HEMATOLOGIC METRICS FROM OLROG'S GULL (LARUS ATLANTICUS) DURING THE NONBREEDING SEASON IN ARGENTINA.

Ecophysiology and conservation studies often require the prior establishment of baseline physiologic metrics. For instance, expected reference intervals for health metrics are valuable tools for veterinarians and conservationists who monitor the health status of endangered populations and species. This study establishes reference intervals for hematologic metrics in free-ranging Olrog's gull (Larus atlanticus) during the nonbreeding season. Fifty-six gulls (immature and adults) were captured and studied in Mar del Plata and neighboring coastal areas (Buenos Aires, Argentina) during the winter of 2018 (n = 22) and 2019 (n = 34). Hematocrit, red blood cells (erythrocytes), hemoglobin, mean cell volume, mean cell hemoglobin (MCH), MCH concentration, white blood cells (WBC; leukocytes), heterophils, lymphocytes, eosinophils, monocytes, and basophils were analyzed. Additionally, the variability of hematologic metrics according to body weight, sex, age, and calendar year was examined. Hematologic metrics were in line with those reported in other seabird species. Males had greater body weight and MCH than females. The heterophil to lymphocyte ratio and lymphocyte levels were higher in adults than in immatures. Hematocrit, WBC, heterophils, and basophils also varied significantly between calendar years. The results highlight the importance of appropriate metrics and reference intervals for monitoring the health status of this threatened species, and it is recommended to implement such comparative assessments among populations.

Rapid determination of thyroid hormones in blood plasma from Glaucous gulls and Baikal seals by HybridSPE®-LC-MS/MS.

A rapid hybrid solid phase extraction (HybridSPE®) protocol tailored to liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) analysis, was developed for the determination of four thyroid hormones, L-Thyroxine (T4), 3,3',5-triiodo-L-thyronine (T3), 3,3',5'-triiodo-L-thyronine (rT3) and 3,3'-diiodo-L-thyronine (T2) in blood plasma from Glaucous gulls (Larus hyperboreus) and Baikal seals (Phoca sibirica). The use of target analyte specific 13C internal standards allowed quantification to be performed through the standard solvent calibration curves and alleviated the need to perform quantification with matrix match curves. The relative recoveries were 100.0-110.1 % for T4, 99.1-102.2 % for T3, 100.5-108.0 % for rT3, and 100.5-104.6 % for T2. The matrix effects ranged from -1.52 to -6.10 %, demonstrating minor signal suppression during analysis. The method intra-day precision (method repeatability, RSD %, N = 5, k = 1 day) and inter-day precision (method reproducibility, RSD %, N = 10, k = 2 days) at the 1 ng/mL concentration of fortification were 8.54-15.4 % and 15.4-24.8 %, respectively, indicating acceptable chromatographic peak stabilities for all target THs even at trace level concentrations. The method limit of detection (LOD) for T4, T3, rT3 and T2 was 0.17, 0.16, 0.30 and 0.17 ng/mL, respectively. The HybridSPE® protocol was simple and rapid (<1 min) upon application, while the HybridSPE® cartridge did not require (as in classical SPE cartridges) any additional equilibration nor conditioning step prior sample loading. A total of 46 blood plasma samples, 30 samples collected from Glaucous gulls and 16 samples collected from Baikal seals, were analyzed for thyroid hormones to demonstrate the applicability of the developed method in these wildlife species. The concentrations of T4 and T3 in blood plasma from the Glaucous gulls were 5.95-44.2 and 0.37-5.61 ng/mL, respectively, whereas those from Baikal seals were 3.57-46.5 and 0.45-2.07 ng/mL, respectively. In both species, rT3 demonstrated low detection rate, while T2 was not detected. Furthermore, cross-array comparison between the HybridSPE®-LC-MS/MS protocol and an established routine radioimmunoassay (RIA) kit-based method was performed for T4 and T3 concentrations from selected Baikal seal plasma samples.

Relationship between blood test values and blood lead (Pb) levels in Black-headed gull (Chroicocephalus ridibundus: Laridae).

