Exposure of Peregrine Falcons to Halogenated Flame Retardants: A 30 Year Retrospective Biomonitoring Study across North America.

Compared to aquatic ecosystem, terrestrial systems have been subjected to fewer investigations on the exposure to halogenated flame retardants (HFRs). Our study utilized peregrine falcon eggs collected from multiple habitats across North America to retrospectively explore both spatial distribution and temporal changes in legacy (e.g., polybrominated diphenyl ethers) and alternative HFRs over a 30 year period (1984-2016). The results reveal intensive HFR exposure in terrestrial ecosystems and chemical-specific spatiotemporal distribution patterns. The correlations between egg levels of the selected HFRs and human population density clearly illustrated a significant urban influence on the exposure of this wildlife species to these HFRs and subsequent maternal transfer to their eggs. Temporal analyses suggest that, unlike aquatic systems, terrestrial ecosystems may undergo continual exposure to consistently high levels of legacy HFRs for a long period of time. Our findings collectively highlight the effectiveness of using peregrine eggs to monitor terrestrial exposure to HFRs and other bioaccumulative chemicals and the need for continuous monitoring of HFRs in terrestrial ecosystems.

Characterizing the Movement of Per- and Polyfluoroalkyl Substances in an Avian Aquatic-Terrestrial Food Web.

The movement of per- and polyfluoroalkyl substances (PFAS) through linked aquatic-terrestrial food webs is not well understood. Tree swallows (Tachycineta bicolor) in such systems may be exposed to PFAS from multiple abiotic and/or biotic compartments. We show from fatty acid signatures and carbon stable isotopes that tree swallow nestlings in southwestern Ontario fed on both terrestrial and aquatic macroinvertebrates. The PFAS profiles of air, terrestrial invertebrates, and swallows were dominated by perfluorooctanesulfonic acid (PFOS). Short-chain perfluoroalkyl acids (PFAAs) were largely restricted to air, surface water, and sediment, and long-chain PFAAs were mainly found in aquatic invertebrates and tree swallows. PFOS, multiple long-chain perfluorocarboxylic acids [perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), perfluorotridecanoic acid (PFTrDA)] and perfluorooctane sulfonamide precursors were estimated to bioaccumulate from air to tree swallows. PFOS bioaccumulated from air to terrestrial invertebrates, and PFOS, PFDA, and perfluorooctane sulfonamidoacetic acids (FOSAAs) bioaccumulated from water to aquatic invertebrates. PFOS showed biomagnification from both terrestrial and aquatic invertebrates to tree swallows, and PFDA and FOSAAs were also biomagnified from aquatic invertebrates to tree swallows. The movement of PFAS through aquatic-terrestrial food webs appears congener- and compartment-specific, challenging the understanding of PFAS exposure routes for multiple species involved in these food webs.

The influence of environmental and ecological factors on the accumulation and distribution of short- and long-chain perfluoroalkyl acids in a mid-trophic avian insectivore.

Perfluoroalkyl acids (PFAAs) include perfluorooctane sulfonic acid (PFOS), perfluorooctanoic acid (PFOA), perfluorohexane sulfonic acid (PFHxS), and perfluorodecane sulfonic acid (PFDS), as well as increasingly used alternative short-chain perfluorosulfonic acids (PFSAs) and short- and long-chain (≥C9) perfluorocarboxylic acids (PFCAs). In the present study, tissues of tree swallows (Tachycineta bicolor) from two sites in southern Ontario, Canada, were analyzed for 17 individual PFAAs and showed egg and nestling tissue (liver, carcass) profiles dominated by PFOS (57-66%). The remaining PFAAs contributed ≤7% each, although collectively the long-chain PFCAs comprised 21-29% of the PFAAs. The short-chain PFSAs and PFCAs were among the lowest concentrations, suggesting that despite increased production and use of these alternative PFAAs, they are not accumulated to the same extent as the long-chain PFSAs and PFCAs. PFOS, PFDS, and some long-chain PFCAs were significantly higher in eggs than in livers and carcasses, whereas PFOA and the two short-chain PFCAs were significantly higher in nestling tissues than in eggs. For the two short-chain PFSAs, concentrations were similar among tissues. Tree swallow tissues at the site near a wastewater treatment plant (WWTP) outfall showed higher concentrations of PFOS, PFDS, PFHxS, and some long-chain PFCAs than tree swallows sampled at the nearby reference site; however, the influence of the WWTP was more equivocal for PFOA, other long-chain PFCAs, and short-chain PFSAs and PFCAs. Carbon stable isotopes (δ13C) and fatty acid signatures indicated that the diets of the WWTP swallows were more terrestrial than the reference swallows. Nonetheless, models considering environmental and ecological variables indicated that site was often the primary driver of PFAA variation among the swallows, with less or no influence of dietary patterns, or sex or body condition, revealing that of WWTP effluent can be an important environmental source of the major PFAAs in tree swallows.

