Incubation temperature and PCB-126 exposure interactively impair shorebird embryo and post-hatch development.

In oviparous wildlife, many critical physiological and behavioural components are strongly influenced by the embryonic and early post-hatch developmental environment. As such, early life stages in these species are highly vulnerable to both natural and anthropogenic stressors. For example, in birds, incubation temperature may influence the rate of egg development while also affecting contaminant metabolism and absorption in body tissues, resulting in potentially multiplicative impacts on embryonic and posthatch development. We tested the hypothesis that cumulative effects of early contaminant exposure and temperature stress can negatively affect avian development and may have interactive effects that are more detrimental than either stressor individually. Using a controlled egg injection and incubation study on killdeer (Charadrius vociferous), eggs were exposed to a known endocrine disruptor, 3,3',4,4',5-pentachlorobiphenyl (PCB-126) and incubated at either low (36 °C), intermediate (37.5 °C), or high (39 °C) temperatures. Our results indicated that eggs incubated at low temperature had earlier detection of heartbeat, longer incubation length, lower growth rate post-hatch, and higher post-hatch mortality, compared to eggs incubated under intermediate temperatures. Higher incubation temperatures resulted in shorter incubation length, earlier detection of heart rate and faster righting time. As predicted, embryo and chick mortality were greater in the PCB-dosed birds incubated at intermediate and high temperatures. Incidence of distended yolk sacs (%) also increased with PCB exposure in all temperature groups, with the largest increase in the high temperature group. Overall, our results show that low incubation temperature can cause greater adverse effects than PCB-126 exposure alone, but that negative effects of PCB-126 exposure are exacerbated by high incubation temperatures. These findings suggest that in natural settings, shorebird embryos may be more susceptible to contaminant exposure when incubated at temperatures either below or above the apparent optimum.

Continuous exposure to mercury during embryogenesis and chick development affects later survival and reproduction of zebra finch (Taeniopygia guttata).

Methylmercury (MeHg) is a global environmental contaminant that bioaccumulates and has multiple toxic modes of action. Aquatic species have traditionally been the focus of wildlife toxicological research on mercury, but terrestrial organisms, including passerine birds, can be exposed to similarly elevated levels of MeHg. In this study we exposed a model passerine, the zebra finch (Taeniopygia guttata), to MeHg in ovo, as chicks only, or with a combined 'in ovo + chick' treatment. We isolated exposure to specific developmental stages through the use of egg injections (3.2 µg Hg/g egg) and controlled oral dosing of chicks (0.24 µg Hg/g bw/day from day 1 to day 30). In ovo exposure to MeHg reduced hatching success, but there was no effect of MeHg on chick growth. We found that in ovo only or chick only exposure did not have long-term effects, but there was some evidence for longer-term effects of combined 'in ovo + chick' exposure on post-fledging survival and potentially sex-biased survival which resulted in very few 'in ovo + chick' exposed females surviving to breed. These females also had lower overall breeding productivity that was mainly due to lower hatching success of their offspring, not lower chick-rearing success. We found no effect of treatment on clutch size or latency to laying among females that did lay eggs. Our study suggests that combined embryonic and nestling MeHg exposure has compounding latent effects on productivity, likely through a mechanism that influences the ability of females to lay fertile eggs that hatch.

In ovo exposure to brominated flame retardants Part I: Assessment of effects of TBBPA-BDBPE on survival, morphometric and physiological endpoints in zebra finches.

