Anticoagulant rodenticides (ARs) are used to control pest rodent species but can result in secondary poisoning of non -target animals, especially raptors. In the present study, differences in AR sensitivity among avian species were evaluated by comparing in vivo warfarin pharmacokinetics and effects, measuring cytochrome P450s (CYPs) expression involved in AR metabolism, and conducting in vitro inhibition assays of the AR target enzyme Vitamin K 2,3 -epoxide reductase (VKOR). Oral administration of warfarin at 4 mg/kg body weight did not prolong prothrombin time in chickens (Gallus gallus), rock pigeons (Columba livia), or Eastern buzzards (Buteo japonicus). Rock pigeons and buzzards exhibited shorter plasma half-life of warfarin compared to chickens. For the metabolite analysis, 4 '-hydroxywarfarin was predominantly detected in all birds, while 10-hydroxywarfarin was only found in pigeons and raptors, indicating interspecific differences in AR metabolism among birds likely due to differential expression of CYP enzymes involved in the metabolism of ARs and variation of VKOR activities among these avian species. The present findings, and results of our earlier investigations, demonstrate pronounced differences in AR sensitivity and pharmacokinetics among bird species, and in particular raptors. While ecological risk assessment and mitigation efforts for ARs have been extensive, AR exposure and adverse effects in predatory and scavenging wildlife continues. Toxicokinetic and toxicodynamic data will assist in such risk assessments and mitigation efforts.

The vast majority of knowledge related to the question "To what extent do pharmaceuticals in the environment pose a risk to wildlife?" stems from the Asian vulture crisis (>99% decline of some species of Old World vultures on the Indian subcontinent related to the veterinary use of the nonsteroidal anti-inflammatory drug [NSAID] diclofenac). The hazard of diclofenac and other NSAIDs (carprofen, flunixin, ketoprofen, nimesulide, phenylbutazone) to vultures and other avian species has since been demonstrated; indeed, only meloxicam and tolfenamic acid have been found to be vulture-safe. Since diclofenac was approved for veterinary use in Spain and Italy in 2013 (home to ~95% of vultures in Europe), the risk of NSAIDs to vultures in these countries has become one of the principal concerns related to pharmaceuticals and wildlife. Many of the other bodies of work on pharmaceutical exposure, hazard and risk to wildlife also relate to adverse effects in birds (e.g., poisoning of scavenging birds in North America and Europe from animal carcasses containing pentobarbital, secondary and even tertiary poisoning of birds exposed to pesticides used in veterinary medicine as cattle dips, migratory birds as a vector for the transfer of antimicrobial and antifungal resistance). Although there is some research related to endocrine disruption in reptiles and potential exposure of aerial insectivores, there remain numerous knowledge gaps for risk posed by pharmaceuticals to amphibians, reptiles, and mammals. Developing noninvasive sampling techniques and new approach methodologies (e.g., genomic, in vitro, in silico, in ovo) is important if we are to bridge the current knowledge gaps without extensive vertebrate testing. Environ Toxicol Chem 2024;43:595-610. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