Hepatic Gene Expression Profiling of American Kestrels (Falco sparverius) Exposed In Ovo to Three Alternative Brominated Flame Retardants.

A number of brominated flame retardants (BFRs) have been reported to interfere with the thyroid signaling pathway and cause oxidative stress in birds, yet the underlying shifts in gene expression associated with these effects remain poorly understood. In this study, we measured hepatic transcriptional responses of 31 genes in American kestrel (Falco sparverius) hatchlings following in ovo exposure to one of three high-volume alternative BFRs: 1,2-bis(2,4,6-tribromophenoxy) ethane (BTPBE), bis(2-ethylhexyl)-2,3,4,5-tetrabromophthalate (TBPH), or 2-ethylhexyl-2,3,4,5-tetrabromobenzoate (EHTBB). Hatchling kestrels exhibited shifts in the expression of genes related to oxidative stress (CYP, GSTA, SOD, and GPX1), thyroid hormone metabolism and transport (DIO1, DIO2, and TTR), lipid and protein metabolism (PPAR, HMGCR, FAB1, and LPL), and cytokine-mediated inflammation (TLR3, IL18, IRF7, STAT3, RACK1, and CEBPB). Male and female hatchlings differed in which genes were differentially expressed, as well as the direction of the effect (up- vs. downregulation). These results build upon our previous findings of increased oxidative stress and disrupted thyroid signaling pathway in the same hatchlings. Furthermore, our results indicate that inflammatory responses appear to occur in female hatchlings exposed to BTBPE and EHTBB in ovo. Gene expression analysis revealed multiple affected pathways, adding to the growing evidence that sublethal physiological effects are complex and are a concern for birds exposed to BTBPE, EHTBB, or TBPH in ovo.

Mercury, legacy and emerging POPs, and endocrine-behavioural linkages: Implications of Arctic change in a diving seabird.

Arctic species encounter multiple stressors including climate change and environmental contaminants. Some contaminants may disrupt hormones that govern the behavioural responses of wildlife to climatic variation, and thus the capacity of species to respond to climate change. We investigated correlative interactions between legacy and emerging persistent organic pollutants (POPs), mercury (Hg), hormones and behaviours, in thick-billed murres (Uria lomvia) (N = 163) breeding in northern Hudson Bay (2016-2018). The blood profile of the murres was dominated by methylmercury (MeHg), followed by much lower levels of sum (∑) 35 polychlorinated biphenyls (PCBs), hexachlorobenzene (HCB) and p,p'-dichlorodiphenyltrichloroethylene (DDE), polybrominated diphenyl ethers (PBDEs) BDE-47, -99 and BDE-100; all other measured organochlorine pesticides and replacement brominated flame retardants had low concentrations if detected. Inter-annual variations occurred in MeHg, circulating triiodothyronine (T3), thyroxine (T4), and the foraging behaviours of the murres, identified using GPS-accelerometers. Compared to the 50-year mean date (1971-2021) for 50% of sea-ice coverage in Hudson Bay, sea-ice breakup was 1-2 weeks earlier (2016, 2017) or comparable (2018). Indeed, 2017 was the earliest year on record. Consistent with relationships identified individually between MeHg and total T3, and T3 and foraging behaviour, a direct interaction between these three parameters was evident when all possible interactions among measured chemical pollutants, hormones, and behaviours of the murres were considered collectively (path analysis). When murres were likely already stressed due to early sea-ice breakup (2016, 2017), blood MeHg influenced circulating T3 that in turn reduced foraging time underwater. We conclude that when sea-ice breaks up early in the breeding season, Hg may interfere with the ability of murres to adjust their foraging behaviour via T3 in relation to variation in sea-ice.