Tetrabromobisphenol A bis(2,3-dibromopropyl) ether (TBBPA-BDBPE) is an additive flame retardant used in polyolefins and polymers. It has been detected in biota, including in avian eggs, yet little is known of its effects. We assessed the pattern of TBBPA-BDBPE concentrations in songbird eggs over the incubation period, and the effects of embryonic exposure to TBBPA-BDBPE in a model songbird species, the zebra finch (Taeniopygia guttata). To assess concentrations during embryo development, eggs were injected on the day they were laid with the vehicle control (safflower oil) or 100 ng TBBPA-BDBPE/g egg, and whole egg contents were collected throughout embryonic development on day 0 (unincubated), 5, 10 and 13. To evaluate effects of embryonic exposure to TBBPA-BDBPE, eggs were injected at Hamburger-Hamilton stage 18 (∼80 h after initiation of incubation) with safflower oil only, 10, 50 or 100 ng TBBPA-BDBPE/g egg (albumin injection volume 1 µl/g). Eggs were monitored for hatching success, and nestlings were monitored for growth and survival. At 15 days post-hatch, tissues were collected to assess physiological effects. TBBPA-BDBPE was incorporated into the egg as the embryo developed, and concentrations started declining in late incubation, suggesting biotransformation by the embryo. There were no effects on hatching success, nestling survival, growth, organ somatic indices, or thyroid hormone homeostasis; however, there was evidence that body condition declined in a dose-dependent manner towards the end of the rapid nestling growth phase. This decreased body condition could be a delayed effect of early developmental exposure, or it may be the result of increased exposure to biotransformation products of TBBPA-BDBPE produced over the nestling period, which are predicted to be more bioaccumulative and toxic than the parent compound.

In ovo exposure to brominated flame retardants Part II: Assessment of effects of TBBPA-BDBPE and BTBPE on hatching success, morphometric and physiological endpoints in American kestrels.

Tetrabromobisphenol A bis(2,3-dibromopropyl ether) (TBBPA-BDBPE) and 1,2-bis(2,4,6-tribromophenoxy)ethane (BTPBE) are both brominated flame retardants (BFRs) that have been detected in birds; however, their potential biological effects are largely unknown. We assessed the effects of embryonic exposure to TBBPA-BDBPE and BTBPE in a model avian predator, the American kestrel (Falco sparverius). Fertile eggs from a captive population of kestrels were injected on embryonic day 5 (ED5) with a vehicle control or one of three doses within the range of concentrations that have been detected in biota (nominal concentrations of 0, 10, 50 or 100 ng/g egg; measured concentrations 0, 3.0, 13.7 or 33.5 ng TBBPA-BDBPE/g egg and 0, 5.3, 26.8 or 58.1 ng BTBPE/g egg). Eggs were artificially incubated until hatching (ED28), at which point blood and tissues were collected to measure morphological and physiological endpoints, including organ somatic indices, circulating and glandular thyroid hormone concentrations, thyroid gland histology, hepatic deiodinase activity, and markers of oxidative stress. Neither compound had any effects on embryo survival through 90% of the incubation period or on hatching success, body mass, organ size, or oxidative stress of hatchlings. There was evidence of sex-specific effects in the thyroid system responses to the BTBPE exposures, with type 2 deiodinase (D2) activity decreasing at higher doses in female, but not in male hatchlings, suggesting that females may be more sensitive to BTBPE. However, there were no effects of TBBPA-BDBPE on the thyroid system in kestrels. For the BTPBE study, a subset of high-dose eggs was collected throughout the incubation period to measure changes in BTBPE concentrations. There was no decrease in BTBPE over the incubation period, suggesting that BTBPE is slowly metabolized by kestrel embryos throughout their ∼28-d development. These two compounds, therefore, do not appear to be particularly toxic to embryos of the American kestrel.

Ecologically-relevant exposure to methylmercury during early development does not affect adult phenotype in zebra finches (Taeniopygia guttata).

