A Florida Perspective on Host Preference, Early Detection, and Identification of the Brown Marmorated Stink Bug, Halyomo...

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Title:
A Florida Perspective on Host Preference, Early Detection, and Identification of the Brown Marmorated Stink Bug, Halyomorpha Halys (Stal)
Physical Description:
1 online resource (82 p.)
Language:
english
Creator:
Poplin, Ashley V
Publisher:
University of Florida
Place of Publication:
Gainesville, Fla.
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Thesis/Dissertation Information

Degree:
Master's ( M.S.)
Degree Grantor:
University of Florida
Degree Disciplines:
Entomology and Nematology
Committee Chair:
HODGES,AMANDA C
Committee Co-Chair:
CAPINERA,JOHN LOWELL
Committee Members:
BALDWIN,REBECCA W
EGER,JOSEPH E
SMITH,TREVOR R

Subjects

Subjects / Keywords:
brown -- bug -- choice -- field -- florida -- guide -- host -- invasive -- marmorated -- no-choice -- preference -- species -- stink
Entomology and Nematology -- Dissertations, Academic -- UF
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Entomology and Nematology thesis, M.S.
bibliography   ( marcgt )
theses   ( marcgt )
government publication (state, provincial, terriorial, dependent)   ( marcgt )
born-digital   ( sobekcm )
Electronic Thesis or Dissertation

Notes

Abstract:
The brown marmorated stink bug (Halyomorpha halys (Stål)) is an invasive agricultural and urban pest in the United States with over 300 recorded host plants. Since its first detection in the late 1990s, this species has become established in at least twelve states and the distribution continues to expand. Although it is not known to be established in Florida, increasing our understanding of the pest potential for H. halys in Florida may allow for early detection and provide informed, economical management recommendations. To determine host specificity and potential impact of H. halys, eight plant species commonly found in Florida’s agronomic crops and landscapes were tested using choice and no-choice tests. The results suggested that H. halys preferred to feed on woody hosts, such as ligustrum and hibiscus, as opposed to Citrus spp. and avocado. This research will be continued using a variety of other plant species in order to yield more conclusive results.   A Florida-specific stink bug field guide and invasive species workshop trainings targeted to Florida State Park Service personnel were developed to increase identification and monitoring efforts of H. halys, and ultimately assist with management efforts if and when it establishes. The efficacy of the stink bug field guide was assessed through an identification activity, and only 18% of participants correctly identified H. halys. Pre- and post-surveys used to assess the efficacy of the workshop trainings illustrated an overall positive impact. Future training sessions will target Florida Master Gardeners, nurserymen, small farm producers, and public gardens personnel.
General Note:
In the series University of Florida Digital Collections.
General Note:
Includes vita.
Bibliography:
Includes bibliographical references.
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Description based on online resource; title from PDF title page.
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This bibliographic record is available under the Creative Commons CC0 public domain dedication. The University of Florida Libraries, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.
Statement of Responsibility:
by Ashley V Poplin.
Thesis:
Thesis (M.S.)--University of Florida, 2013.
Local:
Adviser: HODGES,AMANDA C.
Local:
Co-adviser: CAPINERA,JOHN LOWELL.

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UFRGP
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Applicable rights reserved.
Classification:
lcc - LD1780 2013
System ID:
UFE0046364:00001


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1 A FLORIDA PERSPECTIVE ON HOST PREFERENCE, EARLY DETECTION, AND IDENTIFICATION OF THE BROWN MARMORATED STINK BUG, Halyomorpha halys (Stl) By ASHLEY V POPLIN A THESIS PRESENTED TO THE GRADUATE SCHOOL OF THE UNIVERSITY OF FL ORIDA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE UNIVERSITY OF FLORIDA 2013

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2 2013 Ashley V Poplin

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3 To my mom and dad, without whom I would not be where I am or who I a m today. To Ben, Alex, Beth, Renee, and the rest of my family and friends. Thank you all for being a source of constant support and motivation.

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4 ACKNOWLEDGMENTS First, I would like to express my thanks to Dr. Amanda Hodges for her overwhelming guidanc e and support. I would also like to thank Dr. Rebecca Baldwin, Dr. John Capinera, Dr. Joe Eger, and Dr. Trevor Smith for their assistances as supervisory committee members and contribution to this research. This work was supported in part by funds from the University of Florida Steinmetz funds for biosecurity research. The Invasive Species Workshop Series was supported by USDA APHIS PPQ Farm Bill 10201, Protect U.S., Southern Plant Diagnostic Network, USDA NIFA, and Sentinel Plant Network. I would like to thank Dr. Julio Medal and Andrew Santa Cruz for their assistance in rearing the H. halys laboratory colony, Dr. Trevor Smith for providing space in the FDACS DPI quarantine facility, Phillip Lake for providing assistance with quarantine permit questions, N ik laus Hostettler for efforts and assistance with the University of Florida quarantine facility, and Dr. Mark Abney from the University of Georgia and Kathleen Tattman from USDA ARS BIIRU for providing specimens. I would also like to thank the UF Biosecuri ty Research and Extension Lab for all of their support with bioassays, plant maintenance, and extension education support; Dr. Gurpreeet Brar, Carla Burkle, Hashmet Eke, Abby Griffin, Sarahlynne Guerrero, Dr. Jennifer Hamel, Eric Le V een, Annika Minott, Ste phanie Stocks, and Kay Weigel. I wish to thank the Florida State Park Service biologists and staff for hosting the Invasive Species workshops, and for the guest speakers whom delivered presentations at the workshop; Bradley Danner, Brian Saunders, and Dr Leroy Whilby from Florida CAPS Program; Lyle Buss; Dr. Teresa Cooper; Dr. James Cuda; Dr. Jiri Huclr; and Sedonoia Steininger. I also wish to thank the Florida County Extension Offices and the

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5 Florida Master Gardeners for participating in the assessment of Identifying Stink Bugs and Similar True Bugs of Florida, in addition to Dr. Joe Eger and Lyle Buss for providing assistance in the development of the field guide, and Kay Weigel for her graphic design work.

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6 TABLE OF CONTENTS page ACKNOWLEDGMENTS ................................ ................................ ................................ .. 4 LIST OF TABLES ................................ ................................ ................................ ............ 8 LIST OF FIGURES ................................ ................................ ................................ .......... 9 ABSTRACT ................................ ................................ ................................ ................... 10 CHAPTER 1 IMPORTANCE OF INVASIVE SPECIES AND THE BROWN MARMORATED STINK BUG IN FLORIDA ................................ ................................ ....................... 12 Introduction ................................ ................................ ................................ ............. 12 Literature Review of the Bro wn Marmorated Stink Bug ................................ .......... 15 Life History ................................ ................................ ................................ ....... 15 Natural Enemies ................................ ................................ ............................... 17 Co ntrol Methods ................................ ................................ ............................... 17 Host Plants ................................ ................................ ................................ ....... 18 Monitoring Methods ................................ ................................ .......................... 19 Identifica tion ................................ ................................ ................................ ..... 21 Brown Marmorated Stink Bug Extension Education ................................ ............... 22 Research Objectives ................................ ................................ ............................... 23 2 LABORATORY HOST PREFERENCE OF THE BROWN MARMORATED STINK BUG ( HALYOMORPHA HALYS (STL)) ON PLANT SPECIES RELATIVE TO FLORIDA ................................ ................................ ........................ 30 Introduction ................................ ................................ ................................ ............. 30 Materials and Methods ................................ ................................ ............................ 32 Laboratory Colony ................................ ................................ ............................ 32 Plant Sources ................................ ................................ ................................ ... 33 No choice Tests ................................ ................................ ............................... 34 Choice Tests ................................ ................................ ................................ .... 34 Statistics ................................ ................................ ................................ ................. 36 No choice Tests ................................ ................................ ............................... 36 Choice Tests ................................ ................................ ................................ .... 36 Results ................................ ................................ ................................ .................... 37 No choice Tests ................................ ................................ ............................... 37 Choice Tests ................................ ................................ ................................ .... 37 Discussion ................................ ................................ ................................ .............. 38 3 EVALUATION O F THE INVASIVE SPECIES WORKSHOP PROGRAM FOR FLORIDA PARK SERVICE PERSONNEL ................................ .............................. 46

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7 Introduction ................................ ................................ ................................ ............. 46 Materials and Methods ................................ ................................ ............................ 48 Target Audience ................................ ................................ ............................... 48 Workshop Presentations ................................ ................................ .................. 48 Evaluation Instruments ................................ ................................ ..................... 49 Statistics ................................ ................................ ................................ ................. 51 Results ................................ ................................ ................................ .................... 51 Demographics ................................ ................................ ................................ .. 51 Pre test and Post test Evaluation ................................ ................................ ..... 51 Post workshop Pest Surveys ................................ ................................ ............ 52 Discussion ................................ ................................ ................................ .............. 52 4 ASSESSMENT OF IDENTIFYING STINK BUGS AND SIMILAR TRUE BUGS OF FLORIDA FIELD GUIDE ................................ ................................ ................... 60 Introduction ................................ ................................ ................................ ............. 60 Materials and Methods ................................ ................................ ............................ 61 Development of Field Guide ................................ ................................ ............. 61 Target Audience ................................ ................................ ............................... 62 Evaluation Instruments ................................ ................................ ..................... 62 Identification Activity ................................ ................................ ......................... 63 Delivery of Materials and Assessme nts ................................ ............................ 64 Statistics ................................ ................................ ................................ ................. 64 Results ................................ ................................ ................................ .................... 65 Demographics ................................ ................................ ................................ .. 65 Identification Activity ................................ ................................ ......................... 65 Post test Responses ................................ ................................ ........................ 65 Discussion ................................ ................................ ................................ .............. 66 5 IMPLICATIONS AND FUTURE DIRECTIONS FOR H. halys RESEARCH ............ 72 LIST OF REFERENCES ................................ ................................ ............................... 75 BIOGRAPHICAL SKETCH ................................ ................................ ............................ 82

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8 LIST OF TABLES Table page 1 1 Reported hosts of Halyomorpha halys ................................ ............................... 25 2 1 All combinations of plant species tested in choice tests of adult Halyomorpha halys ................................ ................................ ................................ .................. 42 2 2 Mean number of Halyomorpha halys individuals recovered in no choice tests. 43 4 1 Correct responses in the identification activity for the assessment of Identifying Stink Bugs and Similar True Bugs of Florida. ................................ .... 67

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9 LIST OF FIGURES Figure pag e 2 1 Mean (standard error of the mean, SEM) number of adult Halyomorpha halys present or feeding on test plant species in choice tests. Means with the same letter are not significant ly different (Tukey Kramer, P=0.05). .............................. 44 2 2 Mean (standard error of the mean, SEM) number of adult Halyomorpha halys feeding on any test plant at different times of day. Means with the same let ter are not significantly different (Tukey Kramer, P=0.05). ................................ ....... 45 3 1 Pre test for the Invasive Species Workshop Series. ................................ ........... 54 3 2 Front page of the post test for the Invasive Species Workshop Series. ............. 55 3 3 Back page of post test for the Invasive Species Workshop Series. .................... 56 3 4 Overall mean question responses from post test. A) Mean question responses for question numbers 1 18. B) Mean question responses for question numbers 19 36. The responses were assigned numerical values: strongly disagree (SD) = 1; disagre e (D) = 2; uncertain (U) = 3; agree (A) = 4; and strongly disagree (SA) = 5. ................................ ................................ .......... 57 3 5 Differences in mean change of question responses from post test between participant levels of education. Mean s with the same letter are not ................................ ............ 58 3 6 Differences in mean change of question responses from post test between participant experience with invasive species workshops. Means with the .............. 59 4 1 Overall percentage of correct responses from the identification acti vity used for assessing Identifying Stink Bugs and Similar True Bugs of Florida. Participants classified each specimen as herbivore or predator, as well as identify the species of each specimen. ................................ ............................... 68 4 2 Pre test for assessment of Identifying Stink Bugs and Similar True Bugs of Florida. ................................ ................................ ................................ ............... 69 4 3 Identification activity for the assessment of Identifying Stink Bugs and Similar True Bugs of Florida. ................................ ................................ .......................... 70 4 4 Post test for the assessment of Identifying Stink Bugs and Similar True Bugs of Florida. ................................ ................................ ................................ ........... 71

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10 Abstract of Thesis Presented to the Graduate Scho ol of the University of Florida in Partial Fulfillment of the Requirements for the Degree of Master of Science A FLORIDA PERSPECTIVE ON HOST PREFERENCE, EARLY DETECTION, AND IDENTIFICATION OF THE BROWN MARMORATED STINK BUG, Halyomorpha hal ys (Stl) By Ashley V Poplin December 2013 Chair: Amanda C Hodges Major: Entomology and Nematology The brown marmorated stink bug ( Halyomorpha halys (Stl)) is an invasive agricultural and urban pest in the United States with over 300 recorded host plants. Since its first detection in the late 1990s, this species has become established in at least twelve states and the distribution continues to expand. Although it is not known to be established in Florida, increasing our understanding of the pest pot ential for H. halys in Florida may allow for early detection and provide informed, economical management recommendations. To determine host specificity and potential impact of H. halys eight ndscapes were tested using choice and no choice tests. The results suggested that H. halys preferred to feed on woody hosts, such as ligustrum and hibiscus, as opposed to Citrus spp. and avocado. This research will be continued using a variety of other pla nt species in order to yield more conclusive results. A Florida specific stink bug field guide and invasive species workshop trainings targeted to Florida State Park Service personnel were developed to increase identification and monitoring efforts of H. halys and ultimately assist with management

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11 efforts if and when it establishes. The efficacy of the stink bug field guide was assessed through an identification activity, and only 18% of participants correctly identified H. halys Pre and post surveys u sed to assess the efficacy of the workshop trainings illustrated an overall positive impact. Future training sessions will target Florida Master Gardeners, nurserymen, small farm producers, and public gardens personnel.