Potential disruption of thyroid hormones by perfluoroalkyl acids in an Arctic seabird during reproduction.

Arctic marine ecosystems are experiencing rapid change, such as ocean warming and enhanced pollutants. Perfluoroalkyl acids (PFAAs) arriving via long-range transport have been detected in Arctic wildlife, including seabirds which are considered sentinels of marine ecosystem health. There is evidence that PFAA exposure leads to the disruption of thyroid hormones (THs), such as thyroxine (T4) and triiodothyronine (T3), which play important roles in metabolism, incubation, and thermoregulation in seabirds. Here, we investigated relationships between PFAAs and THs [total T4 (TT4), free T4 (FT4), total T3 (TT3) and free T3 (FT3)] in blood plasma collected from 63 thick-billed murres (Uria lomvia) at a colony located in northern Hudson Bay (2016-2018). We then tested if PFAAs and TH levels were related to fitness-associated reproductive traits, such as body mass and hatch dates. PFUdA, PFOS, and PFTrDA were the dominant PFAAs in murre blood, accounting for approximately 77% of ∑PFAA. Females had higher PFAAs than males, possibly due to higher trophic feeding. While FT3 increased with PFOS, PFNA, PFDA, PFDoA, PFTeDA, ∑PFCA7, and ∑PFAA in murres, TT3 decreased with PFOS, PFDoA, and PFTeDA in males, but not females, suggesting thyroid disruption. TT3 increased with body mass, whereas several long-chain PFAAs were negatively correlated with body mass. Negative relationships between PFNA, PFDoA, PFTrDA, PFTeDA, and ∑PFAA with hatch dates may be the result of a disruption in incubation behaviour, resulting in earlier hatch dates. Consequently, TT3 concentrations were highest in males and females in 2018, a year in which PFAAs were lowest and hatch dates were delayed relative to 2017. As an Arctic seabird experiencing several indirect effects of climate change, the interaction of PFAAs on thyroid activity may cause additional stress to murres.

Climate change and mercury in the Arctic: Biotic interactions.

Global climate change has led to profound alterations of the Arctic environment and ecosystems, with potential secondary effects on mercury (Hg) within Arctic biota. This review presents the current scientific evidence for impacts of direct physical climate change and indirect ecosystem change on Hg exposure and accumulation in Arctic terrestrial, freshwater, and marine organisms. As the marine environment is elevated in Hg compared to the terrestrial environment, terrestrial herbivores that now exploit coastal/marine foods when terrestrial plants are iced over may be exposed to higher Hg concentrations. Conversely, certain populations of predators, including Arctic foxes and polar bears, have shown lower Hg concentrations related to reduced sea ice-based foraging and increased land-based foraging. How climate change influences Hg in Arctic freshwater fishes is not clear, but for lacustrine populations it may depend on lake-specific conditions, including interrelated alterations in lake ice duration, turbidity, food web length and energy sources (benthic to pelagic), and growth dilution. In several marine mammal and seabird species, tissue Hg concentrations have shown correlations with climate and weather variables, including climate oscillation indices and sea ice trends; these findings suggest that wind, precipitation, and cryosphere changes that alter Hg transport and deposition are impacting Hg concentrations in Arctic marine organisms. Ecological changes, including northward range shifts of sub-Arctic species and altered body condition, have also been shown to affect Hg levels in some populations of Arctic marine species. Given the limited number of populations and species studied to date, especially within Arctic terrestrial and freshwater systems, further research is needed on climate-driven processes influencing Hg concentrations in Arctic ecosystems and their net effects. Long-term pan-Arctic monitoring programs should consider ancillary datasets on climate, weather, organism ecology and physiology to improve interpretation of spatial variation and time trends of Hg in Arctic biota.

The influence of global climate change on accumulation and toxicity of persistent organic pollutants and chemicals of emerging concern in Arctic food webs.

