Doctor of Philosophy / Department of Entomology / Franklin Arthur / James R. Nechols / The Indianmeal moth, Plodia interpunctella Hubner, is capable of infesting a number of different commodities including a wide variety of grains, nuts and finished stored products. Therefore, control of the Indianmeal moth is especially needed in areas where food is being stored for human consumption. Increased concerns of consumers and producers regarding the impact of conventional insecticides on the environment and on human health has prompted scientists and the agricultural chemical industry to search for insecticides that do not affect mammalian nervous systems and have limited effects on non-target organisms. One group of insecticides with reduced risks is insect growth regulators (IGRs), which are substances that mimic insect hormones essential to normal development and reproduction. Currently methoprene, a juvenile hormone analog, is labeled for direct application to stored grains, as well as a contact insecticide and as an aerosol application inside mills, warehouses, and indoor food storage facilities. Surface treatments and aerosol space applications can be effective ways to treat the interior surfaces and storage areas of warehouses and food processing facilities. There is little recent research with large-scale aerosol applications in storage sites; furthermore, there are no published references in the scientific literature regarding efficacy of using methoprene alone in aerosol form. Therefore, the purpose of this research was to evaluate the use of surface and aerosol applications of methoprene on finished stored-product packaging materials and facilities for the control of P. interpunctella. Results of this research showed that while methoprene has good residual activity, and efficacy is unaffected by temperature, surface applications of methoprene on packaging materials is not as effective for control of P. interpunctella as aerosol applications of methoprene. Aerosol methoprene is highly effective alone and in combination with conventional chemicals for control of eggs and wandering-phase larvae. Simulations with a population growth model make it possible to estimate impact of insecticide treatments at different temperatures and application times on populations of P. interpunctella. Aerosol treatments are also economically viable as part of an overall integrated pest management program.

Doctor of Philosophy / Department of Entomology / Larry L. Buschman / Kun Yan Zhu / Genomic information for lepidopteran insects, particularly agricultural pest species, is very limited but urgently needed due to their economic importance and biodiversity. The huge economic losses ($ 1-2 billons / year) caused by the European corn borer (Ostrinia nubilalis, Hübner, ECB) makes this insect species one of the major pests of corn in the United States and western world. Management of ECB by conventional methods is limited but has had a great success by transgenic Bt (Bacillus thuringiensis) corn, which targets insect gut. However, the widespread use of Bt corn may lead to the development of Bt resistance in ECB. Knowledge of genes expressed in the insect gut is considered crucial for understanding basic physiology of food digestion, their interactions with Bt toxins and pathogens, and for discovering new targets for pest management. A large database of 15,000 expressed sequence tags (ESTs) was established from the ECB larval gut. To our knowledge, this database represents the largest gut-specific EST database from a lepidopteran pest. Analysis of 10 aminopeptidase-like genes between Cry1Ab–resistant and –susceptible ECB larvae revealed that aminopeptidase P-like (OnAPP) gene is a strong candidate for its role in Bt toxicity and resistance. The RNA interference mediated reduction in the transcript level of OnAPP gene in ECB larvae resulted in their reduced susceptibily to Cry1Ab. Analysis of the chitinase-like gene (OnCht) revealed its essential role in regulating chitin content of peritrophic membrane (PM). Our results suggest that OnCht may influence food digestion, nutrient absorption or movement of digestive enzymes through the PM and can be an important target for insect management. We also identified and characterized six genes involved in the innate immune defense response in ECB and showed that the expression of these genes were induced when challenged with bacteria. In addition to these results, this research generated significant genomic information for the development of microarray from the larval gut of ECB. The establishment of the feeding-based RNA interference technique could potentially help in delivering dsRNA orally to ECB for high throughput screening of effective genes to be targeted for insect pest management.

Doctor of Philosophy / Department of Entomology / Ludek Zurek / Stable flies (Stomoxys calcitrans L.) are blood-feeding insects with a great negative impact on livestock resulting in annual losses around $2 billion in the USA alone. One of the main reasons for such an impact is because stable fly management is very difficult. Stable fly larvae develop primarily in animal manure and live bacteria were shown to be essential for stable fly development. I hypothesized that the microbial community of horse manure changes over time and plays an important role in stable fly fitness. Two-choice bioassays were conducted using 2 week old horse manure (control) and aging horse manure (fresh to 5 week old) to evaluate the effect of manure age on stable fly oviposition. The results showed that fresh manure did not stimulate oviposition and that the attractiveness increased as manure aged but started to decline after 3 weeks. Stable fly eggs artificially placed on 1, 2, and 3 week old manure resulted in significantly higher survival and heavier adults comparing to those developing in fresh, 4, and 5 week old manure. Analysis of the bacterial community of aging horse manure by 454-pyrosequencing of 16S rDNA revealed a major shift from strict anaerobes (e.g. Clostridium, Eubacterium, Prevotella, Bacteroidales) in fresh manure to facultative anaerobes and strict aerobes (e.g. Rhizobium, Devosia, Brevundimonas, Sphingopyxis, Comamonas, Pseudomonas) in 1-5 week old manure. Identified volatile compounds emitted from 2 and 3 week old horse manure included phenol, indole, p-cresol, and m-cresol. However, none of them stimulated stable fly oviposition in two-choice assays. In conclusion, the microbial community of 2 and 3 week old horse manure stimulates stable fly oviposition and provides a suitable habitat for stable fly development. Manure at this stage should be the main target for disrupting the stable fly life cycle. Volatile compounds acting as oviposition stimulants/attractants and their specific bacterial origin remain to be determined. Better understanding of stable fly microbial ecology is critical for development of novel management strategies based on alteration of the microbial community of stable fly habitat to generate a substrate that is non-conducive to fly oviposition and/or larval development.

