Induction of hepatic microsomal hydroxylative enzymes is an important aspect of detoxication of fat-soluble toxicants. The magnitude of induction depends on numerous factors, such as the nature and dose of toxicant as well as dietary factors. Research was conducted on (1) endrin tolerance in rats, (2) preliminary comparisons of inductive effects of various organochlorine pesticides in rats to select compounds for further study in guinea pigs, (3) general effects of various dietary factors on induction, and (4) effects of ascorbic acid deficiency on induction of hepatic microsomal hydroxylative enzymes by organochlorine pesticides in guinea pigs. Measurements were made of body weight gain, feed consumption, liver weight, in vivo and in vitroenzyme activities, and body levels of pesticides and vitamins. Tolerance developed in rats fed 25 ppm endrin in the diet. There was severe intoxication during the first week but complete recovery of rate of body weight gain and feed consumption occurred during the second week, in spite of continued ingestion of the endrin-containing diet. Induction of endrin-degrading microsomal enzymes was proposed as the mechanism for tolerance. Pretreatment with the potent inducer, dieldrin, diminished the severity of endrin intoxication. However, pretreatment with another inducer, phenobarbital, afforded less protection in proportion to the extent of microsomal enzyme induction. Organochlorine pesticides, tested for their inductive capacity in rats, in decreasing order of effectiveness then fed as 25 ppm of the diet for 15 days, were heptachlor epoxide, dieldrin, endrin, 1,1-Bis-(p-chlorophenyl)-2,2,2-Trichloroethane (DDT), Ovex, gamma-chlordane, and lindane. Of these, DDT and dieldrin were compared over the range of 1 to 50 ppm of the diet. Dieldrin was a more potent inducer at all dietary levels. At low doses both compounds produced greater induction when measured by hexobarbital sleep time than by the in vitro enzyme procedures. At high doses, the in vitro 0-ethyl 0-p-nitrophenyl phenyl-phosphonothioate (EPN) detoxication was a more responsive measure. Dieldrin, DDT, and lindane were fed to guinea pigs at 25 ppm of the diet for 15 days, but only dieldrin stimulated a significant level of induction. DDT antagonism of dieldrin storage seen in rats by Street (Sci. 146:1580, 1964) did not occur in guinea pigs, but rather dieldrin antagonized DDT storage. Some general dietary factors affecting induction in rats were observed. A semipurified diet lowered the baseline microsomal enzyme activity but supported induction as effectively as a conventional diet. Vitamin A at very high dietary levels induced enzyme activity; this induction was apparently additive to that produced by 1 ppm dieldrin. Other fat-soluble Vitamins produced inconsistent responses. Ascorbic acid deficiency in guinea pigs impaired induction by dieldrin. Impairment was seen by the second day on the deficient diet. However, dieldrin was able to produce a small amount of induction at all stages of deficiency. In frank scurvy, induction by DDT and lindane was completely blocked, but there was a moderate level of induction by dieldrin. Maintenance of maximum induction was related to dietary rather than liver levels of ascorbic acid; 50 ppm ascorbic acid in the diet was grossly inadequate while 200 ppm supported about 80% of the induction produced by feeding 2000 ppm. It was concluded that (1) microsomal enzyme induction is important in resistance to organochlorine intoxication, (2) factors found in the normal diet can induce microsomal enzyme activity, (3) high dietary levels of ascorbic acid are necessary to support maximum induction, and (4) dieldrin is an inducer of such high potency that it can stimulate a limited amount of induction in spite of ascorbic acid deficiency.

No description available.

No description available.