This review summarizes current understanding of how climate change-driven physical and ecological processes influence the levels of persistent organic pollutants (POPs) and contaminants of emerging Arctic concern (CEACs) in Arctic biota and food webs. The review also highlights how climate change may interact with other stressors to impact contaminant toxicity, and the utility of modeling and newer research tools in closing knowledge gaps on climate change-contaminant interactions. Permafrost thaw is influencing the concentrations of POPs in freshwater ecosystems. Physical climate parameters, including climate oscillation indices, precipitation, water salinity, sea ice age, and sea ice quality show statistical associations with POPs concentrations in multiple Arctic biota. Northward range-shifting species can act as biovectors for POPs and CEACs into Arctic marine food webs. Shifts in trophic position can alter POPs concentrations in populations of Arctic species. Reductions in body condition are associated with increases in levels of POPs in some biota. Although collectively understudied, multiple stressors, including contaminants and climate change, may act to cumulatively impact some populations of Arctic biota. Models are useful for predicting the net result of various contrasting climate-driven processes on POP and CEAC exposures; however, for some parameters, especially food web changes, insufficient data exists with which to populate such models. In addition to the impact of global regulations on POP levels in Arctic biota, this review demonstrates that there are various direct and indirect mechanisms by which climate change can influence contaminant exposure, accumulation, and effects; therefore, it is important to attribute POP variations to the actual contributing factors to inform future regulations and policies. To do so, a broad range of habitats, species, and processes must be considered for a thorough understanding and interpretation of the consequences to the distribution, accumulation, and effects of environmental contaminants. Given the complex interactions between climate change, contaminants, and ecosystems, it is important to plan for long-term, integrated pan-Arctic monitoring of key biota and ecosystems, and to collect ancillary data, including information on climate-related parameters, local meteorology, ecology, and physiology, and when possible, behavior, when carrying out research on POPs and CEACs in biota and food webs of the Arctic.

The potential of aerial insectivores for monitoring microplastics in terrestrial environments.

Limited research has been conducted on microplastics in terrestrial ecosystems and biota, despite being some of the most ubiquitous environmental pollutants. We investigated the presence of microplastics (over 125 µm) in tree swallow (Tachicyneta bicolor) chicks (10 d. o.), an aerial insectivore whose diet involves terrestrial and/or freshwater sources. Swallows nested immediately downstream (300 m) of the discharge pipe of a large, urban wastewater treatment plant (WWTP) or at a rural conservation area (40 km apart). Anthropogenic microparticles (including microplastics) were identified in nearly all WWTP chicks (90%; N = 20) and reference chicks (83%; N = 20). All microparticles were fibers (100%) in the gastro-intestinal (GI) tracts of WWTP nestlings, whereas unexpectedly, they were more diverse in the GI tracts of reference chicks, with ~15% characterized as pre-production plastic pellets. The fecal sacs of most nestlings (90%) contained microparticles, and all were characterized as fibers suggesting their excretion by tree swallows. Compared to WWTP chicks, the reference chicks had more microparticles in their fecal sacs and larger particles (length, width) in their GI tracts, likely reflecting the more aquatic-based diet of the reference chicks fed insects caught adjacent to the nearby dam, compared to the more terrestrial-based diet of the WWTP chicks. The numbers of microparticles were not correlated between GI tracts and fecal sacs, nor with the chicks' condition or size (weight, organs, feathers). We recommend sampling macroinvertebrate prey to permit stronger conclusions regarding WWTPs as possible sources of microplastics for swallows, and to determine if such macroinvertebrates may be a non-lethal method to characterize microparticle diversity ingested by birds as presently identified in chicks' GI tracts. We conclude that sampling fecal sacs only, while not indicative of the diversity of microplastics ingested by terrestrial passerines (e.g., tree swallows), is useful for determining their exposure to microparticles.

Establishment of baseline cytology metrics in nestling American kestrels (Falco sparverius): Immunomodulatory effects of the flame retardant isopropylated triarylphosphate isomers.

