Toxicokinetics of the neonicotinoid insecticide imidacloprid in rainbow trout (Oncorhynchus mykiss).

Studies were conducted to determine the distribution and elimination of imidacloprid (IMI) in rainbow trout. Animals were injected with a low (47.6 µg/kg), medium (117.5 µg/kg) or high (232.7 µg/kg) dose directly into the bloodstream and allowed to depurate. The fish were then sampled to characterize the loss of IMI from plasma and its appearance in expired water (all dose groups) and urine (medium dose only). In vitro biotransformation of IMI was evaluated using trout liver S9 fractions. Mean total clearance (CLT) values determined by non-compartmental analysis of plasma time-course data were 21.8, 27.0 and 19.5 mL/h/kg for the low, medium and high dose groups, respectively. Estimated half-lives for the same groups were 67.0, 68.4 and 68.1 h, while fitted values for the steady-state volume of distribution (VSS) were 1.72, 2.23 and 1.81 L/kg. Branchial elimination rates were much lower than expected, suggesting that IMI is highly bound in blood. Renal clearance rates were greater than measured rates of branchial clearance (60% of CLT in the medium dose group), possibly indicating a role for renal membrane transporters. There was no evidence for hepatic biotransformation of IMI. Collectively, these findings suggest that IMI would accumulate in trout in continuous waterborne exposures.

Effects of the herbicide triclopyr on metamorphic northern red-legged frogs.

Aquatic herbicides are used to manage invasive emergent plants in and around wetlands. Metamorphic frogs that emerge during the aquatic weed management season may be at risk of herbicide exposure. Metamorphic northern red-legged frogs (Rana aurora) were exposed to a triclopyr tank mix labeled for control of broadleaf emergent aquatic weeds such as invasive purple loosestrife (Lythrum salicaria). The tank mix consisted of Renovate® 3 (triclopyr triethylamine salt 44.4%), the modified vegetable oil surfactant Competitor®, and the marker dye Hi-Light® . Metamorphs were exposed to the tank mix and a clean-water control for 96 h, and then reared in clean water for 60 d. Exposure to the tank mix resulted in no treatment-related mortalities, no effects on behavior immediately post exposure, and no effects on body or liver condition indices. Exposure to the tank mix resulted in lethargy during exposure and a 1-d delay in completion of metamorphosis. Deformities present in the rearing population confounded results for some endpoints. Observed effects were minimal, especially compared with the potential for ecological impacts from unmanaged invasive plants. Environ Toxicol Chem 2017;36:2316-2326. © 2017 SETAC.

Plasma cholinesterase activity as a biomarker for quantifying exposure of green sturgeon to carbaryl following applications to control burrowing shrimp in Washington State.

Willapa Bay (Washington State, USA) has been 1 of the rare intertidal locations where large-scale pesticide applications occur. Until recently, carbaryl was applied to control burrowing shrimp that decrease commercial oyster productivity. The bay is a critical habitat for green sturgeon (Acipenser medirostris), an anadromous species listed as threatened under the US Endangered Species Act. However, the hazard that carbaryl poses is unknown. Surrogate seawater-acclimated white sturgeon (A. transmontanus) were exposed to 0 µg L(-1) , 30 µg L(-1) , 100 µg L(-1) , 300 µg L(-1) , 1000 µg L(-1) , and 3000 µg L(-1) carbaryl for 6 h, and brain acetylcholinesterase (AChE) and plasma butyrylcholinesterase (BChE) activities were measured. Enzyme recovery was measured in an additional cohort exposed to 1000 µg L(-1) carbaryl for 6 h. Activity of AChE was reduced (p ≤ 0.001) at concentrations ≥ 100 µg L(-1) with recovery in the 1000 µg L(-1) cohort by 72 h. Surprisingly, BChE activity was greater than controls at concentrations ≥ 300 µg L(-1) (p > 0.05), a finding confirmed in additional fish exposed to 3000 µg L(-1) for 6 h (+30%, p < 0.001) with apparent recovery by 48 h. Plasma samples were collected from free-living green sturgeon before and 4 d to 5 d after application of carbaryl in Willapa Bay. Activity of BChE after application was reduced 28% (p < 0.001), indicating exposure to the pesticide. However, the lack of congruence between BChE and AChE activity in captive white sturgeon exposed to carbaryl indicates that further studies are needed to better understand the risk carbaryl exposure poses to green sturgeon. Environ Toxicol Chem 2016;35:2003-2015. © 2015 SETAC.