Few studies have evaluated immunosuppression due to lead accumulation below the overt toxicity threshold. If low levels of lead accumulation cause immunosuppression in birds, those birds could become more susceptible to pathogens. We aimed to determine if low levels of lead accumulation lead to immunosuppression in Black-headed gulls (Chroicocephalus ridibundus). Gulls were captured in Tokyo-bay and Mikawa-bay from January to April 2019. Their blood samples were analyzed for eight items. The data were analyzed to evaluate the correlation between lead concentrations and the variables from each bay. Lead was positively correlated with the percentage of heterophils and heterophil and lymphocyte ratio and negatively with lymphocytes. Thus, low lead accumulation levels may induce changes in percentage of the heterophils and lymphocyte.

Moving Beyond p < 0.05 in Ecotoxicology: A Guide for Practitioners.

Statistical inferences play a critical role in ecotoxicology. Historically, null hypothesis significance testing (NHST) has been the dominant method for inference in ecotoxicology. As a brief and informal definition of NHST, researchers compare (or "test") an experimental treatment or observation against a hypothesis of no relationship (the "null hypothesis") using the collected data to see if the observed values are statistically "significant" given predefined error rates. The resulting probability of observing a value equal to or greater than the observed value assuming the null hypothesis is true is the p value. Criticisms of NHST have existed for almost a century and have recently grown to the point where statisticians, including the American Statistical Association (ASA), have felt the need to clarify the role of NHST and p values beyond their current common use. These limitations also exist in ecotoxicology. For example, a review of the 2010 Environmental Toxicology & Chemistry (ET&C) volume that found many authors did not correctly report p values. We repeated this review looking at the 2019 volume of ET&C. Incorrect reporting of p values still occurred almost a decade later. Problems with NHST and p values highlight the need for statistical inferences besides NHST, something long known in ecotoxicology and the broader scientific and statistical communities. Furthermore, concerns such as these led the executive director of the ASA to recommend against use of "statistical significance" in 2019. In light of these criticisms, ecotoxicologists require alternative methods. We describe some alternative methods including confidence intervals, regression analysis, dose-response curves, Bayes factors, survival analysis, and model selection. Lastly, we provide insights for what ecotoxicology might look like in a post-p value world. Environ Toxicol Chem 2020;39:1657-1669. Published 2020. This article is a U.S. Government work and is in the public domain in the USA.

Isotopic Discrimination (δ15N, δ13C) in Captive and Wild Common Murres (Uria aalge) and Atlantic Puffins (Fratercula arctica).

Studying the diet of consumers using stable isotopes provides insight into the foraging ecology of individuals and species. To accurately reconstruct the integrated diet of animals using stable isotope values, we must quantify diet-tissue discrimination factors (DTDFs), or the way in which stable isotopes in prey are incorporated into the tissues of consumers. To quantify DTDFs, controlled experiments are needed, whereby consumers are fed a constant diet. However, relatively few controlled-diet studies have been conducted for seabirds. In this study, captive adult Atlantic puffins (Fratercula arctica) and common murres (Uria aalge) were fed a two-source diet of capelin (Mallotus villosus) and Atlantic silverside (Menidia menidia) to determine the DTDFs for the cellular component of blood and plasma for both δ15N and δ13C. The DTDFs for the cellular component (Δ15N: 2.80±0.28; Δ13C: 1.21±0.22) and plasma (Δ15N: 1.72±1.03; Δ13C: -0.18±0.56) of puffins were similar to those for the cellular component (Δ15N: 2.91±0.18; Δ13C: 1.09±0.23) and plasma (Δ15N: 2.18±0.77; Δ13C: -0.70±0.18) of murres. We reconstructed the diet of wild murres and puffins breeding on the northeastern coast of Newfoundland using previously published DTDFs and estimated DTDFs from our feeding experiment. Reconstructed dietary proportions supported a priori knowledge of diet, although outputs were sensitive to the DTDF used. Despite the similarity of our DTDFs for puffins and murres, along with the similarity of our DTDFs with those of other seabird species, our sensitivity analysis revealed considerable differences among resultant dietary contributions from mixing models, further highlighting the importance of using species- and tissue-specific DTDFs to enhance knowledge in the foraging ecology of seabirds using stable isotopes.

Contaminants, prolactin and parental care in an Arctic seabird: Contrasted associations of perfluoroalkyl substances and organochlorine compounds with egg-turning behavior.