Avian populations must mount effective immune responses upon exposure to environmental stressors such as avian influenza and xenobiotics. Although multiple immune assays have been tested and applied to various avian species, antibody-mediated immune responses in non-model avian species are not commonly reported due to the lack of commercially available species-specific antibodies. The objectives of the present study were to advance methods for studying wild bird immune responses and to apply these to the evaluation of cytological responses after exposure of American kestrels, Falco sparverius, to a commercial flame retardant mixture containing isopropylated triarylphosphate isomers (ITP). Hatchlings were gavaged daily with safflower oil or 1.5 ug/g bw/day of ITP suspended in safflower oil, then bled on days 9, 17, and 21. The ITP treatment group (n = 18) and a subset of controls (Poly I:C treatment group; n = 10) were injected on days 9 and 15 with a synthetic analog of viral double-stranded RNA, polyinosinic:polycytidylic acid (Poly I:C), a toll-like receptor ligand and synthetic viral mimic, and responses compared to a sham injected control group (n = 8). The hypotheses tested whether kestrels showed immunological differences among treatment groups, genetic sex, and/or white blood cell (WBC) subpopulation type over time. A flow cytometry (FCM) gating strategy categorized heterophils (H), lymphocytes (L), and monocytes (M) and their proportions, and measured relative fluorescence in response to anti-chicken CD4 binding. Fluorescent cell surfaces and some granular/vacuolar inclusions were visualized by epifluorescence microscopy. A fourth subpopulation with higher levels of granularity than M but less than H became increasingly apparent with time and was gated along with the H subpopulation; its frequency of occurrence was lowest in the ITP group (P = 0.0023). The percentages of cells differed among treatment groups, days, and sexes (P = 0.0001). For both sexes, percentages of H and L were higher than M in control and Poly I:C. In the ITP group, L percentages were higher than H and M (P = 0.0457), and H and L were higher than M on days 9 and 21 (P = 0.0001). The ratios of H:L and H:WBC, indicators of robust immunity, were also higher on days 9 and 21 than on 17 (P = 0.0079). For each sex, the highest levels of activity measured by FCM geometric means (GEO) of fluorescence (indicative of antibody binding) were observed on day 9 (P = 0.0001 female, and P = 0.0011 male) in H over both L and M (P < 0.0001 for each). In males, GEO of the Poly I:C group was higher than that of the ITP group (P = 0.0374), with no difference observed among females over all days. By using a FCM algorithm for population comparisons of fluorescence to investigate binding within H, the T(x) scores indicated higher fluorescence in control and Poly I:C groups over ITP (P = 0.0001). Unlike chickens, Gallus gallus, which express CD4 primarily on L, kestrels bound the commercial antibody primarily within the gated H subpopulation, suggesting an immunophenotypic difference between taxa, despite a ~60% identity of Falco CD4 amino acid sequences with chicken CD4. The emergent cell subset within the gated H presented dendritic-like cell (DLC) morphological and functional properties, apparently serving as an effector cell. This study adds interpretive context to ecological investigations of infection and of potential immunomodulation by emerging compounds, whereby the early innate responses are mediated by the various cell subsets serving as useful quantitative markers of immunological condition. Data showed that dietary exposure to ITP was immunosuppressive for male and female kestrels over the course of the experiment, reducing DLC frequency compared to the Poly I:C controls. Heterophils and DLC were important in facilitating innate immunological responses.

Thyroid disruption and oxidative stress in American kestrels following embryonic exposure to the alternative flame retardants, EHTBB and TBPH.

Brominated flame retardant chemicals, such as 2-ethylhexyl-2,3,4,5-tetrabromobenzoate (EHTBB) (CAS #: 183658-27-7) and bis(2-ethylhexyl)-2,3,4,5-tetrabromophthalate (TBPH) (CAS #: 26040-51-7), have been detected in avian tissues and eggs from remote regions. Exposure to EHTBB and TBPH has been shown to cause oxidative stress and altered thyroid function in rodents and fish, yet no controlled studies have examined potential adverse effects of exposure in birds. Because flame retardants have been detected in wild raptors, we used American kestrels (Falco sparverius) as a model raptor to determine whether in ovo exposure to EHTBB or TBPH affected growth, hatching success, oxidative stress, or thyroid function. We exposed kestrel embryos to nominal concentrations (10, 50, or 100 ng g-1 egg weight) of EHTBB and TBPH via egg-injection on embryonic day 5. Embryonic exposure (~23 d) to EHTBB increased thyroid gland mass, reduced glandular colloid and total thyroxine (T4) in hatchling males and females, whereas deiodinase enzyme activity increased in males but decreased in females. Hatchlings exposed to TBPH in ovo exhibited reduced colloid and increased oxidative stress. Although exposure to EHTBB and TBPH caused several physiological effects (e.g., heart and brain mass), only exposure to 50 ng g-1 EHTBB appeared to reduce hatching success. Our results suggest these flame retardants may be hazardous for predatory birds. Future research should evaluate long-term survival and fitness consequences in birds exposed to these chemicals.

