Designed for the protection of major agricultural crops, neonicotinoids are the fastest-growing class of insecticides used against a broad spectrum of insect pests. Although neonicotinoid toxicity toward non-target organisms (e.g., bees, aquatic insects) has been well-studied, less is known about their distribution of use, transport, and fate in North American agroecosystems. This is especially true of neonicotinoid interactions with wetlands in the Canadian Prairies. Between 2009 and 2012, neonicotinoid use as a seed treatment increased by 30% across the Canadian Prairies. During spring 2012 to spring 2013, I sampled water and sediment from 136 wetlands situated in a range of crop types across central Saskatchewan to determine the extent of neonicotinoid contamination. Wetlands situated in oat, canola, and barley fields consistently contained higher neonicotinoid concentrations in water than in grasslands, but no single crop influenced overall detections. Neonicotinoid detections in water varied from 16% (fall 2012) to 91% (spring 2013) with peak concentrations up to 3110 ng/L found in summer. I found numerous detections of neonicotinoids in spring, after ice-off, but before seeding. Through sampling snow, snow meltwater, and soil particulates from previously treated (clothianidin) and untreated fields, meltwater showed the strongest relationship to initial spring concentrations in wetland water. Neonicotinoid concentrations increased with time in shallow temporary wetlands which appeared most at risk for annual contamination. While snowmelt contamination influenced water concentrations in spring, peak concentrations in wetlands were consistently found during summer sampling completed in 2012-2014. Rapid wetland assessments completed on 238 wetlands (summer of 2012 and 2013) revealed key ecological, hydrological and landscape features that influenced neonicotinoid detections and peak concentrations in Prairie wetlands. The results of my exploratory analysis indicated that plant community composition is a key indicator and/or driver of both detection and concentration of neonicotinoids in Prairie wetlands. In particular, specific shallow marsh plants were commonly associated with either higher (e.g., Scirpus validus) or lower (e.g., Mentha arvensis) neonicotinoid concentrations in natural wetlands suggesting wetland macrophytes in this zone may be either indicators of agricultural disturbance intensity or differentially capable of accumulating the insecticide in its tissue. Therefore, in 2014, I conducted an outdoor microcosm experiment to evaluate thiamethoxam uptake from water by Typha latifolia and Alisma triviale using two concentrations over a 7-day period. Experimental results found some trace positive detections but no quantifiable accumulation of the insecticide in plant tissues. This is despite the fact that results of my 2015 field study found species of Typha, Alisma and Equisetum had neonicotinoids more frequently detected in their tissues, at concentrations ranging from 1.01-8.44 ug/kg. My findings demonstrate that neonicotinoid distribution and fate in Canadian Prairie agroecosystems is driven by interactions between ecological, hydrological, and landscape characteristics. Consequently, these drivers regulate neonicotinoid exposure and persistence in ecologically important regional wetlands. In order to effectively conserve these critical waterbodies, conservation planning should consider the importance of maintaining naturally diverse vegetation zones to mitigate insecticide exposure to wetland-dependant organisms.

