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The tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), is considered the most economically important pest of midsouth cotton. This study was designed to understand the impact of tarnished plant bug populations in the later weeks of flowering cotton and residual effects of novaluron with subsequent applications. Experiments evaluated dynamic threshold approaches in the later flowering period of midsouth cotton, the impacts of novaluron plus subsequent applications at the 3rd week of square, as well as residual properties in a laboratory experiment. Results suggest that a dynamic late season approach can reduce the amount of insecticide applications targeting tarnished plant bug with no penalty to yield. Populations were observed at lowest densities where applications of novaluron were tank mixed with an adulticide (acephate) at the 3rd week of square. Results of this experiment will be important in refining seasonal management recommendations for tarnished plant bug in midsouth cotton production systems.

Harlequin bug (HB), Murgantia histrionica (Hahn), is a pest of cole crops. Alternative control strategies were investigated for control of HB, including trap cropping and systemic neonicotinoid insecticide applications. Potential trap crops, mustard (Brassica juncea "Southern Giant Curled" ), rapeseed (B. napus "Athena"), rapini (B. rapa) and arugula (Eruca sativa) were preferred over collard (B. oleracea "Champion"), and a non-brassica control, bean (Phaseolus vulgaris "Bronco") in field-cage choice tests. Harlequin bug could not complete development on bean, developed poorly on arugula but was found to complete development on mustard, collard, rapeseed and rapini. In the field, mustard was found to be an effective trap crop for reducing HB feeding injury on collard at three experimental sites in 2010 and 2011. Augmentation of the mustard trap crop with a systemic, neonicotinoid insecticide did not increase the level of control of harlequin bug for the duration of the ten week growing period. In olfactometer choice tests, male HB responded to plant volatiles of bean, collard and mustard, but preferred Brassica volatiles over those from bean. Female response to plant volatiles alone was weak and inconsistent. Both males and females preferred volatiles from other males feeding on Brassica host plant over plant volatiles alone, and were deterred by volatiles from males feeding on bean versus the plant alone. Laboratory toxicity assays revealed that the neonicotinoid insecticides imidacloprid, thiamethoxam, dinotefuran, and clothianidin were toxic to HB nymphs; LC50 = 0.57, 0.52, 0.39, and 0.39 mg ai/liter, respectively. Field experiments were conducted to evaluate the efficacy of these insecticides over time when applied as a one-time drench, and all were found to provide significantly higher mortality of HB for at least 14 days after application. / Ph. D.

For the first time in many decades, a hemp pilot program was initiated in Virginia in 2016. Outdoor surveys were conducted in the 2017 and 2018 field seasons to record insect presence and feeding injury to plants. Multiple insect pests were present, including corn earworm (Helicoverpa zea [Boddie]) (Lepidoptera: Noctuidae), brown marmorated stink bug (Halyomorpha halys [Stål]) (Hemiptera: Pentatomidae), and cannabis aphid (Phorodon cannabis) (Hemiptera: Aphididae). In 2019, indoor production surveys revealed that cannabis aphid, twospotted spider mite (Tetranychus urticae Koch) (Acari: Tetranychidae), and hemp russet mite (Aculops cannabicola [Farkas]) (Acari: Eriophyidae) would likely cause production issues. Very little is known about the impact of insect defoliation in hemp so studies were conducted in 2018-2020 to determine impacts on yield and cannabinoid content of grain and cannabinoid variety hemp due to leaf surface area loss. In Virginia over two growing seasons, manual removal of leaf tissue in grain and CBD cultivars did not significantly impact observable effects on physical yield (seed or bud weight) or cannabinoid content (CBD or THC) at time of harvest. Corn earworm is the major pest of hemp produced outdoors and studies occurred to evaluate monitoring and management strategies. Pheromone traps may be valuable in determining when corn earworm moths are present in the vicinity of hemp fields but are not useful in predicting larval presence in buds or final crop damage. Larval presence and final crop damage are related. Brown marmorated stink bug does not appear to be a concern in hemp, at least at this time. / Doctor of Philosophy / For the first time in many decades, a hemp pilot program was initiated in Virginia in 2016. Outdoor surveys were conducted in the 2017 and 2018 field seasons to record insect presence and feeding injury to plants. Multiple insect pests were present, including corn earworm, brown marmorated stink bug, and cannabis aphid. In 2019, indoor production surveys revealed that cannabis aphid, twospotted spider mite, and hemp russet mite would likely cause production issues. Very little is known about the impact of leaf area loss due to insect feeding in hemp so studies were conducted in 2018-2020 to determine impacts on yield and cannabinoid content of grain and cannabinoid variety hemp due to leaf surface area loss. In Virginia over two growing seasons, manual removal of leaf tissue in grain and CBD cultivars did not significantly impact observable effects on physical yield (seed or bud weight) or cannabinoid content (CBD or THC) at time of harvest. Corn earworm is the major pest of hemp produced outdoors and studies occurred to evaluate monitoring and management strategies. Pheromone traps may be valuable in determining when corn earworm moths are present in the vicinity of hemp fields but are not useful in predicting larval presence in buds or final crop damage. Larval presence and final crop damage are related. Brown marmorated stink bug does not appear to be a concern in hemp, at least at this time.

