Following Regulation, Imidacloprid Persists and Flupyradifurone Increases in Nontarget Wildlife.

After regulation of pesticides, determination of their persistence in the environment is an important indicator of effectiveness of these measures. We quantified concentrations of two types of systemic insecticides, neonicotinoids (imidacloprid, acetamiprid, clothianidin, thiacloprid, and thiamethoxam) and butenolides (flupyradifurone), in off-crop nontarget media of hummingbird cloacal fluid, honey bee (Apis mellifera) nectar and honey, and wildflowers before and after regulation of imidacloprid on highbush blueberries in Canada in April 2021. We found that mean total pesticide load increased in hummingbird cloacal fluid, nectar, and flower samples following imidacloprid regulation. On average, we did not find evidence of a decrease in imidacloprid concentrations after regulation. However, there were some decreases, some increases, and other cases with no changes in imidacloprid levels depending on the specific media, time point of sampling, and site type. At the same time, we found an overall increase in flupyradifurone, acetamiprid, thiamethoxam, and thiacloprid but no change in clothianidin concentrations. In particular, flupyradifurone concentrations observed in biota sampled near agricultural areas increased twofold in honey bee nectar, sevenfold in hummingbird cloacal fluid, and eightfold in flowers after the 2021 imidacloprid regulation. The highest residue detected was flupyradifurone at 665 ng/mL (parts per billion [ppb]) in honey bee nectar. Mean total pesticide loads were highest in honey samples (84 ± 10 ppb), followed by nectar (56 ± 7 ppb), then hummingbird cloacal fluid (1.8 ± 0.5 ppb), and least, flowers (0.51 ± 0.06 ppb). Our results highlight that limited regulation of imidacloprid does not immediately reduce residue concentrations, while other systemic insecticides, possibly replacement compounds, concurrently increase in wildlife. Environ Toxicol Chem 2024;00:1-12. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Anticoagulant Rodenticide Toxicity in Terrestrial Raptors: Tools to Estimate the Impact on Populations in North America and Globally.

Anticoagulant rodenticides (ARs) have caused widespread contamination and poisoning of predators and scavengers. The diagnosis of toxicity proceeds from evidence of hemorrhage, and subsequent detection of residues in liver. Many factors confound the assessment of AR poisoning, particularly exposure dose, timing and frequency of exposure, and individual and taxon-specific variables. There is a need, therefore, for better AR toxicity criteria. To respond, we compiled a database of second-generation anticoagulant rodenticide (SGAR) residues in liver and postmortem evaluations of 951 terrestrial raptor carcasses from Canada and the United States, 1989 to 2021. We developed mixed-effects logistic regression models to produce specific probability curves of the toxicity of ∑SGARs at the taxonomic level of the family, and separately for three SGARs registered in North America, brodifacoum, bromadiolone, and difethialone. The ∑SGAR threshold concentrations for diagnosis of coagulopathy at 0.20 probability of risk were highest for strigid owls (15 ng g-1) lower and relatively similar for accipitrid hawks and eagles (8.2 ng g-1) and falcons (7.9 ng g-1), and much lower for tytonid barn owls (0.32 ng g-1). These values are lower than those we found previously, due to compilation and use of a larger database with a mix of species and source locations, and also to refinements in the statistical methods. Our presentation of results on the family taxonomic level should aid in the global applicability of the numbers. We also collated a subset of 440 single-compound exposure events and determined the probability of SGAR-poisoning symptoms as a function of SGAR concentration, which we then used to estimate relative SGAR toxicity and toxic equivalence factors: difethialone, 1, brodifacoum, 0.8, and bromadiolone, 0.5. Environ Toxicol Chem 2024;43:988-998. © 2024 His Majesty the King in Right of Canada and The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC Reproduced with the permission of the Minister of Environment and Climate Change Canada.

Quantifying phenology and migratory behaviours of hummingbirds using single-site dynamics and mark-detection analyses.

