L’utilisation non raisonnée d’insecticides a des conséquences sur l’environnement et la santé humaine mais aussi sur le développement de résistances chez les insectes nuisibles. Dans ce contexte, ce travail de thèse porte sur le développement d'une technique de nanoencapsulation d'un agent synergisant chimique, la deltaméthrine (pyréthrinoide), associé un pro-insecticide l’indoxacarbe (oxadiazine). Sur la base d’études in vitro sur cellules neurosecrétrices de blattes (Periplaneta americana), une action synergique entre la deltaméthrine et le DCJW (métabolite actif de l’indoxacarbe) a été démontrée via un mécanisme intracellulaire original dépendant du calcium. Des études toxicologiques in vivo sur des blattes adultes ont permis de confirmer l’effet synergique entre la deltaméthrine et l’indoxacarbe. Pour optimiser la formulation de l’association de ces composés, des nanocapsules lipidiques contenant la deltaméthrine utilisée comme agent synergisant ont été produites. Les études de toxicité sur blattes ont permis de déterminer les doses effectives les plus faibles de l’association deltaméthrine nanoencapsulée/indoxacarbe et d’obtenir une optimisation de l’effet synergique avec la deltaméthrine nanoencapsulée. Cet effet synergique est plus important que celui du piperonyl butoxyde, composé utilisé dans les formulations d'insecticides classiques. Les résultats indiquent que la deltaméthrine, protégée des estérases par la nanoencapsulation, permet d’optimiser l’efficacité du traitement tout en réduisant les doses d’indoxacarbe. Cette nouvelle stratégie est une première étape dans le développement d'une formulation phytosanitaire efficace contre les insectes nuisibles. / The over-use of pesticides has represented a concern not only for its consequences against the environment but also for the increase in resistance mechanisms in pest insects. In this context, our research project is focused on the development of a nanoencapsulation technique of a deltamethrin (pyrethroid), used as synergistic agent, combined with a pro-insecticide indoxacarb (oxadiazine). Based on in vitro studies performed on cockroach (Periplaneta americana) neurosecretory cells, synergistic effect between deltamethrin and DCJW (active metabolite of indoxacarb) has been characterized, occurring through an original calcium-dependent intracellular mechanism. In vivo toxicological studies on adult cockroaches have confirmed these previous results. To optimize the formulation including the synergistic agent and indoxacarb, lipid nanocapsules (LNCs) containing deltamethrin have been developed. It has been possible to determine the lower effective doses of LNC-deltamethrin/indoxacarb mixture producing the synergistic effect in whole insects. The synergism obtained is more important than that of obtained with piperonil butoxide (PBO), the well-known synergist used in classical insecticide formulations. The results also indicate that LNC-deltamethrin, protected from esterase-induced detoxification enhances the toxicity of indoxacarb while reducing doses. This novel strategy is a first step for the development of a novel formulation more efficient against pest insects.

Stink bugs are major pests of agricultural crops throughout Virginia and much of the United States. Knowledge of the biology, the species complex, and insecticide susceptibility can improve management. A survey was conducted in Virginia to determine the species complex in soybean and cotton and to monitor for nonnative species. Seven stink bug species were identified. Acrosternum hilare (Say) and Euschistus servus (Say) were the most common. Two sampling methods, the sweep net and the beat sheet, were assessed in soybean and cotton. There was less variability with the sweep net method compared to the beat sheet method. Laboratory bioassays and field trials were conducted to evaluate the toxicity and efficacy of selected conventional and organic insecticides against A. hilare and E. servus. In bioassays with conventional insecticides, A. hilare adults and nymphs were susceptible to all pyrethroids tested. Generally, the neonicotinoids, dinotefuran and clothianidin, were more toxic to A. hilare, while thiamethoxam and acetamiprid were more toxic to E. servus. In soybean field efficacy trials, dinotefuran performed comparably to the organophosphates and pyrethroids. Laboratory bioassays with organic insecticides resulted in moderate to high levels of mortality, and in antifeedant and repellency responses. Likewise, soybean field trials indicated that a single application can reduce stink bug numbers for up to two days after treatment; however in tomato field trials multiple weekly applications did not result in significant reductions in stink bug damage. A weather model to predict abundance of A. hilare adults was developed using weekly black light trap catch data collected from 1990 to 2007 at a single location. The two weather variables that resulted in a significant model were days below freezing and mean monthly precipitation from January to April. The model was validated by correlating five independent data sets to predicted weekly trap catch. Mean trap catch plotted over time showed three peaks. In accordance with A. hilare developmental rates, the peaks indicated that two generations and a partial third occur in Virginia. Cumulative trap catch estimated from the 18-yr trap catch mean showed that 10, 50, and 90% of the total seasonal catch should occur by 153, 501, and 1066 degree days, respectively. / Ph. D.

Diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae) is a serious pest of crucifer vegetables (Brassica sp.) worldwide because of a lack of effective natural enemies in certain regions and because of insecticide resistance. In 2003, laboratory and field studies were initiated in Virginia to better understand P. xylostella, its primary natural enemies, and their susceptibilities to insecticides in order to develop an economically and environmentally sound integrated pest management program for collards in the state. Ecological life table studies of P. xylostella immature stages on collards located on the Eastern Shore and on Kentland Farm, near Blacksburg at the New River Valley, VA revealed that most (98 to 99%) of P. xylostella died from natural causes. Mortality factors varied between the two regions. Neonates, small larvae, and large larvae disappearing were major mortality factors. Rainfall, predation, and dispersal probably contributed the most to this mortality. Egg mortality played a bigger role at the New River Valley compared with the Eastern Shore. Three parasitoid species were found that contributed to the mortality of P. xylostella: Diadegma insulare (Cresson) (Hymenoptera: Ichneumonidae); Oomyzus sokolowskii (Kurdjumov) (Hymenoptera: Eulophidae); and Microplitis plutellae (Muesebeck) (Hymenoptera: Braconidae). Additional studies conducted in the laboratory using leaf-dip bioassays revealed that P. xylostella collected from the Eastern Shore of Virginia, showed significant tolerance levels to esfenvalerate, acetamiprid, methomyl, methoxyfenozide, indoxacarb, and acephate compared with a susceptible strain of P. xylostella. The highest tolerance ratio (1,876 fold) was to esfenvalerate, a commonly-used pyrethroid. All of the insecticides tested in this study were quite toxic to the adult stage of the parasitoids, D. insulare and O. sokolowskii. The insect growth regulator, methoxyfenozide was considerably less toxic than other insecticides such as esfenvalerate, methomyl, acephate, spinosad, indoxacarb, and emamectin benzoate at field-rate and 1% of field-rate concentrations. The aforementioned insecticides as well as some other insecticides were evaluated several times in the field for efficacy against P. xylostella as well as other pests of collards. The most efficacious insecticides over five field experiments included acephate, emamectin benzoate, esfenvalerate, methomyl, methoxyfenozide, novaluron, indoxacarb, and spinosad. These insecticides were followed in relative efficacy by Bt kurstaki, acetamiprid, and azadirachtin, which provided relatively inconsistent control of lepidopteran larvae over the experiments. Effective insecticide options in collards that are less toxic to natural enemies and that can fit well into integrated pest management programs include indoxacarb, spinosad, novaluron, emamectin benzoate, methoxyfenozide, and Bacillus thuringiensis subsp. kurstaki. / Ph. D.

1. The inheritance mechanism or resistance to lindane in the German cockroach, Blattella germanica (L.) was studied by determining the toxicity of lindane to the following populations ot cockroaches: a) lindane.resistant and non-resistant strains, b) the F1 and F2 progeny of reciprocal crosses between the two parental strains, c) the F1 and F2 progeny of backcrosses between the reciprocal cross F1 males. and resistant and non-resistant strain females, d) the progeny of pair matings within resistant and non-resistant strains and e) eight successive generation of a resistant strain isolated from further insecticide selection. 2. The method. employed in rearing the cockroaches and obtaining toxicological data on the different populations, as well as the genetical and statistical procedures, are described. 3. The toxicological data tor the mass matings are presented 1n the form of regression lines fitted by appropriate statistical analyses. 4. The toxicological data. for the pair matings, transformed into "indices of resistance," are presented in tabular form. 5.. Appropriate conclusions are drawn from the data to explain the inheritance mechanism of lindane resistance in the German cockroach. / Ph. D.

Dosage mortality data by a topical application method for eight organophosphorus compounds are given for two laboratory-selected .strains of Blattella p:ermanica ( L.), one resistant to malathion and the other resistant to diazinon. Comparison of this data with similar data for a normal susceptible strain of the same species reveals that development of resistance to malathion in this species is highly specific imparting no effective cross resistance to any of the other seven compounds evaluated. Selection with diazinon induces a low level of tolerance to all the organophosphates studied, but there are indications that in most cases this is due to vigor tolerance rather than to true resistance. There is reason to believe that if carboxyesterase activity is responsible for the resistance phenomena to malathion exhibited by this insect, its action is blocked or its detoxifying ability rendered ineffective by the presence of a vinyl group in an organophosphorus compound, such as GC 3707. / Master of Science