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12 CHAPTER 1 IMPORTANCE OF INVASIVE S PECIES AND THE BROWN MARMORATED STINK BUG IN FLORIDA Introduction An invasive pest is defined as a plant, animal or pathogen species that is not native to the ecosystem that causes or is likely to cause economic or environmental harm or harm to human anim al, or plant health (ISAC 2006). Invasions by non native species in the United States cost an estimated $137 billion annually. Arthropod species account for $20 billion of this total due to environmental damages, lost agricultural productivity, and increas ed health problems (Pimentel et al. 2010). The introduction and establishment of an invasive species often leads to expensive eradication and control efforts and can severely impact the trade industry in addition to the direct environmental and agricultura l damage caused by pest species. Because invasive species are commonly introduced through accidental human involvement or immigration, it is no surprise that Florida is considered a high risk state for invasive species due to multiple pathways of introduc tion, suitable climate conditions, and a large agriculture industry. With 12 international airports and 14 deepwater seaports, Florida is a major destination for global trade especially for agricultural commodities (Silagyi 2005). In fact, Miami Internatio nal Airport is the epicenter of United States cut flower, and fruit and vegetable imports, receiving 88% and 55% of shipments respectively. Only a small percentage of imported plant materials are sampled and inspected at ports of entry by the Department of Animal and Plant Health Inspection Service (APHIS) for exotic plant pests, diseases, pathogens, and noxious weeds (USDA APHIS PPQ 2012).

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13 Although safegua rding systems have been established and implemented by state and federal agencies to prevent, detect, eradicate, and mitigate damage of invasive pests, an average of two new non native arthropod species are currently detected in Florida per month. The Flor ida Cooperative Agricultural Pest Survey (CAPS) program also facilitates the prevention of pest introduction through import inspections at high risk interstate interdiction stations. In 2012, the Florida CAPS Interdiction Survey team collaborated with FDAC S Agricultural Law Enforcement and detected 116 exotic pest species (Russel 2012). Of the 116 species, an important invasive species commonly known as the brown marmorated stink bug ( Halyomorpha halys (Stl) (Hemiptera: Pentatomidae)) was intercepted at le ast three times. Additional interceptions of this invasive species were from tourist vehicles travelling to Florida from states infested with this pest (Halbert and Hodges 2011). Significance of Halyomorpha halys Halyomorpha halys the brown marmorated s tink bug, is an invasive pentatomid species found in North America. This highly polyphagous species has over 300 host plants recorded (Nielsen et al. 2008a), including host plants of agricultural importance such as vegetable crops, woody shrubs, and fruit trees. Initially, H. halys was not known to be a significant agricultural pest in the United States. Since 2006, H. halys has become a major pest of fruit trees and soybean in the Northeastern and Mid Atlantic U.S. Nielsen and Hamilton (2009a) recorded 25 80 % damage by H. halys to apples and pears in Pennsylvania and New Jersey. In the early season, H. halys may cause catfacing, dimpling, or concave surface damages. If H. halys feeds on the fruit in mid to late season, necrotic areas are seen where the

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14 p roboscis was inserted. As the distribution of H. halys has expanded and numbers increased, it has become a pest of economic concern in the United States. Halyomorpha halys is native to East Asia and is found in China, Japan, Korea, Myanmar, Taiwan, and Vi etnam (Yu and Zhang 2007; Yang et al. 2009). Although the means of introduction are not certain, H. halys is thought to have been accidentally brought into the United States by hitchhiking on imported cargo from China, Japan, or Korea (Hoebeke and Carter 2 003). In 2001, H. halys was detected in Allentown, Pennsylvania and identified by Karen M. Bernhard, an extension entomologist with the Lehigh County Cooperative Extension office. Although this was the first report of this species in the Western hemisphere H. halys is believed to have immigrated to the United States as early as 1996. Since 2001, H. halys has become established in the following states: llinois, Maryland, Michigan, Minnesota, New Jersey, North Carolina, Ohio, Oregon, Pennsylvania, Tennessee Virginia, and Washington (NAPIS 2013). Halyomorpha halys has also been detected or intercepted, but is not known to be established, in the following states: Colorado, Connecticut, Florida, Georgia, Indiana, Kentucky, Maine, Massachusetts, New Hampshire, Rhode Island, South Carolina, Vermont, West Virginia, and Wisconsin (Halbert, personal communication; NAPIS 2013). Halyomorpha halys has also spread to Europe. In 2007, H. halys was reported in Zurich, Switzerland. Wermelinger et al. (2008) described five separate occurrences of H. halys in Switzerland three reports of single individuals, one report of egg masses, and one report of infestation. At this time, it is unknown how the species was introduced to Zurich.

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15 Literature Review of the Brown Marmorated Stink Bug Life History As with most pentatomid species, Halyomorpha halys (Stl) completes five nymphal instars before reaching the adult stage. At an optimal temperature of 25 o C, egg ecolsion occurs after six days from oviposition (Nielsen et al. 2008a). First instar nymphs burst through a black triangular structure (egg burster) and exit through a circular operculum surrounded by micropylar processes at the top of the egg (Yang et al. 2009). Nymphs tend to aggregate around the egg mass until ecdysis, or molting, from first to second instar occurs. Second instar nymphs disperse and begin feeding on leaves, stems, and occasionally the fruit of host plants. Developmental time at an optimal temperature of 25 o C for first second third fourth and fifth i nstar nymphs is 5, 10, 7, 7, and 11 respectively with a total developmental time (egg incubation to adult) of 50 days (Nielsen et al. 2008a). Although development can occur between 17 and 33 o C, temperature significantly effects developmental time and morta lity. Adult H. halys emerge mid March to April and are active until October in Pennsylvania (Nielsen and Hamilton 2009b). Females must complete reproductive maturation before copulation can occur. Adult females will oviposit between June and September in the northeast. Egg masses are deposited in clusters on the underside of leaves with an average of 28 eggs per mass (Nielsen et al. 2009b; Leskey et al. 2012b; Medal et al. 2012). The eggs are small (about 1mm diameter), pale green to white in color, and sp herically shaped (Gyeltshen et al. 2011). From mid September to early spring in its current U.S. distribution, adults reach a reproductive diapause and exhibit overwintering behavior. Males release pheromones for aggregation purposes, although the pheromo ne has not been identified, and large

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16 numbers of aggregated adults will retreat to nearby buildings and structures (Yu and Zhang 2007). In contrast, other native stink bugs (i.e. Euschistus spp., Brochyamena spp. and Parabrochyamena spp.) diapause in leaf litter of wooded areas. Since effective methods for control or long term management are not available, homeowners are being overwhelmed with the quantity of H. halys invading their homes (Jacobs 2009; Gyeltshen et al. 2011; Halbert and Hodges 2011). Haly omorpha halys has been known to produce one or two generations per year, both in northern China and mid Atlantic regions of the United States (Nielsen et al. 2008a; Nielsen and Hamilton 2009b). However, H. halys has produced up to six generations per year in southern regions of China (Hoffman 1931) and a similar trend may occur in southern states such as Florida. According to a potential geographic distribution map produced using ecological niche modeling, the northern regions of Florida are highly suitable for H. halys establishment while central and southern regions of Florida may be less suitable for establishment (Zhu et al. 2012). Ecological niche modeling identifies environmentally suitable areas for invasive species by comparing climatic conditions in their native range to areas where the species have or could have been introduced. Zhu et al. (2012) considered six bioclimatic variables (e.g. temperature, precipitation, sunshine, and elevation) to produce the potential geographic distribution of H. haly s Although ecological niche modeling is useful for detecting potential areas of invasion and generates opportunities for exclusion, it does not consider biotic interactions (i.e., predators, parasitoids, pathogens, etc.) or dispersal ability of a species. For that reason, all areas of predicted suitability as suggested by ecological niche modeling may not be suitable for establishment.

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17 Natural Enemies There are many natural enemies of H. halys with some being more effective than others. Predators and para sitoids of H. halys may attack the eggs, nymphs, or adults and can include parasitic microhymenoptera and flies, ants, earwigs, lacewings, assassin bugs, other stink bugs, spiders, and birds. Six egg parasitoids of H. halys have been documented in China, each with varying rates of parasitism. These are: Trissolcus flavipes Thomon, Telenomus mitsukurii (Ashmead), Anastatus spp., Acroclosoides spp., Telenomus podisi Ashmead, and Trissolcus halyomorphae Yang (Yu and Zhang 2007; Yang et al. 2009). Of the six s pecies mentioned, T. halyomorphae has the highest parasitism rate and seems to be the most effective egg parasitoid for biological control (Yang et al. 2009). Control Methods Nielsen et al. 2008b tested the toxicity of nine insecticides on H. halys in the laboratory: one organophosphate (phosmet); three neonicotinoids (dinotefuran, cyfluthrin, cyfluthrin, cyhalot hrin, and neonicotinoids, dinotefuran and thiomethoxam, were the most effective insecticides against H. halys populations (Nielsen et al. 2008b). However, the insecticides were only tested in a laboratory setting and field results have not yet been reporte d. Although insecticides may provide minor relief in and around homes, they have not been successful at controlling populations of H. halys for long term pest management (Jacobs 2009). Instead, natural enemies have been the focus for potential biological control. In the U.S., no known naturally occurring specialist predators of H.

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18 halys currently exist. In China, Trissolcus halyomorphae is a native solitary egg parasitoid of H. halys with a high parasitism rate of 20 70% (Yang et al. 2009). In northern r egions of China, T. halyomorphae can respond rapidly to increases in numbers of H. halys because T. halyomorphae can produce up to ten generations per year while H. halys produces one or two generations per year. It is unknown whether this species would b e an effective biological control agent or negatively impact native stink bugs in the United States. The Agricultural Research Service (ARS) program of the United States Department of Agriculture (USDA) is collaborating with several states in a research pr oject to determine the suitability of T. halyomorphae as a biological control agent of H. halys Testing of Trissolcus halyomorphae is currently taking place in Delaware, Michigan, Mississippi, Oregon, and Florida (Smith, personal communication). In Florid a, the Bureau of Methods Development and Biological Control at the Florida Department of Agriculture and Consumer Services Division of Plant Industry (FDACS DPI) is currently researching the host preference of T. halyomorphae by exposing the parasitoid to native and established stink bugs of Florida. If the results of the research are favorable, T. halyomorphae may be considered for release in the United States to control H. halys Host Plants Halyomorpha halys is a polyphagous plant feeder with a wide rang e of host plants. In its native habitat, H. halys reportedly feeds on at least 45 different host plants (Table 1 1). These include fruit trees such as apples ( Malus spp.), oranges ( Citrus spp.), pears ( Pyrus spp.), and plums ( Prunus domestica L.) (Yu and Z hang 2007). Vegetables and other crops serve as hosts as well. For example, H. halys has been

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19 known to feed on cotton ( Gossypium hirsutum L.), soybeans [ Glycine max (L.)], and wheat ( Triticum aestivum L. ) (Yu and Zhang, 2007) (Table 1 1). Populations of H. halys have been reported on the following hosts in the U.S. and China: apple, bean, celosia, corn, grape, hibiscus, lilac, pear, princess tree, soybean, and sunflower (Table 1 1). In Pennsylvania, host plants with the largest amount of H. halys activity consist of highbush cranberry ( Viburnum opulus L. var. americananum Aiton), Siberian pea shrub ( Caragana arborescens Lamarck), and princess tree [ Paulownia tomentosa (Thunberg)] (Nielsen et al. 2008a). Since its detection in Switzerland in 2007, infestat ions of nymphs and egg masses of H. halys have been documented on eight host plants including asparagus ( Asparagus spp.), blue bean shrub ( Decaisnea fargesii Franchet), Japanese angelica tree [ Aralia elata (Miquel)] Japanese stewartia ( Stewartia pseudocame llia Maximowicz), and nasturtium ( Tropaeolum majus L.) (Wermelinger et al. 2008). Two hosts of H. halys in common with the U.S. and Switzerland include butterfly bush and maple (Hoebeke and Carter 2003; Wermelinger et al. 2008). Much of the literature has reported host plants of H. halys through observations based on presence or feeding on these plants. However, some sources have not thoroughly documented the ability of H. halys to survive and reproduce on a given host. In fact, several documented hosts may be alternative hosts when preferred hosts are not available. Further research on the host preferences of H. halys will distinguish the known hosts from alternative hosts. Monitoring Methods Monitoring and identification of both commonly occurring and exo tic species can be an important component of a successful integrated pest management (IPM)