Several epidemiological studies have suggested a role for environmental chemicals in the etiology of neurodegenerative diseases such as Parkinson's disease and Alzheimer's disease. Endosulfan (an organochlorine) and zineb (zinc-ethylene-bis-dithiocarbamate) are used as pesticides on a variety of crops worldwide and pose potential health risks to humans and animals. Both endosulfan and zineb are known to affect nervous system. Because the dopaminergic system continues to develop postnatally, we hypothesized that developmental exposure to endosulfan or zineb alone or in combination would result in alteration of nigrostrial neurotransmitters and would render the nigrostrial dopamine system more susceptible to chemical challenge later in life. The objectives of this study were (1) to determine the effects of endosulfan and zineb individually and in combination on dopaminergic or cholinergic pathways in vivo, (2) to investigate the effects of exposure to endosulfan, zineb and their mixtures administered in early life (during brain development) on subsequent exposure to these pesticides on the dopaminergic and cholinergic systems, in vivo, (3) to investigate the mechanism(s) of induction of neuronal cell death caused by these pesticides using human neuroblastoma SH-SY5Y cells in culture, (4) to define the role of oxidative stress in pesticide-induced neuronal cell death in vitro. Male C57Bl/6 mice of 7-9 months old exposed to zineb (50 and 100 mg/kg), endosulfan (1.55, 3.1 and 6.2 mg/kg) and their mixtures every other day over a 2-week period exhibited higher levels of dopamine accumulation in the striatum. Both pesticide-treated groups displayed significantly lower norepinephrine levels in the striatum (Ï ≤ 0.05) than the controls. The developmental exposure to zineb, endosulfan and their combination enhanced the vulnerability to subsequent neurotoxic challenges occurring later in life. Thus, C57BL/6 mice exposed to zineb, endosulfan and their mixtures as juveniles (postnatal days 5 to 19) and re-exposed at 8 months of age showed a significant depletion of striatal dopamine, to 22%, 16%, and 35% of control, respectively. Acetylcholinesterase activity in the cerebral cortex was found to be significantly increased in all pesticide treated groups. Mice given mixtures of pesticides also showed significantly increased levels of normal and aggregated alpha-synuclein, a hallmark of neurodegenerative diseases such as Parkinson's disease and Alzheimer's disease. The results of these studies indicate that exposure to these pesticides as neonates and re-exposure as adults could result in neurochemical changes that did not reveal at adulthood when the exposure was at juvenile age only. We further investigated the mechanism(s) of activation of pesticide-induced neuronal cell death in vitro. The characteristic of cell death in SH-SY5Y human neuroblastoma cells was examined. These cells are known to retain catecholaminergic phenotype. Cells were exposed to endosulfan, zineb and mixtures of two pesticides, in concentrations ranging from 50 μM to 400 μM. These exposures caused both apoptotic and necrotic cell death in SH-SY5Y cells as evaluated by lactate dehydrogenase release, 7-aminoactinomycin-D and Annexin-V/PI assays. Exposure to mixtures of the pesticides enhanced both the early apoptosis and late apoptosis/necrosis compared to either chemical alone. Visual evaluation using DNA ladder assay and fluorescence Annexin V/PI assay confirmed the contribution of both apoptotic and necrotic events. Furthermore, endosulfan and zineb alone and in combination altered the caspase-3 activity indicating that both pesticides exposure exert their apoptotic effect via the caspase-3 pathway. Because there has been increasing evidence of the role of reactive oxygen species (ROS) and oxidative stress in pesticide-induced neuronal cell death (apoptosis and necrosis), the levels of ROS and antioxidant enzymes were examined. Cells treated with pesticides were found to enhance the generation of superoxide anion and hydrogen peroxide both in a dose- and time-dependent manner. Mixture of pesticides significantly enhanced the production of these reactive oxygen species compared to cells exposed to individual pesticide. Cells treated with pesticides showed a decrease in superoxide dismutase, glutathione peroxidase, and catalase levels. These pesticides also induced lipid peroxides (thiobarbituric acid reactive products) formation in SH-SY5Y cells. Furthermore, cells exposed to these pesticides were found to have increased in the expression of NFkappaB activity in the nucleus. These data support the hypothesis that oxidative stress was induced in neuronal cells by exposing to these pesticides in vitro. Taken together, the results of this study support the above hypothesis and suggest that the cytotoxicity of endosulfan and zineb and their combinations may, at least in part, be associated with the generation of ROS. Furthermore, mice exposed at early age and re-exposed at adulthood become more susceptible to alteration of neurotransmitter levels compared to mice exposed to these pesticides only as juveniles. These findings could add to the growing body of knowledge on the mechanism of pesticide-induced dopaminergic neuronal cell death and could hold tremendous implication for the future understanding of the possible involvement of environmental risk factors in the pathogenesis of Parkinson's disease. / Ph. D.