Doctor of Philosophy / Department of Entomology / Raymond A. Cloyd / The life history of the rove beetle, Atheta coriaria (Kraatz) (Coleoptera:Staphylinidae), predation against the fungus gnat Bradysia sp. nr. coprophila (Lintner) and compatibility with pesticides and plant growth regulators was investigated under laboratory conditions using Sunshine LC1 Professional Growing Mix as a substrate. Duration of life stages was 2.2, 7.1, and 7.8 days for egg, larva and pupa respectively, at 26°C, whereas total development time from egg to adult was 17.0 days. In addition, A. coriaria male and female adult longevity was 60.3 and 47.8 days. Average fecundity was 90.2 eggs per female and the number of adults produced per female was 69.1. There were no significant differences in prey consumption when using second and third instar fungus gnat larvae as prey and starved and non-starved rove beetles. Overall, predation efficacy in Petri dishes was high (70 to 80%) as fungus gnat larval density increased with 3.9, 7.0, 11.1, and 15.3 larvae consumed in 24 hours after exposure of 5, 10, 15 and 20 fungus gnat larvae to one rove beetle adult. However, lower predation rates were found at different predator:prey ratios when using 1 to 5 rove beetles and growing medium as a substrate. The direct and indirect effects of pesticides and plant growth regulators on A. coriaria were investigated under laboratory conditions. Rove beetle survival was consistently higher when adults were released 24 hours after rather than before applying pesticides. Acetamiprid, lambda-cyhalothrin, and cyfluthrin were directly harmful to rove beetle adults, whereas Beauveria bassiana, azadirachtin and organic oils were compatible with A. coriaria. Similarly, the plant growth regulators acymidol, paclobutrazol and uniconazole were not harmful to rove beetle adults. In addition, Beauveria bassiana, azadirachtin, kinoprene, organic oils, and the plant growth regulators did not negatively affect A. coriaria development. However, Beauveria bassiana did negatively affect rove beetle prey consumption. This study demonstrated that A. coriaria is not compatible with the pesticides acetamiprid, lambda-cyhalothrin and cyfluthrin, whereas there is compatibility with organic oils, Beauveria bassiana, azadirachtin, and the plant growth regulators. As such, these compounds may be used in combination with A. coriaria in greenhouse production systems.

The establishment of exotic species in novel environments is a major environmental concern, however, few long-term studies have examined the effects of these species on their host environment and community, especially in forest ecosystems. The arrival and subsequent spread of the greater European pine shoot beetle, Tomicus piniperda (L.) (Coleoptera: Curculionidae, Scolytinae), into southern Ontario pine forests provided a natural experiment to assess biotic interactions between an exotic species and its new community over several years. Reproductive success of Tomicus piniperda colonies of various ages was studied between 2001 and 2004. The size and composition of competitor and natural enemy complexes present in these communities were also quantified over time. The impact of the natural enemy complexes on T. piniperda reproduction was assessed, Brood production (no. eggs and galleries/female) by T. piniperda populations rapidly approached those reported from its native range in Europe, with lower densities of parental adults. Thus, reproduction remained consistently above the replacement level for this beetle over all four years of study suggesting that these recently-introduced populations were growing rapidly and at a greater rate than in their country of origin. Tomicus piniperda successfully integrated into a large bark beetle community, and appeared to be capable of displacing native beetles to more marginal bark habitats, however, these competitors were not eliminated during the course of the study. The long-term effect of this marginalization on populations of native beetles is uncertain. Tomicus piniperda rapidly acquired natural enemies in the introduced areas, however, natural enemy-caused mortality did not show a regulating effect on its populations. It seems that intraspecific competition, rather than predation, regulates T. piniperda populations following introduction. The implications of these findings for the establishment and spread of exotic species in forest systems are examined, especially with reference to a prominent theory for success, the Enemy Release Hypothesis. In short, the Enemy Release Hypothesis is not applicable to an exotic species that is not regulated by natural enemies in its native range, and assessments of the Enemy Release Hypothesis should always include a determination of enemy regulation of the exotic in its native range.