Assessing the risk to green sturgeon from application of imidacloprid to control burrowing shrimp in Willapa Bay, Washington--Part II: controlled exposure studies.

The activities of 2 species of burrowing shrimp have a negative impact on the growth and survival of oysters reared on intertidal mudflats in Willapa Bay and Grays Harbor, Washington (USA). To maintain viable harvests, oyster growers proposed the application of the neonicotinoid insecticide imidacloprid onto harvested beds for the control of burrowing shrimp. In test applications, water column concentrations of imidacloprid were relatively low and dissipated rapidly. The foraging activities of the green sturgeon (listed in the US Endangered Species Act) could result in exposure to higher, more sustained imidacloprid concentrations within sediment porewater and from the consumption of contaminated shrimp. Controlled experiments were conducted using surrogate white sturgeon to determine acute and chronic effect concentrations, to examine overt effects at more environmentally realistic concentrations and durations of exposure, and to assess chemical depuration. The 96-h median lethal concentration was 124 mg L(-1) , and the predicted 35-d no-observed-adverse-effect concentration was 0.7 mg L(-1) . No overt effects were observed following environmentally relevant exposures. Imidacloprid half-life in plasma was greater than 32 h. Measured concentrations of imidacloprid in porewater were significantly lower than the derived acute and chronic effect concentrations for white sturgeon. Exposure risk quotients were calculated using the effect concentrations and estimated environmental exposure. The resulting values were considerably below the level of concern for direct effects from either acute or chronic exposure to an endangered species.

Assessing the risk to green sturgeon from application of imidacloprid to control burrowing shrimp in Willapa Bay, Washington-Part I: exposure characterization.

Willapa Bay and Grays Harbor (WA, USA) comprise the largest region of commercial oyster cultivation on the Pacific Coast. The activities of 2 species of burrowing shrimp impair growth and survival of oysters reared on the intertidal mudflats. To maintain viable harvests, the oyster growers have proposed controlling the shrimp by applying the insecticide imidacloprid onto harvested beds. Green sturgeon (listed in the Endangered Species Act) forage on burrowing shrimp and could be exposed to imidacloprid in the sediment porewater and through consumed prey. Studies were conducted to evaluate the likelihood that green sturgeon would be exposed to imidacloprid and to characterize the subsequent environmental exposure. Comparisons between treated and untreated control beds following test application of the insecticide suggested that green sturgeon fed opportunistically on imidacloprid-impaired shrimp. The highest interpolated imidacloprid residue concentrations in field samples following chemical application were 27.8 µg kg(-1) and 31.4 µg kg(-1) in porewater and shrimp, respectively. Results from modeled branchial and dietary uptake, based on conservative assumptions, indicated that the porewater exposure route had the greatest contribution to systemic absorption of imidacloprid. The highest average daily uptake from porewater (177.9 µg kg(-1) body wt) was 9.5-fold greater than total dietary uptake (18.8 µg kg(-1) body wt). Concentrations and durations of exposure would be lower than the levels expected to elicit direct acute or chronic toxic effects.

Growth and survival of pacific coho salmon smolts exposed as juveniles to pesticides within urban streams in western Washington, USA.

Pesticides are frequently detected in urban streams, with concentrations often exceeding those reported in surface waters within agricultural areas. The authors studied growth, survival, and return rates of coho salmon (Oncorhynchus kisutch) smolts exposed to a pesticide mixture ("cocktail") representative of the pesticides most frequently reported within urban streams in western Washington State, USA, in fall through early spring. Exposure concentrations were selected to represent a reasonable worst-case scenario based on field monitoring data. Smolts were continuously exposed to pulses of the cocktail either from fertilization through swim-up (2007-2008) or from fertilization through smoltification (2007-2008 and 2008-2009), coded wire tagged, and released in 2008 and 2009. Pre-release endpoints (growth, survival, sex ratio, brain acetylcholinesterase activity, and gonado- and hepatosomatic indices) were not affected. However, the number of returning adults exposed to the cocktail to swim-up (0.90%, n = 42) was more than double that of unexposed controls (0.38%, n = 26) in 2008, whereas in 2009, fish exposed through smoltification returned in lower numbers (0.15%, n = 18) than controls (0.37%, n = 30). Variability in return rates among treatments between years was comparable to that observed in previous whole life cycle studies with Pacific salmon and other contaminants. Results suggest that exposure to pesticides in urban streams does not directly impair early life stages of coho salmon, and that additional studies incorporating releases of larger numbers of smolts across several years are necessary to adequately quantify effects on return rates.