Edamame (Glycine max (L.) Merr.), also known as vegetable soybean, is primarily grown and consumed in Asia. In recent years, the demand for edamame in the United States has risen due to its health benefits as an alternative, plant-based protein. Due to the lack of domestic production, most edamame is imported from Asia. In an attempt to increase domestic production, research efforts have begun in Virginia and other regions to develop cultivars and best management practices for growing edamame in the mid-Atlantic region. Beginning in 2018, edamame trials examining breeding lines and cultivars were conducted to look at their suitability for this region. These varieties were sampled and evaluated for insect and disease complexes as well as their implications on plant yield and quality. Most of the insects and diseases that were found were very similar to pest complexes commonly found in cultivated soybeans in Virginia. However, due to edamame being marketed as a vegetable, insects and diseases that caused unsightly blemishes or damage to the pods or seeds were most concerning. Multiple insects and diseases were present but some of the most important insects and diseases we observed from 2018-2020 were pod feeding stink bug (Hemiptera: Pentatomidae), as well as the diseases like purple seed stain, Cercospora kikuchii, and bacterial pustule, Xanthomonas axonopodis pv. glycines. From 2019-2021 an integrated pest management study was conducted to determine best management practices for minimizing insecticide applications while applying them at thresholds to control key pests. In three growing seasons, I was able to determine that pesticides can be limited prior to flowering while insecticide inputs will need to be increased after flowering to protect the pods from pests, specifically stink bugs. Additionally, from 2019-2021 insecticide and fungicide field trials were conducted to test different pesticides on their efficacy against pod damaging pests and diseases. The growing seasons between 2019 and 2021 resulted in varying insect and disease pressure that led to inconsistent results. However, insecticides such as cyclaniliprole and sulfoxaflor performed well compared to other treatments. Lastly in 2020-2021, corn earworm (Helicoverpa zea [Boddie], Lepidoptera: Noctuidae), an important soybean pest and most likely a major pest of edamame, was tested for pyrethroid susceptibility and resistance across the state using a bean-dip bioassay. Pyrethroid efficacy to control this pest across Virginia seemed to vary by location and year, however, the bean-dip bioassay proved to be a time-efficient strategy for further monitoring these populations in the coming years. / Doctor of Philosophy / Edamame, also known as vegetable soybean, is primarily grown and consumed in Asia. Due to the lack of domestic production, most edamame is imported from overseas. Edamame trials were conducted in Virginia beginning in 2018, to look at production practices suitable for growing this crop in the region. Scientists observed edamame to document insect and disease complexes as well as their implications on yield and quality. Most of the insects and diseases that were found were very similar to what is known to already occur in soybeans. However, due to edamame being marketed as a vegetable, insects and diseases that left blemishes or damage to the pod were most concerning. Multiple insects and disease were present but some of the most important insects and diseases we observed were pod feeding stink bug species as well as the diseases purple seed stain and bacterial pustule. In 2019-2021 a pest management study was conducted to determine best management practices for minimizing insecticide applications while applying them at thresholds to control key pests. We were able to determine that pesticide usage can be limited prior to flowering while they will need to be increased after flowering to protect the pods. Additionally, in 2019-2021 insecticide and fungicide field trials were conducted to determine the efficacy of materials against pod damaging pests and diseases. The growing seasons between 2019 and 2021 resulted in varying insect and disease pressure, however, several insecticides with reduced ecological impacts out preformed others. Lastly in 2020-2021, corn earworm, a major pest of soybean and presumable of edamame, was tested for pyrethroid susceptibility and resistance across the state using a bean-dip bioassay. Pyrethroid efficacy across Virginia seemed to vary by location and year but the bean dip bioassay method proved to be a time-effective strategy for monitoring the states corn earworm populations against insecticides.

The immunotoxic effects of multiple pesticide exposure were evaluated. C57BL/6 mouse thymocytes were exposed to lindane, malathion, and permethrin, either separately or in mixtures of two pesticides, in concentrations ranging from 37.5 uM to 1mM. These exposures caused both apoptotic and necrotic cell death in thymocytes as evaluated by 7-aminoactinomycin-D, Annexin-V/PI, and lactate dehydrogenase release assays. When cells were exposed to lindane+malathion, or lindane+permethrin, a significantly greater-than-additive cytotoxicity was observed. The pesticide exposure caused DNA ladder formation with increased laddering in mixtures. Further, the effect of these pesticides on thymocyte oxidative stress was investigated. Thymocytes treated with any of these pesticides generated superoxide and H2O2. The lindane + malathion caused more-than-additive increase in superoxide production compared to single treatments of these pesticides. However, the effect of the lindane + permethrin was not significantly different from individual components of this mixture. The effects of pesticides on antioxidant enzymes were also investigated and only mixtures were found to have significant effects. Alteration in transcription factor NFkB level was measured as an indicator of oxidative stress in thymocytes following 12 h pesticide exposure, in vitro. Only lindane + malathion was found to increase the protein level. Furthermore, the effects of pesticides and their mixtures on immune functions of mice were studied in vivo. Animals (8-12 week old, male mice) were randomly divided into groups of six and injected intraperitoneally with three different doses (one-half, one-third, one-fourth, or one-eight of LD50) of individual pesticides. Exposure to individual pesticides did not alter the thymus/body or spleen/body weight ratios, thymic or splenic cell counts, or CD4/CD8 or CD45/CD90 ratios. However, anti-sRBC plaque forming cell (PFC) counts were significantly lowered with all treatments. Two other groups of animals were injected with lindane + malathion or lindane + permethrin at one-third of the LD50 of each pesticide. Exposure to pesticide mixtures did not alter the CD4/CD8 or CD45/CD90 ratios. However, the thymus/ and spleen/body weight ratios, thymic and splenic cell counts, and PFC counts were significantly lowered. These data indicate that lindane, malathion, and permethrin are immunotoxic and their mixtures can cause higher toxicity compared to individual exposures. In addition, these data support the hypothesis that oxidative stress were induced in thymocytes by exposure to these pesticides in vitro. / Ph. D.

