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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; δ 15 N), 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., Competing Interests: Declaration of competing interest The authors declare that there are no intellectual or financial conflicts of interest., (Crown Copyright © 2021. Published by Elsevier B.V. All rights reserved.)

In the present study, concentrations of legacy and emerging contaminants were determined in three non-destructive matrices (plasma, preen oil and body feathers) of northern goshawk (Accipiter gentilis) nestlings. Persistent organic pollutants (POPs), including polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs) and polybrominated diphenyl ethers (PBDEs), together with emerging pollutants, including per- and polyfluorinated alkyl substances (PFASs), novel brominated flame retardants (NBFRs), phosphorus flame retardants (PFRs) and Dechlorane Plus isomers (DPs) were targeted. Plasma, preen oil and feather samples were collected from 61 goshawk nestlings in Norway (Trøndelag and Troms) in 2015 and 2016, and pollutant concentrations were compared between the three matrices. In plasma, PFASs were detected in the highest concentrations, ranging between 1.37 and 36.0 ng/mL, which suggests that the nestlings were recently and continuously exposed to these emerging contaminants, likely through dietary input. In preen oil, OCPs (169-3560 ng/g) showed the highest concentrations among the investigated compounds, consistent with their high lipophilicity. PFRs (2.60-314 ng/g) were the dominant compounds in feathers and are thought to originate mainly from external deposition, as they were not detected in the other two matrices. NBFRs and DPs were generally not detected in the nestlings, suggesting low presence of these emerging contaminants in their environment and/or low absorption. Strong and significant correlations between matrices were found for all POPs (r s  = 0.46-0.95, p < 0.001), except for hexachlorobenzene (HCB, r s  = 0.20, p = 0.13). Correlations for PFASs were less conclusive: linear perfluorooctane sulfonate (PFOS), perfluoroundecanoate (PFUnA), perfluorododecanoate (PFDoA) and perfluorotetradecanoate (PFTeA) showed strong and significant correlations between plasma and feathers (r s  = 0.42-0.72, p < 0.02), however no correlation was found for perfluorohexane sulfonate (PFHxS), perfluorononanoate (PFNA) and perfluorotridecanoate (PFTriA) (r s  = 0.05-0.33, p = 0.09-0.85). A lack of consistency between the PFAS compounds (contrary to POPs), and between studies, prevents concluding on the suitability of the investigated matrices for PFAS biomonitoring., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

Understanding the levels and drivers of contamination in top predators is important for their conservation and eventual use as sentinels in environmental monitoring. Therefore, metals and trace elements were analyzed in feathers of Bonelli's eagles (Aquila fasciata) from southern Portugal in 2007-2013, where they are believed to be exposed to a wide range of contamination sources such as agricultural land uses, urban areas, active and abandoned mines and a coal-fired power plant. We focused on concentrations of aluminum (Al), arsenic (As), copper (Cu), chromium (Cr), mercury (Hg), lead (Pb), selenium (Se) and zinc (Zn), as these contaminants are potentially associated with those sources and are known to pose a risk for terrestrial vertebrates. Stable isotope values of nitrogen (δ 15 N: 15 N/ 14 N), carbon (δ 13 C: 13 C/ 12 C) and sulphur (δ 34 S: 34 S/ 32 S) were used as dietary proxies to control for potential effects of prey composition on the contamination pattern. The spatial distribution of potential contamination sources was quantified using geographic information systems. Concentrations of Hg in the southern part of the study area were above a reported toxicity threshold for raptors, particularly in territories closer to a coal-fired power plant at Sines, showing that contamination persisted after a previous assessment conducted in the 1990s. Hg and Se levels were positively correlated with δ 15 N, which indicates biomagnification. Concentrations of As, Cr, Cu, Pb and Zn were generally low and unrelated to mining- or industrial activities, indicating low environmental background concentrations. Al was found at higher concentrations in the southernmost areas of Portugal, but this pattern might be related to external soil contamination on feathers. Overall, this study indicates that, among all elements studied, Hg seems to be the most important contaminant for Bonelli's eagles in southern Portugal, likely due to the power plant emissions and biomagnification of Hg in terrestrial food webs., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Persistent organic pollutants (POPs) are still globally distributed and some have been shown to interact with the endocrine system of birds. However, the relationship between POPs and the stress response mediated by the hypothalamic-pituitary-adrenal (HPA) axis is still poorly understood. Raising concerns are now focused on the toxic properties of emergent organophosphate ester flame retardants (OPEs), but whether OPEs interact with the