Corn is a dominating feature in the landscape of the Midwestern United States. Associated with this crop are a plethora of products from fertilizers to pesticides that help farmers maximize grain yield while minimizing costs. A widely accepted form of protection from major pests in the United States is genetically modified corn that has been altered so that it contains genetic material from another species, a soil bacterium, Bacillus thuringiensis Berliner (Bt corn), in order to produce proteins that are toxic to some insect pests. Additional insecticides that are commonly used to protect corn include a neonicotinoid seed coating, clothianidin, and a pyrethroid insecticide applied in-furrow, tefluthrin. The goal of my dissertation was to measure the environmental fate of the Cry1Ab Bt corn protein, clothianidin, and tefluthrin and determine if measured concentrations could be causing effects to non-target species. The Cry1Ab proteins persisted only during the corn growing season in soil, runoff water and sediment with the highest concentrations measured during pollination. Clothianidin was detected in all matrices and remained persistent throughout the year in soil pore water. Tefluthrin was consistently detected in soil, runoff water, and runoff sediment during the corn growing season, but was not found in groundwater or soil pore water. No acute toxicity was observed to any species when exposed to the Cry1Ab protein in corn plant leaf tissue, therefore little risk was anticipated. Non-target species exposed to clothianidin resulted in limited ecological risk from field exposures. Tefluthrin results indicated elevated risk for non-target species exposed to tefluthrin at concentrations that were measured in the field study. There was no increase in toxicity to tefluthrin when non-target species were exposed to a combination of all three insecticides. In summary, the genetically modified corn insecticidal proteins and clothianidin were not found at environmental concentrations exceeding benchmark values for ecological effects, but tefluthrin was consistently detected in the environment at levels that could be causing toxicity to non-target species, especially if it is able to travel off-site.

Thiamethoxam and clothianidin are two neonicotinoids used in seed coatings for crops such as corn and soybeans. Both neonicotinoids have high solubility in water, so they are prone to transport via leaching and runoff. This thesis is comprised of two studies that evaluated potential solutions to mitigate neonicotinoid transport from fields. The first study examined the relationship between soil organic carbon content and neonicotinoid transport in a field planted in soybeans. Soils with increased organic carbon leached less thiamethoxam and clothianidin during early growing season leaching peaks; however, at the end of the season, higher organic carbon content only decreased leached mass of clothianidin. The second study was to determine neonicotinoid uptake of different ground covers used as cover crops or edge-of-field buffer strips, as well as the partitioning of thiamethoxam and clothianidin throughout the plants. Ground covers, such as crimson clover, had the highest recovery of applied thiamethoxam, meaning that it may be a good candidate to retain this pesticide in fields. Thiamethoxam and clothianidin concentrations were higher in leaf tissues than in stems or roots, indicating that above-ground biomass removal may be an effective way to reduce neonicotinoid loading in the environment. From these studies, I concluded 1) practices that raise the amount of organic carbon in the soil may help decrease early-season neonicotinoid transport, resulting in lower concentrations in surrounding waterways, and 2) careful selection of plant species, such as crimson clover, may help reduce neonicotinoid transport in the environment, while potentially reducing exposure to beneficial insects. / Master of Science / Pesticides called neonicotinoids are commonly applied to seeds in row crops, such as corn and soybeans, before they are planted. These pesticides are highly soluble in water, which can lead to them exiting fields through runoff or leaching. This thesis is comprised of two studies that examined several potential solutions for decreasing the amount of neonicotinoids available for transport. The first study examined the relationship between organic carbon in the soil and neonicotinoids, and whether this relationship helps to retain neonicotinoids in a soybean field. Soils with high organic carbon content decreased the amount of neonicotinoids exiting the field during early growing season storms; however, at the end of the season, high organic carbon content only decreased losses for one of the pesticides studied. The second study was to determine which of six plant species and two mixes used as cover crops or buffers were the most effective at removing neonicotinoids from soil, as well as where in the plant these neonicotinoids go after uptake. Ground covers, such as crimson clover, had the highest recovery of applied neonicotinoids, meaning they would be good candidates for planting around fields. Ultimately, neonicotinoid accumulation was higher in leaves than in stems or roots, meaning that removing and disposing of leaves in an environmentally safe way could be an effective way to decrease neonicotinoid pollution. From these two studies, I found that 1) increasing organic matter in the soil can stop neonicotinoids from exiting the area it was applied in, and 2) careful consideration of plant species in or around the field may help intercept neonicotinoids before they exit the field.