Influence of perfluoroalkyl acids and other parameters on circulating thyroid hormones and immune-related microRNA expression in free-ranging nestling peregrine falcons.

Exposure to certain perfluoroalkyl acids (PFAAs) can have considerable effects on the endocrine and immune systems, although such effects remain largely uncharacterized in wildlife. Using an apex avian predator, we investigated possible relationships of thyroid hormones (THs), specifically free (F) and total (T) thyroxine (FT4; TT4) and triiodothyronine (FT3; TT3), and the expression of an immune-related microRNA biomarker (i.e., miR-155), with the concentrations of 11 PFAAs in nestling peregrine falcons (Falco peregrinus). Nestling peregrines (n = 56; usually two chicks of each sex per nest) were blood sampled when 23 ± 4 days old in urban and rural regions of the Laurentian Great Lakes Basin (Ontario, Canada) in 2016 and 2018. The circulating concentrations of several PFAAs were significantly associated with THs and estimated thyroid gland activity (TT3:TT4; FT3:FT4), including PFHxS (FT3; FT3:FT4), PFDS (TT3; TT3:TT4), PFOA (TT4; FT3:FT4), PFTeDA (TT4; FT3:FT4), PFHxDA (TT4; TT3:TT4) and ΣPFCAs (TT4). Our novel evaluation of miR-155 in peregrine nestlings identified significantly negative relationships of plasma miR-155 counts with PFHxS and PFOA concentrations, indicating potential down-regulation of miR-155 expression and impaired immunity. Several PFAA homologues significantly predicted the variation in THs and miR-155 in conjunction with year (e.g., inter-annual differences in weather, ambient temperature, rainfall), region (urban/rural), nestling age, and/or diet (trophic position; δ15N), which suggests that multiple environmental and biological stressors, including PFAA exposure, influenced thyroid activity and immune function in these nestlings. Further research is warranted to identify the mechanisms and additional impacts of PFAA-related thyroid and immune disruption on the growth, development, and health risks in developing birds.

Interspecies comparisons of brominated flame retardants in relation to foraging ecology and behaviour of gulls frequenting a UK landfill.

This study quantifies and compares concentrations and profiles of legacy and alternative (alt-) brominated flame retardants (BFRs) in the eggs of three gull (Laridae) species of international/UK conservation concern - great black-backed gulls (Larus marinus; n = 7), European herring gulls (L. argentatus; n = 16) and lesser black-backed gulls (L. fuscus; n = 11) in relation to their foraging ecology and behaviour in order to investigate potential exposure pathways at a remote landfill in western Scotland, UK. Egg concentrations of sum (∑) polybrominated diphenyl ethers (∑8PBDEs) in all three species exceeded those for most reported avian species using landfill, except for those in North America. Despite relatively high detection frequencies of ∑hexabromocyclododecanes (∑3HBCDDs) (94-100%), concentrations of ∑8PBDEs exceeded ∑3HBCDDs and ∑5alt-BFRs, with ∑8PBDE levels similar in all three species. Egg carbon isotopic (δ13C) values highlighted a greater marine dietary input in great black-backed gulls that was consistent with their higher BDE-47 levels; otherwise, dietary tracers were minimally correlated with measured BFRs. ∑3HBCDD egg concentrations of herring gulls markedly exceeded those reported elsewhere in Europe. Decabromodiphenylethane (DBDPE) was the only alt-BFR detected (6-14% detection rate), in a single egg of each species. The great black-backed gull egg contained the highest concentration of DBDPE measured in biota to date globally and provides strong evidence for its emerging environmental presence as a BDE-209 replacement in UK wildlife. Correlations between δ13C (dietary source) and some measured BFRs in eggs suggest multiple routes of BFR exposure for gulls frequenting landfill through their diet, behaviour, preening, dermal exposure and likely inhalation. The frequent use of landfill by herring gulls and their increased egg BFR burdens suggest that this species may be an important bioindicator of BFR emissions from such sites.

A review of 1,2-dibromo-4-(1,2-dibromoethyl)cyclohexane in the environment and assessment of its persistence, bioaccumulation and toxicity.