Sexual conflict and condition-dependent trait expression have emerged as major themes in sexual selection. There is now considerable evidence suggesting that both conflict and condition-dependence can drive the evolution and expression of sexual traits; still, important questions remain concerning the extent to which conflict shapes sexual traits and the role of condition in mediating conflict. Here, I address these two themes in studies of a ladybird mating system. One set of studies investigates the function and economics of potentially antagonistic traits – nuptial gifts and female mating resistance – while another examines condition-dependence in mating resistance and male ejaculate composition. Nuptial gifts are often considered beneficial to females, but recent thinking suggests they may also allow males to manipulate females, raising the possibility of conflict. I demonstrate that male ladybirds benefit from nuptial feeding by their mates through reduced re-mating frequency. Benefits to female reproduction and lifespan, however, are weak or non-existent. These results show that although males gain from transferring gifts that influence female behaviour, females experience neither harm nor benefit. I next tested the hypothesis that nuptial feeding is maintained – despite an absence of benefits – because female foraging is generally elevated after mating. However, although females indeed display strongly increased foraging after mating, this response did not increase nuptial feeding. Recent studies suggest that individual condition may affect the economics of mating and extent of conflict. Female ladybirds vigorously resist mating, and I show that (1) resistance is condition-dependent, with low-condition females displaying more resistance, and (2) resistance functions to minimize superfluous matings (sexual conflict), rather than to select among males (indirectly benefiting females). Resistance generates selection favouring large males; thus, this work demonstrates that ecological circumstances, through influencing condition, affect the strength of sexual selection. Finally, male condition may influence investment in ejaculate components, but condition-dependence in ejaculate composition is currently poorly understood. I show that, in agreement with theory, males in poor condition transfer smaller ejaculates that nonetheless contain more sperm, but less seminal fluid. Taken together, this work highlights both the value of economic studies in evaluating sexual conflict, and the significance of condition-dependence for sexual selection.

The establishment of exotic species in novel environments is a major environmental concern, however, few long-term studies have examined the effects of these species on their host environment and community, especially in forest ecosystems. The arrival and subsequent spread of the greater European pine shoot beetle, Tomicus piniperda (L.) (Coleoptera: Curculionidae, Scolytinae), into southern Ontario pine forests provided a natural experiment to assess biotic interactions between an exotic species and its new community over several years. Reproductive success of Tomicus piniperda colonies of various ages was studied between 2001 and 2004. The size and composition of competitor and natural enemy complexes present in these communities were also quantified over time. The impact of the natural enemy complexes on T. piniperda reproduction was assessed, Brood production (no. eggs and galleries/female) by T. piniperda populations rapidly approached those reported from its native range in Europe, with lower densities of parental adults. Thus, reproduction remained consistently above the replacement level for this beetle over all four years of study suggesting that these recently-introduced populations were growing rapidly and at a greater rate than in their country of origin. Tomicus piniperda successfully integrated into a large bark beetle community, and appeared to be capable of displacing native beetles to more marginal bark habitats, however, these competitors were not eliminated during the course of the study. The long-term effect of this marginalization on populations of native beetles is uncertain. Tomicus piniperda rapidly acquired natural enemies in the introduced areas, however, natural enemy-caused mortality did not show a regulating effect on its populations. It seems that intraspecific competition, rather than predation, regulates T. piniperda populations following introduction. The implications of these findings for the establishment and spread of exotic species in forest systems are examined, especially with reference to a prominent theory for success, the Enemy Release Hypothesis. In short, the Enemy Release Hypothesis is not applicable to an exotic species that is not regulated by natural enemies in its native range, and assessments of the Enemy Release Hypothesis should always include a determination of enemy regulation of the exotic in its native range.