Incubating eggs represents a trade-off for parent birds between spending enough time fasting to take care of the clutch and to get enough nutrients for self-maintenance. It is believed that the pituitary hormone prolactin plays an important role in such allocation processes. Incubation does not solely imply the active warming of the eggs but also the active egg-turning to facilitate absorption of albumen by the embryo, reduce malposition and prevent the embryo from adhering to the inner shell membrane. However, how prolactin secretion is related to egg-turning behaviors is presently poorly addressed. In addition, several environmental contaminants can affect parental care behaviors through their endocrine disrupting properties but the effects of such contaminants on egg-turning behaviors remain so far unexplored. Using artificial eggs equipped with miniaturized data loggers, we investigated the relationships between egg-turning behaviors, prolactin secretion and contaminants burden in Arctic black-legged kittiwakes (Rissa tridactyla). Specifically, we examined the relationships between blood concentrations of poly- and perfluoroalkyl substances (PFASs), organochlorines (OCs), mercury (Hg), plasma prolactin levels and both egg-turning frequency and angular change. We also incorporated baseline corticosterone levels since this glucocorticoid is known to affect parental care. Plasma prolactin levels were positively related to angular change in female kittiwakes while corticosterone was not related to egg-turning behaviors in either sex. Hg was not related to egg-turning behaviors in either sex. We found contrasting associations between OCs and PFASs, since polychlorinated biphenyls (PCBs) were negatively associated with angular change in females, contrary to linear perfluorooctanesulfonate (PFOSlin) and perfluoroalkyl carboxylic acids (PFCAs) which were positively related to egg-turning frequency and angular change in both sexes. Additionally, PFASs concentrations were positively related to prolactin levels in female kittiwake. The possible stimulation of prolactin secretion by PFASs could therefore make adult kittiwakes to allocate more time taking care of their eggs, and thus possibly modify the trade-off between spending enough time caring for the clutch and obtaining enough nutrients at sea.

Melatonin and corticosterone profiles under polar day in a seabird with sexually opposite activity-rhythms.

The 24 h geophysical light-dark cycle is the main organizer of daily rhythms, scheduling physiology and behavior. This cycle attenuates greatly during the continuous light of summer at polar latitudes, resulting in species-specific and even individual-specific patterns of behavioral rhythmicity, but the physiological mechanisms underlying this variation are poorly understood. To address this knowledge gap and to better understand the roles of the hormones melatonin and corticosterone in rhythmic behavior during this 'polar day', we exploited the behavior of thick-billed murres (Uria lomvia), a charadriiform seabird with sexually opposite ('antiphase') activity-rhythms that have a duration of 24 h. Melatonin concentration in the plasma of inactive males was unexpectedly high around midday and subsequently fell during a sudden decrease in light intensity as the colony became shaded. Corticosterone concentration in plasma did not vary with time of day or activity in either sex. While the reasons for these unusual patterns remain unclear, we propose that a flexible melatonin response and little diel variation of corticosterone may be adaptive in thick-billed murres, and perhaps other polar birds and mammals, by stabilizing glucocorticoids' role of modulating energy storage and mobilization across the diel cycle and facilitating the appropriate reaction to unexpected stimuli experienced across the diel cycle while attending the colony.

Prolactin concentrations predict parental investment and nest survival in a free-living shorebird.

The hormonal milieu that exists during reproduction is one of the key factors influencing the trade-off between reproductive investment and self-maintenance. Much previous work in birds has focused on prolactin as a physiological mediator since prolactin is involved in the onset and maintenance of parental care. However, how prolactin relates to reproductive success in terms of altering parental behavior in wild bird populations is not fully understood. Here, we report prolactin concentrations in breeding Kentish plovers (Charadrius alexandrinus), a small shorebird with variable mating systems and parental care, as an ecological model of mating system evolution. Throughout the breeding season, we estimated the circulating prolactin concentrations in male and female plovers during incubation. In addition, we monitored parental behavior and determined the fate of nests. We found that prolactin concentrations decreased during incubation but increased with clutch completion date. In addition, males and females with high prolactin concentrations spent more time on incubation than those with low prolactin concentrations. Importantly, higher prolactin concentrations in either males or females predict higher nest survival. Our results suggest that prolactin is an indicator of parental behavior in a wild shorebird population, although additional studies including experimental manipulation of prolactin concentrations are necessary to verify this relationship.

SERUM BIOCHEMISTRY VALUES FOR THE OLROG'S GULL (LARUS ATLANTICUS): A STUDY BASED ON FREE-LIVING INDIVIDUALS.

