Evaluation of anticoagulant rodenticide sensitivity by examining in vivo and in vitro responses in avian species, focusing on raptors.

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

A premature stop codon in the CYP2C19 gene may explain the unexpected sensitivity of vultures to diclofenac toxicity.

The unintended environmental exposure of vultures to diclofenac has resulted in the deaths of millions of old-world vultures on the Asian subcontinent. While toxicity has been since associated with a long half-life of elimination and zero order metabolism, the actual constraint in biotransformation is yet to be clarified. For this study we evaluated if the evident zero order metabolism could be due to defects in the CYP2C9/2C19 enzyme system. For this, using whole genome sequencing and de-novo transcriptome alignment, the vulture CYP2C19 open reading frame was identified through Splign analysis. The result sequence analysis revealed the presence of a premature stop codon on intron 7 of the identified open reading frame. Even if the stop codon was not present, amino acid residue analysis tended to suggest that the enzyme would be lower in activity than the equivalent human enzyme, with differences present at sites 105, 286 and 289. The defect was also conserved across the eight non-related vultures tested. From these results, we conclude that the sensitivity of the old-world vultures to diclofenac is due to the non-expression of a viable CYP2C19 enzyme system. This is not too dissimilar to the effects seen in certain people with a similar defective enzyme.

Sub-lethal impacts of lead poisoning on blood biochemistry, immune function and delta-aminolevulinic acid dehydratase (δ-ALAD) activity in Cape (Gyps coprotheres) and white-backed (G. africanus) Vulture chicks.

Although the prevalence of lead poisoning in southern Africa's Gyps vultures is now well-established, its finer physiological effects on these endangered species remain poorly characterised. We evaluated the sub-lethal impact of acute lead exposure on Cape and White-backed Vulture chicks from two breeding colonies in South Africa, by analysing its possible effects on key blood biochemistry parameters, immune function, packed cell volume and δ-aminolevulinic acid dehydratase (δ-ALAD) activity. All 37 White-backed Vulture nestlings sampled displayed elevated lead levels (>10 µg/dL), and seven had blood [Pb] >100 µg/dL. Eight of 28 Cape Vulture nestlings sampled had blood [Pb] exceeding background exposure, with one showing blood [Pb] >100 µg/dL. Delta-aminolevulinic acid dehydratase (δ-ALAD) activity was significantly and negatively related to blood [Pb] in nestlings from both species, with 50% inhibition of the enzyme predicted to occur at blood [Pb] = 52.8 µg/dL (White-backed Vulture) and 18.8 µg/dL (Cape Vulture). Although no significant relationship was found between % packed cell volume (PCV) and blood [Pb], the relatively lower mean PCV of 32.9% in White-backed Vulture chicks, combined with normal serum protein values, is likely indicative of depression or haemolytic anaemia. The leukogram was consistent in both species, although the presence of immature heterophils suggested an inflammatory response in White-backed Vulture chicks with blood [Pb] >100 µg/dL. Values for cholesterol, triglycerides, total serum protein, albumin, globulin, albumin/globulin ratio, alanine aminotransferase (ALT) and gamma-glutamyl transferase (GGT) were consistent with values previously reported. Calcium and phosphorus concentrations suggested no adverse effects on bone metabolism. A significant decrease in urea: uric acid (U:UA) ratio at blood [Pb] >100 µg/dL in White-backed Vulture chicks, brought about by a decrease in urea production, raises the possibility of hepatic abnormality. These results suggest that δ-ALAD activity may serve as a sensitive biomarker of lead toxicity in both species, while highlighting the need to better understand the significant variability in sensitivity that is observed, even between closely related members of the same genus.

Reproductive performance of Peregrine falcons relative to the use of organochlorine pesticides, 1946-2021.