Following the ban of many historically-used flame retardants (FRs), numerous replacement chemicals have been produced and used in products, with some being identified as environmental contaminants. One of these replacement flame retardants is 1,2-dibromo-4-(1,2-dibromoethyl)-cyclohexane (DBE-DBCH; formerly abbreviated as TBECH), which to date has not been identified for risk assessment and potential regulation. DBE-DBCH technical mixtures consist largely of α- and ß-diastereomers with trace amounts of γ- and δ-DBE-DBCH. The α- and ß-isomers are known contaminants in various environmental media. While current global use and production volumes of DBE-DBCH are unknown, recent studies identified that DBE-DBCH concentrations were among the highest of the measured bromine-based FRs in indoor and urban air in Europe. Yet our mass balance fugacity model and modeling of the physical-chemical properties of DBE-DBCH estimated only 1% partitioning to air with a half-life of 2.2 d atmospherically. In contrast, our modeling characterized DBE-DBCH adsorbing strongly to suspended particulates in the water column (~12%), settling onto sediment (2.5%) with minimal volatilization, but with most partitioning and adsorbing strongly to soil (~85%) with negligible volatilization and slow biodegradation. Our modeling further predicted that organisms would be exposed to DBE-DBCH through partitioning from the dissolved aquatic phase, soil, and by diet, and given its estimated logKow (5.24) and a half-life of 1.7 d in fish, DBE-DBCH is expected to bioaccumulate into lipophilic tissues. Low concentrations of DBE-DBCH are commonly measured in biota and humans, possibly because evidence suggests rapid metabolism. Yet toxicological effects are evident at low exposure concentrations: DBE-DBCH is a proven endocrine disruptor of sex and thyroid hormone pathways, with in vivo toxic effects on reproductive, metabolic, and other endpoints. The objectives of this review are to identify the current state of knowledge concerning DBE-DBCH through an evaluation of its persistence, potential for bioaccumulation, and characterization of its toxicity, while identifying areas for future research.

Perfluoroalkyl acids and sulfonamides and dietary, biological and ecological associations in peregrine falcons from the Laurentian Great Lakes Basin, Canada.

Perfluoroalkyl substances (PFAS) are a large, diverse group of chemicals and several perfluoroalkyl acids (PFAAs) are known environmental contaminants. Wildlife exposure to PFAAs and precursors has been shown, but less is known regarding replacements such as shorter-chain PFAS. In the present study, exposure to a suite of PFAAs and associations with dietary, biological and ecological factors were investigated in populations of a sentinel apex species - the peregrine falcon (Falco peregrinus). Nestling blood (n = 57) and sibling eggs (n = 9) were sampled in 2016 and 2018 from nests in rural and urban regions across the Laurentian Great Lakes Basin, Canada. PFSAs (perfluorinated sulfonic acids) including PFHxS, PFOS, and PFDS were detected in most egg and plasma samples, whereas 11 PFCAs (perfluorinated carboxylic acids; C5-C14, C16) compared to eight PFCAs (C8-C14, C16) were detected in most eggs and plasma, respectively. Shorter-chain C8-C10 PFCAs were more dominant in plasma and longer-chain C12-C14 PFCAs in eggs, but profiles were similar for PFOS, PFDS, PFUdA and PFHxDA. The exposure to PFAAs in peregrine falcons is likely mediated by dietary factors such as foraging location (δ13C and δ34S) and trophic position (δ15N) given the associations observed in eggs and nestling plasma, respectively. Moreover, significant relationships were observed for circulating ΣPFCAs and region (rural/urban), and nestling body condition after adjusting for sampling year and dietary tracers, suggesting that compared to rural nestlings, urban nestlings may be more exposed to ΣPFCAs and prone to their potential physiological impacts. Our findings highlight the importance of integrating dietary, biological and ecological factors when studying PFAS exposure in birds.

Assessment of the effects of early life exposure to triphenyl phosphate on fear, boldness, aggression, and activity in Japanese quail (Coturnix japonica) chicks.