The tarnished plant bug (TPB), Lygus lineolaris (Palisot de Beauvois), has been a devastating insect pest in cotton agroecosystems in mid-southern cotton for several decades. Similar to related Heteropteran insect pests, L. lineolaris uses piercing-sucking mouthparts to penetrate harvestable fruiting structures on the cotton plant (i.e., squares, flowers, small bolls) leading to significant reductions in lint yield when infestations are above economic injury levels. Economically damaging infestations of L. lineolaris have occurred in Virginia and North Carolina cotton since 2013. In response, cotton area sprayed with broad-spectrum insecticides for L. lineolaris has increased in these states. As such, this dissertation research sought to answer a variety of questions to address this issue including 1) mapping the spatiotemporal abundance of L. lineolaris across Virginia as well as creating models to predict landscape and climate factors increasing infestation risk, 2) using a systems approach to provide growers with cost-effective and sustainable integrated pest management (IPM) solutions including economic spray thresholds, 3) investigating the incidence and transmission of fungal pathogens by L. lineolaris increasing Fusarium hardlock disease, further impacting yield loss at harvest, and 4) conducting insecticide resistance monitoring of L. lineolaris across the Southeast including biochemical lab assays to determine possible mechanisms of insecticide resistance. These findings will help form compressive IPM and insect resistant management (IRM) plans to equip southeastern cotton growers with practical management tools as well as useful information on "high-risk" areas to invest management resources, increasing economic returns while minimizing environmental impacts of over-spraying foliar insecticides. / Doctor of Philosophy / The tarnished plant bug (Lygus lineolaris) is an important pest in cotton from July through September that causes reduced yields and lower quality of lint and seed. Tarnished plant bugs cost the cotton industry hundreds of millions of dollars in losses annually. However, tarnished plant bug infestations were rare and sporadic in Virginia and North Carolina before 2013. In recent years, tarnished plant bug infestations have nearly doubled in these states, and research is needed to better understand how to manage recent pest outbreaks to avoid substantial economic losses. To address this issue, this dissertation research investigated a variety of questions including 1) the distribution of tarnished plant bugs in this region and the effects of geography and climate on infestation risk, 2) best practices for managing this pest in cotton fields, 3) possible associations between tarnished plant bugs and cotton boll diseases further impacting yield, and 4) whether tarnished plant bugs in this region are resistant to frequently used broad-spectrum insecticides as well as potential mechanisms driving resistance levels. Findings from this research will equip southeastern cotton growers with regionally specific recommendations for managing and monitoring tarnished plant bugs that minimize management costs and increase the quality and quantity of cotton yields.

Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), brown marmorated stink bug, is an invasive, polyphagous pest that has disrupted integrated pest management programs in Mid-Atlantic tree fruit orchards since 2010. The amount of damage to fruit caused by H. halys feeding is highly variable, even within adjacent orchard blocks, and the influence of orchards and wild host plants nearby on H. halys relative abundance is unknown. Pheromone-based trapping studies conducted along woodland-to-orchard transects, along woodland borders at sites with and without adjacent orchards, and within the mid-canopy of several common wild host trees in woodland borders were used to investigate biotic factors underlying the variability of H. halys populations among sites within a geographically proximate area. Additionally, the retention duration of H. halys on wild and cultivated tree hosts was used as a proxy for host acceptability throughout the season. All trapping studies yielded significantly greater H. halys captures during the late season, and in the transect study, pyramid traps deployed at orchard and woodland edges during this period yielded the highest captures. The presence of orchards or open fields adjacent to woodlands had no significant impact on H. halys captures in sticky traps deployed along wooded edges, but captures varied significantly among orchard and open field sites. Among 11 common wild host plant genera surveyed at these sites, five genera, Elaeagnus, Fraxinus, Lonicera, Ailanthus, and Sassafras, were significantly related to H. halys relative densities. Conversely, studies measuring captures in small pyramid traps within the canopy of wild tree hosts revealed no significant differences among hosts at any point in the season, suggesting that this approach might not be appropriate for assessing host plant suitability. Finally, harmonic radar trials assessing host plant retention as a proxy for host suitability revealed that adults released on peach and pheromone-baited apple trees had significantly greater retention durations than those released on non-host, grass, and that adult retention duration increased from the early to late season. In combination, results indicate that the risk posed by H. halys to cultivated orchard crops varied across locations and seasonally, and may be related to the species composition of wild host trees in adjoining woodlands. / Doctor of Philosophy / Brown marmorated stink bug (BMSB), Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), is an invasive insect that has inflicted feeding damage on fruit grown in Mid-Atlantic orchards since 2010. The agricultural landscape in the Mid-Atlantic region complicates BMSB management, as many orchards are bordered by woodlots containing a variety of BMSB host plants. The amount of BMSB feeding injury at harvest can vary greatly among nearby orchards, and even within individual orchards, but it is unknown why these differences exist. Several BMSB trapping studies using pheromone-baited traps revealed that BMSB populations were greatest in the late-season, confirming previous work. Furthermore, captures in traps positioned in lines intersecting orchards and adjacent woodlots revealed late-season BMSB presence was greatest at the orchard and woodland edges. Captures in traps next to woodlots adjacent to orchards and open field sites indicated that the presence of orchards had no impact on captures, but that captures varied significantly among orchard sites and open field sites. A survey of the woody plants in the woodlots adjacent to these sites revealed that five host plant genera were strongly related to BMSB abundance. A study examining BMSB captures in small pyramid traps deployed in the canopy of several common wild tree hosts did not yield differences in captures among hosts at any point in the season, and thus was not a suitable method for assessing seasonal differences in BMSB host use. Finally, harmonic radar technology was used to assess how long BMSB remained on several wild and cultivated host plants at different points in the growing season, as a proxy for host suitability. Peach and pheromone-baited apple showed significantly greater adult retention durations than non-host, grass, and adults remained on host plants longer in the early than late season. Ultimately, these results should aid in the monitoring and management of BMSB by providing a broader understanding of factors underlying the presence and abundance of BMSB on a localized geographic scale.

