Bee Pollinator Mortality Due to Pesticide-Laden Particulate Matter from Beef Cattle Feedyards.

Wild and managed bees are critical for the stability of trophic webs, angiosperm reproduction, and agricultural productivity. Unfortunately, as many as 40% of crop pollinators are in a steep decline due to habitat loss and exposure to agrochemicals. Pyrethroids, neonicotinoids, and macrocyclic lactones are among the many agrochemicals toxic to pollinating insects that are used extensively in industrial beef cattle feeding operations throughout the world. Fugitive feedyard particulate matter (PM) transports agrochemicals into the surrounding environs. To determine the impact of agrochemical-laden feedyard particulate matter on bee pollinators, we conducted in situ experiments wherein honeybees and mason bees were placed downwind and upwind of feedyards (N = 40). Concurrent, colocated total suspended particulate matter samples contained multiple insecticides and parasiticides including pyrethroids, neonicotinoids, and macrocyclic lactones, in significantly higher concentrations downwind of feedyards (bifenthrin, 8.45 ± 4.92; permethrin, 1032.34 ± 740.76; clothianidin, 3.61 ± 1.48; imidacloprid, 73.32 ± 47.52; thiamethoxam, 5.81 ± 3.16; abamectin, 0.45 ± 0.29; ivermectin, 8.88 ± 5.06 ng/g). Honeybees and mason bees sited downwind of feedyards always experienced higher mortality than those correspondingly sited upwind, and male mason bees experienced significantly higher mortality compared to females when both were sited downwind. Bees occurring downwind of beef cattle feedyards for 1 h are 232-260% more likely to die than those occurring upwind. Thus, agrochemicals used on and emitted from beef cattle feedyards are significant threats to bee pollinators.

A novel laboratory method for simulating pollinator exposure to agrochemical-laden particulate matter.

Environmental transport and deposition of particulate matter (PM) associated with toxic chemicals has begun to receive attention as a source of risk to pollinators. For example, dust arising from manipulations of insecticide-treated seed has potential to exert toxic effects among non-target insects. Similarly, synthetic steroid growth promoters, antibiotics and multiple insecticides and parasiticides detected in fugitive beef cattle feedyard PM may also negatively impact pollinators since many of these chemicals have been detected on wildflowers and pollinators collected near beef cattle feedyards. Therefore, there is a need to assess risk to pollinators posed by deposition of agrochemical-laden PM, both in the field and the laboratory. Unfortunately, established laboratory methods for simulating PM exposure or toxicity associated with contaminated PM are few and highly situation-specific. Herein we describe development and use of a PM circulation system that can be employed to evaluate toxicity of agrochemical-contaminated PM in the laboratory under controlled conditions. Two model organisms (honeybees (Apis mellifera) and mason bees (Osmia lignaria)) were exposed to agrochemical-free PM in the circulator system, and post-exposure mortality was compared with controls. No significant differences in mortality between exposed and control bees were observed. Next, honeybees and mason bees were exposed to PM spiked with an insecticide known to exert toxic effects to pollinators (thiamethoxam). Bees experienced significantly higher mortality when exposed to thiamethoxam-laden PM at environmentally relevant concentrations as compared to bees exposed to agrochemical-free PM. These results confirm the validity of these methods for use in controlled laboratory PM toxicity tests and offer a source of positive and negative control groups for laboratory and field experiments examining exposure of pollinators to potentially toxic agrochemical-laden PM. This method facilitates generation of more realistic toxicity data than standard contact toxicity tests when pollinator exposure scenarios involve particulate-based agrochemicals or other toxic chemicals.

Assessment of risks to listed species from the use of atrazine in the USA: a perspective.