Methylmercury causes behavioural and reproductive effects in adult mammals via early developmental exposure. Similar studies in birds are limited and mostly focussed on aquatic systems, but recent work has reported high blood mercury concentrations in terrestrial, passerine songbirds. We used the zebra finch (Taeniopygia guttata) as a model to explore the long-term effects of early developmental exposure to methylmercury exposure. Chicks were dosed orally with either the vehicle control, 0.0315 µg Hg/g bw/day, or 0.075 µg Hg/g bw/day throughout the nestling period (days 1-21 post-hatching). We then measured (a) short-term effects on growth, development, and behaviour (time to self-feeding, neophobia) until 30 days of age (independence), and (b) long-term effects on courtship behaviour and song (males) and reproduction (females) once methylmercury-exposed birds reached sexual maturity (90 days post-hatching). High methylmercury treated birds had mean blood mercury of 0.734 ± 0.163 µg/g at 30 days post-hatching, within the range of values reported for field-sampled songbirds at mercury contaminated sites. However, there were no short-term effects of treatment on growth, development, and behaviour of chicks, and no long-term effects on courtship behaviour and song in males or reproductive performance in females. These results suggest that the nestling period is not a critical window for sensitivity to mercury exposure in zebra finches. Growing nestlings can reduce blood mercury levels through somatic growth and depuration into newly growing feathers, and as a result they might actually be less susceptible compared to adult birds receiving the same level of exposure.

Catbirds are the New Chickens: High Sensitivity to a Dioxin-like Compound in a Wildlife Species.

Dioxins and dioxin-like compounds (DLCs) are highly toxic and persistent global pollutants with extremely large differences in sensitivity across taxonomic groups. The chicken has long been considered uniquely sensitive to DLCs among avian species; but DLC toxicity in nondomesticated birds is largely untested, and the relevance of the chicken as an ecological model is uncertain. New approaches that use genotyping of the AHR1 ligand binding domain to screen for DLC sensitivity among avian species predicted that the gray catbird, a relevant wildlife species, is also highly sensitive. We tested this prediction using egg injections of a dioxin-like PCB (PCB-126) and found that the catbird is at least as sensitive as the chicken to DLCs, based on both embryotoxicity and mRNA induction of phase I metabolizing enzymes (CYP1A4/5). This study is the first to confirm that there are wildlife species as sensitive as the chicken and demonstrates how using predictive genotyping methods and targeted bioassays can focus toxicity assessments on ecologically relevant species.

The Flame-Retardant Tris(1,3-dichloro-2-propyl) Phosphate Represses Androgen Signaling in Human Prostate Cancer Cell Lines.

The effects of six organophosphate flame retardants (OPFRs) tris(2-butoxyethyl) phosphate, tris(2-chloroethyl) phosphate, tris(1-chloro-2-propyl) phosphate, tris(methylphenyl) phosphate, tris(1,3-dichloro-2-propyl) phosphate (TDCIPP), and triethyl phosphate on the activities of androgen receptor (AR), estrogen receptor (ER), and aryl hydrocarbon receptor (AhR) were assessed in human prostate and endometrial cancer cells. OPFRs had no effect on ER or AhR target gene activation in ECC-1 cells. The effect of TDCIPP on mRNA and protein accumulation of AR target genes was examined further. AR-inducible gene and protein expression were significantly altered by TDCIPP exposure and repressed PSA levels in conditioned media of prostate cancer cells. We demonstrated that TDCIPP has no affinity for the AR ligand binding domain (AR-LBD) and exerts its antiandrogenic effects in a noncompetitive fashion. Thus, the clinical relevance of TDCIPP exposure on prostate cancer detection and progression to a therapeutically refractile state ought to be investigated further.

An assessment of the developmental toxicity of BDE-99 in the European starling using an integrated laboratory and field approach.

