Cage studies were performed to determine if the current thresholds for stink bugs in soybean (one bug per 0.3 row m) need to be adjusted based on current soybean production practices and species present. Several soybean development stages were infested by two native stink bug species for three weeks using small cages in a field of double crop soybean at sites in Virginia, Maryland, and Delaware during 2007-2009. Large field cages were infested by an introduced species for two weeks in 2010-2011. Results showed that Euschistus servus Say and Acrosternum hilare Say adults or nymphs did not cause different levels of injury to soybean seed quality or effects on yield. Both A. hilare and the introduced Halyomorpha halys Stål injured soybean seed in a similar fashion at threshold-level densities. Full flowering R2 stage soybean were least affected by stink bug feeding, and full pod and beginning seed R4-R5 stage soybean were slightly more sensitive to injury than R6 although not at the Maryland 2011 site. Several sites had increased seed injury and decreased yield at threshold density populations. Finally, visual observations of stink bug vertical distribution inside soybean canopies were taken several times per day and compared with ambient and within-canopy temperature and relative humidity. Results indicated that these conditions did not influence the percentage of stink bugs below the top 38 cm sweep net intercept zone. In both years of observations, between 15 and 20% of stink bugs were observed below the 38 cm sweep net zone. / Master of Science in Life Sciences

The brown marmorated stink bug (BMSB), Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), is an invasive insect from Asia that has recently become a major pest of agricultural crops and a nuisance to home and business owners in the Mid-Atlantic U.S. Since 2010, H. halys has been reported in many vineyards in Virginia, but the pest significance in this crop is unknown. Sampling was conducted in four commercial vineyards across Virginia in 2012 and 2013 to study the seasonal phenology and pest status of H. halys in vineyards. Adults moved into vineyards as early as May and laid eggs usually on the undersurface of leaves, but occasionally on the berry or the rachis. Grapevines were an early season reproductive host for H. halys. A vineyard adjacent to a sub-urban area with homes and buildings in proximity had an early season peak of H. halys as compared to vineyards adjacent to woods. However, populations declined sharply in late season due to the possible movement of bugs to more preferable host plants such as soybean and corn. In contrast, H. halys was recorded throughout the grape growing period in a vineyard that was surrounded by forests. Significantly more H. halys were recorded from border than interior section of vineyards. A degree-day model suggested that there were enough degree-days to complete a generation of H. halys in Virginia vineyards. H. halys caused direct injury to the grape berries at veraison and pre-harvest berries. Injury expression in the veraison berry can be described as an appearance of a small necrotic spot at the site of the stylets insertion. The spot gradually increased in size and the berries became deformed. H. halys is an economic pest of raspberry, causing direct injury to the berries. Sampling of stink bugs in raspberry plantings in southwestern Virginia showed that the Euschistus species were the most abundant stink bugs in 2008, 2009, 2011 and 2012. However, H. halys became the most abundant in 2013. / Master of Science in Life Sciences

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 (BMSB), Halyomorpha halys (Stål), has become an important pest of soybean in the Mid-Atlantic US. To assess the influence of tree borders on BMSB infestations in soybean, twelve fields were sampled weekly using five 3-min visual counts of BMSB on tree of heaven (TOH) (Ailanthus altissima) and other host trees along a wooded border, on the adjacent soybean edge, 15 m and 30 m into the soybean field. At all locations, BMSB densities increased on TOH wooded borders in July, then, gradually moved into adjacent soybean borders later in the summer. BMSB did not move far from the field edge, with approximately half as many bugs being present at 15 m into the field and very few being detected 30 m into the field. These results validate the use of border sprays for BMSB control in soybean. Additional studies conducted in 2013 and 2014 compared a visual plant inspection method with a standard sweep net strategy for sampling BMSB. Overall, the two methods were highly correlated with a correlation coefficient of R=0.83. Visual inspection appears to be an effective method for assessing BMSB populations in soybean. One of the major factors affecting the distribution and establishment of invasive species is climate. The CLIMEX modeling software uses climatic and biological factors of species to predict the geographic risk for pest outbreaks. A climate simulation model was run with CLIMEX to determine the potential distribution of BMSB in Virginia based on temperature. To develop a more accurate model, factors like resource availability and source population would need to be considered. / Master of Science in Life Sciences