Populations of some fish- and meat-eating birds suffered dramatic declines globally following the introduction of organochlorine pesticides during the late 1940s and 1950s. It has been hypothesised that these population declines during the 1950s-1970s were largely driven by a combination of reproductive failure due to eggshell-thinning, egg breakage and embryonic death attributable to DDT and its metabolites, and to enhanced mortality attributable to the more toxic cyclodiene compounds such as aldrin and dieldrin. Using 75 years (1946-2021) of Peregrine falcon (Falco peregrinus) monitoring data (315 unique nest-sites monitored for 6110 nest-years), we studied the breeding performance of a resident Peregrine population in southern Scotland relative to the spatiotemporal pattern of organochlorine pesticide use. We show that (i) Peregrine breeding success and measures of breeding performance increased substantially following the reduction in, and subsequently a complete ban on, the use of organochlorine pesticides; (ii) improvements in Peregrine breeding performance were more dramatic in southeastern Scotland where agriculture was the predominant land use than in southwestern Scotland where there was less arable and more forested land; (iii) Peregrines nesting closer to the coast generally had higher fledging success (that is, a higher proportion of clutches that produced at least one fledgeling) than those nesting inland farther away from the coast; (iv) low temperatures and excessive rain in May negatively affected Peregrine fledging success; and (v) Peregrine abundance increased in parallel with improvements in reproductive performance following the reduction and then complete ban on the use of organochlorine pesticides in the UK. However, recovery was gradual and occurred over four decades, and rate of recovery varied among measures of reproductive performance (egg, nestling and fledgeling production). Our results suggest that the temporal pattern of organochlorine pesticide use strongly influenced Peregrine reproductive parameters but that the pattern of influence differed regionally. Overall results are consistent with the hypothesis that reproductive failure caused by organochlorine pesticides was an important driver of the decline in the south Scottish Peregrine population, and that improvements in all measures of breeding performance following a reduction and eventual ban on organochlorine use facilitated the observed increase in this population.

Anticoagulant rodenticides are associated with increased stress and reduced body condition of avian scavengers in the Pacific Northwest.

Anticoagulant rodenticides (AR) have been used globally to manage commensal rodents for decades. However their application has also resulted in primary, secondary, and tertiary poisoning in wildlife. Widespread exposure to ARs (primarily second generation ARs; SGARs) in raptors and avian scavengers has triggered considerable conservation concern over their potential effects on populations. To identify risk to extant raptor and avian scavenger populations in Oregon and potential future risk to the California condor (Gymnogyps californianus) flock recently established in northern California, we assessed AR exposure and physiological responses in two avian scavenger species (common ravens [Corvus corax] and turkey vultures [Cathartes aura]) throughout Oregon between 2013 and 2019. AR exposure was widespread with 51% (35/68) of common ravens and 86% (63/73) of turkey vultures containing AR residues. The more acutely toxic SGAR brodifacoum was present in 83% and 90% of AR exposed common ravens and turkey vultures. The odds of AR exposure in common ravens were 4.7-fold higher along the coastal region compared to interior Oregon. For common ravens and turkey vultures that were exposed to ARs, respectively, 54% and 56% had concentrations that exceeded the 5% probability of toxicosis (>20 ng/g ww; Thomas et al., 2011), and 20% and 5% exceeded the 20% probability of toxicosis (>80 ng/g ww; Thomas et al., 2011). Common ravens exhibited a physiological response to AR exposure with fecal corticosterone metabolites increasing with sum ARs (ΣAR) concentrations. Both female common raven and turkey vultures' body condition was negatively correlated with increasing ΣAR concentrations. Our results suggest avian scavengers in Oregon are experiencing extensive AR exposure and the newly established population of California condors in northern California may experience similar AR exposure if they feed in southern Oregon. Understanding the sources of ARs across the landscape is an important first step in reducing or eliminating AR exposure in avian scavengers.

Metabolism of aceclofenac to diclofenac in the domestic water buffalo Bubalus bubalis confirms it as a threat to Critically Endangered Gyps vultures in South Asia.