Developmental exposure of wildlife to anthropogenic contaminants can have long-term effects that are difficult to assess in field monitoring studies, and may not be evident in laboratory studies that lack ecological components. The objective of this study was to assess the long-term effects of early exposure to contaminants under ecological conditions in a model passerine species, the European starling (Sturnus vulgaris). We selected 2,2',4,4',5-pentabromodiphenyl ether (BDE-99) as a representative contaminant, as it is one of the major constituents of the commercial penta-BDE flame retardant mixture, and has been reported in avian egg and tissue samples worldwide. We developed a novel approach to assess the developmental toxicity of BDE-99 in starlings by combining aspects of laboratory and field studies. We dosed free-living nestlings living in natural broods in the field with environmentally relevant concentrations of BDE-99 (0-173.8 ng/g bw/day) for the duration of the nesting cycle. To simulate monitoring of long-term effects we brought birds into captivity just prior to fledging and used photoperiod manipulations to induce reproductive development. We assessed a range of physiological and development measures such as hematocrit, oxidative stress, thyroid hormones, neuroanatomy, growth, molt rate, bill color, and testes development. We found some evidence of thyroid hormone disruption, but there were no effects on any other measures of physiology or development. The European starling could serve as a valuable model species for assessing early exposure and long-term effects of anthropogenic contaminants in terrestrial wildlife using this combined field/laboratory approach.

Wildlife ecological risk assessment in the 21st century: Promising technologies to assess toxicological effects.

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.

Toxicological effects assessment for wildlife in the 21st century: Review of current methods and recommendations for a path forward.

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.

Characterizing imidacloprid and metabolites in songbird blood with applications for diagnosing field exposures.

Neonicotinoids are the most widely used insecticides globally, but their rapid metabolism in vertebrates makes diagnosing wildlife exposure challenging. More detailed information on the pattern of imidacloprid metabolites over time could be used to better approximate the timing and level of exposure. Here, we applied recently developed sensitive analytical methods to measure imidacloprid (IMI) parent compound along with an expanded suite of metabolites (5-OH-IMI, IMI-olefin, desnitro-IMI, IMI-urea, 6-chloronicotinic acid, 5-AMCP, 6-OH nicotinic acid) and six other neonicotinoids in adult red-winged blackbirds (Agelaius phoeniceus) that were experimentally exposed to one of two field-realistic concentrations of imidacloprid (0.8 or 6.9 mg/kg bw). We measured concentrations in small (25 µL) plasma samples collected pre-exposure and at 1-, 6-, 24- and 48-h post-exposure. Imidacloprid was rapidly absorbed and metabolized within 48 h at both doses, with the largest decrease within 6 h post-exposure. The average proportion of parent IMI decreased from 68% of total detectable residues at 1-h to 34% at 6-h post-exposure. Two primary metabolites in blood were 5-OH-IMI and IMI-olefin, and 5-OH-IMI was the most persistent marker of exposure at 48-h. Desnitro-IMI was consistently detected following very recent (≤ 1-h) IMI exposure, and a higher ratio of parent IMI to metabolites also indicated recent exposure. Other metabolites were only detected in the higher dose group, and could be used as indicators of exposure to higher IMI concentrations. This sensitive analytical method and the observed metabolite patterns could be used to inform a growing body of field studies linking neonicotinoid exposure and effects in free-living birds.

A neonicotinoid insecticide reduces fueling and delays migration in songbirds.

Neonicotinoids are neurotoxic insecticides widely used as seed treatments, but little is known of their effects on migrating birds that forage in agricultural areas. We tracked the migratory movements of imidacloprid-exposed songbirds at a landscape scale using a combination of experimental dosing and automated radio telemetry. Ingestion of field-realistic quantities of imidacloprid (1.2 or 3.9 milligrams per kilogram body mass) by white-crowned sparrows (Zonotrichia leucophrys) during migratory stopover caused a rapid reduction in food consumption, mass, and fat and significantly affected their probability of departure. Birds in the high-dose treatment stayed a median of 3.5 days longer at the site of capture after exposure as compared with controls, likely to regain fuel stores or recover from intoxication. Migration delays can carry over to affect survival and reproduction; thus, these results confirm a link between sublethal pesticide exposure and adverse outcomes for migratory bird populations.

Part-per-trillion LC-MS/MS determination of neonicotinoids in small volumes of songbird plasma.