The common bed bug, Cimex lectularius L, has been a persistent pest of humans. Conventional pyrethroid-formulation insecticides are known to be ineffective for controlling modern bed bug populations. This study evaluates alternative treatment methods such as whole-home heat, a biological control agent (Beauveria bassiana), and the fumigant sulfuryl fluoride for control of insecticide-resistant bed bugs. Three heat systems with different energy sources (propane, electric, and glycol) were evaluated to determine attributes contributing to heat treatment efficacy. The glycol system produced the most mortality of the three systems, killing all nymphs, eggs, and most adults. Heat treatment duration and achieving lethal temperatures in complex environments were found to be the most important factors for treatment efficacy. These factors were directly correlated with technician diligence, specifically regarding monitoring surface temperatures and repositioning equipment. A formulation of B. bassiana was evaluated in the laboratory to determine its ability to infect bed bugs under varying conditions of temperature (15°C, 21°C, and 32°C) and humidity (30%, 50%, and 70%). It was found that humidity conditions (30%-50%) at ≈21°C produced the greatest bed bug mortality and the shortest bed bug median survival time. The fumigant sulfuryl fluoride was evaluated for its ability to eliminate bed bugs from motor vehicles and cargo trailers filled to 85% capacity. This study was the first to document that sulfuryl fluoride fumigation at the 1.9X dosage factor can kill all pyrethroid-resistant bed bug life stages (including eggs) in motor vehicles as well as in chambers filled with personal items. / Master of Science in Life Sciences / Since the world-wide bed bug (Cimex lectularius L.) resurgence began in the late 1990s, populations of the common bed bug have been evaluated, but most have been found to be highly resistant to pyrethroid insecticides. This resistance has been particularly troublesome due to the fact that the 1996 US Food Quality Protection Act has eliminated the use of many existing chemistries from indoor use and has inadvertently limited the development of new active ingredients for indoor use due to the cost of the required animal testing to document the No Observable Effects Level (NOEL). Due to the lack of novel chemistry for addressing modern bed bug infestations, pest management professionals have had to identify new methods for applying existing products (chemical and non-chemical) for bed bug control. This study evaluated gaseous, mechanical, and biological control methods for bed bug elimination to determine which factors contribute the most to their efficacy, as well as how these methods might be applied in novel ways for control of bed bugs in homes, personal belongings, and even vehicles. At the turn of the 20th century heat treatments were used for controlling bed bug infestations in homes and other structures. Today, mechanical heating systems are again being used to control bed bug infestations in homes and apartments. This study investigated the utility of three commercial heating systems for their ability to control existing infestations in apartment units. The three heat systems utilized different energy sources, different types of delivery equipment, and required different set up and take down procedures in apartments of different cubic footage and clutter levels. Overall, the Assault glycol heating system was found to be the most effective in its ability to get (almost) all of our hidden (in hard to heat locations) temperature sensors up to bed bug lethal temperature. Interestingly, our statistical analysis determined that getting the hard to heat locations up to lethal temperature was directly correlated with heat technician activity. The more times that the technician entered the home to monitor the treatment and adjust the equipment positions, the greater the chance of getting the hidden sensors up to bed bug lethal temperature (and killing the bed bugs). However, it was also found that none of the heat systems tested killed all of the sentinel bed bugs in every replicate. Therefore, we learned heat treatments cannot be expected to eliminate each and every bed bug in a home, and that supplemental control products such as desiccants dusts should be applied after every heat treatment. When attempting to determine which heat system was the most effective for killing bed bugs, it was determined that regardless of which heat systems was being used, the attention and activity (monitoring temperatures and adjusting equipment) of the heat treatment technician was the most important factor contributing to bed bug mortality as well as the achievement of bed bug lethal temperatures in cracks and crevices for all three systems. Our second study examined the environmental factors that influenced fungal growth after bed bug exposure to the insecticidal product Aprehend® (active ingredient Beauveria bassiana). Over the last several decades, this fungus has been widely used to control multiple insect pests. Recently, it has been labelled for bed bug control in indoor environments. This second study was intended to determine the atmospheric conditions (temperature and humidity) under which Aprehend sporulation was most effective for killing bed bugs. It was found that humidity conditions of 30-50% combined with temperatures of ~21°C produced the highest frequency of fungal infection and the shortest median bed bug survival time. This result was surprising because it was originally hypothesized that the 70% humidity condition would be equally effective for promoting fungal growth. However, at all of the temperatures tested, bed bug fungal infection rates at 70% humidity were not as high as those observed when the humidity was within the 30-50% range. Therefore, this study was able to document that the temperatures and