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20 program. Implementing pest monitoring as a part of IPM programs may allow for early detection of exotic species and improved overall pest management and/or eradicati on efforts. Various methods have been used to monitor populations of H. halys including visual pyramid traps, pheromone baited pyramid traps, beat sheet sampling, sweep net sampling, and light traps. Visual pyramid traps have commonly been used to monitor for several stink bug species, such as Chinavia hilare (Say), Euschistus spp., Nezara viridula (Linnaeus), and Oebalus pugnax (Fabricius) (Leskey and Hogmire 2005; Rashid et al. 2006; Kamminga et al. 2012). Generally, a yellow base pyramid trap design is used to trap stink bugs with or without a pheromone bait due to its attractiveness across species (Leskey and Hogmire 2005; Mizell 2008; Kamminga et al. 2012). However, significantly greater numbers of H. halys nymphs and adults were captured with black py ramid traps than any other colored traps with visual stimuli of green, yellow, clear, or white (Leskey et al. 2012b). The most effective results were obtained when the traps were baited with 450 mg of methyl (2E, 4E, 6Z) decatrienoate. Pyramid traps baite d with the aggregation pheromone of Plautia stali Scott (Hemiptera: Pentatomidae), methyl (2E,4E,6Z) decatrienoate have been reported to yield more H. halys individuals than any other type of monitoring method (Khrimian et al. 2008; Leskey et al. 2012a; Le skey et al. 2012b). This pheromone attracts both male and female adults as well as nymphs, and the number of individuals captured is dependent on increasing amounts of methyl (2E,4E,6Z) decatrienoate (Leskey et al. 2012a). However, this lure is only reliab le during late season population peaks from July to the

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21 first frost (Nielsen et al. 2011). Although it is currently being investigated, the identification of the true aggregration pheromone for H. halys is unknown. Active sampling techniques to monitor fo r H. halys include beat sheet sampling and sweep net sampling. Beat sheet sampling has been used to collect adults and nymphs from fruit trees and woody shrubs (Nielsen et al. 2008b; Nielsen and Hamilton 2009a; Nielsen and Hamilton 2009b; Leskey et al. 201 2), while sweep net sampling is more commonly used for field crops such as soybean (Nielsen et al. 2011). These methods were less effective than pyramid traps in the early crop season (Nielsen et al. 2011). Higher capture rates of nymphs and adults were ob tained with these methods late in the growing season. Black light traps are commonly used in the field to monitor the distribution and abundance of H. halys primarily because these traps are effective, easily positioned in the field, and fairly inexpensi ve. Like many other pentatomid species, H. halys is more active at dusk and in the evening. Although these traps are non specific, they are a highly reliable and effective monitoring tool for early season and low density populations of H. halys (Nielsen et al. 2013) unlike pheromone or pyramid traps. When used independently, each collection technique has its own limitations during different seasons and times of day as well as varying levels of cost and benefits. However, applying multiple detection methods would be a highly effective approach for monitoring H. halys populations. Identification Adult H. halys have a dark brown marbled color on the dorsal side, a pale color on the ventral side, and are typically 12 17mm long. Alternating light and dark bands occur along the lateral edges of the abdomen. The shoulders of the pronotum are

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22 rounded and smooth. The most distinguishing feature is the light and dark banding on the fourth and fifth segments of the antennae. The same banding is often present on the leg s as well. Synonyms of H. halys in the literature include H. picus (F.), H. brevis (Walker), and H. mista (Walker) (Hoebeke and Carter 2003). Many other Hemipteran species may be mistaken for H. halys Species of similar appearance in the United States in clude the brown stink bug [ Euschistus servus servus (Say)], bark stink bugs ( Brochymena spp. and Parabrochymena spp.), spined soldier bug ( Podisus maculiventris Say), Holcstethus spp., and late instar nymphs of leaf footed bugs [ Leptoglossus phyllopus (L.) ] (Hoebeke and Carter 2003; Sargent et al. 2010; Halbert and Hodges 2011). The brown stink bug ( E. servus ) may be confused with H. halys because it has similar coloring and alternate light and dark banding on the edges of the abdomen. The two species may b e distinguished by the pale marks on the edge of the abdomen H. halys has triangular markings and the markings on E. servus are more quadrate. Brochymena and Parabrochymena species may also have similar coloring and banding; however, these bugs have dent iculate anterolateral margins on the pronotum. In addition, the shoulders of the pronotum and the membrane of the forewing has dark markings outlining reticulate veins, whereas the anterolateral margins on H. halys are smooth and the veins are pale with a few darker spots. Podisus maculiventris has spinose shoulders on the pronotum, and both P. maculiventris and Holcostethus spp. are smaller in size compared to H. halys Brown Marmorated Stink Bug Extension Education In 2011, Florida reached a record numbe r of 87.3 million visitors and an estimated value of $67 billion for the tourism industry making this state the top travel H. halys

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23 has been intercepted multiple times in Flor ida at agricultural interdiction stations and on vehicles traveling from infested states. Although established populations have not been detected, H. halys populations are likely to be introduced to Florida via tourist vehicles or luggage. Efforts have b een made to educate the Florida county extension agents about H. halys through state and UF IFAS outreach and extension materials. Even with significant efforts, limited Florida State Park personnel educational activities have occurred for this species. Fl orida State Parks receive an average of over 20 million visitors traveling from Florida, other U.S. states, and international countries annually. As many reported hosts for H. halys include woody shrubs and trees, the potential establishment of undetected populations in Florida State Parks is a significant possibility. Delivery of effective extension education and monitoring tools to Florida State Park personnel is important for early detection of H. halys in Florida. Research Objectives Over the past ten years, published research on H. halys has increased significantly and a majority of current research is focused on monitoring and control methods as established populations become crop pests in the United States. Major gaps in the understanding of host pr eferences for this invasive pest exist. As the distribution increases, it is important to research the potential behaviors and impacts of H. halys agriculture prior to establishment. Increasing our understanding of the pest potential for H. halys in Florida will assist in providing informed and economical management recommendations to extension clientele. In addition, communicating this knowledge to state employees, extension agents, a nd the general public may allow for early detection

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24 of H. halys and create general awareness about the invasive pest. My research objectives include: 1. Determining host specificity of H. halys through host range and host preference tests on economically impo landscape. 2. Assessment of effective invasive species workshop training delivery to Florida State Park Service personnel. 3. Development and assessment of a Florida specific stink bug identification c ard deck to be used by the general public.

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25 Table 1 1. Reported hosts of Halyomorpha halys Common Name Scientific Name Location of Host Status Detection Method Reference Angelica tree, Japanese Aralia elata (Miquel) Seemann Switzerland Heavy infestati on of nymphs. Wermelinger et al. 2008 Apple Malus spp. China Literature Yu and Zhang 2007; Sargent et al. 2010 Apple Malus domestica Linnaeus China Hoebeke and Carter 2003 Apple, paradise Malus pumila Mill. China Literature Yu and Zhang 2007 Apricot Prunus armeniaca (L.) China Literature Yu and Zhang 2007 Apricot, Japanese Prunus mume Siebold and Zuccarini China Hoebeke and Carter 2003 Arborvitae, oriental Platycladus orientalis (L.) China Nymphs or adults observed on plant. Yu and Zhang 2007 Argy i wormwood Artemisia argyi H.Lv and Vaniot China Nymphs or adults observed on plant. Yu and Zhang 2007 Asparagus Asparagus spp. Switzerland Two egg masses collected; observed nymphs feeding. Wermelinger et al. 2008 Basswood Tilia spp. United States Hoebeke and Carter 2003 Bean Phaseolus spp. China; United States Literature Yu and Zhang 2007; Sargent et al. 2010 Beet Beta vulgaris L. China Literature Yu and Zhang 2007 Black nightshade Solanum nigrum L. China Nymphs or adults observed on plant. Hoeb eke and Carter 2003; Yu and Zhang 2007 Bushkiller Cayratia japonica Thunberg China Nymphs or adults observed on plant. Yu and Zhang 2007 Butterfly bush Buddleia spp. United States Hoebeke and Carter 2003; Sargent et al. 2010 Butterfly bush Buddleia dav idii Franchet Switzerland Slight infestation of nymphs; three nymphs collected from gardens and reared into adult stage. Wermelinger et al. 2008

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26 Table 1 1. Continued. Common Name Scientific Name Location of Host Status Detection Method Reference Campho r tree Cinnamomum camphora (L.) China Literature Yu and Zhang 2007 Catalpa Catalpa spp. United States Hoebeke and Carter 2003; Sargent et al. 2010 Celosia Celosia argentea L. China; United States Hoebeke and Carter 2003; Sargent et al. 2010 Chaste tr ee Vitex negundo L. China Nymphs or adults observed on plant. Yu and Zhang 2007 Cherry Prunus avium L. China Hoebeke and Carter 2003; Sargent et al. 2010 Cherry, Black Prunus serotina Ehrhart United States Sargent et al. 2010 Cherry, Chinese sour Prun us pseudocerasus L. China Literature Yu and Zhang 2007 Cherry, Yoshino Prunus x. yedoensis Matsumura United States Sargent et al. 2010 Corn Zea mays L. China; United States Nymphs or adults observed on plant. Yu and Zhang 2007; Sargent et al. 2010 Cot ton, upland Gossypium hirsutum L. China Nymphs or adults observed on plant. Yu and Zhang 2007 Eastern Redbud Cercis canadensis L. United States Sargent et al. 2010 Elm Ulmus spp. United States Sargent et al. 2010 Elm, Siberian Ulmus pumila L. China Li terature Yu and Zhang 2007 Fig Ficus carica L. China Literature Hoebeke and Carter 2003; Yu and Zhang 2007; Sargent et al. 2010 Grape Vitis vinifera L. China; United States Literature Yu and Zhang 2007; Sargent et al. 2010 Hawthorne Crataegus pinnatifid a Bunge China Literature Yu and Zhang 2007 Hibiscus Hibiscus rosa sinensis L. China; United States Hoebeke and Carter 2003; Sargent et al. 2010

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27 Table 1 1. Continued. Common Name Scientific Name Location of Host Status Detection Method Reference Holl yhock Althaea rosea L. China Nymphs or adults observed on plant. Yu and Zhang 2007 Honeysuckle Lonicera spp. United States Hoebeke and Carter 2003; Sargent et al. 2010 Hop, Japanese Humulus scandens (Lour.) Merr. China Nymphs or adults observed on plant Yu and Zhang 2007 Jujube Ziziphus jujuba (L.) China Literature Yu and Zhang 2007 Lilac Syringa spp. China; United States Literature Yu and Zhang 2007; Sargent et al. 2010 Locust, black Robinia pseudoacacia L. China Literature Yu and Zhang 2007 Locus t, honey Gleditsia triacanthos L. United States Sargent et al. 2010 Maple Acer spp. United States Hoebeke and Carter 2003; Sargent et al. 2010 Maple, sycamore Acer pseudoplatanus L. Switzerland One nymph observed feeding on maple seed. Wermelinger et al. 2008 Mulberry Morus spp. China Hoebeke and Carter 2003; Sargent et al. 2010 Mum Chrysanthemum morifolium (Ramat.) Kitam. China Nymphs or adults observed on plant. Yu and Zhang 2007 Nasturtium Tropaeolum majus L. Switzerland Heavy infestation of ny mphs. Wermelinger et al. 2008 Oak Quercus spp. United States Sargent et al. 2010 Orange, mandarine Citrus reticulata Blanco China Literature Yu and Zhang 2007 Orchid Brassia spp. China Literature Yu and Zhang 2007 Parasol tree, Chinese Firmiana plata nifolia (L.) China Literature Yu and Zhang 2007

