Blood transcriptome analysis of common kestrel nestlings living in urban and non-urban environments.

Urbanisation is one of the main anthropogenic forms of land cover affecting an ever-increasing number of wild animals and their habitats. Physiological plasticity represents an important process through which animals can adjust to the novel conditions of anthropogenic environments. Relying on the analysis of gene expression, it is possible to identify the molecular responses to the habitat conditions and infer possible environmental factors that affect the organismal physiology. We have quantified for the first time the blood transcriptome of common kestrel (Falco tinnunculus) nestlings living in urban sites and compared it to the transcriptome of kestrel nestlings inhabiting rural and natural environments. We found mild differences in the expression of genes among sites, indicating adaptability or acclimation of the birds to the urban habitat. We identified 58 differentially expressed genes between urban and natural kestrels, and 12 differentially expressed genes between urban and rural kestrels. The most striking differences among sites involved inflammatory-immunological, metabolic, apoptosis, DNA repair and development genes. In particular, we found that (i) urban kestrel nestlings had higher expression of genes linked to inflammation, repair of DNA damage, or apoptosis than natural kestrel nestlings, and (ii) natural and rural kestrel nestlings had higher expression of genes linked to the development and activation of immune cells, type I interferon response, or major histocompatibility complex than urban kestrel nestlings. Finally, the KEGG enrichment analysis identified the insulin signalling as the main pathway that differed between natural and urban kestrel nestlings. This is one of a limited number of studies on vertebrates that revealed habitat-associated differences in the transcriptome. It paves the way for further in-depth studies on the links between physiological variation and habitat structure at different spatial and temporal scales.

Blood mercury concentrations in four sympatric gull species from South Western France: Insights from stable isotopes and biologging.

Mercury (Hg) is a toxic trace element widely distributed in the environment, which particularly accumulates in top predators, including seabirds. Among seabirds, large gulls (Larus sp) are generalist feeders, foraging in both terrestrial and marine habitats, making them relevant bioindicators of local coastal Hg contamination. In the present study, we reported blood Hg concentrations in adults and chicks of four different gull species breeding on the French Atlantic coast: the European herring gull (Larus argentatus), the Lesser black-backed gull (L. fuscus), the Great black-backed gull (L. marinus) and the Yellow-legged gull (L. michahellis). We also investigated the potential role of foraging ecology in shaping Hg contamination across species, using the unique combination of three dietary tracers (carbon, nitrogen and sulfur stable isotopes) and biologging (GPS tracking). A high concentration of Hg was associated with high trophic position and a marine diet in gulls, which was corroborated by birds' space use strategy during foraging trips. Adults of all four species reached Hg concentrations above reported toxicity thresholds. Specifically, adults of Great black-backed gulls had a high trophic marine specialized diet and significantly higher Hg concentrations than the three other species. Blood Hg was 4-7 times higher in adults than in chicks, although chicks of all species received mainly marine and high trophic position prey, which is expected to be the cause of blood Hg concentrations of toxic concern. By using both stable isotopes and GPS tracking, the present study provides compelling insights on the main feeding habits driving Hg contamination in a seabird assemblage feeding in complex coastal environments.

Controlled expression of the migratory phenotype affects oxidative status in birds.

High caloric intake can increase production of reactive oxygen and nitrogen species. We examined whether the emergence of the migratory phenotype, primarily signalled by increased food intake and fuelling, is accompanied by changes in oxidative status. We induced autumn migration followed by a non-migratory wintering phase in common quails (Coturnix coturnix). We compared three markers of oxidative status - oxidative damage to lipids expressed as thiobarbituric acid reactive substances (TBARS); superoxide dismutase (SOD); and glutathione peroxidase (GPx) - between birds sampled during the migratory and non-migratory phase. We found that the emergence of the migratory phenotype was associated with: (i) higher levels of TBARS in the liver; (ii) lower levels of SOD in red blood cells and, marginally, in the liver; (iii) higher levels of GPx in the pectoral muscle; and (iv) sex-specific changes in red blood cells and liver. We found no link between food intake and variation in markers of oxidative status in any of the tissues examined, despite food intake being higher in the migratory birds. However, the increase in body mass was positively correlated with muscle GPx activity as birds entered the pre-migratory fattening phase, while the amount of decrease in body mass was negatively correlated with muscle GPx as birds transitioned to the non-migratory phase. Such correlations were absent in red blood cells and liver. Our work suggests that during the emergence of the migratory phenotype, birds might strategically displace oxidative costs on the liver in order to safeguard the pectoral muscles, which have a fundamental role in successfully completing the migratory flight.

Detection and Phylogenetic Characterization of a Novel Herpesvirus in Sooty Terns Onychoprion fuscatus.

Since 2005, we have recorded annual episodes of alphaherpesvirus outbreaks in chicks of magnificent frigatebird Fregata magnificens on the Ile du Grand Connétable Nature Reserve in French Guiana. In 2009, we found sooty terns, Onychoprion fuscatus, that live sympatrically with frigatebirds, with visible clinical signs of a potential viral infection. To determine if the symptoms observed in sooty terns could be associated with an alphaherpesvirus previously identified in frigatebirds, we carried out molecular screening of samples collected from seven individuals. We identified and characterized a novel viral sequence from five birds. BLAST searches, pairwise nucleotide, and amino acid sequence comparisons, as well as phylogenetic analyses confirmed that the sequence belonged to the Herpesviridae family, of the Alphaherpesvirinae subfamily. We observed that it clustered with strains isolated from Podargidae (Caprimulgiformes), Columbiformes, and Falconiformes, but was distinct from the frigatebird herpesvirus. We have tentatively named it Onychoprion fuscatus alphaherpesvirus 1, (OfusAHV1). These two sequences, although found syntopic on the Ile du Grand Connétable, belong to two distinct alphaherpesvirus strains. Thus, the clinical symptoms showed by sooty terns do not likely result from a cross-species transmission event. Future work is needed to better characterize the virus and to investigate herpesvirus prevalence in healthy, free-ranging sooty terns, and to assess the impact of the virus on population viability.