Neonicotinoids are the most widely used class of insecticides in the world, and there are increasing concerns about their effects on non-target organisms. Analytical methods to diagnose exposure to neonicotinoids in wildlife are still very limited, particularly for small animals such as songbirds. Blood can be used as a non-lethal sampling matrix, but the sample volume is limited by body size. Neonicotinoids have a low bioaccumulation potential and are rapidly metabolized, therefore, sensitive assays are critically needed to reliably detect their residues in blood samples. We developed an efficient LC-MS/MS method at a part-per-trillion (pg/ml) level to measure eight neonicotinoid related insecticides (acetamiprid, clothianidin, dinotefuran, flonicamid, imidacloprid, nitenpyram, thiacloprid and thiamethoxam) plus one metabolite (6-chloronicotinic acid) in small volumes (50 µL) of avian plasma. The average recovery of target compounds ranged from 95.7 to 101.3%, and relative standard deviations were between 0.82 and 2.13%. We applied the method to screen blood samples from 36 seed-eating songbirds (white-crowned sparrows; Zonotrichia leucophrys) at capture, and detected imidacloprid in 78% (28 of 36), thiamethoxam in 22% (8 of 36), thiacloprid in 11% (4 of 36), and acetamiprid in 11% (4 of 36) of wild-caught sparrows. 6 h after capture, birds were orally dosed with 0 (control), 1.2 or 3.9 mg of imidacloprid/kg bw, test results using this method indicated that plasma imidacloprid was significantly elevated (low 26-times, high 316-times) in exposed groups. This is the first study to confirm neonicotinoid exposure in small free-living songbirds through non-lethal blood sampling, and to demonstrate that environmentally realistic doses significantly elevate circulating imidacloprid concentrations. This sensitive method could be applied to characterize exposure to neonicotinoids in free-living wildlife and in toxicological studies.

Embryonic exposure to environmentally relevant concentrations of a brominated flame retardant reduces the size of song-control nuclei in a songbird.

Environmental contaminants have the potential to act as developmental stressors and impair development of song and the brain of songbirds, but they have been largely unstudied in this context. 2,2',4,4',5-Pentabromodiphenyl ether (BDE-99) is a brominated flame retardant congener that has demonstrated endocrine disrupting effects, and has pervaded the global environment. We assessed the effects of in ovo exposure to environmentally relevant levels of BDE-99 on the neuroanatomy of the song-control system in a model songbird species, the zebra finch (Taeniopygia guttata). Embryos were exposed via egg injection to a vehicle control (DMSO), 10, 100, or 1000 ng BDE-99/g egg on the day the egg was laid. Chicks were raised to sexual maturity to investigate long-term effects of BDE-99 on the adult male brain. Three key song-control nuclei (Area X, HVC, RA) all showed a dose-dependent trend toward decreasing volume as BDE-99 concentration increased, and birds exposed to 1000 ng/g in ovo BDE-99 had significantly smaller song-control nuclei volume compared to control birds. High environmental concentrations of BDE-99 in avian tissues can be within that range and thus could affect development of the song-control system in birds, and potentially other processes. We previously found that BDE-99 exposure during the nestling period had no effect of on the song-control system, although it did have significant effects on some behaviural endpoints. Taken together, these results suggest that exposure to polybrominated diphenyl ether (PBDEs) during critical developmental windows can significantly alter neurological development. © 2018 Wiley Periodicals, Inc. Develop Neurobiol, 2018.

Imidacloprid and chlorpyrifos insecticides impair migratory ability in a seed-eating songbird.

Birds that travel long distances between their wintering and breeding grounds may be particularly susceptible to neurotoxic insecticides, but the influence of insecticides on migration ability is poorly understood. Following acute exposure to two widely used agricultural insecticides, imidacloprid (neonicotinoid) and chlorpyrifos (organophosphate), we compared effects on body mass, migratory activity and orientation in a seed-eating bird, the white-crowned sparrow (Zonotrichia leucophrys). During spring migration, sparrows were captured, held and dosed by gavage daily for 3 days with either the vehicle control, low (10% LD50) or high (25% LD50) doses of imidacloprid or chlorpyrifos and tested in migratory orientation trials pre-exposure, post-exposure and during recovery. Control birds maintained body mass and a seasonally appropriate northward orientation throughout the experiment. Imidacloprid dosed birds exhibited significant declines in fat stores and body mass (mean loss: -17% low, -25% high dose) and failed to orient correctly. Chlorpyrifos had no overt effects on mass but significantly impaired orientation. These results suggest that wild songbirds consuming the equivalent of just four imidacloprid-treated canola seeds or eight chlorpyrifos granules per day over 3 days could suffer impaired condition, migration delays and improper migratory direction, which could lead to increased risk of mortality or lost breeding opportunity.