Nuanced understanding of seasonal movements of partially migratory birds is paramount to species and habitat conservation. Using nascent statistical methods, we identified migratory strategies of birds outfitted with radio-frequency identification (RFID) tags detected at RFID feeders in two sites in California, USA. We quantified proportions of migrants and residents and the seasonal phenology for each movement strategy in Allen's and Anna's hummingbirds; we also validated our methodology by fitting our model to obligate migratory black-chinned hummingbirds. Allen's and Anna's hummingbirds exhibited characteristics of facultative migratory behaviour. We also quantified apparent annual survival for each migratory strategy and found that residents had significantly higher probabilities of apparent survival. Low survival estimates for migrants suggest that a high proportion of birds in the migrant group permanently emigrated from our study sites. Considered together, our analyses suggest that hummingbirds in both northern and southern California sites partake in diverse and highly plastic migratory behaviours. Our assessment elucidates the dynamics underlying idiosyncratic migratory behaviours of two species of hummingbirds, in addition to describing a framework for similar assessments of migratory behaviours using the multi-state open robust design with state uncertainty model and single-site dynamics.

Evaluation of minimally invasive sampling methods for detecting Avipoxvirus: Hummingbirds as a case example.

Avian pox is a common avian virus that in its cutaneous form can cause characteristic lesions on a bird's dermal surfaces. Detection of avian pox in free-ranging birds historically relied on observations of visual lesions and/or histopathology, both which can underestimate avian pox prevalence. We compared traditional visual observation methods for avian pox with molecular methods that utilize minimally invasive samples (blood, toenail clipping, feathers, and dermal swabs) in an ecologically important group of birds, hummingbirds. Specifically, avian pox prevalence in several species of hummingbirds were examined across multiple locations using three different methods: (1) visual inspection of hummingbirds for pox-like lesions from a long-term banding data set, (2) qPCR assay of samples from hummingbird carcasses from wildlife rehabilitation centers, and (3) qPCR assay of samples from live-caught hummingbirds. A stark difference in prevalences among these three methods was identified, with an avian pox prevalence of 1.5% from banding data, 20.4% from hummingbird carcasses, and 32.5% from live-caught hummingbirds in California. This difference in detection rates underlines the necessity of a molecular method to survey for avian pox, and this study establishes one such method that could be applied to other wild bird species. Across all three methods, Anna's hummingbirds harbored significantly higher avian pox prevalence than other species examined, as did males compared with females and birds caught in Southern California compared with Northern California. After hatch-year hummingbirds also harbored higher avian pox prevalences than hatch-year hummingbirds in the California banding data set and the carcass data set. This is the first study to estimate the prevalence of avian pox in hummingbirds and address the ecology of this hummingbird-specific strain of avian pox virus, providing vital information to inform future studies on this charismatic and ecologically important group of birds.

Bioaccumulation and effects of selenium from surface coal mining in an aquatic songbird.

Open-pit mining operations are hailed for safe working conditions for miners as well as economically and logistically favourable outcomes for mining companies. However, ecological impacts of these operations may persist for decades. Expansions of open-pit coal mining in British Columbia of Western Canada are planned. Governmental regulation of background contaminants leached from these mines into nearby water systems were established to mitigate environmental impacts associated with these operations. We analyzed water, periphyton, invertebrate, and American dipper (Cinclus mexicanus) blood, egg, and feather samples for selenium and mercury exposure related to mining operations in the Elk Valley. We also quantified effects associated with exposure on clutch size and hatching success. Selenium concentrations in water, periphyton, and invertebrates were significantly higher downstream of mines compared to reference sites within and outside of the Elk River watershed. Selenium concentrations in water from exposed sites exceeded current regulatory levels established to protect wildlife by up to 20 times. Mercury concentrations were below toxic levels for birds in all media and did not factor into determination of selenium exposure or effects. Egg selenium concentrations were on average 0.9 times the regulatory threshold. Our stable isotope analyses showed that diets of nesting females included a negligible proportion of higher order consumers. We did not detect a significant effect of selenium on the rate of hatching success or clutch size of dippers nesting in exposed sites. We conclude that at the time of the investigation, selenium exposure was not impairing hatching of American dippers at the sampled locations.