Vulture declines in South Asia were caused by accidental poisoning by the veterinary non-steroidal anti-inflammatory drug (NSAID) diclofenac. Although veterinary use of diclofenac has been banned, other vulture-toxic NSAIDs are legally available, including aceclofenac, which has been shown to metabolise into diclofenac in domestic cattle. We gave nine domestic water buffalo the recommended dose of aceclofenac (2 mg kg-1 body weight), collected blood at intervals up to 48 h, and carried out a pharmacokinetic analysis of aceclofenac and its metabolite diclofenac in plasma. Aceclofenac was rapidly converted to diclofenac, and was barely detectable in plasma at any sampling time. Diclofenac was present within 20 min, and peaked 4-8 h after dosing. Aceclofenac is a prodrug of diclofenac, and behaves similarly in domestic water buffalo as it did in domestic cattle, posing the same risk to vultures. We recommend an immediate ban on the veterinary use of aceclofenac across vulture-range countries.

Reference intervals for Cu, Mg, and Zn in captive gyrfalcons and other falcon species in the United Arab Emirates.

Falconry is a long-standing tradition in the United Arab Emirates (UAE), and in 2016, falconry was included by UNESCO in the list of intangible cultural heritage of humanity. The health and wellness of falconry raptors is a priority for the local culture; plasma chemistry analysis plays an important role in monitoring, assessing, and managing diseases in avian patients. Imbalances of Cu, Mg, and Zn have been linked to several diseases in avian species; therefore, determining the reference intervals (RIs) of these minerals has important implications in the clinical management of falcons. We determined the RIs in plasma for Cu, Mg, and Zn in captive (falconry-managed) falcons according to the Quality Assurance and Laboratory Standards Committee of the American Society for Veterinary Clinical Pathology, and the Clinical Laboratory Standards Institute guidelines. Blood chemistry analyses were carried out on 252 clinically healthy falcons examined at the Dubai Falcon Hospital in the UAE: 124 gyrfalcons (Falco rusticolus) and 128 falcons of other species. We observed no significant variation in RIs of Cu (1.5-7.0 µmol/L), Mg (0.49-0.78 mmol/L), or Zn (11.8-34.1 µmol/L) based on different species or sex.

Integrated exposure assessment of northern goshawk (Accipiter gentilis) nestlings to legacy and emerging organic pollutants using non-destructive samples.

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 (rs = 0.46-0.95, p < 0.001), except for hexachlorobenzene (HCB, rs = 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 (rs = 0.42-0.72, p < 0.02), however no correlation was found for perfluorohexane sulfonate (PFHxS), perfluorononanoate (PFNA) and perfluorotridecanoate (PFTriA) (rs = 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.

The perfect threat: Pesticides and vultures.

Probably the most important threat currently affecting vultures worldwide is exposure to pesticides, both accidentally and through deliberate abuse. This is of special concern since around 70% of vulture species are threatened by human activities. However, information about this threat is sparse and geographically biased. We compiled existing knowledge about pesticide exposure in vulture species globally, providing unifying criteria to mitigate this problem with a joint global effort. Most information available about accidental exposure to pesticides in vultures is related to organochlorine pesticides. Non-lethal exposure to these compounds occurs on every continent that vultures inhabit. While concentrations of organochlorine pesticides reported in different samples appear to be too low to produce health impacts, some studies show vultures with levels compatible with health impacts. In addition, there are some reports of vultures contaminated accidentally by anticoagulant rodenticides and external antiparasitic drugs used in veterinary practices. Deliberate abuse of pesticides to poison wildlife also occurs on every continent where vultures live, affecting most (78%) vulture species. However, little information is available for some regions of America, Asia and Europe. The exact number of vultures killed due to deliberate poisoning with pesticides is not well known, but the available figures are alarming (e.g. up to 500 individuals in a single event). The most widely used pesticides affecting vulture populations, and associated with deliberate poisoning, are carbamates and organophosphorus compounds. Of particular concern is the fact that massive poisoning events with these compounds occur, in some cases, within protected areas. This suggests that if this situation is not reversed, some vulture populations could disappear. A combination of measures such as banning pesticides, controlling their distribution-acquisition and environmental education could produce better results that banning pesticides alone. If poisoning with pesticides is not stopped, this threatened avian group could inadvertently go extinct very soon.