Analysis of serum parameters provides information about body condition, nutritional state, and health status of individuals/species, and has broad application in ecological research and veterinary diagnosis. This study establishes baseline values for serum chemistries of the Olrog's gull (Larus atlanticus). Glucose, urea, uric acid, total protein, globulin, albumin, cholesterol, triglyceride, calcium, and phosphorus concentrations were determined, as was the activity of the following enzymes: alkaline phosphatase, creatine phosphokinase, aspartate aminotransferase, and alanine aminotransferase. Thirty nonbreeding gulls (juvenile and subadult individuals) were captured and studied in Mar Chiquita Reserve (Buenos Aires, Argentina) during the wintering periods 2016 (n = 17) and 2017 (n = 13). In general terms, most values for the parameters reported were in line with those previously described for other seabirds. The year had a significant effect on several of the biochemical parameters evaluated, and the sex had a significant effect on the alkaline phosphatase and calcium. This study has defined the serum biochemical reference signatures for free-ranging Olrog's gulls during the nonbreeding period, and contributes to the knowledge of the overall health status of this threatened and endemic species.

The effect of long-range transport, trophic position and diet specialization on legacy contaminant occurrence in great skuas, Stercorarius skua, breeding across the Northeast Atlantic.

High levels of halogenated organic contaminants (HOCs) have been found in the marine predatory seabird great skua (Stercorarius skua) from breeding colonies in the Northeastern Atlantic, with large unexplained inter-colony variation. The present study aimed at analyzing if the HOCs occurrence in breeding great skuas in remote colonies was explained by local baseline food web exposure determined by long-range transport, or by ecological factors such as diet specialization and relative trophic position in the breeding area. The occurrence of organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), and polybrominated diphenyl ethers (PBDEs) was analyzed in plasma of 204 adult great skuas collected over two years (2008 and 2009) and 5 colonies across the North-Atlantic from Shetland to Svalbard. The ΣHOCs levels in plasma ranged across two orders of magnitude, from 40 to 7600 ng/g (wet weight) and differed significantly across the great skua colonies. The variation in contaminant occurrence among colonies did not reflect long-range transport through a latitudinal or remoteness gradient, as the second northernmost colony (Bjørnøya), had the highest contaminant concentrations. No latitudinal or remoteness gradient was evident in the contaminant pattern among the colonies. The contaminant levels increased significantly with increasing δ15N values, and regurgitated pellets of undigested prey suggested that great skuas with higher δ15N values had a higher proportion of bird prey in their diet, mostly seabirds. In contrast, great skuas from colonies with lower δ15N and lower contaminant level fed mostly on fish. The enrichment of δ13C increased with decreasing δ15N and lower contaminant levels. Therefore, individual behavior of great skuas, such as migration strategies and diet specialization, rather than long-range transport and thus baseline food web exposure, explain among and within colony variance in contaminant occurrence.

Sex- and breeding stage-specific hormonal stress response of seabird parents.

Changes in corticosterone (CORT) and prolactin (PRL) levels are thought to provide complementary information on parental decisions in birds in the context of stressful situations. However, these endocrine mechanisms have yet to be fully elucidated, appearing to vary among avian species without any clear pattern. Here, we examined CORT and PRL stress responses in a small Arctic seabird, the little auk (Alle alle). We analysed the levels of these hormones (baseline, and stress response, i.e. the change in the baseline in response to stress) with respect to the breeding phase (mid incubation and mid chick rearing) and the sex of the birds. Baseline CORT concentrations were similar during both breeding phases but baseline PRL levels were higher during incubation than chick rearing. The CORT and PRL stress responses were stronger during incubation than chick rearing (although with respect to CORT the effect was only marginally significant). There were also some sex-specific baseline levels and stress responses for both hormones (during the incubation period males compared to females exhibited higher CORT stress response and lower baseline PRL; during the chick rearing period males exhibited higher PRL stress response). Our results suggest that in the case of the little auk, both the incubation and the chick rearing periods may represent similar levels of physiological stress. However, the birds may be more sensitive to stress during incubation than during chick rearing, possibly because of inter-phase differences in predation pressure. The sex differences suggest differential exposure of males and females to stressors.

Evidence for exposure to selenium by breeding interior snowy plovers (Charadrius nivosus) in saline systems of the Southern Great Plains.