First posted August 11, 2023 For additional information, contact: Director, Columbia Environmental Research CenterU.S. Geological Survey4200 New Haven RoadColumbia, MO 65201Contact Pubs Warehouse Lead mining in the Southeast Missouri Lead Mining District began in the 1700s and continued for nearly 300 years; the waste piles associated with smelting, mining, and milling of lead ores have released metal residues that have contaminated soil and water in the region. Previous studies in the district have indicated potential harm to wildlife, including birds, because of elevated lead concentrations associated with mining. Exposure to soil-borne lead was correlated with elevated lead concentrations in tissues, inhibition of δ-aminolevulinic acid dehydratase (δALAD), and renal lesions in birds foraging on ground-dwelling invertebrates at contaminated sites (compared to reference sites) in the Southeast Missouri Lead Mining District.This study assessed reproductive outcomes for songbirds exposed to soil-borne lead in the district, examined the relation between lead concentrations in soils and in tissues of ground-feeding birds and prey species, and compared the results to literature-based toxicity thresholds for lead that are associated with negative effects in birds. Three lead-contaminated sites and three reference sites (with background concentrations of lead and no known mining inputs) were compared in two ways: individually to all other sites or by site type. Additional effects of lead exposure were evaluated by examining concentrations of biomarkers (oxidative stress, lipid peroxidation, and deoxyribonucleic acid damage) in liver tissues, δALAD inhibition, and renal and hepatic microscopic lesions in birds from lead-contaminated and reference sites.Lead concentrations in soil were site-dependent and were also generally heterogeneous within the lead-contaminated sites. Between 17 and 74 percent of all soil samples at contaminated sites had lead concentrations that exceeded a threshold (1,000 milligrams per kilogram [mg/kg] lead in soil) previously associated with adverse physiological effects in birds in the Southeast Missouri Lead Mining District. Lead concentrations in mixed invertebrates from lead-contaminated sites (282 to 2,230 mg/kg dry weight [dw]) indicated that consuming soil-dwelling prey species is a potential exposure pathway for adult birds and their broods. At lead-contaminated sites, lead concentrations in 40.5 percent of blood samples (adults and their broods) were within a subclinical effects range (0.9 to 2.3 mg/kg dw), and 18.7 percent of samples had lead concentrations that exceeded clinical effects criteria (greater than 2.3 mg/kg dw). In contrast, only 2.6 percent of blood samples from reference sites were within the subclinical effects range for lead; all other blood samples from the reference sites had lead concentrations representative of background concentrations (less than 0.9 mg/kg dw). Subclinical and clinical threshold exceedances for lead concentrations in livers and kidneys were similarly more prevalent at the contaminated sites compared to the reference sites.Lead concentrations in blood were positively correlated with lead concentrations in soil, livers, and kidneys. Lead concentrations in blood were negatively correlated with δALAD activity; greater than 50 percent of the birds collected at lead-contaminated sites exhibited injury via greater than 50 percent inhibition of δALAD in blood compared to birds at reference sites. Birds with elevated lead concentrations in tissues also exhibited enhanced oxidative stress. Microscopic lesions in the livers and kidneys of birds had similar rates of occurrence at the contaminated and reference sites, and lesion prevalence could not be directly linked to lead exposure. Reproductive success was monitored at 585 nests, and 3 out of 5 species had reduced nest success associated with elevated lead concentrations in soil; habitat measures did not help explain nest success. Reduced nest success may have resulted from greater nest predation resulting from neurological and behavioral effects of lead exposure. Ultimately, these lines of evidence indicate that bird health and reproduction have been negatively affected by exposure to lead-contaminated soils in the Southeast Missouri Lead Mining District.

Although there is extensive evidence of declines in the American Kestrel (Falco sparverius) population across North America, the cause of such declines remains a mystery. One hypothesized driver of decline is anticoagulant rodenticide (AR) exposure, which could potentially cause mortality or reduced fitness. We investigated AR exposure in wild American Kestrels in Utah, USA. We collected and tested for AR residues in liver samples (n = 8) from kestrels opportunistically encountered dead and in blood samples (n = 71) from live wild kestrels, both nestlings and adults. We found high detection rates in both tissues. Adult kestrels were more likely to exhibit exposure than juveniles sampled in nests. Three-quarters (six of eight) of tested liver samples from adult kestrels exhibited evidence of AR exposure. Additionally, liver samples (n = 19) opportunistically collected from seven species of raptors within our study area had detectable levels of AR residues, with seven of eight raptor species evidencing exposure; across all raptors, five ARs were detected in liver samples, with brodifacoum the most prevalent, being found in over half (14 of 27) of samples. Over half (7 of 12) of the blood samples from adult kestrels had detectible levels of ARs, while only one of 59 juvenile nest samples tested positive. The difference in exposure rates between adults and juveniles could indicate differential exposure pathways by age class. Based on these findings, we recommend that ARs be further investigated as a potential cause of kestrel declines. Future research could focus on expanding sampling to provide sufficient sample sizes to test for potential nonlethal effects of AR exposure (e.g., fecundity, nesting success), identifying potential exposure pathways, and developing methods for passive sampling of ARs in excreta.