In ovo tris(2-butoxyethyl) phosphate concentrations significantly decrease in late incubation after a single exposure via injection, with no evidence of effects on hatching success or latent effects on growth or reproduction in zebra finches.

Zebra finch (Taeniopygia guttata) eggs were injected with the organophosphate triester flame retardant tris(2-butoxyethyl) phosphate (TBOEP) at 0 µg/g, 0.01 µg/g, 1 µg/g, 10 µg/g, or 50 µg/g egg. Subsets of high-dose eggs were collected throughout incubation to measure TBOEP, which started declining in late incubation and then decreased rapidly to 28% of injected concentration by hatching. The authors found no effects of TBOEP on survival, growth, or reproduction even at very high doses. Environ Toxicol Chem 2017;36:83-88. © 2016 SETAC.

Assessment of neuroanatomical and behavioural effects of in ovo methylmercury exposure in zebra finches (Taeniopygia guttata).

Methylmercury (MeHg) readily crosses the blood brain barrier and is a known neuro-toxicant. MeHg accumulation in the brain causes histopathological alterations, neurobehavioral changes, and impairments to cognitive motor functions in mammalian models. However, in birds the neurotoxic effects of MeHg on the developing pre-hatching brain and consequent behavioral alterations in adult birds have not received much attention. Moreover, passerine birds are poorly represented in MeHg neurotoxicology studies in comparison to other avian orders. Hence in this study, we used the egg injection method to investigate the long term effects of in ovo MeHg exposure on brain histopathology and courtship behavior in a model songbird species, the zebra finch (Taeniopygia guttata). Egg treatment groups included: a low MeHg dose of 0.2µg Hg g-1 egg, a high MeHg dose of 3.2µg Hg g-1 egg, and a vehicle control (water). No adverse effects of in ovo MeHg treatment were detected on courtship song quality or on mating behavior in experimental males at sexually maturity which would suggest that observable neurobehavioral effects of MeHg exposure may depend on the timing of exposure during offspring development. However, neuroanatomical analysis indicated an increase in telencephalon volume with increased MeHg concentrations which may suggest a prolonged inflammatory response in this region of the brain.

Acute embryotoxic effects but no long-term reproductive effects of in ovo methylmercury exposure in zebra finches (Taeniopygia guttata).

Mercury bioaccumulates in terrestrial ecosystems as methylmercury (MeHg), yet little is known about its effects on terrestrial organisms, including songbirds. The authors used a model songbird species, the zebra finch (Taeniopygia guttata), to assess short-term embryotoxic effects of in ovo MeHg exposure on hatching success and posthatching growth and nestling survival, as well as longer-term effects on mating behavior and reproduction. Egg treatment groups included a low-MeHg dose of 0.2 µg Hg g(-1) egg (n = 36), a high-MeHg dose of 3.2 µg Hg g(-1) egg (n = 49), and a control (n = 34). Doses were dissolved in nanopure filtered water and injected into the albumen on the day eggs showed signs of viability (3 d incubation). In ovo exposure to MeHg significantly reduced hatching success (53% in the high-MeHg dose group vs 94% in vehicle controls). Among hatched chicks, however, no effects of MeHg on growth, hematological variables, or nestling survival were detected. While the in ovo injection method resulted in a dose-dependent pattern of MeHg concentrations in blood of surviving chicks at 15 d and 30 d posthatching, there was evidence of rapid excretion of MeHg with nestling age during that growth period. At reproductive maturity (90 d of age), no long-term effects of in ovo exposure to MeHg on female mating behavior, reproductive effort (egg or clutch size), or growth and survivorship of offspring were observed. Environ Toxicol Chem 2016;35:1534-1540. © 2015 SETAC.