Stink bugs, including native brown stink bug (Euschistus servus) and invasive brown marmorated stink bug (Halyomorpha halys), cause damage to a variety of crops including field corn (Zea mays). Frequency and size of stink bug infestations have increased in corn fields in the Mid-Atlantic U.S., and there are growing concerns that these infestations may contribute to reductions in grain quality including increased mycotoxin concentrations. Prior research on native and invasive stink bugs has focused on understanding their biology, the damage they cause, and elucidating effective and economic management strategies. However, few studies examined the potential for stink bugs to facilitate fungal infection and mycotoxin contamination of corn grain. Thus, the objectives of this research were to: 1) assess the relationship between invasive brown marmorated stink bug (H. halys) feeding injuries and fumonisin contamination of field corn in the Mid-Atlantic U.S., 2) determine if stink bugs are a vector for mycotoxigenic Fusarium spp. in corn, and 3) evaluate the efficacy of pesticides for mitigating stink bug feeding injury and associated mycotoxin contamination in field corn. A correlation between H. halys feeding injury and fumonisin concentrations was identified, and the ability of H. halys to increase F. verticillioides infection and fumonisin concentrations in corn was demonstrated in field experiments. Fusarium species including fumonisin-producing F. verticillioides and F. proliferatum were isolated from field-collected stink bugs, and in laboratory experiments, E. servus was able to transmit F. verticillioides to non-infected corn ears after feeding on F. verticillioides-infected corn. In field studies, both fungicide and insecticide reduced stink bug-associated mycotoxin concentrations in corn, but levels of control were inconsistent. Thus, additional tactics that target both the stink bug and Fusarium should be implemented to mitigate risks of mycotoxin contamination in corn. / Doctor of Philosophy / Native and invasive stink bugs can severely damage crops including field corn. Frequency and size of stink bug infestations in Mid-Atlantic U.S. corn fields have increased, and there is growing concern that this may contribute to reductions in grain quality. Insect feeding injury is a risk factor for fungal infection and mycotoxin contamination in corn. Mycotoxins are toxic chemicals produced by certain fungi that have detrimental health effects on animals including livestock and humans. The relationship between stink bug feeding injuries and mycotoxin contamination in corn grain is not well understood, and management strategies to minimize the risk of mycotoxin contamination in corn need to be identified. The main goal of this research was to characterize interactions between stink bugs and mycotoxin-producing fungi and identify tactics for controlling both the insect pest and pathogen. Specific objectives were to: 1) assess the relationship between invasive brown marmorated stink bug (H. halys) feeding injuries and fumonisin contamination of field corn in the Mid-Atlantic U.S., 2) determine if stink bugs are a vector for mycotoxin-producing Fusarium spp. in corn, and 3) evaluate the efficacy of pesticides for mitigating stink bug feeding injury and associated mycotoxin contamination in field corn. Results from this work indicated that stink bugs have the ability to cause feeding injuries which facilitate invasion of mycotoxin-producing Fusarium species, leading to increases in mycotoxin concentrations in corn grain. Studies also demonstrated that stink bugs can vector Fusarium species during feeding and increase Fusarium infection of corn resulting in subsequent mycotoxin contamination. Field studies indicated that pesticide applications targeting both the stink bugs and mycotoxigenic fungi may be needed to minimize risk of mycotoxin contamination in corn. However, under low pest pressure, application of pesticides is unlikely to be profitable.

The brown marmorated stink bug, Halyomorpha halys (Stål), is an invasive species that has spread throughout the United States and become a serious agricultural pest. Studies were conducted in Virginia on the survival of H. halys nymphs on different vegetable hosts and on the oviposition preference of H. halys on common tree hosts in southwestern Virginia. To determine the effects of vegetable host on H. halys survival, nymphs were placed on one of five vegetable host plants and their corresponding vegetable fruit. Survival was highest on sweet corn, followed by bell pepper and snap bean. Eggplant and tomato were poor compared to other vegetable hosts tested, potentially due to secondary compounds released by the plant to reduce feeding. Of six common host trees sampled, H. halys laid the most egg masses on catalpa, Catalpa bignonioides, while aggregation lure had no effect on the number of eggs laid. Across host trees, egg masses were more commonly found in trees with fruiting structures present. An additional study was conducted on a red-sternum phenotype of H. halys. Mortality was recorded by sternum phenotype for each sex. There was no difference in survival between groups at two time points. Biochemical tests were used to determine if the red-sternum was associated with the potentially pathogenic red bacteria Serratia marcescens. Only one stink bug's gut contents had a confirmed infection of S. marcescens. Results from these studies offer insight into H. halys development on vegetable hosts and oviposition preferences on different host trees. / MSLFS / The brown marmorated stink bug (BMSB), is an invasive insect that has spread throughout the United States and become a serious pest. Current research is focused on reducing their populations without increasing insecticide use. Studies were conducted to determine if vegetable host type influenced survival and development of this insect. Nymphs were placed on one of five vegetable plants with the corresponding vegetable fruit. Overall, survival was highest on sweet corn, followed by bell pepper and snap bean. Eggplant and tomato were poor hosts, potentially due to chemicals released by the plant to reduce insect feeding. Six common host trees of BMSB were sampled to determine where females lay the most egg masses and the effect of using an attraction lure. Catalpa trees had the most egg masses throughout the season. Use of a lure had no effect on the number of eggs laid on any tree. A red colored abdomen was observed in some adult insects throughout these studies. To determine if this was associated with higher mortality, male and female insects were observed for two weeks. There were no mortality differences at either seven or 14 days between any of the groups. Tests were used to determine if the red abdomen was caused by a red-colored bacterium. Of the insects sampled, there was only one stink bug with a confirmed infection. Results from these studies provides information that increases knowledge of this pest’s ecology in Virginia.