HPA axis response has not yet been investigated. We measured corticosterone concentrations in feathers (CORTf) as a long-term biomarker of the bird HPA axis response and we investigated their relationship with POP and OPE concentrations in down feathers of nestling cinereous vultures (Aegypius monachus). We also examined whether high contaminant burden and high CORTf concentrations impacted the duration of chick development. The most predominant compounds were the following: p,p'-DDE (3.28 ± 0.26 ng g -1 dw) > γ-HCH (0.78 ± 0.09 ng g -1 dw) > BDE-99 (0.73 ± 0.09 ng g -1 dw) > CB-153 (0.67 ± 0.04 ng g -1 dw). The most persistent POP compounds (CB-170, -177, -180, -183, -187, -194 and p,p'-DDE) were associated (P = 0.02) with high concentrations of CORTf (range: 0.55-6.09 pg mm -1 ), while no relationship was found when OPEs were tested (P > 0.05). Later egg-laying was positively associated to high levels of CORTf (P = 0.02) and reduced duration of chick development (P < 0.001), suggesting a beneficial effect of the HPA axis response on the growth of the chicks. In addition, males with high concentrations of the most persistent POP compounds tended to show a reduced duration of the nestling period (P = 0.05) and an equal fledging success than chicks with lower levels. These findings suggest that POPs, but not OPEs, may interact with the HPA axis response of chicks, although levels were not high enough to cause detrimental consequences., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Raptor feathers have been increasingly used to assess pollutants in ecotoxicological monitoring studies. However, the suitability of down feathers to detect pollutants has not yet been investigated. In this study, concentrations of persistent organic pollutants (POPs) and organophosphate ester flame retardants (OPEs) were assessed in down and juvenile contour feathers of Spanish cinereous vulture (Aegypius monachus) nestlings (circa 73 days old) and contaminant concentrations were compared between both types of feathers from the same individuals. Concentrations of polychlorinated biphenyls (PCBs: 1.30-6.16 ng g -1 dw feather), polybrominated diphenyl ethers (PBDEs: 0.23-1.35 ng g -1 dw feather), p,p'-dichlorodiphenyldichloroethylene (pp-DDE: 0.09-6.10 ng g -1 dw feather) and tris (1-chloro-2-propyl) phosphate (TCiPP: 0.86-48.96 ng g -1 dw feather) were significantly higher in down than in contour feathers. In contrast, contour feathers showed higher levels of the more volatile POP, lindane (0.25-3.12 ng g -1 dw feather). Concentrations of hexachlorobenzene (HCB) and OPEs (except TCiPP) were similar between the two types of feathers. By showing high accumulation of the most persistent POPs investigated, down feathers presented a contamination profile similar to that previously described in raptor eggs. As these feathers grow during the first days of a vulture chicks life, they probably reflect the contaminant burden of the chick due to maternal transfer to the egg. Overall, the present study provides the first indication that down feathers may be useful for biomonitoring studies. Further research is needed to confirm whether nestling down feathers reflect the concentrations in the egg., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Information on trace element pollution in the terrestrial environment and its biota is limited compared to the marine environment. In the present study, we collected body feathers and blood of 37 Northern goshawk (Accipiter gentilis) nestlings from Tromsø (northern Norway), Trondheim (central Norway), and Murcia (southeastern Spain) to study regional exposure, hypothesizing the potential health risks of metals and other trace elements. Blood and body feathers were analyzed by a high resolution inductively coupled plasma mass spectrometer (HR-ICP-MS) for aluminum (Al), nickel (Ni), copper (Cu), zinc (Zn), arsenic (As), selenium (Se), cadmium (Cd), mercury (Hg) and lead (Pb). The influence of regional differences, urbanization and agricultural land usage in proximity to the nesting Northern goshawks was investigated using particular spatial analysis techniques. Most trace elements were detected below literature blood toxicity thresholds, except for elevated concentrations (mean ± SD µgml -1 ww) found for Zn (5.4 ± 1.5), Cd (0.00023 ± 0.0002), and Hg (0.021 ± 0.01). Corresponding mean concentrations in feathers (mean ± SD µgg -1 dw) were 82.0 ± 12.4, 0.0018 ± 0.002, and 0.26 ± 0.2 for Zn, Cd and Hg respectively. Multiple linear regressions indicated region was a significant factor influencing Al, Zn, Se and Hg feather concentrations. Blood Cd and Hg concentrations were significantly influenced by agricultural land cover. Urbanization did not have a significant impact on trace element concentrations in either blood or feathers. Overall metal and trace element levels do not indicate a high risk for toxic effects in the nestlings. Levels of Cd in Tromsø and Hg in Trondheim were however above sub-lethal toxic threshold levels. For holistic risk assessment purposes it is important that the concentrations found in the nestlings of this study indicate that terrestrial raptors are exposed to various trace elements., (Copyright © 2017 Elsevier Inc. All rights reserved.)

