paratypes: Seven fe-
males and 6 males. Four females on T heterophylla
from San Juan, PR. 6.vii.2007 (S. Cruz), 2 females
collected with holotype, and 1 female Arecibo, PR.
20.vii.2007 (S. Cruz). Four males from Cayey, PR.
20.ix.2007, (I. Cabrera), and 2 males collected with
holotype. One female and 1 male from Toa Alta,
PR. in collection of L. A. Mound, Australia; 2 fe-
males (San Juan) and 2 males (Cayey) in USNM,
Washington, D.C. Four females and 3 males depos-
ited with holotype at METB.
Other Material Examined: Dominican Repub-
lic: Two females and 2 males, all from Tabebuia
sp. and deposited at METB. U.S.A., Florida: Nine
females and 3 males, 8 from Tabebuia sp., 1 from
Amphitecna latifolia (P. Mill.) A. H. Gentry, 1
from Schefflera actinophylla (Endl.) H.A.T.
Harms, and 2 specimens of unknown hosts ; 2 of
these specimens deposited at METB, the others
deposited in the Division of Plant Industry, Flor-
ida Department of Agriculture and Consumer
Services, Gainesville, FL.

Relationships

This new species agrees with the generic defi-
nition in Mound & Marullo (1996), and in the
identification key to species it will run to the spe-
cies H. inquilinus (Bournier). Unfortunately, that
species is known only from specimens taken from
an unidentified plant on the Research Station of
Duclos in the island of Guadeloupe. Based on
Bournier's description and notes in Mound &






Florida Entomologist 91(2)


13.


n I sty


V-r


14


Fig. 3-6 Holopothrips tabebuia, new species. 3, Female holotype; 4, Female holotype antenna; 5, Female holo-
type head maxillary stylets (mx sty); 6, Female holotype epimeron (epm), long epimeral seta (epm set).


Marullo (1996), H. tabebuia differs from H. in-
quilinus as follows: (1) Metanotum with longitu-
dinal almost striate, reticulation in the former, in-
stead of almost smooth to weakly reticulate in the
latter; (2) Epimera with only 1 pair of major setae
in the former, instead of 2 pairs in the latter. We


have observed variability in the number of forew-
ing cilia (8-12), and in the size of the 4th antenna
segment (30-50 microns). Further in a few speci-
mens the maxillary stylets appear to be retracted
slightly anterior to the postocular setae, and in
some males the glandular area on sternite VI is


June 2008







Cabrera & Segarra: A New Gall-Inducing Species of Holopothrips


." an


a... .... ., r 1 ...


Fig. 7-9. Holopothrips tabebuia, new species. 7, Fe-
male holotype metanotum and median seta (mdn); 8,
Female holotype pelta; 9, Female holotype tube sper-
mateca (sper).


faint. We have chosen tabebuia as the specific ep-
ithet to denote the thrips relationship with these
host plants in the Bignonaceae.


Host plants and Distribution

Holopothrips tabebuia new species has been
collected in Florida (USA), Puerto Rico, and His-
paniola, mainly from Bignonaceae. Most speci-
mens were collected from Tabebuia sp. A few spec-
imens were taken on Crescentia cujete L.,A. lati-
folia, and S. actinophylla, although there is no ev-
idence of any biological association with these
plants because they were lacking galls and repro-
ducing populations were not found.


ACKNOWLEDGMENTS

We thank Dr. Laurence Mound (CSIRO, Australia)
for invaluable collaboration, and for useful suggestions
and revisions to earlier drafts of this manuscript. We
thank Dr. G. B. Edwards (Florida Department of Plant
Industry) for kindly providing specimens from Florida
for examination, and Dr. Adriano Cavalleri (Brazil) for
suggestions on the manuscript.


REFERENCES CITED

CABRERA I., A. RAMiREZ, S. CRUZ, A. VELEZ, L. SAEZ,
AND A. SEGARRA. 2008. Holopothrips sp. (Thysan-
optera: Phlaeothripidae) en Tabebuia heterophylla y
Tabebuia aurea en Puerto Rico. J. Agric. Univ. Pu-
erto Rico. (In press).
CAVALLERI A., AND L. A. KAMINSKI. 2007. A new Holo-
pothrips species (Thysanoptera: Phlaeothripidae)
damaging Mollinedia (Monimiaceae) leaves in
Southern Brazil. Zootaxa 1625: 61-68.
BOURNIER, A. 1993. Thysanopt6res de Martinique et
Guadaloupe. Zoology :J. Pure and Applied Zool. 3:
227-240.
DOBBS T. T., AND D. W. BOYD. 2006. Status and distri-
bution of Montandoniola moraguezi; (Hemiptera:
Anthocoridae) in the continental United States.
Florida Entomol. 89: 41-46.
MOUND, L. A. 2007. Thysanoptera (Thrips) of the
World-a checklist. http://www.ento.csiro.au/thysan-
optera/worldthrips.html
MOUND, L. A., AND R. MARULLO. 1996. The Thrips of
Central and South America: An Introduction. Mem-
oirs on Entomology International 6: 1-488.






Florida Entomologist 91(2)


11


Fig. 10 -12. Holopothrips tabebuia, new species. 10, female holotype tergite retainer seta (ret set) ; 11, male
paratype sternite glands VII and VIII (glg); 12, male paratype tergite gland VIII (glg).


June 2008























1Fi


Fig. 1-2. Damage caused by
Holopothrips tabebuia, new
species. 1, Tabebuia
heterophylla; 2, Tabebuia aurea.


"i~;;"


rc ff
wij


;5




















3
I





mx sty





Fig. 3-6 Holopothrips tabebuia, new
species. 3, Female holotype; 4, Female
holotype antenna; 5, Female holotype
head maxillary stylets (mx sty); 6,
Female holotype epimeron (epm), long
epimeral seta (epm set).












Indn


*. 7









-----lIl





9

Fig. 7-9. Holopothrips tabebuia ,new species. 7,
Female holotype metanotum and median seta
(mdn); 8, Female holotype pelta; 9, Female
holotype tube spermateca (sper).




