Mass balance study of brominated flame retardants in female captive peregrine falcons.

Little is known about brominated flame retardant (BFR) dynamics in birds, especially large molecules such as decabromodiphenyl ether (BDE-209). In particular, bioaccumulation from food and transfer dynamics to eggs are poorly understood. Therefore, an input-output mass balance study of tri-decaBDEs, DBDPE and HBCDD was performed in three female peregrine falcons from a captive breeding program by analyzing their naturally contaminated food (quail, chicken (cockerels)), plasma, feces and eggs. Predominant BFRs in cockerels and quail were BDE-209 and DBDPE, as well as HBCDD in quail. The predominant BFRs found in falcon plasma were BDE-209, -153 and -183, in eggs, HBCDD, BDE-209 and -153 and in feces, BDE-209. Mean absorption efficiencies (AE) for the tetra-octabrominated BDEs ranged from 84-100% and 70% for HBCDD. The AEs for BDE-206, -207, -208 and -209 varied due to the large variability seen for feces fluxes. All egg/plasma ratios for BDEs were similar and greater than one (range 1.1-2.7), including for BDE-209, indicating efficient transfer from females to the eggs. Excretion via egg-laying was approximately 6.0-29% of the initial, pre-breeding body burden of individual penta-decaBDE congeners, (15-45% for BDE-206). HBCDD was not detected in plasma but was found in eggs, also indicating efficient transfer and excretion via eggs. Input fluxes from food exceeded the output fluxes (feces, eggs) indicating considerable metabolism for tetra-octaBDEs, possibly also for the nona-decaBDEs and HBCDD. Bioaccumulation factors calculated from lipid weight concentrations in plasma and food (BAFp) were highest for BDE-208 (31), -153 (23), -209 (19) and -207 (16) and from eggs and food (BAFe), were highest for HBCDD (140), BDE-153 (41), -208 (42), BDE-207 (24) and BDE-209 (21). BAFe and BAFp values were below 10 for BDE-47, -99 and -100. For one falcon, egg results were available from three different years and estimated half-lives were 65 d (BDE-99), 624 d (BDE-153), 31 d (BDE-154), 349 d (BDE-183), 77 d (BDE-196) and 89 d (BDE-197).

Using an apex predator for large-scale monitoring of trace element contamination: Associations with environmental, anthropogenic and dietary proxies.

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 (δ15N: 15N/14N), carbon (δ13C: 13C/12C) and sulphur (δ34S: 34S/32S) 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 δ15N, 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.

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.

Blood and bone lead levels in South Africa's Gyps vultures: Risk to nest-bound chicks and comparison with other avian taxa.

Poisoning, including secondary lead poisoning, is cited as the single most important cause of vulture mortalities in Africa. To evaluate the prevalence of lead poisoning among South Africa's Gyps vultures compared to other, non-scavenging birds, we obtained blood and bone samples from Cape (Gyps coprotheres) and White-backed (G. africanus) vultures. We found that 66% of White-backed Vultures (n = 110, including 85 nest-bound chicks sampled at Dronfield Nature Reserve) and 80% of Cape Vultures (n = 15) had blood [Pb] in excess of 10 µg/dL, the upper limit of background exposure. Average blood [Pb] were 15.4 µg/dL and 29.7 µg/dL for White-backed and Cape vultures, respectively. Bone samples revealed that 12% of White-backed Vultures (n = 18) and 9% of Cape Vultures (n = 75) suffered from subclinical to severe clinical lead poisoning upon their deaths. By contrast, none of the 40 blood, bone or liver samples obtained from non-scavenging bird species were found to exceed background exposure levels. Our results suggest that, unlike non-scavenging birds, the scavenging lifestyle of Gyps vultures subjects them to lead poisoning on a regular basis. Had environmental sources of lead (e.g., dust) been the source of the lead poisoning at the White-backed Vulture breeding colony at Dronfield, all the chicks would have displayed similar blood lead concentrations. Instead the values ranged from barely detectable to very high, leading us to conclude that metallic lead fragments regurgitated by parents during feeding are responsible for the elevated lead levels in some of the chicks at this site. We conclude the likely source of these particles to be fragments of lead ammunition embedded in the carcasses of hunted animals. These results add to a growing body of evidence underscoring the threat posed by the use of lead ammunition and its potential role in the declines of vultures and other scavenging taxa.