Bumble bees and other wild pollinators are crucial to the support of both natural and agricultural ecosystems. However, unprecedented declines of pollinator populations have been observed all over the world, raising concerns of a looming threat to both the human food supply, as well as sustainability of the biodiversity in local ecological niches. Though declines are well described, the cause behind these still evades scientists. Proposed contributors include anthropogenic-mediated environmental stress, including application of xenobiotics for pest control, and increase of pathogen diversity and abundance due to the shipment of infection human-managed colonies. This research examined these theories and attempted to quantify the threats they may pose. Through development of a chronic, oral toxicity experiment, susceptibility of all Bombus impatiens castes to clothianidin exposure was examined. This exposed a substantial increase in vulnerability of male bumble bees to realistic concentrations of neonicotinoid pesticides, highlighting the crucial need to examine all members of wild bumble bee life cycles before determining pesticide regulations. Additionally, sublethal effects on fitness-related foraging behaviors in Bombus impatiens were examined through development of a voluntary task switching assay. The results of this experiment suggest humoral immune stimulation, through pathogenic infection, leads to significant impairment of cognitive flexibility. Taken together, this data suggests that pesticides and pathogens are capable of causing severe detrimental effects, both lethally and sublethally, in wild bumble bees. I hope this data will eventually contribute to reassessment of environmental regulations and establishment of effective conservation strategies in order to sustain the critical populations of wild bumble bees.

Two field trials were conducted to determine the impact of water management on the efficacy of insecticide seed treatments against rice water weevil, Lissorhoptrus oryzophilus Kuschel, in rice at the Delta Research and Extension Center during 2011 and 2012. The performance of thiamethoxam, chlorantraniliprole, and clothianidin was evaluated when the permanent flood was established at different timings (6 and 8 weeks after planting) and the effect of flush number (0, 1, or 2) on seed treatment performance was evaluated. Seed treatment efficacy was not impacted by delayed flooding, but 2 flushes reduced efficacy of some seed treatments. Experiments were also conducted to determine the impact of reduced seeding rates found in hybrid rice production on the efficacy of insecticide seed treatments targeting rice water weevil. Efficacy was similar when comparing currently labeled rates of thiamethoxam, chlorantraniliprole, and clothianidin with higher rates of these products.

<p><!--[if gte mso 9]><xml> <w:WordDocument> <w:View>Normal</w:View> <w:Zoom>0</w:Zoom> <w:PunctuationKerning /> <w:ValidateAgainstSchemas /> <w:SaveIfXMLInvalid>false</w:SaveIfXMLInvalid> <w:IgnoreMixedContent>false</w:IgnoreMixedContent> <w:AlwaysShowPlaceholderText>false</w:AlwaysShowPlaceholderText> <w:Compatibility> <w:BreakWrappedTables /> <w:SnapToGridInCell /> <w:WrapTextWithPunct /> <w:UseAsianBreakRules /> <w:DontGrowAutofit /> </w:Compatibility> <w:BrowserLevel>MicrosoftInternetExplorer4</w:BrowserLevel> </w:WordDocument> </xml><![endif]--><!--[if gte mso 9]><xml> <w:LatentStyles DefLockedState="false" LatentStyleCount="156"> </w:LatentStyles> </xml><![endif]--><!--[if gte mso 10]> <mce:style><! /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-parent:""; mso-padding-alt:0pt 5.4pt 0pt 5.4pt; mso-para-margin:0pt; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:10.0pt; font-family:"Times New Roman"; mso-ansi-language:#0400; mso-fareast-language:#0400; mso-bidi-language:#0400;} --><!