Atrazine is a triazine herbicide used predominantly on corn, sorghum, and sugarcane in the US. Its use potentially overlaps with the ranges of listed (threatened and endangered) species. In response to registration review in the context of the Endangered Species Act, we evaluated potential direct and indirect impacts of atrazine on listed species and designated critical habitats. Atrazine has been widely studied, extensive environmental monitoring and toxicity data sets are available, and the spatial and temporal uses on major crops are well characterized. Ranges of listed species are less well-defined, resulting in overly conservative designations of "May Effect". Preferences for habitat and food sources serve to limit exposure among many listed animal species and animals are relatively insensitive. Atrazine does not bioaccumulate, further diminishing exposures among consumers and predators. Because of incomplete exposure pathways, many species can be eliminated from consideration for direct effects. It is toxic to plants, but even sensitive plants tolerate episodic exposures, such as those occurring in flowing waters. Empirical data from long-term monitoring programs and realistic field data on off-target deposition of drift indicate that many other listed species can be removed from consideration because exposures are below conservative toxicity thresholds for direct and indirect effects. Combined with recent mitigation actions by the registrant, this review serves to refine and focus forthcoming listed species assessment efforts for atrazine.Abbreviations: a.i. = Active ingredient (of a pesticide product). AEMP = Atrazine Ecological Monitoring Program. AIMS = Avian Incident Monitoring SystemArach. = Arachnid (spiders and mites). AUC = Area Under the Curve. BE = Biological Evaluation (of potential effects on listed species). BO = Biological Opinion (conclusion of the consultation between USEPA and the Services with respect to potential effects in listed species). CASM = Comprehensive Aquatic System Model. CDL = Crop Data LayerCN = field Curve Number. CRP = Conservation Reserve Program (lands). CTA = Conditioned Taste Avoidance. DAC = Diaminochlorotriazine (a metabolite of atrazine, also known by the acronym DACT). DER = Data Evaluation Record. EC25 = Concentration causing a specified effect in 25% of the tested organisms. EC50 = Concentration causing a specified effect in 50% of the tested organisms. EC50RGR = Concentration causing a 50% reduction in relative growth rate. ECOS = Environmental Conservation Online System. EDD = Estimated Daily Dose. EEC = Expected Environmental Concentration. EFED = Environmental Fate and Effects Division (of the USEPA). EFSA = European Food Safety Agency. EIIS = Ecological Incident Information System. ERA = Environmental Risk Assessment. ESA = Endangered Species Act. ESU = Evolutionarily Significant UnitsFAR = Field Application RateFIFRA = Federal Insecticide, Fungicide, and Rodenticide Act. FOIA = Freedom of Information Act (request). GSD = Genus Sensitivity Distribution. HC5 = Hazardous Concentration for ≤ 5% of species. HUC = Hydrologic Unit Code. IBM = Individual-Based Model. IDS = Incident Data System. KOC = Partition coefficient between water and organic matter in soil or sediment. KOW = Octanol-Water partition coefficient. LC50 = Concentration lethal to 50% of the tested organisms. LC-MS-MS = Liquid Chromatograph with Tandem Mass Spectrometry. LD50 = Dose lethal to 50% of the tested organisms. LAA = Likely to Adversely Affect. LOAEC = Lowest-Observed-Adverse-Effect Concentration. LOC = Level of Concern. MA = May Affect. MATC = Maximum Acceptable Toxicant Concentration. NAS = National Academy of Sciences. NCWQR = National Center of Water Quality Research. NE = No Effect. NLAA = Not Likely to Adversely Affect. NMFS = National Marine Fisheries Service. NOAA = National Oceanic and Atmospheric Administration. NOAEC = No-Observed-Adverse-Effect Concentration. NOAEL = No-Observed-Adverse-Effect Dose-Level. OECD = Organization of Economic Cooperation and Development. PNSP = Pesticide National Synthesis Project. PQ = Plastoquinone. PRZM = Pesticide Root Zone Model. PWC = Pesticide in Water Calculator. QWoE = Quantitative Weight of Evidence. RGR = Relative growth rate (of plants). RQ = Risk Quotient. RUD = Residue Unit Doses. SAP = Science Advisory Panel (of the USEPA). SGR = Specific Growth Rate. SI = Supplemental Information. SSD = Species Sensitivity Distribution. SURLAG = Surface Runoff Lag Coefficient. SWAT = Soil & Water Assessment Tool. SWCC = Surface Water Concentration Calculator. UDL = Use Data Layer (for pesticides). USDA = United States Department of Agriculture. USEPA = United States Environmental Protection Agency. USFWS = United States Fish and Wildlife Service. USGS = United States Geological Survey. WARP = Watershed Regressions for Pesticides.