Feather mercury concentrations in North American raptors sampled at migration monitoring stations.

We assessed total mercury (THg) concentrations in breast feathers of diurnal North American raptors collected at migration monitoring stations. For 9 species in the Pacific Flyway, we found species and age influenced feather THg concentrations whereas sex did not. Feather THg concentrations µg/g dry weight (dw) averaged (least squares mean ± standard error) higher for raptors that generally consume > 75% avian prey (sharp-shinned hawk Accipiter striatus: n = 113; 4.35 ± 0.45 µg/g dw, peregrine falcon Falco peregrinus: n = 12; 3.93 ± 1.11 µg/g dw, Cooper's hawk Accipiter cooperii: n = 20; 2.35 ± 0.50 µg/g dw, and merlin Falco columbarius: n = 59; 1.75 ± 0.28 µg/g dw) than for raptors that generally consume < 75% avian prey (northern harrier Circus hudsonius: n = 112; 0.75 ± 0.10 µg/g dw, red-tailed hawk Buteo jamaicensis: n = 109; 0.56 ± 0.06 µg/g dw, American kestrel Falco sparverius: n = 16; 0.57 ± 0.14 µg/g dw, prairie falcon Falco mexicanus: n = 10; 0.41 ± 0.13 µg/g dw) except for red-shouldered hawks Buteo lineatus: n = 10; 1.94 ± 0.61 µg/g dw. Feather THg concentrations spanning 13-years (2002-2014) in the Pacific Flyway differed among 3 species, where THg increased for juvenile northern harrier, decreased for adult red-tailed hawk, and showed no trend for adult sharp-shinned hawk. Mean feather THg concentrations in juvenile merlin were greater in the Mississippi Flyway (n = 56; 2.14 ± 0.18 µg/g dw) than those in the Pacific Flyway (n = 49; 1.15 ± 0.11 µg/g dw) and Intermountain Flyway (n = 23; 1.14 ± 0.16 µg/g dw), and Atlantic Flyway (n = 38; 1.75 ± 0.19 µg/g dw) averaged greater than the Pacific Flyway. Our results indicate that raptor migration monitoring stations provide a cost-effective sampling opportunity for biomonitoring environmental contaminants within and between distinct migration corridors and across time.

Is the current-use flame retardant, DBE-DBCH, a potential obesogen? Effects on body mass, fat content and associated behaviors in American kestrels.