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28 Table 1 1. Continued. Common Name Scientific Name Location of Host Status Detection Method Reference Paulownia Paulownia tomentosa (Thunberg) Steudel United States Hoebeke and Carter 2003 Paulownia t ree Paulownia spp. China Literature Hoebeke and Carter 2003; Yu and Zhang 2007 Peach Prunus persica Batsch China, United States Hoebeke and Carter 2003; Sargent et al. 2010 Pear Pyrus spp. China Literature Yu and Zhang 2007; Sargent et al. 2010 Pear Py rus pyrifolia Nakai China Hoebeke and Carter 2003 Pear, Asian Pyrus serotina L. United States Sargent et al. 2010 Persimmon, Japanese Diospyros kaki L. China Literature Hoebeke and Carter 2003; Yu and Zhang 2007; Sargent et al. 2010 Plum Prunus domest ica L. China Literature Yu and Zhang 2007 Plum Prunus spp. China Literature Yu and Zhang 2007 Pomegranate Punica granatum L. China Literature Yu and Zhang 2007 Raspberry Rubus spp. United States Sargent et al. 2010 Rose, Japanese Rosa rugosa Thunberg United States Sargent et al. 2010 Serviceberry Amenlanchier spp. United States Hoebeke and Carter 2003 Serviceberry, juneberry Amelanchier lamarckii F. G. Schroed Switzerland Slight infestation of nymphs. Wermelinger et al. 2008 Shadbush Amelanchi er spp. United States Hoebeke and Carter 2003 Shrub, blue bean Decaisnea fargesii Franchet Switzerland Excessive infestation of nymphs. Wermelinger et al. 2008 Soybean Glycine max (L.) China, United States Literature Hoebeke and Carter 2003; Yu and Zha ng 2007; Sargent et al. 2010 Spider flower Cleome spp. United States Sargent et al. 2010 Spinach, climbing Basella rubra L. China Hoebeke and Carter 2003 Spindle tree, Japanese Euonymus japonicas Thunberg China Nymphs or adults observed on plant. Yu and Zhang 2007

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29 Table 1 1. Continued. Common Name Scientific Name Location of Host Status Detection Method Reference Staghorn sumac Rhus typhina L. China Nymphs or adults observed on plant. Yu and Zhang 2007 Stewartia, Japanese Stewartia pseudocamell ia Maximowicz Switzerland Excessive infestation of nymphs. Wermelinger et al. 2008 Sunflower Helianthus annuus L. China, United States Nymphs or adults observed on plant. Yu and Zhang 2007; Sargent et al. 2010 Sycamore Platanus spp. United States Sargen t et al. 2010 Tea plant Camellia sinensis (L.) China Literature Yu and Zhang 2007 Tea oil camellia Camellia oleifera Abel China Literature Yu and Zhang 2007 Tobacco, Japanese Nicotiana alata Link and Otto China Literature Yu and Zhang 2007 Tulip tree L iriodendron tulipifera L. United States Sargent et al. 2010 Walnut Juglans spp. United States Hoebeke and Carter 2003 Weeping scholar tree Sophora japonica L. China Nymphs or adults observed on plant. Yu and Zhang 2007 Wheat Triticum aestivum L. Chin a Literature Yu and Zhang 2007 White poplar, Chinese Populus tomentosa Carriere China Literature Yu and Zhang 2007 Wisteria, Chinese Wisteria sinensis (Sims) China Nymphs or adults observed on plant. Yu and Zhang 2007 Wolfberry, Chinese Lycium barbarum L. China Literature Yu and Zhang 2007 Yellow wood Cladrastis kentukea (Dumont de Courset) Rudd United States Sargent et al. 2010 Zelkova Zelkova serrata (Thunberg) Makino United States Sargent et al. 2010

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30 CHAPTER 2 LABORATORY HOST PREFERENCE OF TH E BROWN MARMORATED STINK BUG ( HALYOMORPHA HALYS (STL)) ON PLANT SPECIES RELATIVE TO FLORIDA Introduction Halyomorpha halys (Stl), commonly known as the brown marmorated stink bug (BMSB), is an invasive pentatomid species in the United States. Native to A sia, this species was first identified in Allentown, Pennsylvania in 2001. Subsequent to initial detection, H. halys has been detected in 39 states and agricultural problems have been reported in states as far south as North Carolina and Tennessee (Leskey and Hamilton 2012). With over 300 host plants recorded (Nielsen et al. 2008a), H. halys is considered to be highly polyphagous species and a major pest of vegetable crops, woody shrubs, and fruit trees in the Northeastern and Mid Atlantic states. Crop dama ge reports of 25 80% have been reported for apples and pears in Pennsylvania and New Jersey. Host plant preference of H. halys agriculture is unclear from the currently available literature. Florida Citrus paradisi Macfadyen) and orange ( Citrus sinensis (Linnaeus) Osbeck) (NASS 2012). Although H. halys has a reproductive diapause i n the winter months, the subtropical and tropical climate conditions in central and south Florida may result in a shortened or abbreviated diapause period and allow for the occurrence of more generations per year. In southern China, up to six generations p er year of H. halys have been reported (Hoffman 1931). Successful establishment of H. halys in Florida could have significant economic and ecological impacts for previously reported hosts, such as oranges, as well as other crops with unknown host potential such as grapefruit and strawberry. Host specificity tests

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31 relevant to a few Florida agronomic crops and common landscape ornamental plants for H. halys may provide improved information regarding host preference and the overall impact this pest could have in the state. Host specificity experiments test the relative degree of likeliness that an organism would be used as a host by another organism (Van Driesche and Murray 2004). This type of testing could be used to determine the host preference and host ra nge of H. halys on economically important crops in Florida. Host range and host preference are generally assessed by a combination of three tests: no choice, choice, and open field (Capinera 1985; Van Driesche and Murray 2004; Withers and Mansfield 2005). As these testing schemes have developed from year to year, the combination of no choice and choice tests is most commonly used. Choice tests measure the preference or rank of multiple hosts usually through oviposition or feeding preferences while no choice tests measure the suitability of a host for development of an organism. In choice tests, two or more test species are placed in a cage with one or more specimens of the agent for a fixed period of time under standard laboratory conditions. In no choice te sts, a single test species is placed in a cage with one or more specimens of the agent for a fixed period of time under standard laboratory conditions. Our research will focus on the host specificity of H. halys using economically important crops includin g navel orange ( Citrus x sinensis (Linnaeus) Osbeck [ maxima x reticulata] ), strawberry ( Fragaria spp. Linnaeus), ruby red grapefruit ( Citrus x paradisi MacFadden [ maxima x sinensis ]), avocado ( Persea Americana Miller), and key lime ( Citrus x floridana (Ing ram and Moore) Mabberley) with an overall estimated value of at least $1.86 billion (NASS 2012). Other plant species falling into the category of nursery

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32 and greenhouse products, an industry with on overall value of $1.7 billion (NASS 2012), include the fo llowing: elaeagnus ( Elaeagnus angustifolia Linnaeus), hibiscus ( Hibiscus lunariifolius Willdenow), and ligustrum ( Ligustrum spp. Linnaeus). At least three interceptions of H. halys have occured at agricultural interdiction stations (Russel 2012) and on tou rist vehicles travelling to Florida from infested states (Halbert and Hodges 2011). Nevertheless, established populations have not been detected in Florida. Elaegnus, hibiscus, and ligustrum are commonly found in the Florida landscape or along major highwa ys, and could be possible reservoirs for H. halys to establish before migrating to agricultural crops. Materials and Methods Laboratory Colony Adult specimens and egg masses of H. halys were obtained from laboratory colonies at the Florida Department of Agriculture and Consumer Services Division of Plant Industry (FDACS DPI) Florida Biological Control Laboratory; United States Department of Agriculture ARS) Beneficial Insects Introduction Research Unit; and North Caro lina State University. Halyomorpha halys was reared in the quarantine laboratory at the University of Florida Entomology and Nematology Department in Gainesville, FL. The adult colony was maintained on apples, carrots, green beans, okra and water at room t emperature and humidity (27 o C and 60%RH, respectively) with a 16L:8D photoperiod (Nieslen et al. 2008b). Food and water were replaced three times per week. Eggs were collected from adult cages three times per week using a wet acrylic paintbrush. Egg masses were placed on damp

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33 Upon emergence from the eggs, first instar nymphs remained in the same cage and were misted daily with water. After molting, second instar nymphs were removed where they remained until molting into adults. The second to fifth instar nymphs were maintaine d on carrots, green beans, okra and water under room temperature and humidity in a 16L:8D photoperiod. Plant Sources The following plants were purchased in January 2013: eight one gallon pots of key lime navel orange, and ruby red grapefruit purchased f rom Howey in the Hills, Florida; eight three gallon pots of elaeagnus and ligustrum and eight one gallon pots of strawberry and hibiscus purchased from Gainesville, Florida; and eight one gallon pots of avocado purchased from Homestead, FL. The plant suppl y was replenished in May 2013and consisted of eight one gallon pots of key lime navel orange, and ruby red grapefruit purchased from Howey in the Hills, Florida; eight one gallon pots of elaeagnus, hibiscus, ligustrum, and strawberry purchased from Gaines ville, Florida; and eight one gallon pots of avocado purchased from Homestead, FL. The plants remained in the original pots and were kept in an enclosed greenhouse and watered as needed then moved into the quarantine laboratory for the choice test or no choice test trials. When in the quarantine laboratory and not in use, under the pot f or water and the plants were watered as needed.

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34 No choice Tests To determine the potential for early instar nymph development and to test the host range of H. halys eight plant species were tested using no choice tests. An artificial plant, constructed wi th artificial branches, a one gallon black pot, green tested as a negative control. These tests were completed in three week intervals for a total of three replicates from May 2013 July 2013. To reduce experi ence based bias on a plant species, n ewly emerged s econd instar nymphs were chosen for the no choice experiments as this is the first life stage of H. halys to feed on plant material. Newly laid egg masses were reared in separate cages (round clear plasti c second instar nymphs were chosen at random and placed on the top leaf of a test plant using a damp paintbrush. Each test plant species was placed in separate mesh cages (L pot for water. The specimens were allowed to mature for three weeks. A total of four observers were used to record the recovery and mortality of all nymphs. Each obser ver was allotted ten minutes per trial to collect and record data on a standardized datasheet. To minimize observer error, the observations were repeated three times per trial. The data recorded included t he nymphal stage of alive and dead specimens as wel l as molts recovered. Feeding injury and damage to the plant were additional observations recorded. Choice Tests The host preference of H. halys and ranking of host material were tested using an incomplete block design choice test. Two of the eight plant species were presented

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35 simultaneously to four adult specimens of H. halys two males and two females. All possible combinations of the eight plant species a nd negative control (Table 2 1) were analyzed in 24 hour intervals and each combination was replicat ed three times for a total of 108 trials from January 2013 to July 2013. In preparation for each trial, two males and two females were randomly chosen from the laboratory colony at 8:00PM the day prior. Each specimen was marked with a small dot of Testors enamel paint on the center of the pronotum. The following colors trial females; white indicated second trial females; green indicated third trial females; yellow in dicated first trial males; red indicated second trial males; and light blue indicated third trial males. In an effort to reduce experience based bias on a plant species, each individual was used in no more than three trials. After marking, the specimens we re starved for twelve hours and placed in a small cage (round clear plastic began the following day. Between 7:45AM and 8:30AM (M1), the specimens were released in th e bottom from both test plants. Once released, the specimens were observed for 30 minutes. Three additional thirty minute observations were made starting between 1:45PM and 2:3 0PM (A), 7:45PM and 8:30PM (E), and 7:45AM and 8:00AM (M2) the following day, for a total of four observations per trial and an overall total of 432 observations. After each observation, the specimens remained undisturbed in the mesh cage with the two test plants until the trial concluded.