12


Fig. 10-12. Holopothrips tabebuia, new
species. 10, female holotype tergite retainer
seta (ret set); 11, male paratype sternite
glands VII and VIII (glg); 12, male paratype
tergite gland VIII (gig).







Zobar & Genc: Biology of Queen of Spain Fritillary


BIOLOGY OF THE QUEEN OF SPAIN FRITILLARY, ISSORIA LATHONIA
(LEPIDOPTERA: NYMPHALIDAE)


DAMLA ZOBAR1 AND HANIFE GENC1
1Department of Plant Protection, Agricultural Faculty, Canakkale Onsekiz Mart University,
17100 Canakkale, Turkey

ABSTRACT

The biology and the life cycle of Issoria lathonia (Nymphalidae) (Linnaeus 1758) on its host
plant, Viola tricolor L. (Violaceae), are described from laboratory studies. In the laboratory
eggs are laid singly on the host plant leaves as well as on the surfaces of plastic screen cages.
Newly hatched larvae aggregate and feed on the host plant leaves. Later instars disperse on
the plant and continue to feed on leaves and flowers. Head capsule widths, and weight and
size measurements show that larvae develop through 5 instars. The larvae crawl off the host
plant and pupate off the host. The life cycle from egg to adult requires 23-31 d at 26 C, and
16:8 (L:D) photoperiod in the laboratory. The butterfly has been reared continuously in the
laboratory for about 2 years.

Key Words: Issoria lathonia, Nymphalidae, Argynnini, Viola tricolor

RESUME

Se describe la biologia y el ciclo de vida de Issoria lathonia (Linnaeus, 1758) sobre su plant
hospedera, Viola tricolor L. (Violaceae) basado sobre studios de laboratorio. En el laborato-
rio los huevos estan puestos individualmente sobre las hojas de la plant hospedera igual
como sobre la superficie de la tela plastica de lasjaulas. Las larvas reci6n nacidas se agregan
y se alimentan sobre las hojas de la plant hospedero. Los instares posteriores se dispersan
sobre la plant y continuan su alimentaci6n sobre las hojas y flores. Las medidas de la an-
chura de las capsulas de las cabezas, el peso y el tamaio muestran que las larvas pasan por
cinco instares. Las larvas caminan fuera de la plant hospedero y empupan separadas del
hospedero. El ciclo de vida desde el huevo hasta el adulto require 23-31 dias a los 26 C y un
fotoperiodo de 16:8 (L:D) [16 horas de luz y 8 horas de obscuridad] en el laboratorio. La ma-
riposa ha sido criada continuamente en el laboratorio por aproximadamente 2 aios.


The nymphalid genus Issoria Hiibner 1819 in-
cludes 97 species distributed throughout the
Palaearctic region and in parts of Africa, includ-
ing the I eugenia group, I lathonia group and the
African I. smaragdifera group. Issoria lathonia L.
is commonly known as the queen of Spain fritil-
lary across Europe.
Simonsen (2006 a, b) suggested from morpho-
logical studies that two of the African species
(I. hanningtoni and I baumanni) may not belong
to Issoria, but molecular data (Simonsen et al.
2006), indicates that the African species (includ-
ing I. smaragdifera) form a clade within Issoria.
Issoria spp. in the tribe Argynnini (Fritillaries)
have been studied with respect to female abdom-
inal scent organs (Urbahn 1913), glands, muscles
and genitalia (Simonsen 2006c), genitalic mor-
phology and function (Shir6zu & Yamamoto 1959;
Arnold & Fisher 1977; Ockenfels et al. 1998; Si-
monsen 2006a,b), classification, and host plant
records and oviposition sites (Oliveira & Freitas
1991), and host plant relationships (Ackery
1988). There are few studies of the immature
stages and life cycle of Issoria spp., and none for
Issoria lathonia L. the queen of Spain fritillary.


The adults of the queen of Spain fritillary are
characterized by a strong orange and black color-
ing of the upper sides of the wings (Baytas, 2007;
Tolman & Lewington 1997). The shape of the
wings differs from other fritillaries in being
slightly angular. The undersides of the hindwings
have large, oblong silvery patches. The line with
yellow dots (2 bigger and 1-2 smaller) in the api-
cal corners of upper side forewings is very com-
mon. Adult males and females look alike. The
queen of Spain occurs in the Canary Islands, Ma-
deira, North Africa, Southern, Central and East-
ern Europe, Southern Sweden, Finland, and Tur-
key. The species generally is active from Feb to
late autumn, with 2 to 3 generations. It may hi-
bernate in the egg, larval, pupal, or adult stage
(Tolman & Lewington 1997). Viola tricolor L. and
Viola arvensis Murr. (Violaceae) are common lar-
val host plants, but Simonsen (2006 a, b) found
larvae feeding onAnchusa sp., Ribus sp., and On-
obrychis sp. in Europe. The eggs are laid singly on
leaves of the host plants or nearby weeds.
The aims in this paper are to describe the biol-
ogy and immature stages of Issoria lathonia L.
feeding on Viola tricolor L. in the laboratory.







Florida Entomologist 91(2)


MATERIALS AND METHODS

During May and Jun 2006, Issoria lathonia
adults (n = 35) were captured in the vicinity of
Canakkale, Turkey, at about 325 m above sea
level. Adults were kept in screen cages (60 x 60 x
60 cm) with Viola tricolor host plants. A number
of flowering plants (Lantana camera L., Dianthus
sp., and Carduus sp.) were provided in the adult
rearing cages along with 10% honey solution or
Orange Punch Gatorade dispensed on cotton
balls to provide food for the adults. Eggs were laid
on the host plant leaves as well as on the screen
cage, and they were removed daily, counted, and
kept in a Petri dish on moist filter paper. Hatch-
ing larvae were allowed to feed on freshly cut host
plant material obtained as potted plants from lo-
cal garden shops as needed. Larval food was
changed daily by transferring all larvae to new
plants. Pupae were harvested daily and trans-
ferred to a new cage with a potted host plant and
adult food source. The colony was maintained un-
der controlled laboratory conditions at 26 1C,


60% RH and 16:8 (L:D) photoperiod. The number
of instars was determined from data collected
from 15 larvae examined each day. Shed larval
head capsules were collected, measured, and pre-
served in 70% ethyl alcohol. Larvae were
weighted and their length measured daily for the
15 individuals. All biological stages of Issoria la-
thonia were examined and photographed with an
Olympus C7070 wide zoon camera attached to an
Olympus SZX9 binocular stereo zoom microscope.
The LSD test at 0.05 level of significance was
used to determine separation and significance of
means (SAS 1990).