Interior snowy plover (Charadrius nivosus) population declines and deteriorating conditions throughout the Southern Great Plains (SGP) of Texas, New Mexico, and Oklahoma may be linked to environmental contaminants. Concentrations of V, As, Cd, Pb, and Se were quantified in breeding snowy plover blood, feathers (5th primary; P5), and potential prey (tiger beetles [Cicindela circumpicta and C. togata]). Se was (a) most commonly detected relative to other quantified elements and (b) frequently quantified at levels exceeding background or toxicity thresholds. Of samples greater than instrumentation detection limits, 98% of snowy plover blood and 22% of feather samples were greater than Se toxicity thresholds of 1 ppm ww for blood and 5 ppm dw for feathers (blood quantifiable range: 0.83-15.12 ppm; feathers quantifiable range: 1.90-27.47 ppm). Almost all tiger beetle Se concentrations were below reported invertebrate thresholds of 30 ppm dw (quantifiable range: 0.54-45.84 ppm). Snowy plover blood Se concentrations were related to sex, individual body condition, and local tiger beetle Se concentrations, while plover P5 Se concentrations were related to state, sex, and presence of body molt. Tiger beetle Se concentrations were related to individual study sites in Texas. These results provide some of the first evidence of Se exposure risk for interior snowy plovers nesting in saline lake and alkali flat environments of the SGP. Future efforts should focus upon specific Se uptake pathways during breeding and nonbreeding seasons, as snowy plovers breeding in the SGP appear to be exposed to Se throughout their annual cycle.

Effects of El Niño and La Niña Southern Oscillation events on the adrenocortical responses to stress in birds of the Galapagos Islands.

El Niño Southern Oscillation events (ENSO) and the subsequent opposite weather patterns in the following months and years (La Niña) have major climatic impacts, especially on oceanic habitats, affecting breeding success of both land and sea birds. We assessed corticosterone concentrations from blood samples during standardized protocols of capture, handling and restraint to simulate acute stress from 12 species of Galapagos Island birds during the ENSO year of 1998 and a La Niña year of 1999. Plasma levels of corticosterone were measured in samples collected at capture (to represent non-stressed baseline) and subsequently up to 1 h post-capture to give maximum corticosterone following acute stress, and total amount of corticosterone that the individual was exposed to during the test period (integrated corticosterone). Seabird species that feed largely offshore conformed to the brood value hypothesis whereas inshore feeding species showed less significant changes. Land birds mostly revealed no differences in the adrenocortical responses to acute stress from year to year with the exception of two small species (<18 g) that had an increase in baseline and stress responses in the ENSO year - contrary to predictions. We suggest that a number of additional variables, including body size and breeding stage may have to be considered as explanations for why patterns in some species deviated from our predictions. Nevertheless, comparative studies like ours are important for improving our understanding of the hormonal and reproductive responses of vertebrates to large scale weather patterns and global climate change in general.

Mercury concentrations in multiple tissues of Kittlitz's murrelets (Brachyramphus brevirostris).

Mercury (Hg) is a non-essential, toxic metal that is distributed worldwide. Mercury biomagnifies in food webs and can threaten the health of top predators such as seabirds. The Kittlitz's murrelet (Brachyramphus brevirostris) is a seabird endemic to Alaska and the Russian Far East and is a species of conservation concern in the region. We determined Hg concentrations in eggshells, guano, blood, and feathers of Kittlitz's murrelets sampled from four locations in Alaska. Mercury concentrations in eggshells, guano, and blood were low compared to other seabird species. Mean Hg concentrations of breast feathers from Adak Island and Glacier Bay were significantly greater than those from Agattu Island or Icy Bay. Two Kittlitz's murrelets at Glacier Bay and one Kittlitz's murrelet at Adak Island had Hg concentrations above those associated with impaired reproduction in other bird species, and may merit further investigation as a potential threat to individuals and populations.

Hematological indices of injury to lightly oiled birds from the Deepwater Horizon oil spill.