Model species (e.g., granivorous gamebirds, waterfowl, passerines, domesticated rodents) have been used for decades in guideline laboratory tests to generate survival, growth, and reproductive data for prospective ecological risk assessments (ERAs) for birds and mammals, while officially adopted risk assessment schemes for amphibians and reptiles do not exist. There are recognized shortcomings of current in vivo methods as well as uncertainty around the extent to which species with different life histories (e.g., terrestrial amphibians, reptiles, bats) than these commonly used models are protected by existing ERA frameworks. Approaches other than validating additional animal models for testing are being developed, but the incorporation of such new approach methodologies (NAMs) into risk assessment frameworks will require robust validations against in vivo responses. This takes time, and the ability to extrapolate findings from nonanimal studies to organism- and population-level effects in terrestrial wildlife remains weak. Failure to adequately anticipate and predict hazards could have economic and potentially even legal consequences for regulators and product registrants. In order to be able to use fewer animals or replace them altogether in the long term, vertebrate use and whole organism data will be needed to provide data for NAM validation in the short term. Therefore, it is worth investing resources for potential updates to existing standard test guidelines used in the laboratory as well as addressing the need for clear guidance on the conduct of field studies. Herein, we review the potential for improving standard in vivo test methods and for advancing the use of field studies in wildlife risk assessment, as these tools will be needed in the foreseeable future. Integr Environ Assess Manag 2024;20:699-724. © 2023 His Majesty the King in Right of Canada and The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC). Reproduced with the permission of the Minister of Environment and Climate Change Canada. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

In many countries the use of lead-based ammunition is prevalent, and results in exposure and poisoning of waterfowl and other species of birds. In waterfowl hunting areas large quantities of spent shot may be deposited in wetland and terrestrial habitats. These pellets can undergo transformations, which are influenced by various abiotic and biotic factors. In addition to lead (Pb), other elements like antimony (Sb) and arsenic (As) can be leached from Pb shot into the environment. In vitro simulations that included organic matter and microorganisms were utilized to examine elemental leaching from gunshot. We found that leaching efficiency was the greatest in solutions rich in organic matter derived from artificial root exudates (2.69 % for Pb, 1.16 % for Sb, 1.83 % for As), while leaching efficiency was considerably lower in river water (0.04 %). In vitro simulations containing native microorganisms also exhibited greater leaching efficiency (0.49 % for Pb, 0.52 % for Sb, 1.32 % for As) than in ultrapure deionized water and river water. Surface alterations in gunshot included the formation of a weathering crust and secondary phases dominated by carbonates. Spent gunshot is a source of Pb, Sb and As in wetlands that could affect aquatic ecosystems.