Toxic responses of blue orchard mason bees (Osmia lignaria) following contact exposure to neonicotinoids, macrocyclic lactones, and pyrethroids.

Analysis of particulate matter originating from beef cattle feed yards on the High Plains of the United States has revealed occurrence of multiple pesticides believed to potentially impact non-Apis pollinators. Among these pesticides are those that are highly toxic to Apis mellifera (honey bees). However, little non-Apis bee species toxicity data exist; especially pertaining to beef cattle feed yard-derived pesticides. Therefore, we conducted a series of 96-h contact toxicity tests with blue orchard mason bees (Osmia lignaria) using three neonicotinoids, two pyrethroids, and two macrocyclic lactones. Neonicotinoids (thiamethoxam, imidacloprid, and clothianidin) were most toxic with LD50 values ranging from 2.88 to 26.35 ng/bee, respectively. Macrocyclic lactones (abamectin and ivermectin) were also highly toxic to O. lignaria with LD50 estimates of 5.51-32.86 ng/bee. Pyrethroids (permethrin and bifenthrin) were relatively less toxic with LD50 values greater than 33 ng/bee. Sensitivity ratios for each pesticide were calculated to relate O. lignaria LD50 values to existing honey bee toxicity data. All three neonicotinoids were more toxic to O. lignaria than A. mellifera, but pyrethroids and abamectin were relatively less toxic. Additionally, three of seven pesticides (43%) resulted in significantly different mass normalized LD50 values for male and female O. lignaria. These results indicate that non-Apis pollinators may be highly susceptible to pesticides originating from beef cattle feed yards, necessitating consideration of more stringent regulatory protections than those based on A. mellifera pesticide sensitivity.

Pesticides Used on Beef Cattle Feed Yards Are Aerially Transported into the Environment Via Particulate Matter.

Considering the recent discovery of veterinary pharmaceutical aerial transport from industrial cattle feeding operations via particulate matter, the objective of this study is to determine the extent to which insecticides are also transported into the environment by total suspended particulates emanating from beef cattle feed yards. Of 16 different pesticides quantified in particulate matter samples collected from beef cattle feed yards, permethrin was detected most frequently at >67% of particulate matter samples and at a mean concentration of 1211.7 ± 781.0 (SE) ng/m3. Imidacloprid was detected at a mean concentration of 62.8 ± 38.2 (SE) ng/m3 or equivalent to published concentrations in dust from treated seed planting activities. When insecticide concentrations observed in this study are projected to all United States of America feed yards, the resulting particulate matter (669,000 kg) could contain enough insecticides (active ingredient mass basis) to kill over a billion honeybees daily. Furthermore, a novel transport pathway for macrocyclic lactone entry into the environment was identified. These data raise concern that nontarget organisms may be exposed to potentially toxic levels of pesticides from beef cattle feed yards.

Occurrence and Characterization of Steroid Growth Promoters Associated with Particulate Matter Originating from Beef Cattle Feedyards.

Studies of steroid growth promoters from beef cattle feedyards have previously focused on effluent or surface runoff as the primary route of transport from animal feeding operations. There is potential for steroid transport via fugitive airborne particulate matter (PM) from cattle feedyards; therefore, the objective of this study was to characterize the occurrence and concentration of steroid growth promoters in PM from feedyards. Air sampling was conducted at commercial feedyards (n = 5) across the Southern Great Plains from 2010 to 2012. Total suspended particulates (TSP), PM10, and PM2.5 were collected for particle size analysis and steroid growth promoter analysis. Particle size distributions were generated from TSP samples only, while steroid analysis was conducted on extracts of PM samples using liquid chromatography mass spectrometry. Of seven targeted steroids, 17α-estradiol and estrone were the most commonly detected, identified in over 94% of samples at median concentrations of 20.6 and 10.8 ng/g, respectively. Melengestrol acetate and 17α-trenbolone were detected in 31% and 39% of all PM samples at median concentrations of 1.3 and 1.9 ng/g, respectively. Results demonstrate PM is a viable route of steroid transportation and may be a significant contributor to environmental steroid hormone loading from cattle feedyards.