Avian mortality events are common following large-scale oil spills. However, the sublethal effects of oil on birds exposed to light external oiling are not clearly understood. We found that American oystercatchers (area of potential impact n = 42, reference n = 21), black skimmers (area of potential impact n = 121, reference n = 88), brown pelicans (area of potential impact n = 91, reference n = 48), and great egrets (area of potential impact n = 57, reference n = 47) captured between 20 June 2010 and 23 February 2011 following the Deepwater Horizon oil spill experienced oxidative injury to erythrocytes, had decreased volume of circulating erythrocytes, and showed evidence of a regenerative hematological response in the form of increased reticulocytes compared with reference populations. Erythrocytic inclusions consistent with Heinz bodies were present almost exclusively in birds from sites impacted with oil, a finding pathognomonic for oxidative injury to erythrocytes. Average packed cell volumes were 4 to 19% lower and average reticulocyte counts were 27 to 40% higher in birds with visible external oil than birds from reference sites. These findings provide evidence that small amounts of external oil exposure are associated with hemolytic anemia. Furthermore, we found that some birds captured from the area impacted by the spill but with no visible oiling also had erythrocytic inclusion bodies, increased reticulocytes, and reduced packed cell volumes when compared with birds from reference sites. Thus, birds suffered hematologic injury despite no visible oil at the time of capture. Together, these findings suggest that adverse effects of oil spills on birds may be more widespread than estimates based on avian mortality or severe visible oiling. Environ Toxicol Chem 2018;37:451-461. © 2017 SETAC.

Biomonitoring of fluoroalkylated substances in Antarctica seabird plasma: Development and validation of a fast and rugged method using on-line concentration liquid chromatography tandem mass spectrometry.

We report on a fast, accurate and rugged analytical procedure to determine a wide span of perfluoroalkyl and polyfluoroalkyl substances (PFASs) in seabird plasma. The 26 investigated compounds included perfluoroalkyl carboxylates (C5-C14 PFCAs), perfluoroalkyl sulfonates (C4, C6, C7, C8, C10 PFSAs), perfluorooctane sulfonamide (FOSA) and N-alkyl derivatives (MeFOSA, EtFOSA), N-alkyl perfluorooctane sulfonamido acetic acids (MeFOSAA, EtFOSAA), fluorotelomer sulfonates (4:2 FTSA, 6:2 FTSA, 8:2 FTSA), polyfluoroalkyl phosphate diesters (diPAPs) and perfluorooctane sulfonamide phosphate diester (diSAmPAP). The method described herein requires a reduced sample amount (25µL) and involves rapid and simple sample preparation (protein precipitation with acetonitrile but without acidification) prior to analysis by on-line solid phase extraction (Oasis HLB sorbent) coupled to high performance liquid chromatography negative electrospray ionization tandem mass spectrometry. The optimization was conducted using experimental designs to account for potential interactions between variables. Out of the 26 target analytes, 23 compounds showed excellent accuracy (±25% of the expected values). Intermediate precision and matrix effects remained acceptable for most analytes thanks to efficient internal standardization. A human serum standard reference material (NIST SRM 1957) was included in the validation scheme to evaluate method trueness, which proved satisfactory (│Z-scores│<2 for most compounds). Notwithstanding the small initial sample intake, limits of detection as low as 0.003-0.1ngg-1 plasma were obtained. This allowed the determination of 11 target PFASs in Antarctic seabird plasma samples. ΣPFASs in Antarctic seabird plasma ranged from 0.37 to 19ngg-1, with a predominance of PFOS (>54% of ΣPFASs on average). The reduced plasma amount required implies that the present method could also be applied to the analysis of PFASs in the plasma of smaller biological models.

Family-transmitted stress in a wild bird.

Recent data suggest that, in animals living in social groups, stress-induced changes in behavior have the potential to act as a source of information, so that stressed individuals could themselves act as stressful stimuli for other individuals with whom they interact repeatedly. Such form of cross-over of stress may be beneficial if it enhances adaptive responses to ecological stressors in the shared environment. However, whether stress can be transferred among individuals during early life in natural populations remains unknown. Here we tested the effect of living with stressed siblings in a gull species where, as in many vertebrates, family represents the basic social unit during development. By experimentally modifying the level of stress hormones (corticosterone) in brood mates, we demonstrate that the social transfer of stress level triggers similar stress responses (corticosterone secretion) in brood bystanders. Corticosterone-implanted chicks and their siblings were faster in responding to a potential predator attack than control chicks. In gulls, fast and coordinated reactions to predators may increase the chances of survival of the whole brood, suggesting a beneficial fitness value of cross-over of stress. However, our data also indicate that living with stressed brood mates early in life entails some long-term costs. Near independence, fledglings that grew up with stressed siblings showed reduced body size, high levels of oxidative damage in lipids and proteins, and a fragile juvenile plumage. Overall, our results indicate that stress cross-over occurs in animal populations and may have important fitness consequences.

Toxicological and thermoregulatory effects of feather contamination with artificially weathered MC 252 oil in western sandpipers (Calidris mauri).