Despite advances in toxicity testing and the development of new approach methodologies (NAMs) for hazard assessment, the ecological risk assessment (ERA) framework for terrestrial wildlife (i.e., air-breathing amphibians, reptiles, birds, and mammals) has remained unchanged for decades. While survival, growth, and reproductive endpoints derived from whole-animal toxicity tests are central to hazard assessment, nonstandard measures of biological effects at multiple levels of biological organization (e.g., molecular, cellular, tissue, organ, organism, population, community, ecosystem) have the potential to enhance the relevance of prospective and retrospective wildlife ERAs. Other factors (e.g., indirect effects of contaminants on food supplies and infectious disease processes) are influenced by toxicants at individual, population, and community levels, and need to be factored into chemically based risk assessments to enhance the "eco" component of ERAs. Regulatory and logistical challenges often relegate such nonstandard endpoints and indirect effects to postregistration evaluations of pesticides and industrial chemicals and contaminated site evaluations. While NAMs are being developed, to date, their applications in ERAs focused on wildlife have been limited. No single magic tool or model will address all uncertainties in hazard assessment. Modernizing wildlife ERAs will likely entail combinations of laboratory- and field-derived data at multiple levels of biological organization, knowledge collection solutions (e.g., systematic review, adverse outcome pathway frameworks), and inferential methods that facilitate integrations and risk estimations focused on species, populations, interspecific extrapolations, and ecosystem services modeling, with less dependence on whole-animal data and simple hazard ratios. Integr Environ Assess Manag 2024;20:725-748. © 2023 His Majesty the King in Right of Canada and The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC). Reproduced with the permission of the Minister of Environment and Climate Change Canada. This article has been contributed to by US Government employees and their work is in the public domain in the USA.

Anticoagulant rodenticides (ARs) are widespread environmental contaminants that pose risks to scavenging birds because they routinely occur within their prey and can cause secondary poisoning. However, little is known about AR exposure in one of the rarest avian scavengers in the world, the California condor (Gymnogyps californianus). We assessed AR exposure in California condors and surrogate turkey vultures (Cathartes aura) to gauge potential hazard to a proposed future condor flock by determining how application rate and environmental factors influence exposure. Additionally, we examined whether ARs might be correlated with prolonged blood clotting time and potential mortality in condors. Only second-generation ARs (SGARs) were detected, and exposure was detected in all condor flocks. Liver AR residues were detected in 42% of the condors (27 of 65) and 93% of the turkey vultures (66 of 71). Although concentrations were generally low (<10 ng/g ww), 48% of the California condors and 64% of the turkey vultures exposed to ARs exceeded the 5% probability of exhibiting signs of toxicosis (>20 ng/g ww), and 10% and 13% exceeded the 20% probability of exhibiting signs toxicosis (>80 ng/g ww). There was evidence of prolonged blood clotting time in 16% of the free-flying condors. For condors, there was a relationship between the interaction of AR exposure index (legal use across regions where condors existed) and precipitation, and the probability of detecting ARs in liver. Exposure to ARs may complicate recovery efforts of condor populations within their current range and in the soon to be established northern California experimental population. Continued monitoring of AR exposure using plasma blood clotting assays and residue analysis would allow for an improved understanding of their hazard to condors, particularly if paired with recent movement data that could elucidate exposure sources on the landscape occupied by this endangered species.

U.S. regulatory and research agencies use ecotoxicity test data to assess the hazards associated with substances that may be released into the environment, including but not limited to industrial chemicals, pharmaceuticals, pesticides, food additives, and color additives. These data are used to conduct hazard assessments and evaluate potential risks to aquatic life (e.g., invertebrates, fish), birds, wildlife species, or the environment. To identify opportunities for regulatory uses of non-animal replacements for ecotoxicity tests, the needs and uses for data from tests utilizing animals must first be clarified. Accordingly, the objective of this review was to identify the ecotoxicity test data relied upon by U.S. federal agencies. The standards, test guidelines, guidance documents, and/or endpoints that are used to address each of the agencies' regulatory and research needs regarding ecotoxicity testing are described in the context of their application to decision-making. Testing and information use, needs, and/or requirements relevant to the regulatory or programmatic mandates of the agencies taking part in the Interagency Coordinating Committee on the Validation of Alternative Methods Ecotoxicology Workgroup are captured. This information will be useful for coordinating efforts to develop and implement alternative test methods to reduce, refine, or replace animal use in chemical safety evaluations.