RESULTS

Females laid their eggs singly, either on the
host plant or on the mesh screen of the cage. The
eggs are elliptical, pale yellowish, conical, and flat-
tened at the top, with 20 to 22 longitudinal ridges
(Fig. 1A). They are about 0.38 0.01 mm in length
and 0.31 0.01 mm in diameter (n = 17). The color
of the eggs changed from pale yellow to brownish


Fig. 1. Immature stages of Issoria lathonia. (A) An egg; (B) An egg about to hatch; (C) First instar; (D) Second
instar; (E) Fifth instar, (F) Lateral view of pupa; (G) Adult feeds on honey solution (H) Unsuccessful adult emerging
(I) Laboratory colony on Viola tricolor.


June 2008







Zobar & Genc: Biology of Queen of Spain Fritillary


black (Fig. 1B), after about 3-4 d as the mandibles
and head of the larvae became visible through the
chorion. The eggs hatch in 3-4 d at 26C.
We determined that there are 5 instars based
upon weight, length, and head capsule measure-
ments (Table 1). The duration of time spent in the
egg stage, in each of the 5 instars, and the pupal
stage are shown in Table 2.
The first instar body is initially translucent
yellow but changes to pale dark yellow after feed-
ing. There are long setae over the body (Fig. 1C).
The head is dark brown to black and the mouth-
parts are dark yellow. The legs and prolegs are
translucent yellow and tarsal segments are dark
brown to black. First instars aggregate and prefer
to feed on flower parts of the host plant. Second in-
stars are gray-brown in color with gray lateral
bands (Fig. 1D). Each segment contains a row of
long spines. The head is black and the mouthparts
are dark brown black. Longitudinal dorsal and
subdorsal bands are evident. The thoracic legs are
brown with the tarsal claws darkened. Third in-
stars are similar in appearance to second instars,
but each segment contains a row of short, branch-
ing spines. The head is black with light brown
eyes. Third instars no longer aggregate, but dis-
tribute themselves over the plant. The fourth and
fifth instars are similar in appearance to each
other and to third instars (Fig. 1E). Cream-white
patches are clearly evident on the black back-
ground of the body of fourth and fifth instars. A
row of short branching spines are evident, are or-
ange colored at the base. The legs are black but
the prolegs are light brown-orange. Fourth instars
consumed a large quantity of host leaves and flow-
ers (Fig. 11). Mature larvae attached with the cre-
master to a supportive surface and remained in a
crescent shape about 5-6 h. Then, hanging
straight down, they changed within 3-5 minutes
into the characteristic pupal shape and appear-
ance. The pupal color is variable to light brown to
dark brown-black, but shiny, uniform, and brown-
ish with white dorsal patch resembling a bird-
dropping. Pupae are initially very soft and dark
brown (Fig. 1F). They have a black patch area over
the wings, and the labial palpi and antennae were


visible through the cuticle. The pupal abdomen
consists of 10 segments, with the 10th bearing the
cremaster. Female pupae are 1.50 0.01 mm long
and 0.31 0.03 mm wide (dorsoventral measure-
ment in the thoracic region), and weighed an av-
erage of 289.4 46 mg (n = 21). Male pupae are
1.70 0.04 mm long, 0.41 0.03 mm wide, and
weighed an average 313.4 51 mg (n=26). Fe-
males emerged from the pupal stage about a day
earlier than males, on average (Table 2).
Adult males and females were similar in ap-
pearance (Fig. 1G). The wingspan was 39.2 2.90
mm in females and 40.3 1.98 mm in males (n =
10). Mating pairs often rested quietly together 4-
5 h. Females started laying eggs about 2 d after
mating. Adults survived in the laboratory about 2
weeks. The duration from egg to adult was 23-31
d at 26C, 16:8 (L:D) photoperiod. In the labora-
tory, 38% of pupae failed to become adults
(Fig. 1H).

DISCUSSION

This is the first detailed description of the biol-
ogy of Issoria lathonia L. and of the immature
stages. The larvae of the queen of Spain fritillary
feed on several species in the genus Viola in Vio-
lacea, but the main food plants are wild pansy (Vi-
ola tricolor L.) and field pansy (Viola arvensis
Murr.). These plants are larval food plants for
some other nymphalid species, including Boloria
bellona, B. selene, Speyeria aphrodite, S. atlantis,
S. cybele, Argynnis pandora, and A. paphia (Hes-
selbarth et al. 1995). Viola aetolica, V lutea, V bi-
flora, V calcarata, V corsica, and V odorata have
been recorded as larval host plants for Issoria
spp. (Tolman & Lewington 1997), but larvae
refuse to feed on "African violets" (Saintpaulia
spp.) (Tolman & Lewington 1997). Issoria latho-
nia has strong local migratory habits and is an
endangered species (Verovnik 2000; Kotiaho et al.
2005) in many countries. It occurs in a wide vari-
ety of habitats where the larval food plants occur.
In our study, Issoria lathonia adults mated
readily in the laboratory. Availability of the larval
host plant in the adult cages seemed not to be crit-


TABLE 1. MEASUREMENTS OF HEAD CAPSULE, WEIGHT, AND LENGTH OF LARVAL ISSORIA LATHONIA IN EACH INSTAR
(MEAN + SD, N = 15).