humidity combinations that would typically be found within human homes (21oC at 30-50% humidity) were the most effective for producing fungal infections when bed bugs were exposed to the Aprehend product. The final study addressing novel methods for controlling insecticide resistant bed bugs investigated the efficacy of using Vikane gas fumigant (sulfuryl fluoride) at the 1.9× dosage rate for eliminating bed bugs in two challenging infestation situations: personal vehicles, and confined spaces densely packed with personal belongings. The vehicles used in this study were large minivans with seating that folded into the floor. The confined spaces were cargo trailers filled to 85% capacity with books, furniture, and other household items. Each van and trailer was equipped with ~90 sentinel bed bugs consisting of three groups of 9-11 bed bug eggs, 10 nymphs, and 10 adults. The Vikane Fumiguide calculator was used to determine the target dosage (g-h/m3) to apply in each replicate (e.g., one van or trailer). Sulfuryl fluoride concentrations were measured throughout the fumigation process using a Spectros SF-ReportIR. Concentration readings were input into the Fumiguide to determine when the accumulated dosage (g-h/m3) was achieved, and when aeration should be initiated. After aeration was complete, the sentinel bed bugs were removed from the replicates and bed bug nymph and adult mortality was recorded. Bed bug eggs were monitored for 23 d to determine latent mortality. Fumigated bed bug mortality for each replication was 100% regardless of life stage. Latent mortality was observed in a single bed bug egg, where the nymph never fully hatched. This study determined that fumigation with sulfuryl fluoride at the 1.9× dosage factor is an effective method for eliminating insecticide resistant bed bugs from vehicles and personal belongings in densely packed situations. Overall, it was found that the Beauveria bassiana product was most effective when applied under atmospheric conditions that are typically found in indoor home environments. This discovery was very reassuring, because the Aprehend product is one of very few that actually have residual activity with regard to bed bug control. This study also found that whole home heat treatments require rigorous pest technician attention and monitoring to achieve the best results. It was also determined that heat treatments cannot be assumed to be 100% effective on their own, and that they should be supplemented with additional (residual) treatment applications. Not surprisingly, fumigation with sulfuryl fluoride was determined to be 100% effective for eliminating bed bugs in personal belongings that had been packed into treatment chambers. The fumigation process also proved to be 100% effective for eliminating bed bug infestations in transport vehicles which cannot not be adequately treated and are at risk for having electronic components damaged if treated with heat.

The brown marmorated stink bug (Halyomorpha halys) (Stål) is known for overwintering in human homes. Studies were conducted to understand the behavior of adult bugs in response to characteristics of potential overwintering structures including gap sizes, light, and presence of dead conspecifics. In a test where bugs were placed in increasingly tight boxes, most bugs settled in gaps 4.5 - 5.5 mm high. None settled in a space less than 3.5 mm high. In boxes with constant tightness, bugs tended to move to the back, and primarily settled along walls and in corners. In both box types, bugs tended to turn and face the cavity entrance during settling. In tests of responses of disturbed overwintering bugs to common household light bulbs over the course of a year, bugs were taken from shelters and exposed to lit bulbs. Bug responses were seasonal, with attraction to light bulbs in spring and summer, but little or no consistent response in fall or winter. The lack of response occurred more often at night than in afternoons. Because not all bugs survive the winter, corpses can accumulate. Single shelter-seeking H. halys were found not to respond to the presence of dead aggregations from the previous years unless touch was possible, in which case aggregations were joined. Dead aggregations from the same year had a repellent effect. Presence of a single dead bug from the same year did not provide any survivorship advantage to overwintering bugs. Results suggest possible improvements to trap shelters. / Master of Science in Life Sciences / The brown marmorated stink bug (Halyomorpha halys) is known for overwintering in human homes. This research investigated the responses of these bugs to some characteristics of overwintering structures, including crevice tightness preference, light sources, and dead bugs leftover from previous years. Tightness experiments indicated that bugs in crevices with hard walls prefer to settle in spaces between 4.5 and 5.5 mm high. Bugs also tended to stay on the floor, go back as far as they could, and face the entrance of the crevice. These findings will help improve overwintering box designs for collection and research, and will help pest control professionals focus their efforts. When bugs were disturbed and exposed to common household light bulbs, they tended to move toward the bulbs in late summer and in spring, but responded less in fall and winter, and responded more often in the afternoon than at night. Bugs were also exposed to other dead bugs in several ways. When lone bugs were exposed to groups of dead bugs from previous years, they went to them only if they could touch them. Single dead bugs had less of an impact. Exposure to groups of dead bugs from the same year drove live bugs away. Overwintering brown marmorated stink bugs do not appear to eat other bugs that have recently died. These results suggest several possible improvements to the designs of trap shelters used for research and control.