The current-use brominated flame retardant, 1,2-dibromo-4-(1,2-dibromoethyl)cyclohexane (DBE-DBCH), is capable of perturbing sex steroid and thyroid hormone pathways in vitro and in vivo. Chemicals with this capability may also disrupt metabolic processes and are candidate obesogens, but this potential has not yet been determined for DBE-DBCH. Our objective was to examine gross biomarkers of metabolic disruption in captive American kestrels. Birds were exposed by diet to the ß isomer at the environmentally relevant dose of 0.239 ng ß-DBE-DBCH/g kestrel/day, from 30 days (d) prior to pairing through until chicks hatched (82 d) (n = 30 breeding pairs) or for 28 d (n = 16 pre-breeding pairs), and were compared with vehicle-only exposed controls. Body mass was assessed throughout the breeding season at biologically relevant time points, flight and feeding behavior was measured in 5-min samples daily, and plasma triglycerides and cholesterol were assessed at d10 of brood rearing. Treated males were heavier than controls at pairing (p = 0.051), the final week of courtship (p = 0.061), and at d10 (p = 0.012) and d20 of brood rearing (p = 0.051); ß-DBE-DBCH-exposed breeding females were similar in weight to control females. Treated birds tended to have higher plasma triglycerides (p = 0.078), which for females, was positively associated with body mass (p = 0.019). Heavier breeding males had higher plasma concentrations of testosterone and total thyroxine (p ≤ 0.046). Overall, both sexes exposed to ß-DBE-DBCH demonstrated reduced flight behavior and increased feeding behavior during courtship. In the pre-breeding pairs, treated male and female kestrels had a higher percentage of body fat than respective controls (p = 0.045). These results demonstrate that ß-DBE-DBCH elicited inappropriate fat and weight gain in adult American kestrels, consistent with their increased feeding, reduced flight activity and endocrine changes, and suggests that DBE-DBCH may be an obesogen warranting further research to test this hypothesis.

Can variability in corticosterone levels be related to POPs and OPEs in feathers from nestling cinereous vultures (Aegypius monachus)?

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 ±â€¯0.26 ng g-1 dw) > γ-HCH (0.78 ±â€¯0.09 ng g-1 dw) > BDE-99 (0.73 ±â€¯0.09 ng g-1 dw) > CB-153 (0.67 ±â€¯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.

Perfluoroalkyl substances (PFASs) and polychlorinated naphthalenes (PCNs) add to the chemical cocktail in peregrine falcon eggs.

A suite of perfluoroalkyl substances (PFASs) and polychlorinated naphthalenes (PCNs) were determined in 41 peregrine falcon eggs collected in South Greenland between 1986 and 2014. Median concentrations of perfluorinated sulfonic acids (ΣPFSA) and perfluorinated carboxylic acids (ΣPFCA) were 303 ng/g dry weight (dw) (58 ng/g wet weight, ww) and 100 ng/g dw (19 ng/g ww), respectively, which was comparable to other studies. Perfluorooctane sulfonate (PFOS) accounted for 94% on average of all PFSAs, but did not show a significant time trend. Perfluorohexane sulfonate (PFHxS), perfluoroheptane sulfonate (PFHpS) and perfluorodecane sulfonate (PFDS) showed non-linear decreases over the study period, while some long-chain PFCAs increased significantly. The PCN profile was dominated by the penta-, hexa- and tetrachlorinated congeners CN-52/60, CN-66/67 and CN-42. CN-54, an indicator of combustion, accounted for 2.4% of ΣPCN on average. All PCN congeners showed a decreasing tendency, which was significant for lipid-normalized concentrations of CN-53, CN-54 and CN-63. The ΣPCN median concentration was 21 ng/g lipid weight, which is in the high end of concentrations reported for bird eggs. The PCN and PFAS concentrations add to an already high contaminant burden and a complex chemical cocktail in the peregrine falcon population in Greenland, mainly reflecting contaminant exposure during migration and winter stays in Central and South America.

Preliminary Findings of Structure and Expression of Opioid Receptor Genes in a Peregrine Falcon ( Falco peregrinus), a Snowy Owl ( Bubo scandiacus), and a Blue-fronted Amazon Parrot ( Amazona aestiva).