--[endif] --></p><p class="MsoBodyText" style="margin-bottom: 0.0001pt;">U disertaciji je izvr&scaron;ena voltametrijska karakterizacija i određivanje pet&nbsp;neonikotinoidnih insekticida (imidakloprida, tiametoksama, klotianidina, nitenpirama i&nbsp;acetamiprida) na različitim elektrodama na bazi ugljenika, kao &scaron;to su elektroda od&nbsp;ugljenične paste, elektroda od staklastog ugljenika i bizmut-filmom modifikovani&nbsp;staklasti ugljenik. Pored ispitivanja u model-rastvorima, razvijene visokoosetljive&nbsp;diferencijalne pulsne voltametrijskemetode su primenjene za određivanje neonikotinoida&nbsp;u realnim uzorcima. Neonikotinoidi su određivani u uzorcima rečne vode i komercijalnim&nbsp;formulacijama, a praćena je i njihova fotolitička i fotokatalitička razgradnja. Radi&nbsp;potvrđivanja voltametrijskih rezultata, izvedena su komparativna merenja uz primenu&nbsp;tečne hromatografije sa detektorom od niza dioda i infracrvenespektrometrije. Detekcija&nbsp;pojedinih degradacionih proizvoda nastalih u toku ispitivanih fotodegradacionih procesa&nbsp;je praćena tečnohromatografskom metodom sa tandemskom masenospektrometrijskom&nbsp;detekcijom. Proučavana je povr&scaron;inska struktura CR 5 grafitnog praha, elektrode od&nbsp;ugljenične paste na bazi trikrezil-fosfata i elektrode od staklastog ugljenika pomoću&nbsp;skenirajuće elektronske mikroskopije.</p> / <p>This thesis is concerned with the voltammetric characterization and&nbsp;determination of five neonicotinoid insecticides (imidacloprid, thiamethoxam,&nbsp;clothianidin, nitenpyram and acetamiprid) at different carbon-based electrodes such as&nbsp;carbon paste, glassy carbon and bismuth-film modified glassycarbon. In addition to the&nbsp;investigations in model solutions, the developed highly sensitive differential pulse&nbsp;voltammetric methods were applied for the determination of neonicotinoids in real&nbsp;samples. Neonicotinoids were determined in river water samples, commercial&nbsp;formulations, and their photolytic and photocatalytic degradation was also monitored. To&nbsp;confirm the voltammetric results, comparative measurements were performed by liquid&nbsp;chromatography with diode-array detector and infrared spectrometry. Detection of some&nbsp;degradation products formed during the photodegradation processes was monitored using&nbsp;liquid chromatography with tandem mass-spectrometric detection. The surface structure&nbsp;of CR 5 graphite powder, tricresyl phosphate-based carbon paste electrode and glassy&nbsp;carbon electrode was investigated by scanning electron microscopy.</p>

<p>Mayflies are important macroinvertebrate members of steam communities and are highly sensitive to agricultural insecticides. In this dissertation I sought to explore whether Heptageniidae<i> </i>mayflies in Indiana have evolved greater tolerance to clothianidin, a neonicotinoid insecticide that has been widely applied over the predominant corn and soybean crops of the state for the last 20 years. Over the five chapters of this dissertation, I investigate different aspects of the effects of clothianidin on mayfly populations. First, I collected mayfly populations from streams around the state across a gradient of agricultural landscape to see if proximity to corn and soybean crops was associated with tolerance to the insecticide. I found significant variation in tolerance between the populations but only connected it to land use for one species and one insecticide. Second, I investigated the potential for increases in predation rates due to exposure to low concentrations of clothianidin by both a vertebrate and invertebrate predator. Sublethal exposure did increase the risk of predation by the invertebrate predator but not the vertebrate. Third, I collected four populations of mayflies and placed them in a common stream which is heavily impacted by agricultural runoff. The most tolerant of the four populations had the highest survival rates and was native to the polluted stream, suggesting that it was best adapted to the local conditions. Finally, the last chapter is a discussion of the factors influencing the evolution of contaminant tolerance, the implications for biomonitoring, and a series of recommendations for incorporating evolved tolerance into biomonitoring practices. Collectively, this work suggests that the lethal and sublethal effects of clothianidin exposure create conditions suitable for the evolution of tolerance in Heptageniid mayflies. While there are already differences in tolerance between populations, it seems likely that mayfly pesticide tolerance will continue to increase as pesticides continue to impact aquatic ecosystems. </p>