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36 A total of five observers were used to record the specimen behaviors. To minimize observer error, a standardized question based data sheet was used to uniformly record observations. The recorded behaviors included the fo llowing: location of each specimen, feeding activity and feeding locatio n, duration of feeding activity. Additional behaviors, including mating and oviposition, were also recorded. Statistics No choice Tests The mean number of second instars, third insta rs, and exuviae was determined using Microsoft Excel 2010. Due to the small sample size and low recovery of individuals, addition al summary statistics were not performed. Choice Tests For the choice tests, summary statistics for the number of specimens pr esent on each plant and the number of specimens feeding were prepared using the binomial distribution in PROC GLIMMIX (General Linear Model for Mixture Distributions), accounting for non normality and non homogenous variances, and Least Squares Means in SA S 9.3 (SAS Institute Inc., 2002 2010, Cary, NC, USA). The differences between each plant species for the number of specimens present and the number of specimens feeding was analyzed using Tukey Kramer Least Squares Means Adjustment for multiple compariso ns (SAS Institute Inc., 2002 2010, Cary, NC, USA), allowing for adjusted p values for all pairwise comparisons. The correlation between the correlation between the specime spent feeding on a test plant were analyzed using multivariate correlations, pairwise correlations, and non

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37 2010, Cary, NC, USA). For determi ning the difference between the number of specimens feeding by observational time of day, the Poisson distribution in PROC GLIMMIX was used along with Tukey Kramer Least Squares Means Adjustment for multiple comparisons. Results No choice Tests On averag e, more second instar nymphs were able to survive on hibiscus, ligustrum and strawberry in the no choice tests ( Table 2 2 ). The second instar nymphs were also able to molt into the third instar in at least one replication when tested with hibiscus, ligust rum, and elaeagnus. Choice Tests The average number of times adult specimens were present on a test plant ( F =10.73; df=8, 200; P =0.0001) and feeding on a test plant ( F =3.58; df=8, 147; P =0.0008) in the choice tests were significant between test plant spe cies. Adult specimens were present more often on elaeagnus, ligustrum, and orange and differed significantly from all other plant species ( Figure 2 1 ). However, adults fed more on ligustrum than any other plant and fed significantly more on ligustrum than avocado and the negative control (Figure 2 1 ). The Spearmans for the correlation between adult specimens present on a test plant and adult specimens feeding on a test plant was 0.4227 ( P <0.0001). Although, the Spearmans for the correlation between adu lt specimens present on a test plant and percentage of adult specimens present and feeding on a test plant was 0.1389 ( P =0.0817). The average number of times adult specimens were feeding on a test plant ( F =6.05; df=3, 311.4; P =0.0005) in the choice tests was also significant between

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38 observational times of day. More adult specimens were feeding during the evening observations, and differed significantly from both morning observations ( Figure 2 2 ). Discussion Survivorship of multiple generations is importa nt when determining the pot ential host range of an insect. The survivorship of H. halys early instar nymphs were tested in no choice tests. Mo re second instar nymphs survived on ligustrum and strawberry, and significantly more on hibiscus. In addition, at least one nymph was successfully able to molt into the third instar when on hibiscus, ligustrum, and elaeagnus. Therefore, hibiscus and ligustrum could be considered host plants of H. halys However, additional replications are needed to confirm these find ings. Also, less than half of the individuals were recovered in no choice tests with avocado, elaeagnus, grapefruit, key lime, orange, and negative control. It is unclear if the individuals escaped from the cages or were lost in the soil. Furthermore, the current methodology may not be truly indicative of the actual survivorship due to the low recovery rate of individuals. The methods of the no choice tests may need to be modified for a higher recovery rate of individuals. Future modifications may include u tilizing a positive control to test against the unknown hosts for a better understanding of the effects in the no choice experiments. Additional modifications may include a smaller arena for a higher recovery rate of individuals, as well as sufficient humi difiers and thermostats for constant control over the environmental conditions. When given a choice, adult specimens were present more often on elaeagnus, ligustrum, and orange in comparison to all other test plant species. However, the number of adult s pecimens feeding on avocado was lower than elaeagnus, grapefruit, hibiscus, key lime orange, and strawberry and significantly lower than ligustrum. The

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39 discrepancy between the significant presence of H. halys on avocado compared to the low feeding occurre nce supports the Spearmans ( =0.1389; P =0.0817) for the correlation between adults present on a test plant and percentage of adults feeding on a test plant. Even though the correlation is significant at the value shows a very weak relationship between percentage of adult s feeding on a plant and adult specimens present on a plant. Overall, these results suggest that adult presence on a plant may be more indicative of roosting behavior rather than feeding. Furthermore, the adult specimens were observed p robing on the negative control in five trials for an average feeding t ime of 5 minutes and 41 seconds. These results suggest that H. halys may taste a plant before committing to feeding. Consequently the documented host lists of H. halys in older literatu re may inaccurately report occurrences of H. halys on a plant species as hosts if the rate and length of feeding activity was not properly documented. Experiment s in a laboratory setting control abiotic and biotic factors, and possibly eliminate external nuisance factors that could affect the results of the data. Because outside factors are eliminated or reduced in a laboratory setting, host ranges may be more definitely determined. Nonetheless, it is important to consider host experience and host depriva tion when designing choice and no choice tests (Withers and Mansfield 2005). For example, it is best to use nave specimens that are unfamiliar with the test species and avoid rearing specimens on similar plants in order to decrease bias. Due to limited nu mbers in the laboratory colony, some adult specimens in each trial of the choice tests may have been used in previous trials. Thus, some error may have occurred due to bias in previously experience plant species. Host deprivation may also

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40 generate false po sitive if specimens are deprived of food for too long or false negative responses if specimens are fed shortly before the experiment begins. Depriving each specimen in the choice tests of food for 12 hours may or may not have caused error or false positive results. Other sources of error may include insect confinement, causing the insects to feed on a test species due to starvation or feed on a test species that would not have been selected in the field (Capinera 1985; Van Driesche and Murray 2004). More adults fed during the afternoon and evening observations, and the evening observations differed significantly from both morning observations. The increased feeding activity in the afternoon and evening correspond with the increased flight activity of H. ha lys observed at dusk and evening hours (Nielsen et al. 2013). Increased feeding and flight activity throughout the evening may indicate that black lights could be a lower cost, yet highly effective method of monitoring for H. halys in Florida in comparison to visual pyramid (black or yellow) or pheromone baited pyramid traps. Black light traps could be placed along the edges of agricultural crops near forested areas as well as agricultural interdiction stations and major highways. Halyomorpha halys most f requently fed upon ligustrum in the choice tests and more second instar nymphs were able to survive on ligustrum Ligustrum is in the plant family Oleaceae, a botanical family including evergreen and deciduous species, which includes related plants such a s lilacs ( Syringa spp.) and jasmines ( Jasminum spp.). Following ligustrum, H. halys most frequently fed on hibiscus (Malvales: Malvaceae) and second instar nymphs were also able to survive on hibiscus The plant family Malvaceae includes woody ornamentals and, most importantly cotton ( Gossypium

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41 spp.). The preference of H. halys for woody hosts in this study is consistent with the fruit tree damage reported in the Mid Atlantic States. Future host specificity tests and monitoring efforts on Florida specific plants should include the Oleaceae, Malvaceae, and other related families most commonly grown in greenhouses and nurseries. Although reported in the literature as a host (Yu and Zhang 2007), our research suggests that Citrus spp. (Sapindales: Rutaceae) ma y not be preferred compared to other woody hosts, particularly those found in the order Rosales. Further replications are needed to confirm our findings.

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42 Table 2 1. All combinations of plant species tested in choice tests of adult Halyomorpha halys N umber Combinations 1 avocado x hibiscus 2 avocado x key lime 3 avocado x ligustrum 4 avocado x orange 5 avocado x grapefruit 6 avocado x strawberry 7 avocado x elaeagnus 8 hibiscus x key lime 9 hibiscus x ligustrum 10 hibiscus x orange 11 hibisc us x grapefruit 12 hibiscus x strawberry 13 hibiscus x elaeagnus 14 key lime x ligustrum 15 key lime x orange 16 key lime x grapefruit 17 key lime x strawberry 18 key lime x elaeagnus 19 ligustrum x orange 20 ligustrum x grapefruit 21 ligustrum x strawberry 22 ligustrum x elaeagnus 23 orange x grapefruit 24 orange x strawberry 25 orange x elaeagnus 26 grapefruit x strawberry 27 grapefruit x elaeagnus 28 strawberry x elaeagnus 29 negative x avocado 30 negative x hibiscus 31 negative x key lime 32 negative x ligustrum 33 negative x orange 34 negative x grapefruit 35 negative x strawberry 36 negative x elaeagnus

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43 Table 2 2 Mean number of Halyomorpha halys individuals recovered in no choice tests. Plant Species 2nd Instars Recovered 3rd Instars Recovered Exuviae Recovered Avocado 5.667 0 0 Elaeagnus 4.667 0.333 0 Grapefruit 4 0 0 Hibisucs 11 2 1.667 Key lime 1.667 0 0 Ligustrum 8.333 5 4 Orange 6 0 0 Strawberry 8.667 0 0 Negative Control 4.667 0 0

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44 Figure 2 1 Mean (st andard error of the mean, SEM) number of adult Halyomorpha halys present or feeding on test plant species in choice tests. Means with the same letter are not significantly different (Tukey Kramer, P=0.05). b a b b b a a b b bc abc abc ab abc a abc abc c 0 0.05 0.1 0.15 0.2 0.25 0.3 0.35 0.4 0.45 Number of Adults Plant Species Presence Feeding

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45 Figure 2 2 Mean (standard error of the mean, SEM) number of adult Halyomorpha halys feeding on any test plant at different times of day. Means with the same letter are not significantly different (Tukey Kramer, P=0.05). b ab a b 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 Morning 1 Afternoon Evening Morning 2 Number of Adults Plant Species

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46 CHAPTER 3 EVALUATION OF THE INVASIVE SPECIES WORKSHOP PROGRAM FOR FLORIDA PAR K SERVICE PERSONNEL Introduction Invasive species are described as plants, animals, or pathogens that are not native to an ecosystem and cause or are likely to cause economic or environmental harm or harm to human, animal, or plant health (ISAC 2006). The introduction and establishment of an invasive species may result in increased eradication and control costs as well as negatively impact trade and the environment. In the United States, invasions of non native species cost an estimated $137 billion annuall y, and $20 billion of these costs are due to environmental damages, lost agricultural productivity, and increased health problems caused by arthropods species (Pimentel et al. 2010). Preventing the introduction of exotic invasive species is the best defen se against the destructive effects of invasions. Preventing invasive species introductions may not always be possible, especially in a state such as Florida. Florida has 12 international airports and 14 deepwater seaports where plant materials are received However, only a small percentage of imported plant materials are sampled and inspected at ports of ealth Inspection Service (APHIS) for exotic plant pests, diseases, pathogens, and noxious weeds (USDA APHIS PPQ 2012). Because there are multiple pathways for species introduction and suitable climate conditions, Florida is considered a high risk state for invasive species introductions. Early detection and rapid response (EDRR) systems have been developed as a second line of defense to prevent permanent establishment of invasive species through

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47 monitoring and control methods (National Invasive Species Cou ncil 2003). In the United States, EDRR systems have been adopted on the national level by the Federal Interagency Committee for the Management of Noxious and Exotic Weeds, National Invasive Species Council, and the United States Forest Service (FICMNEW 20 03; NISC 2003; USFS 2006). Florida agencies, such as the Florida Invasive Species Partnership and Florida Cooperative Invasive Species Management Areas, have also implemented enforced EDRR programs (FISP 2013). All EDRR systems consist of three actions for reducing impacts of invasive species: early detection, rapid assessment, and rapid response (NISC 2003). For the purposes of this research, we will concentrate on early detection efforts. Early detection of invasive species through monitoring and identifi cation efforts is not only an essential component for successful EDRR systems, but also for integrated pest management (IPM) programs (NISC 2003; USEPA 2012). The Florida Cooperative Agricultural Pest Survey (CAPS) Program is responsible for conducting pes t surveys in high risk areas for species on special watch lists (FDACS DPI 2008). The current and ongoing survey program managed by the Florida CAPS Program may target individual pest species such as red palm mite ( Raoiella indica Hirst), light brown apple moth [ Epiphyas postvittana (Walker)], and rice cutworm [Spodoptera litura (Fabricius)] or may concentrate on specific high risk areas and/or commodities such as crop surveys, imported warehouses, and solid wood packing materials. Due to limited resource, the Florida CAPS Program cannot monitor for invasive species throughout the entire state of Florida. Involving organizations or volunteers in collaborative monitoring efforts, referred to as passive detection networks, is important for expanding early dete ction

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48 efforts. By monitoring for invasive species while conducting other activities in their daily lives, passive detection networks may expand the area covered and monitor for invasive species not occurring on special watch lists (i.e., brown marmorated s tink bug [Halyomorpha halys (Stl)], red palm weevil [ Rhynchophorus ferrugineus (Olivier)], Mexican bromeliad weevil [ Metamasius callizona (Chevrolat)], etc.). Coordinating public outreach and educational extension events is an effective way to distribute information about the importance of invasive species and the benefits of EDRR systems. Professionals and volunteers should be sufficiently trained in detection, collection, and reporting methods for suspected invasive species. Our research will focus on t he development and delivery of Florida specific invasive species education materials to be used in a series of workshop trainings directed toward Florida State Park Service personnel. Materials and Methods Target Audience The Invasive Species Workshop tr aining sessions and corresponding educational extension materials were tailored for Florida State Park Service personnel (e.g. biologists, park rangers, and volunteers), as well as affiliated groups with a general interest in invasive species detection and management. A total of three training sessions were held at separate Florida State Park district offices in the following locations: northwest region (District 1), northeast (District 2), and central (District 3). Workshop Presentations Each workshop co nsisted of PowerPoint presentations developed through the collaborative efforts of the University of Florida (UF) Biosecurity Research & Extension Lab, UF Agricultural Education and Communication Department, USDA APHIS PPQ,