The Chesapeake Bay, along the mid-Atlantic coast of North America, is the largest estuary in the United States and provides critical habitat for wildlife. In contrast to point and non-point source release of pesticides, metals, and industrial, personal care and household use chemicals on biota in this watershed, there has only been scant attention to potential exposure and effects of algal toxins on wildlife in the Chesapeake Bay region. As background, we first review the scientific literature on algal toxins and harmful algal bloom (HAB) events in various regions of the world that principally affected birds, and to a lesser degree other wildlife. To examine the situation for the Chesapeake, we compiled information from government reports and databases summarizing wildlife mortality events for 2000 through 2020 that were associated with potentially toxic algae and HAB events. Summary findings indicate that there have been few wildlife mortality incidents definitively linked to HABs, other mortality events that were suspected to be related to HABs, and more instances in which HABs may have indirectly contributed to or occurred coincident with wildlife mortality. The dominant toxins found in the Chesapeake Bay drainage that could potentially affect wildlife are microcystins, with concentrations in water approaching or exceeding human-based thresholds for ceasing recreational use and drinking water at a number of locations. As an increasing trend in HAB events in the U.S. and in the Chesapeake Bay have been reported, additional information on HAB toxin exposure routes, comparative sensitivity among species, consequences of sublethal exposure, and better diagnostic and risk criteria would greatly assist in predicting algal toxin hazard and risks to wildlife.

An important component of assessing the hazards of anticoagulant rodenticides to non-target wildlife is observations in exposed free-ranging individuals. The objective of this study was to determine whether environmentally realistic, sublethal first-generation anticoagulant rodenticide (FGAR) exposures via prey can result in direct or indirect adverse effects to free-flying raptors. We offered black-tailed prairie dogs (Cynomys ludovicianus) that had fed on Rozol® Prairie Dog Bait (Rozol, 0.005% active ingredient chlorophacinone, CPN) to six wild-caught red-tailed hawks (RTHA, Buteo jamaicensis), and also offered black-tailed prairie dogs that were not exposed to Rozol to another two wild-caught RTHAs for 7 days. On day 6, blood was collected to determine CPN's effects on blood clotting time. On day 7, seven of the eight RTHAs were fitted with VHF radio telemetry transmitters and the RTHAs were released the following day and were monitored for 33 days. Prothrombin time (PT) and Russell's viper venom time confirmed that the CPN-exposed RTHAs were exposed to and were adversely affected by CPN. Four of the six CPN-exposed RTHAs exhibited ptiloerection, an indication of thermoregulatory dysfunction due to CPN toxicity, but no signs of intoxication were observed in the reference hawk or the remaining two CPN-exposed RTHAs. Of note is that PT values were associated with ptiloerection duration and frequency; therefore, sublethal CPN exposure can directly or indirectly evoke adverse effects in wild birds. Although our sample sizes were small, this study is a first to relate coagulation times to adverse clinical signs in free-ranging birds.

A literature review of bioaccumulation and biotransformation of organic chemicals in birds was undertaken, aiming to support scoping and prioritization of future research. The objectives were to characterize available bioaccumulation/biotransformation data, identify knowledge gaps, determine how extant data can be used, and explore the strategy and steps forward. An intermediate approach balanced between expediency and rigor was taken given the vastness of the literature. Following a critical review of > 500 peer-reviewed studies, > 25,000 data entries and 2 million information bytes were compiled on > 700 organic compounds for ~ 320 wild species and 60 domestic breeds of birds. These data were organized into themed databases on bioaccumulation and biotransformation , field survey , microsomal enzyme activity , metabolic pathway , and bird taxonomy and diet . Significant data gaps were identified in all databases at multiple levels. Biotransformation characterization was largely fragmented over metabolite/pathway identification and characterization of enzyme activity or biotransformation kinetics. Limited biotransformation kinetic data constrained development of an avian biotransformation model. A substantial shortage of in vivo biotransformation kinetics has been observed as most reported rate constants were derived in vitro. No metric comprehensively captured all key contaminant classes or chemical groups to support broad-scope modeling of bioaccumulation or biotransformation. However, metrics such as biota-feed accumulation factor, maximum transfer factor, and total elimination rate constant were more readily usable for modeling or benchmarking than other reviewed parameters. Analysis demonstrated the lack of bioaccumulation/biotransformation characterization of shorebirds, seabirds, and raptors. In the study of bioaccumulation and biotransformation of organic chemicals in birds, this review revealed the need for greater chemical and avian species diversity, chemical measurements in environmental media, basic biometrics and exposure conditions, multiple tissues/matrices sampling, and further exploration on biotransformation. Limitations of classical bioaccumulation metrics and current research strategies used in bird studies were also discussed. Forward-looking research strategies were proposed: adopting a chemical roadmap for future investigations, integrating existing biomonitoring data, gap-filling with non-testing approaches, improving data reporting practices, expanding field sampling scopes, bridging existing models and theories, exploring biotransformation via avian genomics, and establishing an online data repository.

