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.

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.

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.

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.

Transferrin receptor 2: evidence for ligand-induced stabilization and redirection to a recycling pathway.

Transferrin receptor 2 (TfR2) is a homologue of transferrin receptor 1 (TfR1), the protein that delivers iron to cells through receptor-mediated endocytosis of diferric transferrin (Fe(2)Tf). TfR2 also binds Fe(2)Tf, but it seems to function primarily in the regulation of systemic iron homeostasis. In contrast to TfR1, the trafficking of TfR2 within the cell has not been extensively characterized. Previously, we showed that Fe(2)Tf increases TfR2 stability, suggesting that trafficking of TfR2 may be regulated by interaction with its ligand. In the present study, therefore, we sought to identify the mode of TfR2 degradation, to characterize TfR2 trafficking, and to determine how Fe(2)Tf stabilizes TfR2. Stabilization of TfR2 by bafilomycin implies that TfR2 traffics to the lysosome for degradation. Confocal microscopy reveals that treatment of cells with Fe(2)Tf increases the fraction of TfR2 localizing to recycling endosomes and decreases the fraction of TfR2 localizing to late endosomes. Mutational analysis of TfR2 shows that the mutation G679A, which blocks TfR2 binding to Fe(2)Tf, increases the rate of receptor turnover and prevents stabilization by Fe(2)Tf, indicating a direct role of Fe(2)Tf in TfR2 stabilization. The mutation Y23A in the cytoplasmic domain of TfR2 inhibits its internalization and degradation, implicating YQRV as an endocytic motif.

Manual small incision cataract surgery in a United Kingdom university teaching hospital setting.

PURPOSE: To assess the complication rates and visual outcomes of manual small incision cataract surgery (MSICS) in a university teaching hospital setting. METHOD: Retrospective audit encompassing a 3-year period looking at the visual outcomes, and intraoperative and postoperative complications of planned MSICS performed on brunescent cataracts. RESULTS: MSICS was performed in 55 eyes of 49 patients. Intraoperatively, zonular dehiscence without vitreous loss occurred in one eye (1.8%), and vitreous loss occurred in one eye (1.8%). Postoperative complications occurred in seven eyes (12.7%). The most common complication was raised intraocular pressure (three eyes, 5.5%). None developed endophthalmitis or dropped nucleus. Mean follow-up duration was 10.0 weeks. In 49 eyes (89.1%), vision improved postoperatively. Sixty five percent achieved a best-corrected vision of 6/12 or better. The mean postoperative astigmatism was significantly higher than preoperatively (1.40D vs. 0.99D, p = 0.02). CONCLUSION: MSICS is safe and effective for dense and brunescent cataracts in a UK university hospital setting. However, it was associated with a statistically significant increase in astigmatism postoperatively.

Increased temperature and lower resource quality exacerbate chloride toxicity to larval Lithobates sylvaticus (wood frog).

A chemical contaminant of growing concern to freshwater aquatic organisms, including many amphibians, is chloride ion. The salinization of freshwater ecosystems is likely caused, in part, by the application of massive amounts of road de-icing salts to roadways during winter months. The issue of freshwater salinization has become the subject of many toxicity studies and is often investigated in conjunction with other chemical stressors. However, few published studies attempt to investigate the interactions of elevated chloride concentration and increased temperature. Further, no studies have investigated the gap between the recommended feeding conditions typically used in standard toxicity tests and those that may exist in natural amphibian habitats. This study addressed the critical issues of elevated chloride, increased temperature, and variation in food quality. We conducted a 96-h acute toxicity test to investigate acute chloride toxicity as impacted by different diets, as well as a chronic toxicity test to investigate the impacts of chloride, temperature, and resource quality on the survival and development of larval Lithobates sylvaticus (wood frogs). Chloride LC50s ± 1 SE were 3769.22 ± 589.05, 2133.00 ± 185.95, and 2644.69 ± 209.73 mg Cl-/L were for non-fed, low-protein diet, and high-protein diet, respectively. For the chronic toxicity study, elevated chloride decreased tadpole survival. Increased temperature, and lower resource quality, were found negatively impacted survival of tadpoles and altered time-to-metamorphosis. This study shows that environmentally relevant concentrations of chloride, temperatures, and the protein content of the diet all exert critical effects on larval wood frogs.

Will temperature increases associated with climate change potentiate toxicity of environmentally relevant concentrations of chloride on larval green frogs (Lithobates clamitans)?

An important challenge in amphibian ecotoxicology and conservation is that amphibian toxicity tests are usually focused on a single chemical while populations experience multiple, simultaneous stressors. For example, about 14 million tons of road de-icing salts are used each year in North America with NaCl accounting for 98% of total salt use and, hence, elevated chloride is an important environmental stressor to aquatic organisms, including amphibians. As well, higher temperature as a result of climate change is becoming an increasingly important environmental stressor. There are no data on the combined effects of chloride and temperature on amphibians hinders conservation efforts. We conducted field studies to characterize chloride concentrations and water temperatures in known amphibian breeding habitats and performed toxicity tests to explore impacts of these two stressors on a common anuran, the green frog (Lithobates clamitans). A 96-hour acute toxicity test was conducted to first determine a chloride LC50 (2587.5 mg Cl-/L) at a single, neutral temperature, which was used to inform the treatment levels of the sub-chronic test, which also included a temperature range. In the sub-chronic study, green frog larvae were exposed to three temperatures (18, 22, and 25 °C), and four concentrations of chloride (0, 500, 1000, and 2000 mg Cl-/L) for 35 days. At all temperatures, tadpoles exposed to 2000 mg Cl-/L had significantly higher mortality. While there was no significant effect of temperature alone on mortality, survival of tadpoles was significantly lower at 1000 mg Cl-/L at the two higher temperatures suggesting a potentiation of chloride ion toxicity with increasing temperature. Comparing toxicity results to field measurements of chloride and temperature suggests green frog tadpoles and other species with similar sensitivity are likely negatively affected. Data on additional species and populations would further increase our understanding of how salt and temperature may shape aquatic communities.

Association of human transferrin receptor with GABARAP.

A yeast two-hybrid screen identified a specific interaction between the cytoplasmic domain of transferrin receptor (TfR) and GABARAP, a 14 kDa protein that binds to the gamma2 subunit of neuronal GABA(A) receptors. The specificity of the TfR-GABARAP interaction was demonstrated by in vitro binding assays with purified proteins and by co-immunoprecipitation of GABARAP with endogenous TfR from HeLa cell lysates. Replacement of the YTRF internalization motif with ATRA within the cytoplasmic domain of TfR reduced interaction with GABARAP in the yeast two-hybrid screen and in vitro binding assays. The intracellular location of GABARAP using chimeric GABARAP-GFP showed that the majority of GABARAP was located in perinuclear vesicles. Our results show that GABARAP plays a more general role outside the confines of neuronal cells and GABA(A) receptors.