Utility of the avian sub-acute dietary toxicity test in ecological risk assessment and a path forward to reduce animal use.

The United States Environmental Protection Agency (USEPA) has long required both avian sub-acute dietary and acute oral studies to inform risk assessments for pesticides. Recently, the USEPA collaborated with People for the Ethical Treatment of Animals to determine whether the results of the acute oral avian toxicity test or the sub-acute dietary toxicity test consistently generated the greatest risk predictions in USEPA tier 1 assessments for pesticides first registered between 1998 and 2017. Their study concluded that in 99% of the cases, risk conclusions were driven by the acute oral study (OPPTS 850.2100, OCSPP 850.2100, or similar) because using these data results in higher risk quotients than sub-acute dietary data. Shortly after publishing these results, the USEPA released a formal memorandum providing guidance for waiving the sub-acute dietary study for most pesticides. The USEPA will, however, retain the option to require sub-acute dietary studies for pesticides with certain chemical properties. However, as the avian sub-acute dietary study has an exposure regimen that is often more representative of how birds are exposed to pesticides under actual use conditions than does the acute oral study (i.e., as part of a dietary item eaten over the course of a day and not a bolus dose), this study can provide useful context for risk assessment on a case-by-case basis. Decision criteria are needed to determine a path forward that both minimizes vertebrate animal testing and positions the avian sub-acute dietary data as an option for risk refinement. Decision criteria are proposed here with recommendations for refining the design of avian sub-acute dietary studies to ensure that the data generated are optimized to support a science-based acute avian risk assessment, supported by a case study demonstrating when and how sub-acute dietary studies may be used in a higher-tier risk assessment. Integr Environ Assess Manag 2022;18:1629-1638. © 2022 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Field evaluation of the potential for avian exposure to clothianidin following the planting of clothianidin-treated corn seed.

The objective of this study was to quantify consumption of clothianidin-treated corn seed by birds following standard planting practices. Based on post-planting seed counts on 21 fields in southwestern Ontario, Canada, between 29 and 813 seeds/ha (mean of 224 ± 167 (SD)) were estimated to remain on the soil surface immediately post planting (i.e., less than one seed per 10 m2). This represents between 0.03 and 1.2% of the total sown seeds. The number of seeds missing on each field on the third day after planting as a result of any process (e.g., removal by foraging birds or mammals or burial as a result of heavy rains) ranged from 0 to 136 seeds/ha (0 to 0.0136 seeds/m2). Behavior monitoring of individual birds and 24 h remote video surveillance were deployed to investigate how much of the treated seed remaining on the soil surface was consumed by birds. Spotting scopes were used to monitor the full duration of the field visits of 596 individual birds during morning hours for three consecutive days after planting on the 21 fields. Two birds were observed consuming treated seeds (one seed each) and three birds consumed seeds for which the treatment status could not be visually confirmed. Additionally, constant (24 h) video surveillance for 2-4 days immediately after planting was deployed at 24 areas where multiple treated seeds were found on the soil surface. Across 1,380 h of collected video footage (including both day and night periods), no birds were observed to consume any treated seeds. This study provides field evidence of two factors that determine exposure of birds to clothianidin-treated corn seeds: (1) standard sowing practices in Ontario are effective at burying treated seeds such that the proportion of sown seeds that remain on the soil surface after planting is low, and (2) birds monitored on these fields consumed very few of the clothianidin-treated corn seeds remaining on the soil surface after planting. As these results are dependent on planting techniques and seed characteristics, they are not necessarily applicable to other types of clothianidin treated seed.

A large-scale field study examining effects of exposure to clothianidin seed-treated canola on honey bee colony health, development, and overwintering success.