Wild raptors are widely used to assess exposure to different environmental contaminants, including anticoagulant rodenticides (ARs). ARs are used on a global scale for rodent control, and act by disruption of the vitamin K cycle that results in haemorrhage usually accompanied by death within days. Some ARs are highly persistent and bioaccumulative, which can cause significant exposure of non-target species. We characterized AR exposure in a heterogeneous sample of dead raptors collected over 12 years (2008-2019) in south-eastern France. Residue analysis of 156 liver samples through LC-MS/MS revealed that 50% (78/156) were positive for ARs, with 13.5% (21/156) having summed second-generation AR (SGAR) concentrations >100 ng/g ww. While SGARs were commonly detected (97.4% of positive samples), first-generation ARs were rarely found (7.7% of positive samples). ARs were more frequently detected and at greater concentration in predators (prevalence: 82.5%) than in scavengers (38.8%). Exposure to multiple ARs was common (64.1% of positive samples). While chlorophacinone exposure decreased over time, an increasing exposure trend was observed for the SGAR brodifacoum, suggesting that public policies may not be efficient at mitigating risk of exposure for non-target species. Haemorrhage was observed in 88 birds, but AR toxicosis was suspected in only 2 of these individuals, and no difference in frequency of haemorrhage was apparent in birds displaying summed SGAR levels above or below 100 ng/g ww. As for other contaminants, 17.2% of liver samples (11/64) exhibited Pb levels compatible with sub-clinical poisoning (>6 μg/g dw), with 6.3% (4/64) above the threshold for severe/lethal poisoning (>30 μg/g dw). Nine individuals with Pb levels >6 μg/g dw also had AR residues, demonstrating exposure to multiple contaminants. Broad toxicological screening for other contaminants was positive for 18 of 126 individuals, with carbofuran and mevinphos exposure being the suspected cause of death of 17 birds. Our findings demonstrate lower but still substantial AR exposure of scavenging birds compared to predatory birds, and also illustrate the complexity of diagnosing AR toxicosis through forensic investigations.

Anticoagulant rodenticides (ARs) are commonly used to control rodent pests. However, worldwide, their use is associated with secondary and tertiary poisoning of nontarget species, especially predatory and scavenging birds. No medical device can rapidly test for AR exposure of avian wildlife. Prothrombin time (PT) is a useful biomarker for AR exposure, and multiple commercially available point-of-care (POC) devices measure PT of humans, and domestic and companion mammals. We evaluated the potential of one commercially available POC device, the Coag-Sense® PT/INR Monitoring System, to rapidly detect AR exposure of living birds of prey. The Coag-Sense device delivered repeatable PT measurements on avian blood samples collected from four species of raptors trapped during migration (Intraclass Correlation Coefficient > 0.9; overall intra-sample variation CV: 5.7%). However, PT measurements reported by the Coag-Sense system from 81 ferruginous hawk (Buteo regalis) nestlings were not correlated to those measured by a one-stage laboratory avian PT assay (r = − 0.017, p = 0.88). Although precise, the lack of agreement in PT estimates from the Coag-Sense device and the laboratory assay indicates that this device is not suitable for detecting potential AR exposure of birds of prey. The lack of suitability may be related to the use of a mammalian reagent in the clotting reaction, suggesting that the device may perform better in testing mammalian wildlife