Instar Head capsule width (mm) Weight (mg) Length (mm)
First 0.186 0.00 a 0.02 0.01 a 2.66 0.61 a
Second 0.338 0.03 b 3.52 0.30 b 4.73 0.79 b
Third 0.508 0.02 c 14.42 1.61 c 7.46 0.99 c
Fourth 0.789 0.02 d 63.49 11.05 d 13.80 2.54 d
Fifth 1.231 0.07 e 213.09 48.67 e 21.66 2.92 e
LSD* 0.0289 0.7237 1.2803

*LSD= Fisher's Least Significant Difference between any 2 means. The means with a column followed by a different letter are
different from each other (P < 0.05).







Florida Entomologist 91(2)


TABLE 2. DEVELOPMENT TIME FOR LABORATORY CUL-
TURE OF ISSORIA LATHONIA AT 26C (MEAN
SD, N = 15).

Stage/instar Days

Egg 3.70 0.7
First 3.00 0.6
Second 2.93 1.1
Third 2.53 0.8
Fourth 2.73 0.7
Fifth 4.00 0.5
Pupae
Female 4.26 0.7
Male 5.30 0.7


ical for adult oviposition. The host plant, Viola tri-
color, is widely available as an ornamental plant
in the local greenhouses and it also can be cul-
tured easily in small pots. During 2 years of rear-
ing the queen of Spain fritillary butterfly we
found some evidence of disease, especially in the
pupal stage. Further studies are necessary to
solve this problem. The migratory status of the
butterfly in Turkey is not known. The ease with
which it can be reared and the availability of food
plants year around may make the queen of Spain
fritillary at attractive species for display in but-
terfly houses, and it is a valuable model butterfly
for further research in genetics, behavior, migra-
tory habits, and physiology.

ACKNOWLEDGMENTS

The authors thank Dr. James L. Nation for useful
comments and suggestions on an earlier version of the
manuscript. This research was based partly on the
M.Sc. Thesis of Damla Zobar, supervised by Hanife
Genc, and supported in part by Canakkale Onsekiz
Mart University Scientific Research Council (Project no:
2005/33).

REFERENCE CITED

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view of nymphalid butterflies. Biol. J.Linnean Soc.
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ARNOLD, R. A., AND R. L. FISCHER. 1977. Operational
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ria (Lepidoptera: Nymphalidae). Ann. Entomol. Soc.
America, 70(4): 455-468.


GENC, H. 2005. Determination of sex in pupae of Phy-
ciodes phaon (Lepidoptera: Nymphalidae). Florida
Entomol. 88(4): 536-537.
HESSELBATH, G., H. VAN ORSSCHOT, AND S. WAGENER.
1995. Die Tagfalter der Tiirkei. Vol 1, 2, 3.Published
by Selbstverlag. Sigbert Wagener, Bocholt. Selb-
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KOTIAHO, J., A. KAITALA, AND J. PAIVINEN. 2005. Pre-
dicting the Risk of Extinction from Shared Ecologi-
cal Characteristics. Proc. Natl. Acad. Sci. USA 6:
102.
OCKENFELS, P., M. BOPPRE, O. W. FISCHER, AND S.
SCHULTZ. 1998. Chemical communication in the sil-
ver-washed fritillary Argynnis paphia (Lepidoptera,
Nymphalidae). Poster from 15th Annual Meeting of
the International Society of Chemical Ecology. URL:
http://www.chemecol.org/meetings/98/ posters.html.
OLIVERIARA, P. S., AND A. V. L. FREITAS. 1991. Host
plant record for Eunica bechnina magnipunctata
(Nymphalidae) and observations on oviposition sites
and immature biology. Journal of Research on the
Lepidoptera 30: 140-141.
SHIROZU, T., AND T. YAMAMOTO. 1959. Morphology of
the male genital organs of Argyronome laodice
japonica M6n6tries (Lepidoptera: Nymphalidae).
Sieboldia 1: 161-168.
SIMONSEN, T. J. 2006a. The male genitalia segments in
fritillary butterflies: Comparative morphology with
special reference to the 'rectal plate' in Issoria (Lep-
idoptera: Nymphalidae). European J. Entomol. 103:
425-432.
SIMONSEN, T. J. 2006b. Fritillary phylogeny, classifica-
tion and larval hostplants: reconstructed mainly on
the basis of male and female genitalic morphology
(Lepidoptera: Nymphalidae: Argynnini). Biol. J. Lin-
nean Soc. 89: 627-673.
SIMONSEN, T. J. 2006c. Glands, muscles and genitalia.
Morphological and phylogenetic implications of his-
tological characters in the male genitalia of Fritil-
lary butterflies (Lepidoptera: Nymphalidae:
Argynnini). Zoologica Scripta, 35, 231-241.
SIMONSEN, T. J., N. WAHLBERG, A. V. Z. BROWER, AND R.
JONG. 2006. Morphology, molecules and Fritillaries:
approaching a stable phylogeny for Argynnini (Lepi-
doptera: Nymphalidae). Insect Systematics and Evo-
lution 37: 405-418.
TOLMAN, T., AND R. LEWINGTON. 1997. Butterflies of Eu-
rope. Princeton University Press. 310 pp.
VEROVNIK, R. 2000. A contribution to the knowledge of
the butterfly fauna (Lepidoptera: Rhopalocera) of
the Cerkljansko-Idrijsko region, west Slowenia, with
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wissenschaft 50: 277-358.