Building design, maintenance, and management can have significant impacts on accessibility and suitability for pest species. The brown marmorated stink bug, Halyomorpha halys (Stål), causes household nuisance pest problems because of its habit of using human homes as winter refuges. Studies were conducted to understand behaviors and characteristics relevant to this problem, including response to gravity, ability to pass through openings, responses to heat, and mortality estimations in wall assemblies. In the lab, winter shelter seeking bugs were shown to exhibit upward movement, and field observations of bugs on building exteriors gave similar results. In experiments testing the size limits on openings through which bugs could pass, height limited tests excluded most females at 4 mm, and all bugs at 3 mm. Pronotum width limited tests excluded most females at 8 mm and nearly all bugs at 7 mm. Accompanying measurements of over 900 bugs found an average female pronotum width of 8.33 mm and height of 4.03 mm, and male pronotum width of 7.47 mm and height of 3.50 mm, with minimum sizes indicating that only a small percentage of bugs will pass the smallest openings tested. Heat response experiments of shelter seeking bugs were first piloted outdoors, and then modified to be a forced choice indoor test. In outdoor tests on a flat plane wall section with alternating heated sections, bugs did not respond to thermal contrast but rather immediately walked off of the wall. In the indoor forced choice test, a box of four cavity walls was used. Bugs did not respond to the heated sections in either the adjacent or opposite configuration. Thermal simulation modeling was used to evaluate the possible effects of varying wall assembly materials and configurations on cold-related mortality of bugs overwintering in the cavity space behind cladding. Simulation results indicated that bugs electing to overwinter in the space between cladding and sheathing were at risk of freezing deaths, with mortality expectations increasing in better insulated buildings. The results of these studies will inform future control measures and impact studies in buildings. / Ph. D.