Brown marmorated stink bug, Halyomorpha halys (Stål), is a crossover pest impacting agriculture and invading urban environments. Studies were conducted to better understand the management of the bug through its physiology, behavior and susceptibility to insecticidal intervention. Halyomopha halys exhibit great variability in overwintering site selection with choices including manmade structures and tree bark. Because of these diverse sites, the bug must have the ability to withstand varying conditions throughout the overwintering cycle. We were able to determine that H. halys is chill intolerant and capable of adapting its tolerance to temperatures by season, sex, and location of acclimation. The mean supercooling point (± SEM) in the winter in Minnesota was -17.06°C ± 0.13° and in Virginia was -13.90°C ± 0.09°. Laboratory experiments conducted in Blacksburg, VA were able to determine baseline lethal high temperatures over time against H. halys adults. To achieve 100% mortality, temperatures fell between 45°C and 50°C, 40°C and 45°C, and 42°C and 45°C, over 15-min, 1-h, and 4-h, respectively. Moving forward, we were able to utilize this information to develop heat treatment guidelines for export shipping cargo infested with overwintering H. halys. In a controlled field experiment, we determined that exposing the coldest areas of an infested vehicle to temperatures greater than 50°C for a minimum of 15 minutes resulted in 100% mortality of overwintering BMSB adults. In 2012 and 2013, citizen scientists were recruited through Virginia Cooperative Extension to assist in evaluating several in home light traps designed to help eradicate overwintering H. halys adults in homes. Over the course of the two year study, fourteen houses participated in the study with 72% of those houses having stink bug activity. It was found that the most effective trap was an aluminum foil pan trap. In 2013, the trap was 19 times more effective at catching stink bug adults than any other trap tested. In September of 2014, a near-field experiment was conducted to determine the residual efficacy of several recommended and labeled insecticides for treatment of homes against invading H. halys adults. This study used constructed window screen bags that were dipped in insecticide solution. After the initial treatment, bugs were exposed to the bags for 24h weekly, up to 54 days after treatment (DAT). It was determined that 2 DAT all insecticides had activity except for indoxacarb. All insecticides lost efficacy after 29 DAT except for lambda-cyhalothrin, beta-cyfluthrin, beta-cyfluthrin + imidacloprid, lambda-cyhalothrin + thiamethoxam, and dinotefuran, which had some measureable activity even after 40 DAT. Each of these insecticides contained a pyrethroid alone or in combination with a neonicotinoid. Laboratory bioassays were conducted to determine the LC50 values of clothianidin, dinotefuran, imidacloprid, and thiamethoxam against H. halys nymphs using a systemic application method. Those LC50 values were found to be 0.077, 0.013, 0.068, and 0.018 ppm, respectively. Field experiments conducted in Virginia in 2012 and 2013 showed a significant reduction of stink bug damage using two soil applications of neonicotinoid insecticides in pepper and tomato. In North Carolina, a single drip irrigation application significantly reduced stink bug damage in 2012 and 2014 using dinotefuran or imidacloprid. / Ph. D.

Although bed bug eggs are a difficult life stage to control with our currently labeled insecticides, few studies have examined how bed bug egg morphology and physiology is potentially related to pesticide resistance in bed bug eggs. Bed bug egg morphological features were examined using scanning electron microscopy (SEM) and the chorion and respiration structures were identified. Scanning electron microscopy photographs and bed bug egg measurements indicated there were no morphological differences between different bed bug egg strains (susceptible and resistant). Bed bug egg respiration rates measured by the amount of oxygen consumed (standard metabolic rate; SMR) also indicated there was no difference in SMR between different bed bug egg strains. Water conservation during respiration is vital to terrestrial insects. Therefore, similar patterns would be expected between egg water loss and respiration rates. However, susceptible strain eggs lost more water than one resistant strain of bed bug eggs, which was dissimilar from the respiration results, indicating that bed bug egg water loss and respiration are not directly related. Dose- response bioassays using two insecticide formulations (Temprid; imidacloprid/β-cyfluthrin, and Transport; acetamiprid/bifenthrin) indicated that bed bug eggs collected from pyrethroid resistant adult bed bug strains are also highly resistant. RNA sequencing of bed bug eggs from two resistant strains indicated that egg resistance may be directly related to the overexpression of multiple genes associated with insecticide resistance. / Master of Science in Life Sciences