Bluebirds Experience Impaired Hatching Success in Conventionally Sprayed Apple Orchard Habitats: A 31-Year Study.

We analyzed the reproductive success of eastern bluebirds (Sialia sialis) nesting in apple orchards and non-orchard nest-boxes in southern Ontario, Canada, from 1988 to 2018. Using data from 2397 nest-boxes monitored at 20 orchard sites and 52 non-orchard sites, we first modeled phenological parameters typically linked to climate change across both site types. We found that the first egg of each brood was laid significantly earlier in the season each year over our 31-year study. Clutch initiation occurred 4 days earlier in the spring in 2018 compared to 1988. Average clutch size in the first or second brood did not change significantly during our 31-year study; however, clutches were significantly smaller in orchards compared to non-orchards (0.10 ± 0.03 fewer eggs between sites). Nests built in orchards were also at 6.1-fold greater risk of parasitism and 2.1-fold greater risk of depredation than nests in non-orchards. After accounting for depredation and nest parasitism, hatching success was still significantly lower in orchards than in non-orchards. Overall, hatching success was 4%-5% lower in orchards. The probability of successfully fledging did not differ significantly between site types. In 2012, a ban on use of the organophosphate insecticide azinphos-methyl in orchards was enacted in Canada. We did not find a difference in hatching or fledging success in orchards after the ban. In our assessment of available data, we conclude that any pesticide effect on hatching success of eastern bluebirds in sprayed orchards is most likely the consequence of long-term exposure to dichlorodiphenyltrichloroethane (DDT) compounds in orchard soils and bioaccumulation in eggs rather than pesticides in use since regulation of DDT in the 1970s. Environ Toxicol Chem 2021;40:3369-3378. © 2021 Her Majesty the Queen in Right of Canada. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. Reproduced with the permission of the Minister of Environment and Climate Change Canada.

Current contrasting population trends among North American hummingbirds.

As pollinators, hummingbirds play a critical role for both the function of ecological communities and in providing ecosystem services for people. To examine the conservation status of North American hummingbirds, we analyzed Breeding Bird Survey data for 8 species and 3 genera from 1970 to 2019 (long-term) and from 2009 to 2019 (short-term, approximately three generations). Among the Selasphorus genus, Allen's, rufous, and broad-tailed hummingbirds have declined since 1970, and the rate of decline increased from 2009 to 2019. Contrasting the trends from the past half-century, ruby-throated hummingbirds of Eastern North America have declined since approximately 2004 throughout most of the species' breeding range. In contrast, Anna's hummingbird populations have increased dramatically since 1970 in their range in western North America. We also tested whether apparent declines might be due to a growing mismatch between the timing of breeding and the timing of BBS surveys. We found no evidence for such an effect, thus supporting the hypothesis that trends reflect true demographic change. Our analyses and geographic modelling highlight the urgent need of regulatory action to conserve hummingbirds uniquely capable of filling their niche in North America.

Neonicotinoid pesticides exert metabolic effects on avian pollinators.

Neonicotinoids are neurotoxic systemic insecticides applied extensively worldwide. The impacts of common neonicotinoids like imidacloprid on non-target invertebrate pollinators have been widely studied, however effects on vertebrate pollinators have received little attention. Here, we describe the first study evaluating the effects of short-term (3 d) exposure to a range of environmentally relevant concentrations ([Formula: see text] to [Formula: see text]Body Weight) of imidacloprid on wild-caught ruby-throated hummingbirds. Within 2 h of exposure, hummingbirds exhibited a significant depression in energy expenditure (up to [Formula: see text]). We did not observe significant effects on foraging behaviour measured in the subsequent 2 h to 4 h, although the effect size estimate was large (0.29). We also analyzed tissues collected 24 h after the final dose and did not observe significant effects on immune response or cholinesterase activity, although this may be related to our small sample size. We determined that hummingbirds excrete imidacloprid quickly (elimination half-life of [Formula: see text]) relative to other bird species. Hummingbirds have high energetic demands and store relatively little energy, especially during migration and breeding seasons. Therefore, changes in their metabolism following exposures to imidacloprid observed herein could bear important survivorship consequences for hummingbirds.