To further knowledge of the physiology of opioid receptors in birds, the structure and expression of the µ-, δ-, and κ-opioid receptor genes were studied in a peregrine falcon ( Falco peregrinus), a snowy owl ( Bubo scandiacus), and a blue-fronted Amazon parrot ( Amazona aestiva). Tissue samples were obtained from birds that had been euthanatized for poor release prognosis or medical reasons. Samples were taken from the brain (telencephalon, thalamus, pituitary gland, cerebellum, pons, medulla oblongata, mesencephalon), the spinal cord and dorsal root ganglions, and plantar foot skin. Messenger RNA was recovered, and reverse transcription polymerase chain reaction (RT-PCR) was performed to generate complementary DNA (cDNA) sequences. Gene structures were documented by directly comparing cDNA sequences with recently published genomic sequences for the peregrine falcon and the blue-fronted Amazon parrot or by comparisons with genomic sequences of related species for the snowy owl. Structurally, the avian µ-opioid receptor messenger RNA (mRNA) species were complex, displaying differential splicing, alternative stop codons, and multiple polyadenylation signals. In comparison, the structure of the avian κ-receptor mRNA was relatively simple. In contrast to what is seen in humans, the avian δ-receptor mRNA structure was found to be complex, demonstrating novel 3-prime coding and noncoding exons not identified in mammals. The role of the δ-opioid receptor merits further investigation in avian species.

Mercury and lead exposure in avian scavengers from the Pacific Northwest suggest risks to California condors: Implications for reintroduction and recovery.

Mercury (Hg) and lead (Pb) are widespread contaminants that pose risks to avian scavengers. In fact, Pb exposure is the primary factor limiting population recovery in the endangered California condor (Gymnogyps californianus) and Hg can impair avian reproduction at environmentally relevant exposures. The Pacific Northwest region of the US was historically part of the condor's native range, and efforts are underway to expand recovery into this area. To identify potential threats to reintroduced condors we assessed foraging habitats, Hg and Pb exposure, and physiological responses in two surrogate avian scavenger species (common ravens [Corvus corax] and turkey vultures [Cathartes aura] across the region between 2012 and 2016. Mercury exposure near the Pacific coast was 17-27-fold higher than in inland areas, and stable carbon and sulfur isotopes ratios indicated that coastal scavengers were highly reliant on marine prey. In contrast, Pb concentrations were uniformly elevated across the region, with 18% of the birds exposed to subclinical poisoning levels. Elevated Pb concentrations were associated with lower delta-aminolevulinic acid dehydratase (δ-ALAD) activity, and in ravens there was an interactive effect between Hg and Pb on fecal corticosterone concentrations. This interaction indicated that the effects of Hg and Pb exposure on the stress axis are bidirectional, and depend on the magnitude of simultaneous exposure to the other contaminant. Our results suggest that condors released to the Pacific Northwest may be exposed to both elevated Hg and Pb, posing challenges to management of future condor populations in the Pacific Northwest. Developing a robust monitoring program for reintroduced condors and surrogate scavengers will help both better understand the drivers of exposure and predict the likelihood of impaired health. These findings provide a strong foundation for such an effort, providing resource managers with valuable information to help mitigate potential risks.

The use of liver slices from the Cape vulture (Gyps coprotheres) to better understand the role of liver toxicity of non-steroidal anti-inflammatory drugs (NSAIDs) in vultures.

Diclofenac, a non-steroidal anti-inflammatory drug (NSAID) was responsible for the death of millions of vultures on the Asian subcontinent, following the consumption of diclofenac contaminated carcasses. The aim of this research was to establish if liver slices could serve as an alternate means of predicting the toxicity of NSAIDs in Gyps vultures. The Cape vulture liver slices was prepared and incubated with four NSAIDs for 6 h. A percent clearance of 1.0 ±â€¯0.253, 0.58 ±â€¯0.153, 0.961 ±â€¯0.312 and 1.242 ±â€¯0.406 (%/h*g) was attained for diclofenac, carprofen, ketoprofen and meloxicam respectively. Both meloxicam and diclofenac exerted toxic effects on the hepatic cells. Protein content indicated that the vulture tissue had lower enzyme levels than expected for an animal of its size. The poor distinction between the ex vivo hepatic percent clearance of meloxicam and diclofenac indicates that liver slices is not an ideal model to investigate NSAIDs toxicity in Cape vulture.