Laboratory Determination of Particulate-Matter-Bound Agrochemical Toxicity among Honeybees, Mason Bees, and Painted Lady Butterflies.

Pollinator population declines are global phenomena with severe consequences for native flora and agriculture. Many factors have contributed to pollinator declines including habitat loss, climate change, disease and parasitism, reductions in abundance and diversity of foraging resources, and agrochemical exposure. Particulate matter (PM) serves as a carrier of toxic agrochemicals, and pollinator mortality can occur following exposure to agrochemical-contaminated PM. Therefore, laboratory-controlled experiments were conducted to evaluate impacts of individual PM-bound agrochemicals. Honeybees (Apis mellifera), blue orchard mason bees (Osmia lignaria), and painted lady butterfly (Vanessa cardui) larvae were exposed to bifenthrin, permethrin, clothianidin, imidacloprid, abamectin, and ivermectin via suspended, airborne PM. Agrochemical concentrations in PM to which pollinators were exposed were based on concentrations observed in fugitive beef cattle feedyard PM including a "mean" treatment and a "max" treatment reflective of reported mean and maximum PM-bound agrochemical concentrations, respectively. In general, pollinators in the mean and max treatments experienced significantly higher mortality compared with controls. Honeybees were most sensitive to pyrethroids, mason bees were most sensitive to neonicotinoids, and painted lady butterfly larvae were most sensitive to macrocyclic lactones. Overall, pollinator mortality was quite low relative to established toxic effect levels derived from traditional pollinator contact toxicity tests. Furthermore, pollinator mortality resulting from exposure to individual agrochemicals via PM was less than that reported to occur at beef cattle feedyards, highlighting the importance of mixture toxicity to native and managed pollinator survival and conservation. Environ Toxicol Chem 2023;42:2642-2650. © 2023 SETAC.

Atmospheric transport of particulate matter and particulate-bound agrochemicals from beef cattle feedlots: Human health implications for downwind agricultural communities.

Beef cattle feedlot particulate matter (PM) is a complex mixture of dust, animal waste, agrochemicals, and bioaerosols. However, no empirical data exist quantifying downwind residential exposure to PM or PM-bound agrochemicals. In the current study, authors investigated transport of PM and co-occurring insecticides and anthelmintics downwind of three feedlots in the Southern Great Plains (SGP) of North America from May-August 2022. PM collected on total suspended particulate (TSP) filters was analyzed via UHPLC-MS/MS for six pyrethroids and five macrocyclic lactones (MLs). Downwind TSP concentrations rapidly declined from 0.01 to ≤1.6 km (Monte Carlo mean ± SEM; 5049 ± 96.1 µg/m3) and stabilized >1.6-12.4 km (1791 ± 9.9; µg/m3). Distance decay >1.6 km indicated downwind PM exceeded levels of safe human exposure during diurnal peak periods. Pyrethroids and MLs were detected >LOQ in 96.2 and 98.1 % of downwind samples. Screening-level cumulative residential exposure indicates elevated pyrethroid risk (LOC = 1; RI = 0.173) to rural children (1-2 yrs) living near cropland operations in the SGP, with disproportionate co-exposure to feedlot PM and legacy pollution in low-income, Hispanic and Latino communities. Frequent occurrence and sustained transport of pyrethroids downwind of feedlots facilitate residue accumulation in outdoor residential areas that must also be quantified to assess the magnitude of daily average and lifetime-adjusted oral and dermal exposure in surrounding communities. Results significantly expand the known distribution of feedlot-derived PM and agrochemicals and highlight human exposure pathways unrecognized in residential human health assessments supporting pesticide registration and feedlot risk evaluation.

Atmospheric deposition of particulate matter from beef cattle feedlots is a likely contributor of pyrethroid occurrence in isolated wetland sediment: Source apportionment and ecological risk assessment.