Assessment of concentrations and effects of organohalogen contaminants in a terrestrial passerine, the European starling.

European starlings (Sturnus vulgaris) are a valuable model species for the assessment of concentrations and effects of environmental contaminants in terrestrial birds. Polybrominated diphenyl ethers (PBDEs) are found in birds throughout the world, but relatively little is known of their concentrations or effects in free-living terrestrial passerines. We used a nest box population of European starlings to 1) measure the variation in egg concentrations of persistent organohalogen contaminants at an agricultural site, and 2) assess whether individual variation in PBDE concentrations in eggs was related to reproductive parameters, as well as maternal or nestling characteristics including body condition, thyroid hormones, oxidative stress, and hematocrit. As PBDEs were the main contaminant class of interest, we only assessed a subset of eggs for other organohalogen contaminants to establish background concentrations. Exposure to organohalogen contaminants was extremely variable over this relatively small study area. Geometric mean wet weight concentrations (range in brackets) of the major contaminants were 36.5 (12-174) ng/g ΣDDT (n=6 eggs) and 10.9 (2-307) ng/g ΣPBDEs (n=14). ΣPCBs at 3.58 (1.5-6.4) ng/g (n=6) were lower and less variable. There were low levels of other organochlorine (OC) pesticides such as dieldrin (2.02 ng/g), chlordanes (1.11 ng/g) and chlorobenzenes (0.23 ng/g). The only form of DDT detected was p,p'-DDE. The congener profiles of PBDEs and PCBs reflect those of industrial mixtures (i.e. DE-71, Aroclors 1254, 1260 and 1262). For all of the contaminant classes, concentrations detected in eggs at our study site were below levels previously reported to cause effects. Due to small sample sizes, we did not assess the relationship between ΣPCBs or ΣOCs and adult or chick condition. We observed no correlative relationships between individual variation in PBDE concentrations in starling eggs and reproductive success, maternal condition, or nestling condition in the corresponding nests.

Amino acid sequence of the AhR1 ligand-binding domain predicts avian sensitivity to dioxin like compounds: in vivo verification in European starlings.

Research has demonstrated that the sensitivity of avian species to the embyrotoxic effects of dioxin-like compounds can be predicted by the amino acid identities at two key sites within the ligand-binding domain of the aryl hydrocarbon receptor 1 (AhR1). The domestic chicken (Gallus gallus domesticus) has been established as a highly sensitive species to the toxic effects of dioxin-like compounds. Results from genotyping and in vitro assays predict that the European starling (Sturnus vulgaris) is also highly sensitive to dioxin-like compound toxicity. The objective of the present study was to test that prediction in vivo. To do this, we used egg injections in field nesting starlings with 3,3',4,4',5-pentachlorobiphenyl (PCB-126), a dioxin-like polychlorinated biphenyl. Eggs were dosed with either the vehicle control or 1 of 5 doses (1.4, 7.1, 15.9, 32.1, and 52.9 ng PCB-126/g egg). A dose-dependent increase in embryo mortality occurred, and the median lethal dose (LD50; 95% confidence interval [CI]) was 5.61 (2.33-9.08) ng/g. Hepatic CYP1A4/5 messenger RNA (mRNA) expression in hatchlings also increased in a dose-dependent manner, with CYP1A4 being more induced than CYP1A5. No effect of dose on morphological measures was seen, and we did not observe any overt malformations. These results indicate that, other than the chicken, the European starling is the most sensitive species to the effects of PCB-126 on avian embryo mortality reported to date, which supports the prediction of relative sensitivity to dioxin-like compounds based on amino acid sequence of the AhR1.