The external contamination of bird feathers with crude oil might have effects on feather structure and thus on thermoregulation. We tested the thermoregulatory ability of western sandpipers (Calidris mauri) in a respirometry chamber with oil applied either immediately prior, or three days before the experiment. The birds were then exposed to a sliding cold temperature challenge between 27°C and -3°C to calculate thermal conductance. After the experiment, a large blood sample was taken and the liver extracted to measure a range of parameters linked to toxicology and oxidative stress. No differences in thermal conductance were observed among groups, but birds exposed to oil for three days had reduced body temperatures and lost more body mass during that period. At necropsy, oiled birds showed a decrease in plasma albumin and sodium, and an increase in urea. This is reflective of dysfunction in the kidney at the loop of Henle. Birds, especially when exposed to the oil for three days, showed signs of oxidative stress and oxidative damage. These results show that the ingestion of externally applied oil through preening or drinking can cause toxic effects even in low doses, while we did not detect a direct effect of the external oil on thermoregulation over the temperature range tested.

Effect of oral exposure to artificially weathered Deepwater Horizon crude oil on blood chemistries, hepatic antioxidant enzyme activities, organ weights and histopathology in western sandpipers (Calidris mauri).

Shorebirds were among birds exposed to Mississippi Canyon 252 (MC252) crude oil during the 2010 Deep Water Horizon (DWH) oil spill in the Gulf of Mexico. The western sandpiper (Calidris mauri) was chosen as one of four species for initial oral dosing studies conducted under Phase 2 of the avian toxicity studies for the DWH Natural Resource Damage Assessment (NRDA). Thirty western sandpipers were assigned to one of three treatment groups, 10 birds per group. The control group was sham gavaged and the treatment groups were gavaged with 1 or 5mL oil kg bw-1 daily for 20 days. Periodic blood samples for hemoglobin measurements were collected during the trial. A final blood sample used to determine hemoglobin concentration in addition to complete blood counts, plasma clinical chemistries, haptoglobin concentration and plasma electrophoresis was collected when birds were euthanized and necropsied on day 21. Tissues were removed, weighed and processed for subsequent histopathological evaluation. There were numerical decreases in hemoglobin concentrations in oil-dosed birds over the 21-day trial, but values were not significantly different compared to controls on day 21. There were no significant differences between controls and oiled birds in complete blood counts, plasma chemistries, haptoglobin concentration, and plasma electrophoresis endpoints. Of the hepatic oxidative stress endpoints assessed, the total antioxidant capacity assessment (Trolox equivalents) for the control group was lower compared to the 1mL oil kg bw-1 group. Absolute liver weights in the 5mL oil kg bw-1 group were significantly greater compared to controls. While not conclusive, the numerical decrease in hemoglobin concentration and significant increase in absolute liver weight are consistent with exposure to oil. Histological changes in the adrenal gland could be considered a non-specific indicator of stress resulting from exposure to oil. It is possible that the quantity of oil absorbed was not sufficient to induce clearly evident hemolytic anemia or that the western sandpiper is relatively insensitive to ingested oil.

Blood transcriptomes and de novo identification of candidate loci for mating success in lekking great snipe (Gallinago media).

We assembled the great snipe blood transcriptome using data from fourteen lekking males, in order to de novo identify candidate genes related to sexual selection, and determined the expression profiles in relation to mating success. The three most highly transcribed genes were encoding different haemoglobin subunits. All tended to be overexpressed in males with high mating success. We also called single nucleotide polymorphisms (SNPs) from the transcriptome data and found considerable genetic variation for many genes expressed during lekking. Among these, we identified 14 polymorphic candidate SNPs that had a significant genotypic association with mating success (number of females mated with) and/or mating status (mated or not). Four of the candidate SNPs were found in HBAA (encoding the haemoglobin α-chain). Heterozygotes for one of these and one SNP in the gene PABPC1 appeared to enjoy higher mating success compared to males homozygous for either of the alleles. In a larger data set of individuals, we genotyped 38 of the identified SNPs but found low support for consistent selection as only one of the zygosities of previously identified candidate SNPs and none of their genotypes were associated with mating status. However, candidate SNPs generally showed lower levels of spatial genetic structure compared to noncandidate markers. We also scored the prevalence of avian malaria in a subsample of birds. Males infected with avian malaria parasites had lower mating success in the year of sampling than noninfected males. Parasite infection and its interaction with specific genes may thus affect performance on the lek.