June 2008







Mankin et al: Sounds of Wood-Boring Beetle Larvae


TEMPORAL AND SPECTRAL FEATURES OF SOUNDS OF WOOD-BORING
BEETLE LARVAE: IDENTIFIABLE PATTERNS OF ACTIVITY ENABLE
IMPROVED DISCRIMINATION FROM BACKGROUND NOISE


R. W. MANKIN', A. MIZRACH2, A. HETZRONI2, S. LEVSKY3, Y. NAKACHE4 AND V. SOROKER3
'US Department of Agriculture, Agriculture Research Service,
Center for Medical, Agricultural, and Veterinary Entomology, Gainesville, FL, USA

Institute of Agricultural Engineering, Bet Dagan, Israel

'Institute of Plant Protection, Bet Dagan, Israel

4Havat Eden, Israel

ABSTRACT

Rhynchophorus (curculionid) larvae produce economic damage to ornamental and date palm
crops that could be mitigated significantly by early detection and treatment. Acoustic tech-
nology enables early detection, but often it is difficult to distinguish insect sounds from back-
ground noise containing energy at the resonant frequencies of stiff, fibrous structures in
trees and other plants. Tests were conducted with currently available acoustic instrumen-
tation and software to assess the capability of these methods to discriminate curculionid, ce-
rambycid, and buprestid larval sounds from background noise in woody structures. An
approach to the discrimination problem is to monitor the temporal patterns of the 3-10-ms
sound impulses produced by locomotory and feeding activities. Playback and computer anal-
yses of larval sounds revealed trains of impulses separated by intervals of less than 500 ms
that experienced listeners frequently use as indications of potential insect sounds. Further
analyses identified a subgroup of trains, denoted as bursts, containing > 6 and < 200 im-
pulses, which occurred frequently when larvae were present but only rarely when larvae
were absent. The incorporation of bursts into the analysis process significantly improved the
capability to distinguish sounds produced by beetle larvae from background noise when
these insects were hidden in stiff, fibrous structures, and likely will be of assistance also in
other applications where consistent activity patterns of hidden pests can be identified.

Key Words: Rhynchophorus ferrugineus, Monochamus titillator, Agrilus dozieri, acoustic de-
tection

RESUME

Las larvas de Rhynchophorus (Curculionidae) produce daio econ6mico a los cultivos orna-
mentales y a la palmer dactilifera que puede ser mitigado significativamente por la detec-
ci6n temprana y tratamiento. La tecnologia actstica permit una detecci6n temprana, pero
a menudo es dificil distinguir los sonidos hechos por los insects del ruido de fondo que con-
tiene energia a las frecuencias resonancia de las estructuras fibrosas presents en los arbo-
les y otras plants. Se realizaron pruebas con instruments acusticos actuales disponibles y
programs de computadora para evaluar la habilidad de estos m6todos para discriminar los
sonidos de larvas de las families Curculionidae, Cerambycidae y Buprestidae del ruido de
fondo de las estructuras de la madera. Un estrategia al problema de discriminaci6n es hacer
un monitoreo de los patrons temporales de los impulses de sonido de 3-10 ms (milisegun-
dos) producidos por las actividades de locomoci6n y alimentaci6n. El andlisis de los sonidos
de las larvas recolectados por la computadora y el repaso de la grabaci6n de sonido revel6
una series de impulses separados por intervalos de menos de 500 ms que los oyentes con ex-
periencia usan frecuentemente como indicadores de sonidos potenciales de insects. Los
andlisis subsiguientes identificaron un subgrupo de series, denotados como brotes repenti-
nos de > 6 y < 200 impulses, que ocurri6 con frecuencia cuando las larvas estaban presents
pero raramente cuando las larvas estaban ausentes. La incorporaci6n de los brotes repenti-
nos en el process de andlisis mejoro significativamente la habilidad para distinguir los soni-
dos producidos por larvas de escarabajos del ruido de fondo cuando estos insects estaban
escondidos en estructuras duras y fibrosas, y probablemente esta incorporaci6n sera de ayu-
dar en otras aplicaciones donde los patrons de actividad consistentes con plagas escondidas
pueden ser identificados.







Florida Entomologist 91(2)


The Rhynchophorus palm weevils (Coleoptera:
Curculionidae) are important worldwide pests of
palm trees, Arecaceae (Palmae) spp., including
economically important coconut (Cocos nucifera
L.), date (Phoenix dactylifera L.), and ornamental
palms (Murphy & Briscoe 1999). The American
palm weevil, R. palmarum L., is destructive in the
Neotropics (Giblin-Davis 1993, Oehlschlager et al.
2002), and the palmetto weevil (PTW),R. cruenta-
tus (F.), in southern Florida. The red palm weevil
(RPW), R. ferrugineus (Olivier), has caused losses
of up to 10-25% or more in coconut palm planta-
tions in Asia and the Middle East (Murphy &
Briscoe 1999). Currently, RPW is spreading in Eu-
ropean Mediterranean countries, endangering
picturesque landscapes attractive to tourists.
Adult females of these species lay eggs in
softer or protected areas of the trees, including in
wounds in the trunks of established trees, at the
base of the palm leaves at tree crowns, and adja-
cent to offshoots. The young larvae penetrate into
the trunk, creating cavities and tunnels (Giblin-
Davis 2001). Several generations may develop
within a single tree. Infested trees suffer from re-
duced productivity, and heavy infestations result
in collapsed trees, leading occasionally to total
loss of a date palm crop (Blumberg et al. 2001).
Adult populations of RPW, PTW, and R. pal-
marum can be efficiently monitored using phero-
mone based traps (Soroker et al., 2005; Oeshlager
et al., 1993, 1995 for RPW and PTW, respec-
tively). However, these methods are unsuitable
for quarantine inspections of planting material.
The larvae are large, but their cryptic tunneling
behavior makes their direct detection by visual
inspection impossible. Consequently, infested
planting material is often transported to a new lo-
cation before the first detectable symptoms of in-
festation appear. There is a need for direct, rapid
and accurate techniques for examination of trans-
ported planting material and suspected trees.
The chewing and locomotory activities of palm
weevil larvae produce distinct sounds, and heavy
infestations can often be heard by humans (Gib-
lin-Davis 2001). However, severe damage to palm
tissue already has occurred by the time the larval
sounds can be detected without electronic assis-
tance. Preliminary studies conducted by Mizrach
et al. (2003), Hetzroni et al. (2004a, b), and Soro-
ker et al. (2004, 2006) demonstrated that sensi-
tive microphones and dedicated amplifiers enable
detection of the movement and feeding sounds of
RPW larvae in palm shoots and trees. They char-
acterized the sounds as short impulses with
strong energy between 2 and 6 kHz, but confirmed
that certain types of background noise could in-
terfere with identification of the larval signal.
The experiments and signal processing analy-
ses reported here were conducted to gain experi-
ence with available acoustic technology and to
evaluate potential methods for improving the ca-


ability to discriminate sounds of beetle larvae
hidden in fibrous plants from incidental back-
ground noises.