Industrial cattle feeding operations (feedlots) have been subject to public scrutiny in recent decades regarding environmental impacts of site runoff and aerial dispersion of agrochemical-laden particulate matter (PM). However, source apportionment of multi-use pesticides is challenging in mixed agricultural settings. Beef cattle on feed and row crop production are heavily concentrated in the Southern Great Plains of North America, where playa wetlands are vulnerable to agrochemical inputs and sedimentation from surrounding land use. In the current study, playa basin sediment (n = 33) was analyzed via UHPLC-MS for 21 agrochemicals spanning eight classes (macrocyclic lactones, neonicotinoids, organophosphates, pyrethroids, triazoles, ß-methoxyacrylates, a carboximide, and phenylpyrazole). Pyrethroids were detected most frequently (75.8% of basins). Sediment pyrethroid concentrations were also significantly correlated (R2 = 0.178, p = 0.007) with feedlot proximity (<1-50 km). Principal component analysis (PCA) of land use metrics extracted three principal components (74.3% of total variance), with principal component regression (PCR) showing the greatest agrochemical occurrence in basins heavily weighted by cropland buffer acreage (≤1 km) and feedlot proximity. Sediment toxicity benchmarks protective of two benthic invertebrates (Hyallela azteca and Chironomus spp.) identified λ-cyhalothrin, fenvalerate, and esfenvalerate as individual compounds exceeding levels of acute (RQ > 0.5) and chronic (RQ > 1) concern in >5% and >50% of cases, respectively. However, additive toxicity of co-occurring pyrethroids represents an acute high risk (RI > 1; median RI; acute = 2.4, chronic = 38.6) to benthic invertebrates in >75% of cases, which may threaten higher-order wetland taxa via bioaccumulation and trophic transfer.

Embryotoxicity of Corexit 9500 in mallard ducks (Anas platyrhynchos).

Embryotoxicity of the oil dispersant Corexit 9500 was examined using fertilized mallard duck eggs. Corexit 9500 was topically applied below the air cell to eggs in volumes ranging from 0 to 100 µL on day 3 of incubation. The highest incidence of mortality occurred at developmental stage 4, one day post-Corexit 9500 application. Hatching success was significantly decreased among eggs treated with ≥ 20 µL of Corexit 9500 as compared to controls (P ≤ 0.047). No egg treated with ≥ 40 µL successfully hatched. The application volume resulting in 50% mortality (corrected for control survival) was determined to be 15.5 µL. Developmental stage at embryo death was also significantly decreased compared to controls in eggs exposed to 40 µL (P = 0.0042) and above.

Agrochemical occurrence on colocated wildflowers and wild bees collected near beef cattle feed yards and row crops.

It is well established that agrochemicals can pose significant threats to native pollinators; however, relatively little is known about pollinator risks associated with agrochemicals that are used on beef cattle feed yards. Recently, feed yard-derived agrochemicals and those from row crop agriculture were quantified on wildflowers growing on the High Plains, USA. To better characterize pollinator risks on the High Plains, we collected colocated wildflowers and foraging bees across three field seasons for analytical determination of residual agrochemicals. Agrochemicals were detected and quantified on the majority of wildflowers (85%) and nearly half of bees (49%). Permethrin was the most frequently detected analyte on wildflowers (32%) and bees (17%). Flower hazard quotients and flower hazard indices were calculated to deterministically evaluate risk to foraging pollinators. Mean flower hazard quotients exceeded one for 5/16 analytes (31%), and flower hazard quotients calculated for 30% of wildflowers were greater than 50. Flower hazard quotients for clothianidin exceeded 400 for 14% of wildflowers, which portends conditions conducive to frequent bee mortalities. Flower hazard indices were greater on wildflowers from mid-July to mid-September as compared with wildflowers collected earlier in the summer, which coincides with row crop planting and increased prevalence of feed yard flies. Hazard quotients and hazard index values calculated from agrochemical residue data suggest that pollinators frequenting wildflowers near beef cattle feed yards and row crops on the High Plains are at risk from both individual sources, and more so when considered in combination. Integr Environ Assess Manag 2022;18:163-173. © 2021 SETAC.

Use of nest bundles to monitor agrochemical exposure and effects among cavity nesting pollinators.