Anticoagulant rodenticides are widely used for rodent control in agricultural and urban settings. Their intense use can sometimes result in accidental exposure and even poisoning of livestock. Can milk, eggs or meat derived from such accidently exposed animals be consumed by humans? Data on the pharmacokinetics of chlorophacinone in milk of accidently exposed ewes were used to estimate the risk associated with its consumption. Three days after accidental ingestion, chlorophacinone was detected in plasma of 18 ewes, with concentrations exceeding 100 ng/mL in 11 animals. Chlorophacinone was detected in milk on day 2 post-exposure and remained quantifiable for at least 7 days in milk of these 11 ewes. Concentrations in milk were much lower than in plasma and decreased quickly (mean half-life of 2 days). This study demonstrated dose-dependent mammary transfer of ingested chlorophacinone. Variation in prothrombin time (PT) on Day 3 suggested that some of the ewes that ingested chlorophacinone may have been adversely affected, but PT did not facilitate estimation of the quantity of chlorophacinone consumed. Using safety factors described in the literature, consumption of dairy products derived from these ewes after a one-week withdrawal period would pose low risk to consumers.

Pesticides coated to the seed surface potentially pose an ecological risk to granivorous birds that consume incompletely buried or spilled seeds. To assess the toxicokinetics of seeds treated with current-use fungicides, Japanese quail (Coturnix japonica) were orally dosed with commercially coated wheat seeds. Quail were exposed to metalaxyl, tebuconazole, and fludioxonil at either a low dose (0.0655, 0.0308, and 0.0328 mg/kg of body weight, respectively) or a high dose (0.196, 0.0925, and 0.0985 mg/kg of body weight, respectively). Fungicides were rapidly absorbed and distributed to tissues. Tebuconazole was metabolized into tert-butylhydroxy-tebuconazole. All compounds were eliminated to below detection limits within 24 h. The high detection frequencies observed in fecal samples potentially offer a non-invasive matrix to monitor pesticide exposure. With the summation of total body burden across plasma, tissue, and fecal samples, less than 9% of the administered dose was identified as the parent fungicide, demonstrating the importance to monitor both active ingredients and their metabolites in biological samples.

A seminal question in ecotoxicology is the extent to which contaminant exposure evokes prolonged effects on physiological function and fitness. A series of studies were undertaken with American kestrels ingesting environmentally realistic concentrations of the second-generation anticoagulant rodenticide (SGAR) brodifacoum. Kestrels fed brodifacoum at 0.3, 1.0, or 3.0 mu g/g diet wet weight for 7 d exhibited dose-dependent hemorrhage, histopathological lesions, and coagulopathy (prolonged prothrombin and Russell's viper venom times). Following termination of a 7-d exposure to 0.5 mu g brodifacoum/g diet, prolonged blood clotting time returned to baseline values within 1 wk, but brodifacoum residues in liver and kidney persisted during the 28-d recovery period (terminal half-life estimates >50 d). To examine the hazard of sequential anticoagulant rodenticide (AR) exposure, kestrels were exposed to either the first-generation AR chlorophacinone (1.5 mu g/g diet) or the SGAR brodifacoum (0.5 mu g/g diet) for 7 d and, following a recovery period, challenged with a low dose of chlorophacinone (0.75 mu g/g diet) for 7 d. In brodifacoum-exposed kestrels, the challenge exposure clearly prolonged prothrombin time compared to naive controls and kestrels previously exposed to chlorophacinone. These data provide evidence that the SGAR brodifacoum may have prolonged effects that increase the toxicity of subsequent AR exposure. Because free-ranging predatory and scavenging wildlife are often repeatedly exposed to ARs, such protracted toxicological effects need to be considered in hazard and risk assessments. Environ Toxicol Chem 2020;39:468-481. (c) 2020 SETAC