Brain acetylcholinesterase activity in shiner perch (Cymatogaster aggregata) and juvenile chinook salmon (Oncorhynchus tshawytscha) after application of carbaryl to control burrowing shrimp within Willapa Bay, Washington.

Carbaryl has been applied in Willapa Bay, Washington, for five decades to control burrowing shrimp (Neotrypaea californiensis and Upogebia pugettensis) on commercial oyster (Crassostrea gigas) beds. Concerns about effects on nontarget species, including fishes, have led to restrictions in use despite a lack of data on in situ exposure. We measured brain acetylcholinesterase (AChE) activity in adult Shiner perch (Cymatogaster aggregata) and juvenile Chinook salmon (Oncorhynchus tshawytscha) after operational applications. We hypothesized that exposure in Shiner perch would be greater than in juvenile Chinook salmon because of their greater site fidelity and benthic foraging. However, Shiner perch exhibited no statistically significant AChE inhibition. Enzyme activity was statistically decreased (≤14 %) in juvenile Chinook salmon after a second spray event; however, inhibition was less than that associated with overt effects and was similar to controls by 48 h after the spray. Diet analyses confirmed that Shiner perch were primarily feeding on benthic invertebrates and that juvenile Chinook salmon were feeding primarily within the water column. Composition of Shiner perch diets and amount of food consumed varied little among channels and time periods; however, Shiner perch on beds consumed more food 6 h after application than those at other time points and locations. There were no consistent differences in the diets of juvenile Chinook salmon within channels among time periods. Results suggest (1) that carbaryl applications pose little hazard to fish in the bay having habitat and dietary preferences similar to those of Shiner perch and juvenile Chinook salmon and (2) that quantification of direct exposure in the field is essential to adequately assess risk.

Pesticides in urban streams and prespawn mortality of Pacific coho salmon.

The listing of several runs of Pacific salmon as threatened or endangered and associated federal, state, and local efforts to restore/enhance salmon habitat in the Pacific Northwest make it imperative that the factors associated with these population declines are understood. Prespawn mortality (PSM) has been documented in coho salmon (Oncorhynchus kisutch) within urban streams in western Washington since the late 1990s and is characterized by a suite of neurological and respiratory symptoms with mortality occurring shortly thereafter. Mortality rates in returning adults have ranged between 17 and 100%. The cause of PSM is not known, but the presence of pesticide residues within urban streams led to a hypothesis that PSM in coho salmon and pesticides in urban streams were linked. We exposed pairs of "green" (unripe) prespawn male and female coho salmon to a pesticide mixture ("cocktail") reported in urban streams in western Washington State, USA. Longevity, ripening in female salmon, and brain acetylcholinesterase were not significantly affected by continuous exposure to the maximum reported concentrations of the pesticides. Fertilization, hatching success, and growth of fry were also not affected when green adults were exposed to these concentrations for 96 h. The absence of effects suggests it is unlikely that pesticides within stormwater are singularly responsible for PSM in coho salmon or that they impair the reproductive capability of exposed adults.

Pesticides in urban streams and early life stages of Pacific coho salmon.