Cavity nesting bees are proficient and important pollinators that can augment or replace honey bee pollination services for some crops. Relatively little is known about specific pesticide concentrations present in cavity nesting insect reed matrices and associated potential risks to cavity nesting bees. Nesting substrates (Phragmites australis reeds in bundles) were deployed in an agriculturally intensive landscape to evaluate colonization and agrochemical exposure among cavity nesting pollinators over two consecutive field seasons. Composition of insect species colonizing reeds within nest bundles varied considerably; those placed near beef cattle feed yards were dominated by wasps (93% of the total number of individuals occupying reed nest bundles), whereas nest bundles deployed in cropland-dominated landscapes were colonized primarily by leaf cutter bees (71%). All nesting/brood matrices in reeds (mud, leaves, brood, pollen) contained agrochemicals. Mud used in brood chamber construction at feed yard sites contained 21 of 23 agrochemicals included in analysis and >70% of leaf substrate stored in reeds contained at least one agrochemical. Moxidectin was most frequently detected across all reed matrices from feed yard sites, and moxidectin concentrations in nonviable larvae were more than four times higher than those quantified in viable larvae. Agrochemical concentrations in leaf material and pollen were also quantified at levels that may have induced toxic effects among developing larvae. To our knowledge, this is the first study to characterize agrochemical concentrations in multiple reed matrices provisioned by cavity-nesting insects. Use of nest bundles revealed that cavity nesting pollinators in agriculturally intensive regions are exposed to agrochemicals during all life stages, at relatively high frequencies, and at potentially lethal concentrations. These results demonstrate the utility of nest bundles for characterizing risks to cavity nesting insects inhabiting agriculturally intensive regions.

Effects of polycyclic aromatic hydrocarbons in northern bobwhite quail (Colinus virginianus).

Polycyclic aromatic hydrocarbons (PAH) are ubiquitous contaminants of aquatic and terrestrial ecosystems, and are known to induce biochemical alterations in exposed organisms. Aside from a variety of adverse physiological effects associated with exposure to petroleum products, oils, and oil sludges, little is known about the effects of individual PAH on birds. Acute toxicity of naphthalene, pyrene, and benz[a]anthracene (BAA) was examined in adult northern bobwhite quail (Colinus virginianus). Additionally, subacute (8 d) and subchronic (60 d) studies were conducted to assess alterations in metabolic enzyme activity. Neither naphthalene, nor pyrene, nor BAA exposure via oral gavage produced acute toxicity up to the limit dose of 2 g/kg body weight. In the subacute study, quail provided feed containing the highest concentration of BAA for 5 d had significantly increased renal ethoxyresorufin O-deeththylase (EROD) activity compared to controls. Following a 3-d recovery period, significant increases between 10 and 100 mg/kg of BAA in feed existed for both hepatic EROD and pentoxyresorufin O-deethylase (PROD) activity compared to controls. Subchronic exposure to BAA (ranging from 0.1 to 10 mg/kg) also resulted in a significant rise of EROD and PROD in both kidney and liver tissue compared to controls. Though the individual PAH used in this study were not acutely toxic, these results confirm that these individual PAH induce alterations in metabolic enzyme activity in northern bobwhite quail.

Exposure of Foraging Bees (Hymenoptera) to Neonicotinoids in the U.S. Southern High Plains.

Exposure to pesticides is a major threat to insect pollinators, potentially leading to negative effects that could compromise pollination services and biodiversity. The objectives of this study were to quantify neonicotinoid concentrations among different bee genera and to examine differences attributable to body size and surrounding land use. During the period of cotton planting (May-June), 282 wild bees were collected from habitat patches associated with cropland, grassland, and urban land cover and analyzed for three neonicotinoids (thiamethoxam, clothianidin, and imidacloprid). Twenty bees among eight genera contained one or more of the neonicotinoid compounds and detections occurred in all landscape types, yet with the most detections occurring in cropland-associated habitats. Apis Linnaeus (Hymenoptera: Apidae), Melissodes Latreille (Apidae), Perdita Smith (Andrenidae), and Lasioglossum Curtis (Halictidae) had multiple individuals with neonicotinoid detections. Two of the largest bees (Apis and Melissodes) had the greatest number of detections within genera, yet the relatively small-bodied genus Perdita had the three highest neonicotinoid concentrations reported. The number of detections within a genus and average generic body mass showed a marginally significant trend towards larger bees having a greater frequency of neonicotinoid detections. Overall, the relatively low percentage of detections across taxa suggests practices aimed at conserving grassland remnants in intensified agricultural regions could assist in mitigating exposure of wild bees to agrochemicals, while differences in bee traits and resource use could in part drive exposure. Further work is needed to address variable agrochemical exposures among pollinators, to support strategies for conservation and habitat restoration in affected landscapes.