Anticoagulant rodenticides (ARs) are part of the near billion-dollar rodenticide industry. Numerous studies have documented the presence of ARs in nontarget wildlife, with evidence of repeated exposure to second-generation ARs. While birds are generally less sensitive to ARs than target rodent species, in some locations predatory and scavenging birds are exposed by consumption of such poisoned prey and, depending on dose and frequency of exposure, exhibit effects of intoxication that can result in death. Evidence of hemorrhage in conjunction with summed hepatic AR residues >0.1-0.2 mg kg(-1) liver wet weight are often used as criteria to diagnose ARs as the likely cause of death. In this review focusing on birds of prey and scavengers, we discuss AR potency, coagulopathy, toxicokinetics and long-lasting effects of residues, and the role of nutrition and vitamin K status on toxicity, and identify some research needs. A more complete understanding of the factors affecting AR toxicity in nontarget wildlife could enable regulators and natural resource managers to better predict and even mitigate risk. Published 2020. This article is a U.S. Government work and is in the public domain in the USA

Statistical inferences play a critical role in ecotoxicology. Historically, null hypothesis significance testing (NHST) has been the dominant method for inference in ecotoxicology. As a brief and informal definition of NHST, researchers compare (or "test") an experimental treatment or observation against a hypothesis of no relationship (the "null hypothesis") using the collected data to see if the observed values are statistically "significant" given predefined error rates. The resulting probability of observing a value equal to or greater than the observed value assuming the null hypothesis is true is the p value. Criticisms of NHST have existed for almost a century and have recently grown to the point where statisticians, including the American Statistical Association (ASA), have felt the need to clarify the role of NHST and p values beyond their current common use. These limitations also exist in ecotoxicology. For example, a review of the 2010 Environmental Toxicology & Chemistry (ET&C) volume that found many authors did not correctly report p values. We repeated this review looking at the 2019 volume of ET&C. Incorrect reporting of p values still occurred almost a decade later. Problems with NHST and p values highlight the need for statistical inferences besides NHST, something long known in ecotoxicology and the broader scientific and statistical communities. Furthermore, concerns such as these led the executive director of the ASA to recommend against use of "statistical significance" in 2019. In light of these criticisms, ecotoxicologists require alternative methods. We describe some alternative methods including confidence intervals, regression analysis, dose-response curves, Bayes factors, survival analysis, and model selection. Lastly, we provide insights for what ecotoxicology might look like in a post-p value world. Environ Toxicol Chem 2020;39:1657-1669. Published 2020. This article is a U.S. Government work and is in the public domain in the USA.

Birds are potentially exposed to neonicotinoid insecticides by ingestion of coated seeds during crop planting. Adult male Japanese quail were orally dosed with wheat seeds coated with an imidacloprid (IMI) formulation at either 0.9 or 2.7 mg/kg body weight (BW) (∼3 and 9% of IMI LD50 for Japanese quail, respectively) for 1 or 10 days. Quail were euthanized between 1 and 24 h postexposure to assess toxicokinetics. Analysis revealed rapid absorption (1 h) into blood and distribution to the brain, muscle, kidney, and liver. Clearance to below detection limits occurred at both dose levels and exposure durations in all tissues within 24 h. Metabolism was extensive, with 5-OH-IMI and IMI-olefin detected at greater concentrations than IMI in tissues and fecal samples. There was no lethality or overt signs of toxicity at either dose level. Furthermore, no evidence of enhanced expression of mRNA genes associated with hepatic xenobiotic metabolism, oxidative DNA damage, or alterations in concentrations of corticosterone and thyroid hormones was observed. Application of the toxicokinetic data was used to predict IMI residue levels in the liver with reasonable results for some field exposure and avian mortality events. It would appear that some affected species of birds are either consuming larger quantities of seeds or exhibit differences in ADME or sensitivity than predicted by read-across from these data.