MATERIALS AND METHODS

Insects and Recording Procedures

Sounds produced in 6 two-year-old potted date
palms infested with RPW larvae were compared
with sounds in 3 uninfested controls. Infestation
was conducted by inserting a 3rd-4th-instar (85.7
31.6 mg) into a 6-mm-diam. hole bored into the
trunk. Sounds of larval activity were recorded
weekly for 10 weeks for 1-2-min periods, once per
week starting one week after infestation, with a
microphone and amplifier (Larven Lauscher,
NIR-Service, Bad Vilbel, Germany) attached to
each tree (Soroker et al. 2004). In addition, labo-
ratory experiments were conducted to examine
patterns of insect feeding and movement sounds.
Second and third instars (20-30 mg) were intro-
duced into 28-33-mm-diam. stalks of sugarcane
(Saccharum officinarum L), cut into lengths of 60-
80 mm, on which they fed readily. The larvae were
inserted into holes drilled parallel to the fibers. A
1-2-min record was obtained from each stalk 62-
69 d after egg-laying. The larvae were weighed
immediately after the recordings had been ob-
tained, and ranged in weight from 0.17 to 1.7 g
(mean + SE, 0.76 + 0.156 g). Recordings also were
obtained from uninfested sugarcane stalks.
For comparisons with other tree-boring in-
sects, sounds of pine sawyer (PSW) larvae, Mono-
chamus titillator (F.), were collected for 1-2-min
periods on several different days from a pine bolt
with the aid of an accelerometer system (Mankin
et al. 2000, 2002). The ages of the larvae were un-
known, but 4 adults emerged over a 1-month pe-
riod, beginning about 2 weeks after the recording.
Sounds of 2 buprestid (BUP) larvae, Agrilus do-
zieri Fisher, were collected similarly from an oak
tree branch about 2 weeks before the adults
emerged, 3 d apart. The experiments with PSW
and BUP were conducted in a laboratory with low
levels of background noise. In all the experi-
ments, subjective evaluation of larvae activity
was conducted by listening to the sounds with a
headset while recording (usually by an author).
Recordings were digitized at 44.1 kHz on a com-
puter and saved in .wav format.

Signal Processing

Digitized signals were analyzed with Raven
1.2 software (Cornell Lab of Ornithology, Ithaca,
NY) and other dedicated signal sampling soft-
ware developed in Matlab (MathWorks, Inc., Nat-
ick, MA), or with a customized software program,
DAVIS (Mankin 1994; Mankin et al. 2000). Sig-
nals were band-pass filtered between 0.2 and 12


June 2008







Mankin et al: Sounds of Wood-Boring Beetle Larvae


kHz to eliminate low-frequency background
noise. Fast Fourier Transforms were calculated
on 256-point time-slices of the waveforms with a
Hamming window, and spectrograms were calcu-
lated from sections with 90 per cent overlap.
Mean spectra (profiles) used for characterizing
and identifying putative insect sound impulses
were calculated based on 512-point time slices
centered on the peak of each impulse in a section
of recording independently verified to contain in-
sect sounds without background noise (see Man-
kin 1994; Mankin et al. 2000, 2007).
In playback and oscillographic analyses of
these signals, it became evident that a consider-
able fraction of the larval sound impulses were of
low amplitude, barely above the peak background
noise levels. Consequently, it was necessary to
systematically adjust the signal processing am-
plitude threshold, Ta (Mankin et al. 2000), to
maintain its level just above background. This
was accomplished by calculating the root mean
square of consecutive samples in 0.186-s (8192-
point) time slices to estimate the peak to peak
background noise in each interval. The Ta was re-
set for each time slice by multiplying the root
mean square by a user-settable factor (3.25 in
these tests), which held Ta just above the local
peak background noise.
The DAVIS program identified and timed
groups (trains) of impulses with interpulse inter-
vals less than a preset duration, I, storing the be-
ginning and end times of these trains in a spread-
sheet along with the numbers of impulses in each
train. Impulses that failed to match one or more
specified insect sound profiles were discarded and
not entered in the spreadsheet. The records con-
tained sometimes as few as 30 or as many as
10,000 impulses, of which about 5%-70% were dis-
carded, depending on the background noise levels.
The beginning of a train was set as the beginning
of the first valid impulse after a period where the
interpulse interval was > I, and the end was set as
the end of the last valid impulse whose interpulse
interval was < I. We analyzed the signals in this
study using the setting, I = 500 ms.

RESULTS AND DISCUSSION

Larval Sound-Impulse Characteristics

Examples of sounds produced by moving and
feeding R. ferrugineus,A. dozieri, and M. titillator
larvae in a palm tree, oak branch, and pine bolt,
respectively, are shown in Fig. 1. Signals pro-
duced by RPW in sugarcane were similar in tem-
poral pattern to those in palm. The beetle larvae
produced variable-amplitude, 3-10-ms impulses
with strong energy between 0.4-8 kHz (see spec-
trograms A, B, and C in Fig. 1). The signals ex-
tend to higher frequencies than the 0.5-1.8 kHz
range of sounds produced in soil because wood


A.




A.

IL
B.




B.


C.

ii


4







8
4

0 ~ ~ L


C.