To date, knowledge of persistent organic pollutant (POP) mediated oxidative stress responses in avian species is rather limited. We therefore investigated whether exposure to polybrominated diphenyl ethers (PBDEs) and organochlorine pesticides (OCPs) in two predatory bird species, namely black kite (Milvus migrans) and spotted owlet (Athene brama), was associated to activities of antioxidant enzymes, such as glutathione peroxidase (GPx), glutathione S-transferase (GST), glutathione reductase (GR) and catalase (CAT), or expression of GPx and superoxide dismutase (SOD) genes. As part of this investigation, we evaluated whether feathers were suitable to reflect internal body burdens and their associated oxidative stress effects. p,p'-DDE was unanimously recorded with highest concentrations in feathers and blood of both species. In general, the non-significant associations reflect that feathers are not always a suitable indicator for internal body burdens of POPs, depending on the feather type and the age of the bird. The activity of GST and GR was significantly higher in spotted owlet whereas GPx and CAT was higher (albeit not significant) in spotted owlet and black kite respectively. In comparison, mRNA expression of GPx, SOD and Cu,ZnSOD was significantly higher in black kite. Regression analysis showed that the activity of GST and GR was significantly associated with p,p'-DDE in blood of spotted owlet. Similarly, activity of CAT and GR was significantly correlated with BDE-100 in feathers of spotted owlet. In comparison, mRNA expression of SOD was found significantly associated with ∑PBDEs in blood of spotted owlet as well as p,p'-DDE in feathers of black kite. Significant associations of various POPs with biological responses may suggest that POP exposure may be contributing to oxidative stress in the studied bird of prey species. This first investigation indicates the necessity for further research on cause-effect relationships between POP exposures and changes in general health of free ranging birds., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Feathers are useful for monitoring contaminants in wild birds and are increasingly used to determine persistent organic pollutants. However, few studies have been conducted on birds with known exposure levels. We aimed to determine how well nestling feather concentrations reflect in ovo exposure to hexabromocyclododecane (α-, β- and γ-HBCDD), and to determine if feather concentrations are related to physiological biomarkers. Captive kestrels (n = 11) were exposed in ovo to maternally transferred HBCDD-isomers at concentrations of 127, 12 and 2 ng/g wet weight of α-, β- and γ-HBCDD (measured in sibling eggs), respectively, and compared to controls (n = 6). Nestling growth was monitored at 5 d intervals and circulating thyroid hormone concentrations assessed at d 20. Tail feathers were collected prior to the first molt and analyzed for HBCDD isomers. The mean ΣHBCDD concentration in feathers was 2405 pg/g dry weight (in exposed birds) and α-, β- and γ-HBCDD made up 32%, 13%, and 55%, respectively of the ΣHBCDD concentrations. This isomer distribution deviated from the typical dominance of α-HBCDD reported in vertebrate samples. Exposed chicks had significantly higher feather concentrations of β- and γ-HBCDD compared with controls (p = 0.007 and p = 0.001 respectively), while α-HBCDD concentrations did not differ between the two groups. Feather concentrations of α-HBCDD were best explained by egg concentrations of β- or γ-HBCDD concentrations (w i  = 0.50, 0.30 respectively), while feather concentrations of β- and γ-HBCDD were influenced by growth parameters (rectrix length: w i  = 0.61; tibiotarsus length: w i  = 0.28). These results suggest that feather α-HBCDD concentrations may reflect internal body burdens, whereas β- and γ-HBCDD may be subject to selective uptake. The α-HBCDD concentrations in the feathers were negatively associated with the ratio of plasma free triiodothyronine to free thyroxine (T 3 :T 4 ; p = 0.020), demonstrating for the first time that feather concentrations may be used to model the effect of body burdens on physiological endpoints., (Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.)