In summer 2012, we initiated a large-scale field experiment in southern Ontario, Canada, to determine whether exposure to clothianidin seed-treated canola (oil seed rape) has any adverse impacts on honey bees. Colonies were placed in clothianidin seed-treated or control canola fields during bloom, and thereafter were moved to an apiary with no surrounding crops grown from seeds treated with neonicotinoids. Colony weight gain, honey production, pest incidence, bee mortality, number of adults, and amount of sealed brood were assessed in each colony throughout summer and autumn. Samples of honey, beeswax, pollen, and nectar were regularly collected, and samples were analyzed for clothianidin residues. Several of these endpoints were also measured in spring 2013. Overall, colonies were vigorous during and after the exposure period, and we found no effects of exposure to clothianidin seed-treated canola on any endpoint measures. Bees foraged heavily on the test fields during peak bloom and residue analysis indicated that honey bees were exposed to low levels (0.5-2 ppb) of clothianidin in pollen. Low levels of clothianidin were detected in a few pollen samples collected toward the end of the bloom from control hives, illustrating the difficulty of conducting a perfectly controlled field study with free-ranging honey bees in agricultural landscapes. Overwintering success did not differ significantly between treatment and control hives, and was similar to overwintering colony loss rates reported for the winter of 2012-2013 for beekeepers in Ontario and Canada. Our results suggest that exposure to canola grown from seed treated with clothianidin poses low risk to honey bees.

Chlorpyrifos residue levels in avian food items following applications of a commercial EC formulation to alfalfa and citrus.

Two 10-day field residue studies were conducted to measure the amount of chlorpyrifos residue found in typical avian food following applications of a commercial 480 g liter(-1) EC (Lorsban 4E) at 1.1 kg AI ha(-1) (1 lb AI acre(-1)) to alfalfa and at 2.3 kg Al ha(-1) (2.0 lb AI acre(-1)) to citrus. Avian food items used in these studies included: crickets (Acheta domestica (L)), earthworms (Lumbricus terrestris L), darkling ground beetle larvae (Tenebrio molitor L), seed heads (Triticum sp), and naturally occurring flying and ground-dwelling insects. The studies incorporated a design involving three main study plots placed within larger treated areas of an alfalfa crop and a mature orange grove. The three main study plots represented three replications and each contained four sub-plots. One sub-plot, on each study plot, was sampled on day 0 (2-h post-application), day 1, day 5 and day 10 post-application. Chlorpyrifos residues were present in all avian food sampled following the application; however, residue levels were lower than estimated residue values typically used by the US EPA to establish expected environmental concentration (EEC) used in screening assessments of risk to terrestrial wildlife.

Acute toxicity of lead, steel, and an iron-tungsten-nickel shot to mallard ducks (Anas platyrhynchos).

Twenty mallards (Anas platyrhynchos) of both sexes were dosed by oral gavage with Heavi-Shot (H-S; Environ-Metal, Inc., Sweet Home, Oregon, USA) pellets, 20 with steel shot, and 10 with lead (Pb) pellets, all of equal size. All pellets were fired from a shotgun into an absorbent material, retrieved, and weighed prior to introduction into the ducks. Birds were fed whole kernel corn and grit and observed for signs of toxicity for 30 days following dosing. Hevi-Shot pellets lost an average of 6.2% of their mass and steel shot pellets lost 57% of their mass in the birds' gizzards. Almost all (90%) of the Pb shot dosed birds died before the end of the study, while no mortality was observed in the steel or H-S dosed groups. Even though total food consumption differed between the H-S and steel shot groups, mean bird weight change was not different. There were no significant morphologic or histopathologic abnormalities of the liver and kidney in the H-S and steel shot groups. Results indicated that mallards dosed orally with eight No. 4 H-S pellets were not adversely affected over a 30-day period, and that H-S provides another environmentally safe nontoxic shot for use in waterfowl hunting.

Avian food selection with application to pesticide risk assessment: are dead and desiccated insects a desirable food source?

Past evaluations of pesticide exposure have been conducted with substantial uncertainty regarding avian consumption of contaminated food items. One question is whether birds consume invertebrates that are killed by a chemical application and that may present an increasing chemical concentration as they desiccate. We addressed the research question in two phases. First, a laboratory study was conducted in which wild-caught birds were individually offered three food choices, i.e., live, fresh-dead, and desiccated insect larvae. Second, these same food choices plus live, fresh-dead, and desiccated crickets were presented in study plots in two agricultural crops, i.e., a cornfield and an orchard. The experimental food items were monitored with videography equipment to determine their fate and to compare laboratory and field results. Laboratory results showed that birds have a strong preference for live and fresh-dead prey over desiccated prey, with live prey taken before fresh-dead prey in most trials. The field study revealed a similar preference for live prey over desiccated prey, with preference for fresh-dead prey intermediate to the two other types.