Toxicity of a glufosinate- and several glyphosate-based herbicides to juvenile amphibians from the Southern High Plains, USA.

Pesticide toxicity is often proposed as a contributing factor to the world-wide decline of amphibian populations. We assessed acute toxicity (48 h) of a glufosinate-based herbicide (Ignite 280 SL) and several glyphosate-based herbicide formulations (Roundup WeatherMAX, Roundup Weed and Grass Killer Super Concentrate, Roundup Weed and Grass Killer Ready-To-Use Plus on two species of amphibians housed on soil or moist paper towels. Survival of juvenile Great Plains toads (Bufo cognatus) and New Mexico spadefoots (Spea multiplicata) was reduced by exposure to Roundup Weed and Grass Killer Ready-To-Use Plus on both substrates. Great Plains toad survival was also reduced by exposure to Roundup Weed and Grass Killer Super Concentrate on paper towels. New Mexico spadefoot and Great Plains toad survival was not affected by exposure to the two agricultural herbicides (Roundup WeatherMAX and Ignite 280 SL) on either substrate, suggesting that these herbicides likely do not pose an immediate risk to these species under field conditions.

Development of resistance to cyfluthrin and naphthalene among Daphnia magna.

In this study, Daphnia magna were exposed to a pyrethroid insecticide (cyfluthrin) or a polycyclic aromatic hydrocarbon (naphthalene) for 12 generations to evaluate development of resistance followed by a 12 generation recovery period. Twenty-four hour old D. magna were exposed to concentrations of each chemical resulting in 50-70% mortality to select for the least sensitive individuals. LC50 values, survival, reproductive output, and time to first brood in stressor-exposed and control D. magna were recorded for each generation. Significant changes in LC50 values were observed after 4 generations and then declined after 6-10 generations post-exposure. D. magna were 5 times less sensitive to cyfluthrin and 3 times less sensitive to naphthalene as compared to controls after 12 generations of exposure. There were no differences in survival, time to first brood, or total number of offspring produced between control and either of the resistant F13 D. magna. Cyfluthrin exposed D. magna exhibited cross-resistance to DDT and methyl parathion, and naphthalene resistant D. magna were less sensitive than controls to both pyrene and benz(a)anthracene. When the cytochrome P450 inhibitor piperonyl butoxide was used in conjunction with cyfluthrin and naphthalene the sensitivity of resistant and control D. magna were equal, suggesting P450s were responsible for conveying resistance. This study demonstrates that life history and organisms' capacity to develop resistance is important to consider ensuring accuracy of ecological risk assessments.

Toxicity of Agrochemicals Among Larval Painted Lady Butterflies (Vanessa cardui).

In the Southern High Plains of the United States, beef cattle feed yards and row crop agriculture are predominant sources of agrochemical usage. Beef cattle feed yards use large quantities of veterinary pharmaceuticals to promote cattle growth and health, along with insecticides to control insect pests, whereas row crop-based agriculture relies on herbicides, fungicides, and insecticides to increase yields. Previous studies have documented the occurrence of agrochemicals beyond feed yard and row crop agriculture boundaries in uncultivated, marginal areas, raising concern that migratory pollinators and pollinators indigenous to the Southern High Plains frequenting these remaining habitat corridors may become exposed to toxic agrochemicals. Larvae of the painted lady butterfly (Vanessa cardui) were used to investigate the potential toxicity of agrochemicals used on feed yards and in row crop agriculture among pollinators. Moxidectin, an antiparasiticide used on beef cattle feed yards, was determined to be extremely toxic to V. cardui larvae, with a lethal dose at which 50% of larvae died of 2.1 ± 0.1 ng/g. Pyraclostrobin, clothianidin, and permethrin all delayed V. cardui development. However, moxidectin was the only chemical that produced significant toxic effects at environmentally relevant concentrations. These results indicate that agrochemicals originating from feed yards have the potential to adversely impact the development of pollinator larvae occurring in the Southern High Plains. Environ Toxicol Chem 2019;38:2629-2636. © 2019 SETAC.