Pest resistance to pesticides can have severe impacts on both commercial agriculture and the environment. But many resistance problems are exacerbated because pest susceptibility is a dynamic, common-property resource subject to inefficient allocation by the market. Theoretically, the impact of resistance can be mitigated through regulatory management of the control technology set. However, the current pesticide regulatory process does not include resistance considerations in its quantitative analyses due to the computational difficulties encountered when trying to optimize complex bioeconomic models. As a result, regulatory efforts may actually promote increased susceptibility depletion and the rapid emergence of resistance. This study overcame these problems by forming a dynamic bioeconomic model that combined: 1) a widely accepted genetic simulator used by entomologists; 2} an aggregate economic surplus model with nationwide regulatory relevance; and 3) an improved simulation optimization algorithm that conserved computational resources. For the purpose of illustration, the bioeconomic model was parameterized to represent the U.S. apple production system. Information generated through optimization of the dynamic bioeconomic model suggested that resistance becomes quantitatively important when planning horizons exceed 10 years, confirming that the economic performance of the production system becomes severely sub-optimal when susceptibility depletion is not incorporated into decision-making. Furthermore, insecticide withdrawals from an initial control technology set led to large additional losses in economic surplus, although the exact magnitude of these impacts varied depending on the characteristics of the insecticide withdrawn. Substantial withdrawal-induced losses in of the planning horizon, and they were accompanied by temporal shifts in insecticide applications. The need to incorporate a dynamic, bioeconomic simulation analysis in the regulatory process was demonstrated by comparing statically optimal and extant insecticide use recommendations with the dynamically̅optimal solutions. Optimal solutions drastically reduced economic surplus losses, although they did lead to increased levels of insecticide use. Ultimately, management of the resistance/regulation nexus requires that both current economic data and the time–dynamics of system biology play a prominent role in the benefits assessment process. This can only be accomplished if an investment is made in the necessary basic research and model development. / Ph. D.

A field scale model for predicting the surface losses of pesticides (Pesticide Losses In Erosion and Runoff Simulator, PLIERS) was developed. PLIERS accounts for pesticide losses by degradation and volatilization, the washoff of pesticides from plant canopy and surface residue, the adsorption and desorption of pesticides to and from soil particles, and the movement of pesticides in the dissolved and adsorbed phases. Hydrologic data are generated by the comprehensive watershed model, FESHM; which contains an extended sediment detachment and transport algorithm. PLIERS uses first order rate equations to describe degradation and volatilization, and pesticide washoff. The adsorption of pesticides to individual particle size classes is estimated using the Freundlich equation. Movement of atrazine and 2,4-D in runoff and sediment was measured on twelve field plots under simulated rainfall. The plots were treated with conventional or no-tillage in combination with one of three residue levels (0, 750, and 1500 kg/ha). Runoff and sediment losses were found to increase with decreasing residue cover for both tillage systems. No-till reduced sediment loss and total runoff volume by 98 and 92 percent, respectively, compared to conventional tillage. Concentrations of atrazine and 2,4-D ir1 runoff and sediment were greater from the no-till plots than from the conventional plots but the total losses were less. Both pesticides were carried predominately in the dissolved phase. Averaged over all plots, the atrazine losses were 2.9 percent of applied amount for conventional tillage and 0.3 percent for no-tillage. The corresponding values for 2,4-D were 0.3 percent and 0.02 percent. PLIERS was validated using data from the rainfall simulator field plot studies. Agreement between predicted and observed data was very good for dissolved pesticide losses and satisfactory for adsorbed pesticide losses. In addition, the effects of tillage type and residue level were reflected in PLIERS predictions. PLIERS shows great potential as a flexible planning tool since it could be used with any comprehensive hydrologic model and is able to predict the losses of pesticides under various field conditions. / Master of Science