4 !
0.5 1.0 1.5
Time (s)

Fig. 1. Oscillograms, and spectrograms of several
sound impulses produced by a (A., A.) RPW, (B., B ) BUP,
and (C., C.) PSW larvae; in the spectrograms, a darker/
lighter color indicates frequencies of greatest/least en-
ergy.


has a lower attenuation coefficient than soil at
high frequencies (Mankin et al. 2000). Other dif-
ferences between the transmission of sound in a
fibrous structure and soil include resonances that
are influenced by the dimensions, mass, and stiff-
ness of the structure (Cremer et al. 1988; Evans
et al. 2005; Hambric 2006). Signals with energy
at a resonance frequency will transmit farther in
structures than nonresonant signals, and the
long-distance transmission of background noises
with components at resonant frequencies can in-
terfere with detection of weak, insect-produced
signals. One such resonance can be seen as a con-
tinuous, faint band near 2.5 kHz in the RPW spec-
trogram (Fig. 1A), and similar resonances can be
seen near 3 kHz in the BUP spectrogram (Fig.
1B ) and near 4.6 kHz in the cerambycid spectro-
gram (Fig. 1C).
The effects of resonance are observable not
only in the background noise spectra but also in
the spectra of larval sound impulses (Fig. 2A-B).
The examples of RPW sounds recorded from sep-
arate palms have peaks near 1.1 and 2.6 kHz in
Fig. 2A. The sounds recorded from sugarcane
have similar peaks. However, an example of palm







Florida Entomologist 91(2)


RPW in palm


Frequency (kHz)
B PSW in pine


it ... .... .... -- -- ---- -- - -- ---- -- ------ ---
8'\

-20 t-

II
Z iBackground BUP in oak
0 2 4 6 8
Frequency (kHz)

Fig. 2. Mean spectra of sounds recorded during a 1-
min period from larval infestations: (A) RPW in palm
trees (dashed line, n = 303; dash-dot-dot line, 348 im-
pulses) and in sugarcane (dash-dot line, n = 139), and
(B) BUP in oak branch (dash-dot line, n = 142) and PSW
in pine bolt (dashed line, n = 131). Example spectrum of
background noise is shown in (A) palm tree and (B) oak
branch (solid lines), with background peak at 1 kHz
marked as BP1, and peak at 2.4 kHz marked as BP2.


tree background noise also has peaks very close to
these values, at 1(marked at BP1 in Fig. 2A) and
2.4 kHz (marked at BP2); consequently, the en-
ergy at these frequencies is not due necessarily to
the larvae alone. This result is unlike what is typ-
ically found for background noise in soil or air
(Mankin 1994; Mankin et al. 2000; Mankin &
Benshemesh 2006), where the energy of most
background noise was found to decrease rapidly
above 0.2 kHz. In previous studies with soil in-
sects, the relatively low energy of background
noise at frequencies above 0.5 kHz enabled con-
struction of insect sound profiles and background
noise profiles that could be used to identify indi-
vidual sound impulses as insect sounds or back-
ground noises (Mankin et al. 2000, 2001, 2007).
For RPW in palm trees, there are differences in
the spectra of larval and background noise be-
tween 3.4 and 6 kHz (Hetzroni et al. 2004a, b, see
also Fig. 2A) that can be used to distinguish RPW
sounds from background noise. Such differences
were used to correctly classify ca. 90% of sounds


produced in infested palm trees (Hetzroni et al.
2004a, b). However, the examples of BUP spectra
and background noise in oak branches present a
more complicated result. In Fig. 2B, the BUP sig-
nal has peaks near 4.7 and 5.5 kHz, but there also
are large peaks near 4.5 and 54 kHz in background
sounds recorded from an uninfested oak branch.
Other examples have been observed, particularly
in urban environments, where background noise
contains peaks of high energy above 0.5 kHz;
which provides impetus to develop another method
in addition to spectral profile analysis to help dis-
tinguish insect sounds from background noises.

Larval Sound Temporal Patterns

An additional method of interest for discrimi-
nating insect sounds from background noise is
the identification of repetitive patterns that may
occur when the larvae move inside their tunnels
or scrape the wood fibers during feeding or tun-
neling activity. An example of such a pattern is
seen in Fig. 3, adapted from Hetzroni et al.
(2004a, b) A group of repeated impulses appear in
Fig. 3Aa that are clearly distinguishable from the
unpatterned impulses that were recorded from
the uninfested palm (Fig. 3Bb). The existence of
identifiable, repeated patterns in records of insect
sounds has been noted previously (Andrieu &
Fleurat-Lessard 1990; Mankin et al. 1997; Zhang
et al. 2003).
Unlike the patterned sounds produced during
insect communication (e.g. Walker et al. 2003),
the RPW larval sound patterns examined in this
study, as well as those produced by the BUP and
PSW larvae, were consistent over only short peri-
ods of time. The most consistent pattern was not
the series of high-amplitude impulses seen in Fig.
3A, but rather a mixture of low- and high-ampli-
tude impulses that occurred in well-defined
trains with short interpulse intervals, separated
by 0.5-s or longer intervals between trains. Expe-
rienced listeners usually recognized these trains
as distinct sounds, and frequently recognized
them as characteristic of insect sounds.
The numbers of impulses per train varied con-
siderably over the records in the palm tree study (1-
99 impulses) and even greater in the sugarcane
tests (1 239 impulses). Playbacks of records sug-
gested that trains with > 6 but < 200 impulses were
most recognizable to experienced listeners as insect
movement and feeding activity. Generally, back-
ground noises in records from uninfested wood or
sugarcane had fewer than 7 impulses per train.
Consequently, we separated out a particular class of
impulse trains, those with > 6 but < 200 impulses
separated by intervals < 500 ms, as potential predic-
tors of insect infestation, denoting them as bursts.
The distributions of impulse trains and bursts
in six recordings from different insects are shown
in Table 1, and examples of several impulse trains


June 2008







Mankin et al: Sounds of Wood-Boring Beetle Larvae


10 20


, i I


* L. 1-*11 .I i


30 40 p
Impulse pattern


,I, .11


'' lr


-ii


wl. 1.wav -44- 1 H; detOction0-1


Time (s)