Triphenyl phosphate (TPHP) is an organophosphate ester (OPE) used as a flame retardant (FR) and plasticizer. TPHP has previously been shown to disrupt behaviour in fish and mammals, but to our knowledge, this is the first study on the behavioural effects of TPHP in birds. Early life stage Japanese quail (Coturnix japonica) were exposed to nominal doses of 0 ng/g (vehicle-control), 5 ng/g (low dose), 50 ng/g (mid dose), and 100 ng/g (high dose) TPHP, both as embryos (via air cell injection prior to incubation) and as chicks (via daily gavage until 5 days post-hatch). The low dose reflects TPHP levels recorded in wild avian eggs, but actual environmental exposure levels may be higher given that TPHP is known to be rapidly metabolized in birds. We previously reported that the chicks exposed to TPHP in this study experienced reduced growth and resting metabolic rate, and sex-specific changes in thyroid function. The current study focuses on behavioural endpoints. We found that high-TPHP chicks exhibited less neophobia than vehicle-controls, and low-TPHP chicks exhibited more aggression towards conspecifics. No differences were observed in the responses of Japanese quail chicks to activity or tonic immobility (fear response) tests. These data add weight of evidence to previous findings suggesting that TPHP, among other OPEs, can disrupt ecologically-relevant behaviours in exposed vertebrates.

Uptake, Deposition, and Metabolism of Triphenyl Phosphate in Embryonated Eggs and Chicks of Japanese Quail (Coturnix japonica).

The toxicokinetics of triphenyl phosphate (TPHP) in vivo including the uptake, deposition, and biotransformation into the metabolite diphenyl phosphate (DPHP) is presently reported in embryonated eggs and chicks of Japanese quail. Quail were dosed with TPHP at 3 concentrations by air cell egg injection on embryonic day 0, followed by daily oral dosing after chicks hatched (5 d). Vehicle-only exposed controls were also used. In dosed eggs, only 33% of the TPHP remained 2 d after injection (no hepatic development); after 10 d (post-hepatogenesis), only 2% remained. The estimated TPHP half-lives in the eggs ranged from 1.1 to 1.8 d for the 3 dosed groups. In all exposed eggs and chicks, DPHP significantly increased with dose (0.001 < p < 0.044). It appears that DPHP is an important metabolite in quail, making up 41 to 74% of all metabolites formed in embryonated eggs. In chicks, at medium and high doses, DPHP concentrations significantly exceeded those of TPHP (p ≤ 0.007), making up 67 and 76% of the total burden, respectively. Our findings suggest that rapid TPHP metabolism occurred in chicks and embryonated quail eggs but that this may vary with the age of the embryonated egg and the stage of embryo development, which should be considered when evaluating concentrations of TPHP and DPHP measured in eggs of wild birds. Environ Toxicol Chem 2020;39:565-573. © 2019 Her Majesty the Queen in Right of Canada. Environmental Toxicology and Chemistry © 2019 SETAC.

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.

Flame retardant concentrations and profiles in wild birds associated with landfill: A critical review.

Given factors such as their persistence and toxicity, legacy brominated flame retardants (BFRs) like polybrominated diphenyl ethers (PBDEs) and hexabromocyclododecane (HBCDD), are designated as persistent organic pollutants (POPs) and are subject to regulation. Waste streams likely represent a substantial reservoir of legacy BFRs given that they were once widely applied to goods which are increasingly likely to be obsolete. Waste streams are also increasingly likely to be a source of emerging flame retardants, in particular, novel BFRs (NBFRs), the halogenated norbornene flame retardant Dechlorane Plus (DDC-CO) and the brominated, chlorinated or non-halogenated organophosphate triester flame retardants (PFRs). Many bird populations rely on landfill and its surrounding land-use for inter alia the opportunities it provides for activities such as foraging and resting. However, studies on captive and wild (free-living) birds have demonstrated deleterious effects of several FRs. Globally, approximately 250 bird species, including many of conservation concern, are reported to use landfill and surrounding habitat (including wastewater treatment operations), thus putting birds potentially at risk of exposure to such chemicals. We synthesise and critically evaluate a total of 18 studies covering eight avian species published between 2008 and 2018 (inclusive) across four continents that report flame retardant (FR) burdens in birds utilising landfill. Several such studies found FRs at among the highest concentrations detected in wild biota to date. We recommend that ongoing research be focused on landfill-associated birds, given that landfill is an important source of FRs and other anthropogenic chemicals, and particularly at sites where species are of conservation concern. We suggest ways in which the comparative power of studies could be enhanced in the future, the reporting of a minimum common suite of key chemicals, and where feasible, standardisation of the tissue compartments (i.e., eggs) to be studied. We conclude by identifying future research directions.