Master of Science / Department of Entomology / Raymond A. Cloyd / Western flower thrips (WFT), Frankliniella occidentalis, is one of the most destructive insect pests of greenhouse production systems because of the direct and indirect damage they cause and their ability to develop resistance to a wide variety of insecticides. A common method of avoiding resistance development is by rotating insecticides that have different modes of action. Entomopathogenic organisms (EPO), such as fungi and bacteria, have modes of action that are very different from standard insecticides. Thus, by incorporating EPO into rotation programs, greenhouse producers may preserve the efficacy of insecticides used for suppression of WFT populations. Therefore, the objectives of this study were to 1) determine the efficacy of entomopathogenic fungi on WFT adults and nymphs, and to assess product effectiveness when used beyond the expiration date; 2) evaluate the efficacy of entomopathogenic fungi against WFT nymphs when combined with the insect growth regulator, azadirachtin; and 3) evaluate different rotation programs that include EPO and standard insecticides commonly used to suppress WFT populations. To satisfy objective one and two, a series of laboratory bioassays were conducted in which WFT nymphs and adults were exposed to three entomopathogenic fungi (Beauveria bassiana, Isaria fumosoroseus, and Metarhizium anisopliae) at two label rates (maximum and minimum), and two product conditions (fresh and expired). Furthermore, a bioassay in which each entomopathogenic fungi was tested with and without azadirachtin was conducted to determine if there was any synergistic effect on WFT nymphal mortality. Results indicated that adults are generally more susceptible to infection by entomopathogenic fungi than nymphs, fresh products resulted in higher mortality than expired products, and azadirachtin, when mixed with the entomopathogenic fungi, did not increase mortality of WFT nymphs except when combined with M. ansiopliae. Insecticide rotation programs that included EPO were evaluated by conducting a series of greenhouse experiments in which chrysanthemum, Dendranthema x morifolium plants were artificially infested with WFT adults. Eight-week rotation programs were applied to each plant and weekly counts of adults captured on yellow sticky cards were recorded. A final quality assessment of damage due to WFT feeding on foliage and flowers (1 to 5 in which 1 = no damage, and 5 = greater than 75% damage) was also recorded. In addition, a cost comparison of each rotation program was determined. Generally, insecticide rotations programs which incorporated EPO resulted in no significant difference in WFT populations compared to standard insecticide rotation programs without EPO. Furthermore, there were no significant differences between any of the rotation programs in regards to foliage and flower quality. Based on the results of the cost comparison, there may be a cost savings associated with using EPO. Therefore, by incorporating EPO into insecticide rotation programs, greenhouse producers may reduce costs as well as reduce selection pressure on WFT populations, which may avoid or delay resistance development.

Typescript (photocopy). / Digitized by Kansas Correctional Industries

Bumble bees provide valuable pollination services to many agricultural crops and wild flower species. Consequently, evidence that wild populations are in decline has caused widespread concern. Among multiple causal factors, some have singled out neonicotinoid pesticides as potentially a major contributor to these declines. Bumble bees are exposed to neonicotinoids, such as imidacloprid and thiamethoxam, whilst foraging for nectar and pollen from treated crops. For neonicotinoids to cause population decline, the typical residues that bumble bees encounter in the field (defined here as between 1–12 μg kg-1) should be capable of reducing colony success by detrimentally impacting demographically relevant endpoints such as reproduction and worker performance. Whether field-realistic neonicotinoids are capable of causing such effects is yet to be fully established. The overall aim of this thesis was to investigate the effects of field-realistic neonicotinoids on endpoints of demographic importance and improve understanding of the effect mechanisms of neonicotinoids in bumble bees. Laboratory experiments were conducted with Bombus terrestris L. exposed to dietary neonicotinoids up to 98 μg kg-1. Results showed that food consumption and production of brood (eggs and larvae) in queenless B. terrestris microcolonies were significantly reduced by the two highest concentrations of imidacloprid and thiamethoxam tested (39, 98 μg kg-1), but only imidacloprid produced a negative effect when concentrations were in the typical field-realistic range. Imidacloprid’s affect on microcolonies was mirrored in queenright colonies where field-realistic concentrations substantively reduced both feeding and brood production. It was postulated that the detrimental effects of imidacloprid on brood production emerge principally from nutrient limitation imposed by the failure of individuals to feed. Removing imidacloprid from the bees’ diet resulted in the recovery of feeding and brood production in queenright colonies, even when previously exposed to high doses (98 μg kg-1). Investigation into the effect mechanisms of imidacloprid in B. terrestris revealed that cytochrome P450 enzymes are not important for metabolism of the neonicotinoid in adult workers. A transcriptomic analysis indicated B. terrestris exhibit a general stress response to imidacloprid, characterised by the alteration in expression of genes involved in, for example, metabolism and storage of energy. The thesis findings raise further concern about the threat of imidacloprid to wild bumble bees. However, they also suggest that some demographically important endpoints are resilient to imidacloprid as a realistic pulsed exposure, and that bumble bees may be less sensitive to field-realistic concentrations of thiamethoxam. Further research, which is required to fully establish the demographic consequences for bumble bees of exposure to neonicotinoids, can be developed based on the foundation of work presented here.

published_or_final_version / Community Medicine / Master / Master of Public Health

4 pp. / Prevention through thinning, irrigating and/or applying preventive insecticides are the best methods of protecting trees from bark beetles. Detecting evidence of infestation is important to determine if spraying will be effective. Correct insecticides when applied properly can be effective. Insecticide injections or systemics have not proven effective against bark beetles.