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49 the Florida Department of Agricu lture and Consumer Services Division of Plant Industry, and the Florida Cooperative Agricultural Pest Survey Program on the following topics: importance of invasive species; federal and state agencies affiliated with invasive species, trapping and monitori ng methods for target pests; sampling and reporting methods for commonly occurring and exotic species. An additional six species specific presentations were delivered at each workshop and included presentations on the following species: the brown marmorate d stink bug cactus moth [ Cactoblastis cactorum (Berg)], emerald ash borer ( Agrilus planipennis Fairmaire), bean plataspid [ Megacopta cribraria (Fabricius)], Mexican bromeliad weevil, pine bark beetles ( Ips spp. and Dendoctronus spp.), redbay ambrosia bee tle ( Xyleborus glabratus Eichhoff), and red palm weevil. Species were selected based on responses from district biologists and for likelihood of establishment in each region. In addition to the PowerPoint presentations, the participants engaged in a han ds on activity. The hands on activity was comprised of six stations with mounted specimens of invasive species discussed at each workshop, as well as native species that are commonly confused with the invasive species. Hand lenses and microscopes were prov ided when necessary. Overall, the order of presentations and activities were uniformly delivered across workshops. Each workshop lasted five hours and included a scheduled thirty minute lunch break. Evaluation Instruments Prior to participating in any educational tests, participants were required to complete and sign an informed consent form. This voluntary agreement is an essential process for any research conducted at the University of Florida utilizing human

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50 participants, and provides participants w ith a description of the planned procedure to record data (UF IRB 02). All data were collected in such a manner so that the participants remained anonymous and disclosure of participant responses did not place the participant at risk. The workshop traini ngs were evaluated using a pre test and post test (Fig ures 3 1 3 3). The purpose of the pre test was to collect demographic information and determine any previous experiences with invasive species workshops or programs. The post test consisted of 43 ques tions and was designed using a four level model to assess participant reaction, learning, change in behavior, and desired results (Kirkpatrick 1996). Questions #1 36 were modeled after Rockwell and Kohn (1989) post then pre evaluation design to measure t perceived change in behavior and reactions. The remaining seven questions assessed participant learning through a set of multiple choice questions. The desired result of this workshop series was to inspire participants to become ac tively involved in early detection of invasive species through monitoring, collecting, and reporting of suspected species. To measure the efficacy of the workshops, all participants were given three pest survey forms and encouraged to survey for any of the invasive species discussed at the training sessions. Completed pest survey forms were mailed to and analyzed by Lyle Buss at the University of Florida Insect Identification Lab. All documents used in thi s research were approved by the Behavioral/NonMedic al Institutional Review Board (IRB02).

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51 Statistics 36 on the post test was summarized with PROC UNIVARIATE in SAS 9.3 (SAS Institute Inc., 2002 2010, Cary, NC, USA). The responses f or questions #1 36 from the post test were assigned numerical values: strongly disagree (SD) = 1; disagree (D) = 2; uncertain (U) = 3; agree (A) = 4; and strongly disagree (SA) = 5. The groups of participants from different workshops, educational backgro unds, and previous experience with invasive species workshops or educational programs were compared in PROC GLM in SAS 9.3 (SAS Institute Inc., 2002 2010, Cary, NC, USA) and analyzed using Tukey Kramer Least Squares Means Adjustment for multiple compari sons (SAS Institute Inc., 2002 2010, Cary, NC, USA) to calculate mean responses for each question and test for significant differences in means (ANOVA) between groups of each category. Results Demographics There were 56 participants, representing the t hree Florida State Park districts. The demographic profile of the participants indicated that 52% ( n =29) were male. The majority of the participants (98%, n=54) were white, while 2% (n=1) were Hispanic and one participant this section blank. The mean age was 42 13 years old. Pre test and Post test Evaluation There was a positive overall change from the workshop for questions #1 36 on the post test ( Figure 3 4 ). There were no significant differences in responses from participants that had (55%) or had not (45%) previously attended workshops on invasive species, or from participants that had (5%) or had not (95%) previously attended a First Detector, Sentinel Plant Network, or Protect US program However, there were

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52 significant differences in responses between levels of education for questions 1, 5, 10, 11, 26, 29, 33, 34, and 36 ( Figure 3 5 ). Participants that completed trade, technical or vocational training (21%) tended to have a higher change in response than participants that completed some post doc toral work (5%) or had a post doctoral degree (40%). There were also significant differences in responses between groups that had or had not previously attended workshops or seminars on invasive species for questions 3, 25, 26, 34, 35, and 36 ( Figure 3 6 ). Participants that had not previously attended workshops or seminars on invasive species (45%) tended to have a higher change in response. Post workshop Pest Surveys Pest survey forms were collected from the beginning of the workshop series in February 2 013 to July 2013. A total of seven pest survey forms were completed by workshop attendees, returned, and analyzed by Lyle Buss at the University of Florida Insect Identification Lab. Two participants collected and correctly identified the bean plataspid [ M egacopta cribraria (Fabricius)], and two new county records of the bean plataspid in Florida have been recorded as a result. One participant collected cactus pads and correctly identified cactus moth [ Cactoblastis cactorum (Berg)] damage. The remaining fou r participants did not find invasive species when surveying for pests in their state parks. Discussion The Invasive Species Workshop Series had a positive overall impact based on the responses to que stion on the post test ( Figure 3 4 ). In some areas, th e workshop had a more positive impact on participants with an intermediate level of education (Figure 3 5) and participants that had not previously attended workshops or seminars focused on invasive species (Figure 3 6) Although Florida State Park Service

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53 personnel have limited training in entomological pests, it may be beneficial to include more technical information in the presentations for those participants with higher levels of education and those participants that have attended similar workshops. S ome invasive species discussed, such as the brown marmorated stink bug ( H. halys ) and the bean plataspid ( M. cribraria ), are considered agricultural pests and may or may not be of ecological concern in Florida State Parks. However, these species may become well establish in forested areas before migrating to agricultural crops at which point the species would be difficult to manage. It is important to have collaborative efforts from forestry and agriculture to mitigate the effects of invasive species on eco nomic and ecological interests. As demonstrated by the post workshop pest survey forms, reaching out to Florida State Park personnel may help with early detection of invasive species and set new county records for invasive species. Additional training s essions will target different audiences, including Florida Master Gardeners, nurserymen, small farm producers, and public gardens personnel. The Invasive Species Workshop Series training sessions are scheduled to be completed by July 2014. As the training sessions conclude, a follow up survey will be conducted with previous attendees for additional feedback. In the future, collaboration with the Florida State Park Service, and even the National Park Service, through workshops and other educational extension materials to increase the probability of detecting invasive species.

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54 Figure 3 1. Pre test for the Invasive Species Workshop Series.

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55 Figure 3 2. Front page of the post test for the Invasive Species Workshop Series.

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56 Figure 3 3. Back page of p ost test for the Invasive Species Workshop Series.

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57 Figure 3 4. Overall mean question responses from post test. A) Mean question responses for question numbers 1 18. B) Mean question responses for question numbers 19 36. The r esponses were ass igned numerical values: strongly disagree (SD) = 1; disagree (D) = 2; uncertain (U) = 3; agree (A) = 4; and strongly disagree (SA) = 5. 0 1 2 3 4 5 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 Response Value Question Number Response Before Workshop Response After Workshop Change in Response 0 1 2 3 4 5 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 Response Value Question Number A B

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58 Figure 3 5. Differences in mean change of question responses from post test between participant levels of education ab ab ab ab ab ab a a ab a ab b ab ab ab ab ab ab ab b ab ab b ab ab b ab ab a a a a a ab ab a b b b ab b ab ab ab b ab b b ab b b b b b b b b b b ab b b b 0 0.5 1 1.5 2 2.5 3 1 5 10 11 26 29 33 34 36 Change in Question Response Question Number Some high school High school graduate Some college Trade school College graduate Some post-grad work Post-grad degree

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59 Figure 3 6. Differences in mean change of question responses from post test between participant experience with invasive species workshops. Means with the same P=0.05). b b b b b b a a a a a a 0 0.5 1 1.5 2 3 25 26 34 35 36 Change in Response Question Number Previously attended workshop Did not previously attend workshop

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60 CHAPTER 4 ASSESSMENT OF IDENTIFYING STINK BUGS AND SIMILAR TRUE BUGS OF FLORIDA FIELD GUIDE Introduction Field guides are a type of reference designed to aid in the identification of plants and animals in their natural environment, and are generally small enough to be carried in the field. Although more technical field guides exist, most field guides are accessible to the general public and include pictures and descriptions of multiple life stages to assist in distinguishing between similar groups or species. There are many field guides available for general insect identification, i.e., Peterson Field Guide to Insects (Borror and White 1998), National Audobon Society Field Guide to Nort h American Insects and Spiders (Milne et al. 1980), and Kaufman Field Guide to the Insects of North America (Eaton and Kaufman 2007). However, there are only a handful of field guides exclusively for stink bug identification. The most common field guides f or stink bug identification include Field Guide to Stink Bugs of Agricultural Importance in the Upper Southern Region and Mid Atlantic States (Kamminga et al. 2009) and Stink Bugs of the Midwest (Jesse et al. 2011). Stink bugs, as well as many other true bug species, are commonly found in crops, gardens, and landscapes throughout the state of Florida. Infestations of herbivorous stink bugs, such as the southern green stink bug [ Nezara viridula (Linnaeus)], may significantly damage plants. However, predaci ous stink bugs are beneficial in controlling plant pests. For instance, the spined soldier bug [ Podisus maculiventris (Say)] preys on larvae of the Colorado potato beetle ( Leptinotarsa decemlineata Say), corn earworm [ Helicoverpa zea (Boddie)], fall armywo rm [ Spodoptera frugiperda (J.E. Smith)], Mexican bean beetle ( Epilachna varivestis

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61 Mulsant), and several more economically important pests (De Clercq 2008). Proper identification of stink bugs and true bugs is essential before determining appropriate pest management options. Until recently, there has not been a Florida specific field guide for stink bug identification. Due to multiple pathways of introduction, suitable climate conditions, and a large scale agricultural production, Florida is considered a high risk state for invasive species. Invasive stink bug species of particular agricultural concern include the brown marmorated stink bug [ Halyomorpha halys (Stl)] and the bagrada bug [ Bagrada hilaris (Burmeister)]. Monitoring for these species may be di fficult because there are species native to Florida with similar markings and coloration. For this reason, a field guide titled Identifying Stink Bugs and Similar True Bug Species in Florida was developed as a reference for identifying Florida specific sti nk bug and related true bug species. My research will focus on assessing the utility of Identifying Stink Bugs and Similar True Bug Species in Florida through a n identification activity of commonly encountered stink bug and true bug species in Florida. Materials and Methods Development of Field Guide Development of Identifying Stink Bugs and Similar True Bugs in Florida occur red from January 2012 until May 2013. This field guide includes photographs and information on 25 species with 68 colored pages. S pecies selected for this field guide are frequently encountered in Florida with the exception of two invasive species of economic concern that are not currently established in Florida, the brown marmorated stink bug and the bagrada bug.