MicroRNAs regulate survival in oxygen-deprived environments.

Some animals must endure prolonged periods of oxygen deprivation to survive. One such extreme model is the northern crayfish (Orconectes virilis), that regularly survives year-round hypoxic and anoxic stresses in its warm stagnant summer waters and in its cold, ice-locked winter waters. To elucidate the molecular underpinnings of anoxia resistance in this natural model, we surveyed the expression profiles of 76 highly conserved microRNAs in crayfish hepatopancreas and tail muscle from normoxic, acute 2 h anoxia, and chronic 20 h anoxia exposures. MicroRNAs are known to regulate a diverse array of cellular functions required for environmental stress adaptations, and here we explored their role in anoxia tolerance. The tissue-specific anoxia responses observed herein, with 22 anoxia-responsive microRNAs in the hepatopancreas and only four in muscle, suggest that microRNAs facilitate a reprioritization of resources to preserve crucial organ functions. Bioinformatic microRNA target enrichment analysis predicted that the anoxia-downregulated microRNAs in hepatopancreas targeted Hippo signalling, suggesting that cell proliferation and apoptotic signalling are highly regulated in this liver-like organ during anoxia. Compellingly, miR-125-5p, miR-33-5p and miR-190-5p, all known to target the master regulator of oxygen deprivation responses HIF1 (hypoxia inducible factor-1), were anoxia downregulated in the hepatopancreas. The anoxia-increased transcript levels of the oxygen-dependent subunit HIF1α highlight a potential critical role for miRNA-HIF targeting in facilitating a successful anoxia response. Studying the cytoprotective mechanisms in place to protect against the challenges associated with surviving in oxygen-poor environments is critical to elucidating the vast and substantial role of microRNAs in the regulation of metabolism and stress in aquatic invertebrates.

Regulation of nuclear factor of activated T cells (NFAT) and downstream myogenic proteins during dehydration in the African clawed frog.

Xenopus laevis, otherwise known as the African clawed frog, undergoes natural dehydration of up to 30% of its total body water during the dry season in sub-Saharan Africa. To survive under these conditions, a variety of physiological and biochemical changes take place in X. laevis. We were interested in understanding the role that the calcineurin-NFAT pathway plays during dehydration stress response in the skeletal muscles of X. laevis. Immunoblotting was performed to characterize the protein levels of NFATc1-4, calcium signalling proteins, in addition to myogenic proteins (MyoD, MyoG, myomaker). In addition, DNA-protein interaction ELISAs were used to assess the binding of NFATs to their consensus binding sequence, and to identify the effect of urea on NFAT-binding. Our results showed that NFATc1 and c4 protein levels decreased during dehydration, and there were no changes in NFATc2, c3, and calcium signalling proteins. However, MyoG and myomaker both showed increases in protein levels during dehydration, thus indicating that the late myogenic program involving myoblast differentiation, but not satellite cell activation and myoblast proliferation, could be involved in preserving the skeletal muscle of X. laevis during dehydration. In addition, we observed that urea seems to reduce NFATc3-binding to DNA during control, but not during dehydration, possibly indicating that NFATc3 is protected from the denaturing effects of urea as it accumulates during dehydration. These findings expand upon our knowledge of adaptive responses to dehydration, and they identify specific protein targets that could be used to protect the skeletal muscle from damage during stress.