Pesticides are frequently detected in urban streams and are believed to be primarily the result of homeowner use. Although concentrations in most cases are low (<1 µg/L), there is concern that pesticide inputs threaten efforts to restore and enhance salmon habitat. The authors exposed early life stages of coho salmon (Oncorhynchus kisutch) to a pesticide mixture ("cocktail") representative of those pesticides most frequently reported in urban streams in western Washington State, USA. Life stages were continuously exposed to pulses of the cocktail simulating those in urban streams in fall and winter when coho salmon eggs and sac fry are present. Nominal concentrations of eight herbicides, two insecticides, a fungicide, and a breakdown product were the maximum detected. Fertilization, hatching success, survival, deformities, and growth of fry were not significantly affected. A reduction in fertilization success (19-25%) was not reproducible even when gametes were exposed to 100 times the maximum concentrations detected. Based on the end points examined in the present study, the results suggest that direct exposure to the pesticides most frequently detected in urban streams in western Washington does not impair early life stages of coho salmon and is not a major factor governing the recovery of salmon populations. The extent to which pesticide exposure would affect smoltification, outmigration, and ocean survival needs to be determined.

Effects of the herbicide imazapyr on juvenile Oregon spotted frogs.

Conflict between native amphibians and aquatic weed management in the Pacific Northwest is rarely recognized because most native stillwater-breeding amphibian species move upland during summer, when herbicide application to control weeds in aquatic habitats typically occurs. However, aquatic weed management may pose a risk for aquatic species present in wetlands through the summer, such as the Oregon spotted frog (OSF, Rana pretiosa), a state endangered species in Washington. Acute toxicity of herbicides used to control aquatic weeds tends to be low, but the direct effects of herbicide tank mixes on OSFs have remained unexamined. We exposed juvenile OSFs to tank mixes of the herbicide imazapyr, a surfactant, and a marker dye in a 96-h static-renewal test. The tank mix was chosen because of its low toxicity to fish and its effectiveness in aquatic weed control. Concentrations were those associated with low-volume (3.5 L/ha) and high-volume (7.0 L/ha) applications of imazapyr and a clean-water control. Following exposure, frogs were reared for two months in clean water to identify potential latent effects on growth. Endpoints evaluated included feeding behavior, growth, and body and liver condition indices. We recorded no mortalities and found no significant differences for any end point between the herbicide-exposed and clean-water control frogs. The results suggest that imazapyr use in wetland restoration poses a low risk of direct toxic effects on juvenile OSFs.

Development of a new method for the determination of residues of the neonicotinoid insecticide imidacloprid in juvenile chinook (Oncorhynchus tshawytscha) using ELISA detection.

The neonicotinoid insecticide imidacloprid (IMI) has been proposed as an alternative to carbaryl for controlling indigenous burrowing shrimp on commercial oyster beds in Willapa Bay and Grays Harbor, Washington. A focus of concern over the use of this insecticide in an aquatic environment is the potential for adverse effects from exposure to non-target species residing in the Bay, such as juvenile Chinook (Oncorhynchus tshawytscha) and cutthroat trout (O. clarki). Federal registration and State permiting approval for the use of IMI will require confirmation that the compound does not adversely impact these salmonids following field applications. This will necessitate an environmental monitoring program for evaluating exposure in salmonids following the treatment of beds. Quantification of IMI residues in tissue can be used for determining salmonid exposure to the insecticide. Refinement of an existing protocol using liquid-chromatography mass spectrometry (LC-MS) detection would provide the low limits of quantification, given the relatively small tissue sample sizes, necessary for determining exposure in individual fish. Such an approach would not be viable for the environmental monitoring effort in Willapa Bay and Grays Harbor due to the high costs associated with running multiple analyses, however. A new sample preparation protocol was developed for use with a commercially available enzyme-linked immunosorbent assay (ELISA) for the quantification of IMI, thereby providing a low-cost alternative to LC-MS for environmental monitoring in Willapa Bay and Grays Harbor. Extraction of the analyte from the salmonid brain tissue was achieved by Dounce homogenization in 4.0 mL of 20.0 mM Triton X-100, followed by a 6 h incubation at 50-55 °C. Centrifugal ultrafiltration and reversed phase solid phase extraction were used for sample cleanup. The limit of quantification for an average 77.0 mg whole brain sample was calculated at 18.2 µg kg(-1) (ppb) with an average recovery of 79%. This relatively low limit of quantification allows for the analysis of individual fish. Using controlled laboratory studies, a curvelinear relationship was found between the measured IMI residue concentrations in brain tissue and exposure concentrations in seawater. Additonally, a range of IMI brain residue concentrations was associated with an overt effect; illustrating the utility of the IMI tissue residue quantification approach for linking exposure with defined effects.