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62 Labeled diagrams of the dorsal and ventral surfaces of a stink bug were incorporated in the field guide, including separate diagrams for the ventral surfaces of herbivorous and predaceous species with distinguishable characteristics highlighted. At the top of each species page, the species name and common name (if applicable) were nymphs, adults, and common hosts were described for each species with corresponding images for each life distribution maps were also included when the information was available for a species. Target A udience The target audience for this study was Florida Master Gardeners, which is made up of citizens of Florida w ith a common interest in providing research based information about gardening to educate Floridians (UF Master Gardener Program 2012). Master Gardeners receive some general training in entomology with an emphasis on insects of horticultural significance. T he following county extension offices in northwest, northeast, and central Florida with active Master Gardener Programs were chosen to participate in the assessment for Identifying Stink Bugs and Similar True Bugs of Florida: Alachua Bay, Citrus, Duval, F lagler, Nassau, Orange, Seminole, St. Lucie, and Washington. A maximum of 20 Florida Master Gardeners from each county were invited to participate in the assessment. Evaluation Instruments Prior to participating in the assessment, participants were requi red to carefully read and sign an informed consent form. This voluntary agreement is an essential part of any research conducted by the University of Florida utilizing human participants, and provides participants with a description of the planned procedur e to record data (UF

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63 IRB 02). All data must be collected in such a manner so that the participants remain anonymous and disclosure of participant responses does not place the participant at risk. For evaluation of the field guide, participants were given a pre survey (Figure 4 2) before and a post survey (Figure 4 4) immediately after the identification activity (Figure 4 3 ) The pre survey was designed to collect demographic information and determine any previous experiences with insect identification. Th e post survey was modeled after the four level approach for evaluations (Kirkpatrick 1996). The front page consisted of the identification activity to assess participant learning. On the back page of the post survey, there were an additional five questions to assess participant reaction and change in behavior as a result of using the field guide (Kirkpatrick 1996). The informed consent form, pre survey, and post survey for this study were approved by the University of Florida Institutional Review Board (P rotocol #2013 U 0596, 2013). Identification Activity The following specimens were left unmarked, placed in a covered box, and respectively numbered 1 8: bean plataspid [ Megacopta cribraria (Fabricius)]; milkweed bug [ Oncopeltus fasciatus (Dallas)]; whe el bug [ Arilus cristatus (Linnaeus)]; rice stink bug [ Oebalus pugnax (Fabricius)]; harlequin bug [ Murgantia histrionica (Hahn)]; leaffooted bug [ Leptoglossus phyllopus (Linnaeus)]; Euschistus quadrator (Rolston); and brown marmorated stink bug (Table 4 1) Each county extension office received a separate collection box of unmarked specimens. Using only the field guide as a resource, the participants were given thirty minutes to identify all unmarked

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64 specimens and classify each specimen as an herbivore or a predator. Hand lenses were also provided for easy viewing of the specimens. Delivery of Materials and Assessments Materials sent to each county extension office included one collection box of unmarked specimens, twenty informed consent forms, twenty ind ividually marked pre test and post tests, and twenty Ziploc bags with a copy of Identifying Stink Bugs and Similar True Bugs of Florida, a hand lens, and a pen for each participant. All pre tests and post tests were placed in manila envelopes to conceal th e questions and responses from each participant. All materials were mailed to each county extension office, with the exception of Alachua and Nassau County, and assessments were completed between June 2013 and July 2013. The county extension agents were given standardized instructions on how to deliver the assessment. Upon completion, the county extension agents were instructed to mail back the informed consent forms along with the pre tests and post tests. In regard to Alachua and Nassau County, we arran ged to personally deliver the assessments to the Master Gardeners. Statistics For the identification activity, the frequency of correct responses for species identification and classification (e.g. herbivore or predator) were analyzed using frequency and contingency tables produced by PROC FREQ in SAS 9.3 (SAS Institute Inc., 2002 differences of responses between the following categorical variables: participants from different co unty extension offices; participants with different educational backgrounds; participants with different backgr ound of entomological training; and previous

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65 experience with identification resources (i.e. Distance Diagnostic and Identification System, Nation al Plant Diagnostic Network, Protect U.S., etc. ). Results Demographics With an overall total of 75 participants, 32% ( n =24) were male and 68% ( n =51) were female. The majority of the participants (91%, n =68) were white, while 3% ( n =2) were Asian, 1% ( n =1) were African American, 1% ( n =1) were American Indian or Alaska native, and 4% ( n =3) did not respond. The mean age was 64 9 years old. Identification Activity Over 70% of participants correctly classified each specimen as herbivore or predator with th e lowest percentage of correct responses for E. quadrator (73%) and H. halys (78%) ( Figure 4 1). For specimen identification, over 70% of participants correctly identified the following species: M. cribraria (88%), O. fasciatus (92%), A. cristatus (88%), a nd M. histrionica (89%). Fewer than 70% of participants correctly identified O. pugnax (67%), L. phyllopus (65%), E. quadrator (33%), and H. halys (18%), and the percentage of correct responses for E. quadrator and H. halys was significantly lower. Post t est Responses After completing the identification activity, the participants were asked 1.) if they would refer to the field guide as an identification resource for stink bugs and true bugs in the future, 2.) if they were confident in distinguishing herbi vores from predators when using the field guide as a reference, and 3.) which specimen they felt least confident identifying. A high percentage of participants strongly agreed (63%) or agreed (29%) that they would use the field guide as an identification r esource. When using the field guide, the majority of participants agreed (55%) or strongly agreed (25%) that they felt

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66 confident distinguishing herbivores from predators. The specimens participants felt least confident identifying were O. pugnax (31%) and E. quadrator (31%), followed by H. halys (11%), M. cribraria (6%), O. fasciatus (6%), and L. phyllopus (5%). Discussion Participants were able to successfully distinguish herbivores from predators when referring to the field guide. However, only four out of eight specimens had a percentage of correct responses above 70%. Halyomorpha halys and E. quadrator had the lowest percentage of correct responses, with 18% and 33% of correct responses respectively. Most participants misidentified these species as E. servus (Say), E. tristigmus (Say), or Podisus maculiventris Say. Both H. halys and E. quadrator have a similar brown marbled coloration with alternating light and dark colored banding along the outer edges of the abdomen. This suggests that stink bug or tr ue bug species with the same coloration, particularly a brown marbled color, are more difficult to identify. In future revisions of the field guide, it would be beneficial to add photographic keys for species with similar appearances and highlight distingu ishable characteristics within the images.

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67 Table 4 1. Correct responses in the identification activity for the assessment of Identifying Stink Bugs and Similar True Bugs of Florida. Specimen number Specimen identification Specimen classification (her bivore or predator) 1 Megacopta cribraria (Fabricius) Herbivore 2 Oncopeltus fasciatus (Dallas) Herbivore 3 Arilus cristatus (Linnaeus) Predator 4 Oebalus pugnax (Fabricius) Herbivore 5 Murgantia histrionica (Hahn) Herbivore 6 Leptoglossus phyllopus (Linnaeus) Herbivore 7 Euschistus quadrator (Rolston) Herbivore 8 Halyomorpha halys (Stl) Herbivore

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68 Figure 4 1. Overall percentage of correct responses from the identification activity used for assessing Identifying Stink Bugs and Similar True Bug s of Florida. Participants classified each specimen as herbivore or predator, as well as identify the species of each specimen. 0 20 40 60 80 100 120 Percentage Specimen Classification Identification

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69 Figure 4 2. Pre test for assessment of Identifying Stink Bugs and Similar True Bugs of Florida.

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70 Figure 4 3. Identificatio n activity for the assessment of Identifying Stink Bugs and Similar True Bugs of Florida.

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71 Figure 4 4. Post test for the assessment of Identifying Stink Bugs and Similar True Bugs of Florida.

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72 CHAPTER 5 IMPLICATIONS AND FUTURE DIRECTIONS FOR H. halys RE SEARCH The introduction and establishment of invasive species often leads to expensive eradication and control efforts, direct environmental and agricultural damage, and can severely impact the trade industry. Once an invasive species becomes established eradication may not be possible and effective control methods can be difficult to achieve. When possible, prevention of invasive species introductions or early detection of new populations is the best defense against the destructive effects of invasions. Halyomorpha halys is an invasive agricultural and urban pest in the United States. Since its detection in the late 1990s, H. halys has become established in at least twelve states and the distribution continues to expand. Most research in the past decad e pertaining to H. halys has been focused on the development of monitoring and control methods. However, little research concentrates on the host preferences of H. halys and the potential impacts it may have on agricultural commodities for uninfested state s. If H. halys were to establish as far south as Florida, the economic and ecological impact of H. halys could be significant. The preference of H. halys for woody hosts is consistent throughout the literature, the fruit tree damage reported in the Mid At lantic States, and this research. Monitoring and early detections programs for H. halys in the southeastern United States, as well as other states without established populations, should set pyramid or black light traps around the perimeter of woody agricu ltural crops, along highways with woody vegetation, or in areas with an abundance of woody landscape plants. Although Citrus reticulata is reported as a host in native regions of China (Yu and Zhang 2007), our research suggests that Citrus spp. may not be preferred and may serve as alternate

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73 hosts. The University of Florida Biosecurity Research and Extension Lab will continue additional replications of the choice and no choice tests and include a larger variety of plant species for a better understanding of the host specificity of H. halys New no choice methodology will also be tested. Following additional replications of choice tests, new choice test methodology may also tested. Early detection of H. halys is essential for mitigati ng the ecological and e conomic effects of this invasive species. Partnerships with federal and state government agencies, researchers, agriculture extension clientele, and the general public increases the likelihood of detecting H. halys and other invasive species. These collabo rative efforts may include workshop trainings, E learning modules, and other educational extension materials. The purpose of the Invasive Species Workshop Series was to bring diverse groups together for a common cause of early detection and invasive specie s awareness. The Invasive Species Workshop Series will include additional training sessions and target a variety of different audiences, including Florida Master Gardeners, nurserymen, small farm producers, and public gardens personnel. The training sessio ns are scheduled to be completed by July 2014. In addition, the brown marmorated stink bug presentation will be developed into an E learning module and be available through the National Plant Diagnostic Network and Protect U.S. websites. The field guide, Identifying Stink Bugs and Other True Bugs, was developed for general public use. The field guide assessment indicated that stink bug and true bug species with similar coloration may be easily misidentified. Only 18% of participants correctly identified H halys which was commonly misidentified as E. servus (Say), E. tristigmus (Say), or Podisus maculiventris Say. Future revisions of the field guide will

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74 include photographic keys for species with similar coloration and highlight distinguishable characteri stics within the images. In order to provide improved public accessibility, an electronic version of the field guide will be available once all revisions have been made. Additional trainings for Florida Master Gardeners and the general public may be useful to better explain how to use the field guide for identifying true bug specimens. Halyomorpha halys research continues to have a significant impact on monitoring strategies and control techniques for this species. Although it seems that the introduction of this invasive species to Florida is inevitable, the host specificity research and extension efforts should continue in hopes that the destructive effects of this species are reduced.

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75 LIST OF REFERENCES BORROR, D. J., AND WHTIE, R. E. 1998. Peterson fi eld guide to insects. Houghton Mifflin Harcourt, New York, New York, USA. CAPINERA, J. L. 1985. Determination of host plant preferences of Hemileuca oliviae (Lepidoptera: Saturniidae), Paropomala wyomingensis (Orthoptera: Acrididae), and Diapheromera vel i (Orthoptera: Phasmatidae) by choice test and crop analysis. J. Kansas Entomol. Soc. 58(3): 465 471. DE CLERCQ, P. 2008. Spined soldier bug, Podisus maculiventris (Say) (Hemiptera: Pentatmoidae, Asopinae). pp. 3508 3510. In Capinera, J. L. (editor) Ency clopedia of Entomology, Vol. 4. Springer, Heidelberg. DEVOS, M., AND JANDER, G. 2008. Choice and no choice assays for testing the resistance of A. thaliana to chewing insects. J. Vis. Exp (15), e683, DOI: 10.3791/683. EATON, E. R., AND KAUFMAN, K. 200 7. Kaufman field guide to insects of North America. Houghton Mifflin Company, New York, New York, USA. FEDERAL INTERAGENCY COMMITTEE FOR THE MANAGEMENT OF NOXIOUS AND EXOTIC WEEDS (FICMNEW). 2003. A national early detection and rapid response system for invasive plants in the United States. Retrieved November 6 2013, from http://www.fws.gov/ficmnew/FICMNEW_EDRR_FINAL.pdf FLORIDA DEPARTMENT OF AGRICULTURE AND CONSUMER SERVICES (FDACS). 20 11. Florida agriculture by the numbers. Retrieved November 6, 2013, from http://www.florida agriculture.com/education.htm FLORIDA DEPARTMENT OF AGRICULTURE AND CONSUMER SERVICES DIVISION O F PLANT INDUSTRY (FDACS DPI). 2008. Cooperative Agricultural Pest Survey program (CAPS). Retrieved November 6, 2013 from http://www.freshfromflorida. com/Divisions Offices/Plant Industry/Bureaus and Services/CAPS FLORIDA DEPARTMENT OF ENVIRONMENTAL PROTECTION (DEP). 2011. Millions Retrieved November 6, 2013, from http://content.govdelivery.com/bulletins/gd/FLDEP c4328 FLORIDA INVASIVE SPECIES PARTNERSHIP (FISP). March 2013. Cooperative Invasive Species Management Areas (CISMAs). Retrieved November 6 2 013, from http://www.floridainvasives.org/cismas.html

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76 GYELTSHEN, J., BERNON, G., HODGES, A., STOCKS, S., AND BRAMBILA, J. 2011. Brown marmorated stink bug, Halyomorpha halys (Stl). Featured Cr eatures Retrieved November 6, 2013 from http://entnemdept.ufl.edu/creatures/veg/bean/brown_marmorated_stink_bug.htm HALBERT, S. 2011. Personal communication. F lorida Department of Agriculture and Consumer Services, Division of Plant Industry. HALBERT, S., AND HODGES, G. S. 2011. The brown marmorated stink bug, Halyomorpha halys (Stl). Retrieved November 6 2013, from http://www.freshfromflorida.com/content/download/23899/486244/halyomorpha halys.pdf HAMILTON, G. C. 2009. Brown marmorated stink bug. American Entomol 55(1): 19 20. HOEBEKE, E. R., AND CARTER, M E. 2003. Halyomorpha halys (Stal) (Heteroptera: Pentatomidae): a polyphagous plant pest from Asia newly detected in North America. Proc. Entomol. Soc. Wash 105(1): 225 237. HOFFMAN, W. E. 1931. A pentatomid pest of growing beans in South China. Peking Nat. Hist. Bull. 5(1):25 26. INVASIVE SPECIES ADVISORY COMMITTEE (ISAC). 2006. Invasive species definition clarification and guidance white paper. Submitted by the Definitions Subcommittee of the Invasive Species Advisory Committee. Retrieved November 6 2013, from http://www.invasivespecies.gov/global/ISAC/ISAC_documents/ISAC%20Definititi ons%20White%20Paper%20%20 %20F INAL%20VERSION.pdf JACOBS, S. 2009. Brown marmorated stink bug, Halyomorpha halys. Insect Advice from Extension, Pennsylvania State University Retrieved November 6, 2013 from http://ento.psu.edu/extension/factsheets/brown marmorated stink bug (Revised April 2011). Midwest. Iowa State University of Science and Technology. Retrieved November 6 2 013, from http://www.iasoybeans.com/productionresearch/publications/BrownMarmoratedS tinkBug/BrownMarmoratedStinkBug.pdf JONES, J. R., AND LAMBDIN, P. L. 2009. New county and state records for Tennessee of an exotic pest, Halyomorpha halys with potential economic and ecological implications. Florida Entomol 92 (1): 177 179.