Sexual conflict and condition-dependent trait expression have emerged as major themes in sexual selection. There is now considerable evidence suggesting that both conflict and condition-dependence can drive the evolution and expression of sexual traits; still, important questions remain concerning the extent to which conflict shapes sexual traits and the role of condition in mediating conflict. Here, I address these two themes in studies of a ladybird mating system. One set of studies investigates the function and economics of potentially antagonistic traits – nuptial gifts and female mating resistance – while another examines condition-dependence in mating resistance and male ejaculate composition. Nuptial gifts are often considered beneficial to females, but recent thinking suggests they may also allow males to manipulate females, raising the possibility of conflict. I demonstrate that male ladybirds benefit from nuptial feeding by their mates through reduced re-mating frequency. Benefits to female reproduction and lifespan, however, are weak or non-existent. These results show that although males gain from transferring gifts that influence female behaviour, females experience neither harm nor benefit. I next tested the hypothesis that nuptial feeding is maintained – despite an absence of benefits – because female foraging is generally elevated after mating. However, although females indeed display strongly increased foraging after mating, this response did not increase nuptial feeding. Recent studies suggest that individual condition may affect the economics of mating and extent of conflict. Female ladybirds vigorously resist mating, and I show that (1) resistance is condition-dependent, with low-condition females displaying more resistance, and (2) resistance functions to minimize superfluous matings (sexual conflict), rather than to select among males (indirectly benefiting females). Resistance generates selection favouring large males; thus, this work demonstrates that ecological circumstances, through influencing condition, affect the strength of sexual selection. Finally, male condition may influence investment in ejaculate components, but condition-dependence in ejaculate composition is currently poorly understood. I show that, in agreement with theory, males in poor condition transfer smaller ejaculates that nonetheless contain more sperm, but less seminal fluid. Taken together, this work highlights both the value of economic studies in evaluating sexual conflict, and the significance of condition-dependence for sexual selection.

Doctor of Philosophy / Department of Entomology / James F. Campbell / Phillip E. Sloderbeck / Robert "Jeff" J. Whitworth / Knowledge of factors influencing the establishment, persistence and distribution of stored-product pests aids the development of effective Integrated Pest Management (IPM) programs in food storage and processing facilities. This research focused primarily on Tribolium castaneum, which is one of the most important pests of mills. Populations of T. castaneum from different food facilities can potentially be interconnected by either their own dispersal behavior or by human transportation. Population genetic structure analyses based on microsatellites and other insertion-deletion polymorphisms (“indels”) showed that populations from different mills around the US are genetically distinct from each other, but the level of differentiation was not correlated with the geographic distance. A potential source of insect infestation within a food facility is spillage that accumulates outside or movement from bulk storage facilities on site. Results from three facilities showed that most stored-product species were captured both inside and outside buildings, but T. castaneum was rarely captured outside of the facilities. Spatial distribution of all species outside was associated with the proximity of buildings, not necessarily with areas with accumulated spillage. T. castaneum populations inside facilities are potentially exposed to frequent genetic bottlenecks resulting from structural fumigations. Changes in allele frequencies through time, based on the analysis of microsatellites and other indels in individuals collected in a mill, confirmed bottleneck effects. To understand how spatial distribution of T. castaneum within a mill could be influenced by environmental and physical factors, a range of variables were measured at each trap location. There was significant variation among trap locations regarding beetle captures and the variables measured, but increase in beetle captures correlated only with increase in temperature and spillage production. Tribolium castaneum response to visual cues could influence attraction to pheromone and kairomone olfactory cues used in traps. Results of laboratory experiments showed that adults respond to tall narrow black shapes and placing traps in front of these shapes can increase captures. This research provides new insights into factors influencing the spatial distribution of T. castaneum and could help in improving monitoring programs for this important pest of the food industry.

Master of Science / Department of Entomology / David C. Margolies / James R. Nechols / Many crops grown in greenhouses are damaged by the twospotted spider mite, Tetranychus urticae. The predatory mite, Phytoseiulus persimilis, is a commercially-available predator that is commonly used to control twospotted spider mites on greenhouse crops; but its efficacy varies among crops, and it is generally ineffective at low prey densities. In general, predator foraging efficiency depends on how well predators find prey patches, the length of stay in prey patches, and consumption of prey while in prey patches. With respect to P. persimilis, I asked how this predator responds to different prey distributions, as might be encountered at different stages of spider mite infestations. I also asked how components of foraging, namely consumption rate and dispersal tendency, affected predator efficiency. To examine the former, I established T. urticae eggs on 6-leafed cucumber plants in two distributions. To examine the latter, I imposed artificial selection on a population of P. persimilis to create a line that exhibited extremely high consumption and one that demonstrated a greater tendency for dispersal. Subsequently, foraging efficiency was assessed by observing predator oviposition and consumption of twospotted mite eggs on individual leaves of 6-leafed cucumber plants. The number of eggs laid by predators corresponded to the number of prey consumed regardless of predator line. In addition, predators from both lines distributed their eggs proportional to where they fed. However, prey consumption differed between selected lines in response to prey distribution. Predators selected for high consumption fed more on the basal leaf where they were released; whereas prey consumption by the high dispersal and control lines were more evenly distributed throughout the plant. These results contribute to a better understanding of how foraging behavior is modified in plant landscapes under different levels of expression of foraging traits. They also indicate that predator release strategies likely would need to modified in accordance with the kind of foraging trait(s) used in artificial selection programs. In general, my research, when combined with future studies at a broader landscape level, will facilitate decisions by biological control practitioners about whether changes in foraging efficiency resulting from artificial selection justify the cost investment of producing selected lines of P. persimilis