Persistence of elevated concentrations of PM, affiliated pharmaceuticals, and tetracycline resistance genes downwind of feedyards.

Beef cattle feedyards have been identified as sources of large amounts of particulate matter (PM) which may transport affiliated chemicals including steroids, beta agonists, and antibiotics from feedyards into the environment. This study is the first to examine persistence of PM-affiliated pharmaceuticals downwind of feedyards using multiple downwind samples collected at increasing distances from feedyard boundaries (n = 5). Concentrations of antibiotics and ractopamine per gram of PM remained consistent at all downwind locations (out to 4.8 km) whereas concentrations per m3 air decreased significantly at distances between 0.1 and 0.7 km downwind, corresponding to significant decreases in mass of PM. Monensin was present in the highest concentrations of any measured pharmaceutical, with concentrations of 37 µg/g PM (376 ng/m3) air in samples collected within 0.1 km downwind of feedyards. Total copy count of tetracycline resistance genes (tetW, tetQ, tetO, tetM, tetL, and tetB) were also significantly increased in samples collected within 0.1 km downwind of feedyards (106 copies) as compared to samples collected upwind (103 copies) and farther downwind (104 copies) of feedyard boundaries. These results suggest that transport of pharmaceutical-laden PM into the terrestrial environment is occurring primarily via PM deposition within 0.7 km of the feedyard, while aerial transport persists over longer distances (>4.8 km).

Plasma vitellogenin in Morelet's crocodiles from contaminated habitats in northern Belize.

Vitellogenin induction has been widely used as a biomarker of endocrine disruption in wildlife, but few studies have investigated its use in wild reptiles living in contaminated habitats. This study examined vitellogenin induction in Morelet's crocodiles (Crocodylus moreletii) from wetlands in northern Belize contaminated with organochlorine (OC) pesticides. Vitellogenin was measured in 381 crocodile plasma samples using a vitellogenin ELISA previously developed for this species. Vitellogenin was detected in nine samples, all from adult females sampled during the breeding season. Males and juvenile females did not contain detectable levels of vitellogenin; however, many of these animals contained OC pesticides in their caudal scutes, confirming contaminant exposure. The lack of a vitellogenic response in these animals may be attributable to several factors related to the timing and magnitude of exposure to endocrine-disrupting chemicals and should not be interpreted as an absence of other contaminant-induced biological responses.

Effects of HMX exposure upon metabolic rate of northern bobwhite quail (Colinus virginianus) in ovo.

We evaluated the use of the gas exchange rate as an ecologically relevant indicator of chemical stress in avian embryos/eggs. Northern bobwhite quail (Colinus virginianus) were exposed to octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) via feed containing nominal concentrations of 0, 12.5, 50.0, and 125.0 mg kg(-1). Metabolic rates (oxygen consumption) of developing quail eggs were then measured via respirometry to examine potential effects of HMX exposure. Metabolic rates were examined on 5, 9, and 21 d of incubation. Next, concentrations of HMX in embryos/eggs were determined by liquid chromatography-mass spectrometry. Mean (+/-SE) concentrations of HMX in eggs were 21.0+/-5.9, 1113+/-79.0, 3864+/-154.0, and 7426+/-301.1 ng g(-1) in control, low, medium and high dose groups, respectively. There were significant differences in oxygen consumption among the three embryo ages, however differences among the ages were not consistent among dose groups (age x dose group interaction p<0.0001). Oxygen consumption rates did not vary as a function of HMX in embryos (p=0.18). No evidence was observed for alterations of in ovo metabolic rates associated with HMX exposure.