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77 KAMMINGA, K., HERBERT, JR., A., MALONE, S., KAHUR, T. P., AND GREENE, J. 2009. Field guide to stink bugs of agricultural importance in the upper southern region of the Mid Atlantic States. Virginia Integrated Pest Management Center. Retrieved November 6 2013, from http://pubs.ext.vt.edu/444/444 356/444 356_pdf.pdf KAMMINGA, K. L., KOPPEL, A. L., HERBERT, D. A., AND KUHAR, T. P. 2012. Biology and management of the green stink bug. Journal of Integrated Pest Management. 3(3): C1 C8. KHRIMIA N, A., SHEARER, P. W., ZHANG, A., HAMILTON, G. C., AND ALDRICH, J. R. 2007. Field trapping of the invasive brown marmorated stink bug, Halyomorpha halys with geometric isomers of methyl 2,4,6 decatrienoate. J. Agric. Food Chem. 56(1): 197 203. KIRKPATRI level model. Training & Development. 50(1): 54 59. LESKEY, T. C., AND HOGMIRE, H. W. 2005. Monitoring stink bugs (Hemiptera: Pentatmoidae) in mid Atlantic apple and peach orchards. J. E con. Entomol. 98(1): 143 153. LESKEY, T. C., WRIGHT, S. E., SHORT, B. D., AND KHRIMIAN, A. 2012a. Development of behaviorally based monitoring tools for the brown marmorated stink bug (Heteroptera: Pentatomidae) in commercial tree fruit orchards. J. Ento mol. Sci. 47 (1): 76 85. LESKEY, T. C., SHORT, B. D., BUTLER, B. R., AND WRIGHT, S. E. 2012b. Impact of the invasive brown marmorated stink bug, Halyomorpha halys (Stl), in Mid Atlantic tree fruit orchards in the United States: Case studies of commercia l management. Psyche. doi:10.1155/2012/535062 MCPHERSON, J. E., AND MCPHERSON, R. M. 2000. Stink bugs of economic importance in America north of Mexico. CRC Press, Boca Raton, Florida, USA. MEDAL, J., SMITH, T., FOX, A., CRUZ, A. S., POPLIN, A., AND HOD GES, A. 2012. Rearing the brown marmorated stink bug Halyomorpha halys (Heteroptera: Pentatomidae). Florida Entomologist 95(3): 800 802. MILNE, L. J., MILNE, M. J. G., AND RAYFIELD, S. 1980. National Audobon Society field guide to North American insects and spiders. AA Knopf, New York, New York, USA. MIZELL III, R. F. 2008. Monitoring stink bugs with the Florida stink bug trap. Insect Traps and Sampling Retrieved November 6, 2013 from http://nfrec.ifas.ufl.edu/MizellRF/stink_bugs/stink_bugs.htm

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78 NATIONAL AGRICULTURAL PEST INFORMATION SYSTEM (NAPIS). November 6 2013. Purdue University. Survey status of brown marmorated stink bug Halyomorpha halys (All years)." Retrieved No vember 6 2013, from http://pest.ceris.purdue.edu/map.php?code=IQAQQKA&year=3year# NATIONAL AGRICULTURAL STATISTICS SERVICE (NASS). 2012. Florida agriculture overview. Florida Agriculture Statistical Bulletin. Retrieved November 6 2013 from http://www.nass.usda.gov/Statistics_by_State/Florida/Publications/Ann ual_Statist ical_Bulletin/fasb12p.htm NATIONAL INVASIVE SPECIES COUNCIL (NISC). 2003. General guidelines for the establishment and evaluation of invasive species early detection and rapid response systems. Version 1. 16 pp. NIELSEN, A. L., HAMILTON, G C., AND MATADHA, D. 2008a. Developmental rate estimation and life table analysis for Halyomorpha halys (Hemiptera: Pentatomidae). Environ. Entomol 37 (2): 348 355. NIELSEN, A. L. SHEARER, P. W., AND HAMILTON, G. C. 2008b. Toxicity of insecticides to H alyomorpha halys (Hemiptera: Pentatomidae) using glass vial bioassays. J. Econ. Entomol 101 (4): 1439 1442. NIELSEN, A. L., AND HAMILTON, G. C. 2009a. Seasonal Occurrence and Impact of Halyomorpha halys (Hemiptera: Pentatomidae) in Tree Fruit. J. Econ. E ntomol. 102(3): 1133 1140. NIELSEN, A. L., AND HAMILTON, G. C. 2009b. Life history of the invasive species Halyomorpha halys (Hemiptera: Pentatomidae) in northeastern United States. Ann. Entomol. Soc. Am. 102(4): 608 616. NIELSEN, A. L., HAMILTON, G. C., AND SHEARER, P. W. 2011. Seasonal phenology and monitoring of the non native Halyomorpha halys (Hemiptera: Pentatomidae) in soybean. Environ. Entomol. 40(2): 231 238. NIELSEN, A. L., HOLMSTROM, K., HAMILTON, G. C., CAMBRIDGE, J., AND INGERSON MAHAR, J. 2013. Use of black light traps to monitor the abundance, spread, and flight behavior of Halyomorpha halys (Hemiptera: Pentatomidae). J. Econ. Entomol. 106(3): 1495 1502. NORLAND TILBURG, E. V. 1990. Controlling error in evaluation instruments. Journal of Extension. 28(2). Retrieved November 6 2013, from http://www.joe.org/joe/1990summer/tt2.php

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79 PEA, J. E. 2003. Pests of avocado in Florida. Proceedings V World Avocado Congress (Actas V Congreso Mundial del Aguacate). pp. 487 494. PIMENTEL, D., LACH, L., ZUNIGA, R., AND MORRISON, D. 2010. Environmental and economic costs associated with non indigenous species in the United States, pp. 285 303. In Pimentel, D. (ed.), Biological invasions: Economi c and environmental costs of alien plant, animal, and microbe species. CRC Press, Danvers, MA. RAFFA, K. F., HAVILL, N. P., AND NORDHELM, E. V. 2002. How many choices can your test animal compare effectively? Evaluating a critical assumption of behaviora l preference tests. Oecologia 133(3): 422 429. RASHID, T., JOHNSON, D. T., AND BERNHARDT, J. L. 2006. Sampling rice stink bug (Hemiptera: Pentatmoidae) in and around rice fields. Environ. Entomol. 35(1): 102 111. ROCKWELL, S. K., AND KOHN, H. 1989. Po st then pre evaluation. Journal of Extension. 27 (2): 19 21. Retrieved November 6 2013, from http://www.joe.org/joe/1989summer/a5.php RUSSELL, D. 2012. Agricultural interdiction station inspectio n (PowerPoint presentation). Retrieved November 6 2013 from http://freshfromflorida.s3.amazonaws.com/pdf_caps2013w agricultural interdiction stations 2012.pdf SARGENT, C., DIVELY, G., HOOKS, C., RAUPP, M., SARDANELLI, S., SHREWSBURY, P., CLEMENT, D., AND MALINOSKI, M. 2010. Brown marmorated stink bug, Halyomorpha halys Stl (Hemiptera: Pentatomidae). University of Maryland Fact Sheet. Retr ieved November 6 2013, from http://www.pestthreats.umd.edu/content/documents/BMSBFactSheet_10 2010_000.pdf SILAGYI, A. 2005. Introduction of the Cooperative Agricultural Pest Survey Program. Agronomy Seminar, UF Agronomy Department, Gainesville, Florida. Retrieved November 6 2013, from http://santarosa.ifas.ufl.edu/documents/invasive _salagyi.pdf SMITH, T. R. 2012. Personal communication. Florida Department of Agriculture and Consumer Services, Division of Plan Industry, Bureau of Methods Development and Biological Control. STATE OF FLORIDA. 2013. Facts about Florida. 2013. Retri eved November 6 2013, from http://www.stateofflorida.com/Portal/DesktopDefault.aspx?tabid=95

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80 UNITED STATES DEPARTMENT OF AGRICULTURE ANIMAL AND PLANT HEALTH INSPECTION S ERVICE PLANT PROTECTION AND QUARANTINE (USDA APHIS PPQ). 2012. Questions and answers: New risk based sampling protocol and propagative monitoring and release program at plant inspection stations. Retrieved November 6 2013, from http://www.aphis.usda.gov/publications/plant_health/2012/risk_based_sampling.p df UNITED STATES ENVIRONMENTAL PROTECTION AGENCY. 2012. Integrated pest management (IPM) principles. Retr ieved November 6 2013, from http://www.epa.gov/pesticides/factsheets/ipm.htm UNITED STATES FOREST SERVICE (USFS). 2006. Early detection and rapid response. Retrieved November 6 2013, from http://www.fs.fed.us/invasivespecies/earlydetection.shtml UNIVERSITY OF FLORIDA (UF) INSTITUTIONAL REVIEW BOARD (IRB) 02. 2007. Consent/Assent procedures and instructions. Retriev ed November 6 2013 from http://irb.ufl.edu/irb02/ifc.html UNIVERSITY OF FLORIDA (UF) MASTER GARDENER PROGRAM. 2012. About the Master Gardener Program. Retrieved November 6 2013, from http://gardeningsolutions.ifas.ufl.edu/mastergardener/about/index.shtml VAN DRIESCHE, R. G., AND MURRAY, T. J. 2004. Overview of testing schemes and designs used to estimate host range s. In Van Driesche and R. Reardon, Eds.), pp. 68 89. USDA Forest Service, Morgantown, West Virginia, U.S.A. WERMELINGER, B., WYNIGE R, D., AND FORSTER, B. 2008. First records of an invasive bug in Europe: Halyomorpha halys Stal (Heteroptera: Pentatomidae), a new pest on woody ornamentals and fruit trees? Mitteilungen der Schweizerischen Entomologischen Gesellschaft 81(1): 1 8. WITHER S, T., AND MANSFIELD, S. 2005. Choice or no choice tests? Effects of experimental design on the expression of host range. Second International Symposium on Biological Control of Arthropods. Davos, Switzerland, 12 16 September, 2005. (pp. 620 633). United S tates Department of Agriculture, Forest Service. YANG, Z. Q., YAO, Y. X., QUI, L. F., AND LI, Z. X. 2009. A new species of Trissolcus (Hymenoptera: Scelionidae) parasitizing eggs of Halyomorpha Halys (Heteroptera: Pentatomidae) in China with comments on its biology. Ann. Entomol. Soc. Am. 102(1): 39 47.

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81 YU, G., AND ZHANG, J. 2007. The brown marmorated stink bug, Halyomorpha halys (Heteroptera: Pentatomidae) in PR China. International Workshop on Biological Control of Invasive Species of Forests, Beijing, P.R., China, 20 25 September, 2007. (pp. 58 62). United States Department of Agriculture, Forest Service. ZHU, G., BU, W., GAO, Y., AND LIU, G. February 2012. Potential geographic distribution of brown marmorated stink bug invasion ( Halyomorpha halys ). PLoS one 7(2): e31246.

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82 BIOGRAPHICAL SKETCH Ashley Poplin is from Daytona Beach, Florida and graduated from Keystone Heights High School in May 2007. After graduation she attended University of Central Florida and majored in f orensic s cience with a minor in c hemistry and a certificate in c rime s cene i nvestigation. During her senior year at UCF, Ashley was inspired by a lecture in her forensic archeology class and gained an interest in entomology. Ashley was offered a research assistantship under the direction of Dr. Amanda Hodges at the University of Florida beginning August 2011. As part of her assistantship, she collaborated with multiple government agencies to develop ex tension education materials. The appeal of developing these materials motivated her to incorporate extension education as part of her research to complement her project on the brown vestigator and continue her education in forensic entomology.