Submitted by DIEGO FELISBINO FRAGA null (diegofraga@fazu.br) on 2016-03-08T21:00:47Z No. of bitstreams: 1 Tese_Diego_Felisbino_Fraga.pdf: 1816202 bytes, checksum: 645838702bb204f2d81827bcc74d32f0 (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-03-09T18:05:33Z (GMT) No. of bitstreams: 1 fraga_df_dr_jabo.pdf: 1816202 bytes, checksum: 645838702bb204f2d81827bcc74d32f0 (MD5) / Made available in DSpace on 2016-03-09T18:05:33Z (GMT). No. of bitstreams: 1 fraga_df_dr_jabo.pdf: 1816202 bytes, checksum: 645838702bb204f2d81827bcc74d32f0 (MD5) Previous issue date: 2016-02-22 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / O Controle Biológico Natural, ou Conservativo, é uma ferramenta indispensável para o sucesso do Manejo Integrado de Pragas, pois permite uma interação entre os componentes do ambiente, o que o coloca na linha de frente das táticas e estratégias de manejo de pragas em agroecossistemas. Basicamente, esta interação, denominada interação tritrófica, envolve as plantas, os organismos fitófagos e seus inimigos naturais. Neste contexto, diversos fatores são importantes para o controle biológico, tais como condições abióticas e bióticas, destacando-se características das plantas (compostos metabólitos secundários e arquitetura, por exemplo) e potenciais presas/hospedeiros (semioquímicos, etc.). Inimigos naturais podem utilizar estes recursos durante o seu processo de busca e localização do hospedeiro, de modo que o conhecimento sobre o comportamento de inimigos naturais em função da fenologia e arquitetura das plantas, bem como sobre o potencial de voláteis emitidos por plantas e presas é fundamental para um programa de manejo integrado de pragas. Desta forma, o presente trabalho teve por objetivos: a) avaliar a dinâmica populacional e a distribuição vertical de ovos de Chrysodeixis includens (Lepidoptera: Noctuidae) e de seus ovos parasitados por Trichogramma spp. (Hymenoptera: Trichogrammatidae) na cultura da soja; b) determinar a resposta comportamental do percevejo predador Orius insidiosus (Hemiptera: Anthocoridae) para voláteis associados à Halyomorpha halys (Hemiptera: Pentatomidae). Sendo assim, em Jaboticabal, SP, Brasil, durante as safras agrícolas de 2012/13 e 2013/14, foram realizadas coletas de ovos de C. includens presentes em diferentes terços das plantas de soja, durante diferentes estádios fenológicos da cultura. Semanalmente, o número de ovos presentes nas plantas foi anotado e os mesmos eram coletados e levados para laboratório para aguardar a emergência de lagartas ou de adultos de Trichogramma spp. O número de adultos do parasitoide emergido por ovo era registrado, bem como a razão sexual. Os adultos foram identificados ao nível específico, encontrando-se a espécie Trichogramma pretiosum presente na cultura da soja. Fêmeas de C. includens preferiram ovipositar com mais intensidade nos terços médio e inferior das plantas, principalmente durante os estádios reprodutivos da cultura. O parasitismo de ovos por T. pretiosum apresentou uma tendência semelhante, em que a medida que se aumentou a densidade de ovos de C. includens nas plantas, também elevou-se o número médio de ovos parasitados, indicando uma forte correlação positiva entre a praga e o inimigo natural. Para atingir o segundo objetivo desta tese, e fornecer alternativas para o controle biológico de H. halys, um pentatomídeo invasor nos EUA, foram conduzidos experimentos para testar a atratividade de O. insidiosus para voláteis emitidos pela presa e por plantas de feijoeiro atacadas por ela. Foram conduzidos experimentos em casa-de-vegetação, laboratório e em campo, em que foi identificado altas concentrações do composto tridecano. Foi avaliado a atratividade de O. insidiosus para diferentes concentrações deste composto, bem como os efeitos desta atração na predação de ovos. Foi observado que O. insidiosus é atraído por tridecano, no entanto, esta atração não afeta a predação de ovos, em casa de vegetação e no campo. / Natural ou Conservative Biological Control is a valuable tool for the success of Integrated Pest Management as it allows a natural interaction between the components of the environment, which places it at the forefront of the tactics and pest management strategies in agroecosystems. This interaction, so-called tritrophic interaction, involves plants, phytophagous organisms and their natural enemies. In this context several factors are important for biological control, such as biotic and abiotic conditions, highlighting plant characteristics (secondary metabolites compounds and plant architecture, for example) and possible prey/hosts (semiochemicals, etc.). Natural enemies can use this information during the its host foraging behavior, so the knowledge of the behavior of natural enemies according to the phenology and plant architecture as well as potential volatiles emitted by prey plants and potential is key for an integrated pest management program. Thus, this study aimed to: a) assess the population dynamics and vertical distribution of natural biological control of Chrysodeixis includens (Lepidoptera: Noctuidae) eggs by Trichogramma spp. (Hymenoptera: Trichogrammatidae) in soybean field; b) determine the behavioral response of the predator Orius insidiosus (Hemiptera: Anthocoridae) for volatile associated with Halyomorpha halys (Hemiptera: Pentatomidae). Thus, in Jaboticabal, SP, Brazil, during the growing seasons of 2012/2013 and 2013/2014, samples were taken from C. includens eggs present on different thirds of soybean plants during different phenological stages of the crop. Weekly, number of eggs present in plants was recorded and the eggs were collected ant taken to the laboratory to await the emergence of larvae or adult Trichogramma spp. The number of adult Trichogramma spp. emerged per egg was noted, as was the sex ratio. The adults were identified to species level, finding Trichogramma pretiosum as the main species present in soybeans. Females of C. includens intensively preferred to lay eggs in the middle and lower thirds of the plants, especially during the reproductive stages of soybean plants. Egg parasitism by T. pretiosum presented a similar tendency, which as the C. includens eggs density increased in plants, also increased the average number of parasitized eggs, indicating a strong positive correlation between the pest and the natural enemy. To achieve the second objective of this thesis, and provide alternatives for the biological control of H. halys (an invasive pentatomid in the US). experiments were conducted to test the attractiveness of O. insidiosus to volatiles emitted by the prey and bean plants attacked by it. Experiments were conducted under greenhouse, laboratory and field conditions. It was identified that tridecane was the major compound in damage-bean pods as well as on H. halys. The attractiveness of O. insidiosus was evaluated for different concentrations of this compound as well as the effects of this attraction as predators of eggs. It was observed that O. insidiosus is attracted to tridecane, however, this attraction not poisitivamente affected the predation of eggs in the greenhouse and in the field. / CNPq: 164869/2014-0

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