Tuta absoluta, a member of the moth family, causes devastating yield loss to tomato farmers around the world. Its recent migration into the tomato fields of Nepal puts tomato farmers at a high risk of yield loss. In response, chemical pesticide use by Nepali farmers is increasing. Integrated pest management (IPM) practices have been implemented in hopes of reducing the frequency of chemical pesticide use while controlling yield risks. This study examines the extent and determinants of Tuta absoluta IPM adoption and its effect on the frequency of pesticide use for Nepali tomato farmers. Primary data was collected from four-hundred and one households in four districts throughout Nepal. Two levels of IPM practices were assessed, simple and complex, based on the need for additional knowledge and tools. An instrumental variable probit analysis was used to analyze the determinants of IPM adoption. Household distance to nearest agricultural extension office was a significant factor decreasing the likelihood of the adoption of complex practices. Amount of land dedicated to tomato production, membership status of the primary decision maker, IPM training regarding Tuta absoluta practices and severity of Tuta absoluta were found to increase the likelihood of the adoption of complex practices. In order to analyze pesticide use, a simple linear regression was used. Primary decision maker's age, gender, and education level were significant determinates to decrease the amount of expenditures spent on chemical pesticides to control for Tuta absoluta. IPM adoption level, amount of land dedicated to tomato production and severity of Tuta absoluta damage were significant determinates to increase the amount of expenditures spent on chemical pesticides to control for Tuta absoluta. / Master of Science / Tuta absoluta, a member of the moth family, causes devastating yield loss to tomato farmers around the world. Its recent migration into the tomato fields of Nepal puts tomato farmers at a high risk of yield loss. In response, chemical pesticide use by Nepali farmers is increasing. Integrated pest management (IPM) practices have been implemented in hopes of reducing the frequency of chemical pesticide use while controlling yield risks. This study examines the extent and determinants of Tuta absoluta IPM adoption and its effect on the frequency of pesticide use for Nepali tomato farmers. Data was collected from four-hundred and one households in four districts throughout Nepal. Different economic tools were used to conduct the analyses. Results show the farther a household is to the near agricultural extension office, the less likely a household is to adopt complex practices. An increased amount of land dedicated to tomato production, the primary decision maker being a member of a marketing or community organization, the primary decision maker having attended IPM training regarding Tuta absoluta practices and the greater the severity of Tuta absoluta in the household tomato fields, the more likely a household is to adopt complex practices. Contrary to one of the goals of IPM practices, this study shows complex adopters use more chemical pesticides than simple adopters. Results this study shows a need for further education of the relationship between IPM practices and the use of chemical pesticides.

The purpose of this research was to investigate and compare the effects of different abrasion treatments on the liquid-fabric interaction of selected nonwoven barrier fabrics. The abrasion treatments included moderate and severe abrasion, flat and flat/flex abrasion, and dry and wet abrasion. The liquid-fabric interactions included wetting/wicking, retention, and penetration through nonwoven fabrics using water/surfactant solution. Results of this study indicated that abrasion treatments increased the wetting/wicking rate of fabrics. The flat/flex abrasion caused a greater increase in the wetting/wicking rate of fabrics than the flat abrasion. Abrasion treatments also increased liquid penetration. The flat abrasion increased liquid penetration more than flat/flex abrasion. On increasing abrasion severity, there was a significant increase in liquid penetration. There was no consistent effect on liquid retention. It was highly influenced by fabric types. Wet abrasion did not differ significantly from dry abrasion in its effects on liquid/fabric interaction. Six nonwoven fabrics used in this study included a hydroentangled cotton fabric with a fluorochemical finish (HCF), a hydroentangled cotton fabric laminated with a microporous film (HCE), a spunbonded polypropylene with microporous film (PSM), a four layer laminated nonwoven including spunbonded polypropylene, microporous film, hydroentangled cotton layer, and spunbonded polypropylene (PECP), a spun-bonded, melt-blown, spun-bonded polypropylene (SMS), and standard Tyvek®. Among the six fabrics, the cotton fabrics with a fluorochemical finish (HCF) and the cotton fabric with a microporous film (HCE) showed an excellent potential as protective material, since they provided high liquid resistance before and after abrasion. However, there was no consistent trend for microporous film fabrics or for cotton containing fabrics to provide a good liquid protection. In general, it was concluded that abrasion significantly decreased liquid protection of protective fabrics. / Master of Science