Fig. 3. Acoustic identification of RPW larval activity recorded from an infested (A) and uninfested (B) palm trees.
Continuous line indicate original signal and shaded ovals indicate positive identification of potential larvae activity.
Expanded view (Aa) show regularly timed impulses recorded from infested palm, and expanded view (Bb) shows ir-
regularly timed impulses recorded from uninfested palm.


denoted as bursts are shown in Fig. 4. Most of the
computer-identified bursts in the 4 examples, de-
noted by the numbered, shaded areas, correspond
to groups of low-and high-amplitude impulses
separated by periods with only sparsely occurring
impulses. In Fig. 4A1, however, almost all of the
impulses are low-amplitude, which suggests that
the use of burst analysis as a method to augment
profile analysis will be most successful in tests
where sensors can detect very weak signals near
the background noise threshold.
Use of Impulse-Train and Burst Analyses to Assess In-
festation Likelihood

An important goal of insect acoustic detection
is to discriminate whether or not individually
tested samples contain infestations. A successful
approach has been to construct indices of infesta-
tion based on quantitative and/or qualitative char-
acteristics of detected sounds, and assess the like-
lihood of infestation using discretized indicators,
e. g., low, medium, and high likelihood ofinfesta-


tion based on lower and upper thresholds of the in-
dices (Mankin et al. 2007). In 1 example, recording
sites in soil were classified at a low likelihood of
white grub, Phyllophaga crinita (Burmeister), in-
festation if 2 or fewer sounds (impulse-trains) per
min were detected that matched white grub spec-
tral profiles and at high likelihood if greater than

TABLE 1. NUMBERS OF IMPULSE TRAINS AND BURSTS DE-
TECTED IN 1-MIN RECORDINGS FROM LISTED
SOURCE.

Source No. trains No. bursts'

RPW in palm 55 15
RPW in palm 13 12
BUP in oak branch 29 5
BUP in oak branch 28 3
PSW in pine bolt 6 6
PSW in pine bolt 6 5

'No. impulse trains with > 6 and < 200 impulses with inter-
vals <500 ms.





1"- r I '


I I







Florida Entomologist 91(2)


A1l

Z0
E- .


A2
) 0 f3











-c 1 1
.S a




-1
C

7 Tn



2 4 6 8
Time (s)

Fig. 4. Examples of impulse trains identified by com-
puter as potential larval sound bursts (indicated by
numbering and shading) in records from (Al) and (A2)
RPW in palm, (B) BUP in oak branch, and (C) PSW in
pine bolt.



20 impulse-trains per min were detected (Mankin
et al. 2007). Here we considered a similar use of
impulse-train bursts to assess RPW infestations in
the laboratory tests with sugarcane.
Initial analyses of records from infested and
uninfested sugarcane indicated that the RPW-in-
palm and RPW-in-sugarcane spectral profiles in
Fig. 2 readily classified background signals as


noise with DAVIS and correctly identified signals
produced in infested sugarcane as being produced
by RPW. To consider the distribution of impulse-
train bursts in infested and uninfested sugar-
cane, we analyzed one 50-s segment recorded
from each of 21 different infested and 4 unin-
fested stalks, discarding sounds that did not
closely match the profiles (Mankin et al. 2007,
2000). The results are summarized in Table 2,
where the complete data set is separated into 4
categories, 1 for recordings in uninfested sugar-
cane, and 3 for records in infested sugarcane
where the numbers of bursts ranged from 7 to 17,
4 to 6, and 0 to 3, respectively, and the mean lar-
val weights, numbers of trains, and numbers of
impulses per train are listed for each burst cate-
gory. The correlation the numbers of bursts and
the larval weight was not significant (Spearman
= 0.41, P = 0.08 for null hypothesis of p = 0) (Proc
CORR, SAS Institute 2004a). Likewise, the corre-
lation between numbers of impulse trains and
larval weight correlation was not significant
(Spearman p = 0.06, P = 0.79 for null hypothesis
of p = 0) (Proc CORR, SAS Institute 2004a).
The observed distribution of bursts suggested
that the likelihood of infestation (Mankin et al.
2007) in these tests could be set at: Low, n, < 1;
Medium, 1 < nb < 3; High, n, > 3, where n, is No.
bursts per 50-s record. Based on these criteria, 2
of the 4 records from uninfested sugarcane had
Low likelihood of infestation, and 2 had Medium
likelihood of infestation, while 2, 1, and 18 of 21
records from infested sugarcane had Low, Me-
dium, and High likelihood of infestation, respec-
tively. These 2 distributions were significantly
different under the Wilcoxon Two-sample Exact
Test, with sum of scores = 17.0 and P = 0.0025
(Proc NPAR1WAY, SAS Institute, 2004b). Al-
though additional testing will be necessary to es-
tablish the practical efficacy of incorporating
burst analysis into acoustic surveys of RPW in
palm trees and offshoots, these results indicate
that it is possible, using already existing signal
processing software, to extract previously unmea-


TABLE 2. COMPARISONS OF MEAN LARVAL WEIGHTS, NO. IMPULSE TRAINS, AND NO. IMPULSES PER TRAIN FOR RECORD-
INGS IN WHICH LISTED NOS. BURSTS WERE DETECTED IN 50-S PERIOD.

Mean SE

Range of nos. bursts' Larval wt. (g) (n,) No. trains (n,) No. imp. / train

7-17 0.98+ 0.19 (9) 20.10 2.45 (10) 15.00 5.74
4-6 0.69 0.39 (7) 14.14 + 5.59 (7) 40.10 17.27
0-3 0.27 0.05 (3) 1.25 0.75 (4) 47.80 44.36
Uninfested2 6.5 1.71 (4) 5.58 2.93

Records with Nos. bursts in listed range, where a burst is an impulse train with > 6 and < 200 impulses: (n,), No. records in
mean for larval weight in listed burst range; (n,), No. records in means for trains and impulses per train in listed burst range.
Weights not available for 2 records.
In 4 recordings from uninfested sugarcane, 2 had 0 bursts, and 2 had 1 burst.


June 2008




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