Exposure to PFAS is Associated with Telomere Length Dynamics and Demographic Responses of an Arctic Top Predator.

Environmental factors that can influence telomeres are diverse, but the association between telomeres and exposure to environmental contaminants is yet to be elucidated. To date, prior studies have focused on legacy persistent chlorinated pollutants (POPs), while the effects of poly- and perfluoroalkyl substances (PFAS) have been poorly documented. Here, we investigated the associations among PFAS congeners, absolute telomere length (cross-sectional approach), and telomere dynamics (rate of telomere length change over time, longitudinal approach) in one of the most contaminated arctic top predators, the glaucous gull Larus hyperboreus from Svalbard. We further estimated the effect of PFAS on apparent survival rates and re-sighting probabilities using a 10-year capture/recapture dataset (2010-2019). We found that birds exposed to higher concentrations of perfluorononadecanoate (PFNA) (median of 1565 pg/mL of ww in males and 1370 pg/mL of ww in females) and perfluorotetradecanoate (PFTeDA) (median of 370 pg/mL of ww in males and 210 pg/mL of ww in females) showed the slowest rate of telomere shortening. We also found that high blood concentrations of perfluorooctanoate (PFOA) (median of 120 pg/mL of ww in males and 150 pg/mL of ww in females) and perfluorohexanesulfonate (PFHxS) (median of 495 pg/mL of ww in males and 395 pg/mL of ww in females) were positively associated with higher re-sighting probabilities and apparent survival in males but not in females. Our work is the first to report an association between single PFAS compounds and telomeres, and the first to link PFAS exposure with survival probabilities, suggesting that the effect of PFAS exposure might be more tied to the type of compound rather than the total concentration of PFAS.

Mercury exposure in relation to foraging ecology and its impact on the oxidative status of an endangered seabird.

Mercury is a natural element extensively found in the Earth's crust, released to the atmosphere and waters by natural processes. Since the industrial revolution, atmospheric deposition of Hg showed a three-to-five-fold enrichment due to human activities. Marine top predators such as seabirds are recognized valuable bioindicators of ocean health and sensitive victims of Hg toxic effects. Hg negatively affects almost any aspect of avian physiology; thus, birds prove valuable to study the effect of Hg exposure in vertebrates. The Black-vented Shearwater is endemic to the North-Eastern Pacific Ocean, where it forages along the Baja California Peninsula during the breeding period. The area has no industrial settlement and is in the southern portion of the California Current System (CCS). After observing possible contamination effects in eggshells, we decided to quantify the exposure of breeding birds to Hg and test for possible effects on oxidative status of the species. The concentration of Hg in erythrocytes averaged 1.84 µg/g dw and varied from 1.41 to 2.40 µg/g dw. Males and females had similar Hg concentrations. The individual trophic level (reflected by δ15N) did not explain Hg exposure. In contrast, individuals foraging inshore had higher Hg concentrations than those foraging more offshore (reflected by δ13C). Shearwaters having higher concentrations of Hg had lower activity of the antioxidant enzyme glutathione peroxidase and showed lower non-enzymatic antioxidant capacity. Levels of plasma oxidative damage, superoxide dismutase and catalase were not associated with Hg. Our results indicate that (i) the foraging habitat is the factor explaining Hg exposure and (ii) there is some evidence for potential harmful effects of Hg exposure to this seabird species of conservation concern. CAPSULE: The foraging habitat is the factor explaining Hg exposure in seabirds and we observed potential harmful effects of Hg exposure in a seabird species of conservation concern.

Jumping in the Night: An Investigation of the Leaping Activity of the Western Tarsier (Cephalopachus bancanus borneanus) Using Accelerometers.

Accelerometers enable scientists to quantify the activity of free-living animals whose direct observation is difficult or demanding due to their elusive nature or nocturnal habits. However, the deployment of accelerometers on small-bodied animals and, in particular, on primates has been little explored. Here we show the first application of accelerometers on the western tarsier (Cephalopachus bancanus borneanus), a nocturnal, small-bodied primate endemic to the forests of Borneo. The fieldwork was carried out in the Lower Kinabatangan Wildlife Sanctuary, Sabah, Malaysian Borneo. We provide guidelines for the deployment of accelerometers on tarsiers that might also be applied to other primate species. Our collected data on 2 females show levels of leaping activity comparable to those previously described using direct observation of wild or captive individuals. The 2 females showed different patterns of leaping activity, which calls for work to explore individual differences further. Our work demonstrates that accelerometers can be deployed on small primates to acquire body motion data that would otherwise be demanding to collect using classic field observations. Future work will be focused on using accelerometer data to discriminate in more detail the different behaviours tarsiers can display and to address the causes and consequences of individual variations in activity.

Oxidative stress biomarkers are associated with visible clinical signs of a disease in frigatebird nestlings.

Infectious diseases are one of the most common threats for both domestic and wild animals, but little is known about the effects on the physiological condition and survival of wild animals. Here, we have tested for the first time in a wild vertebrate facing a viral disease possibly due to herpesvirus (i) whether nestlings with either low levels of oxidative damage or high levels of antioxidant protection are less susceptible to develop visible clinical signs, (ii) whether the disease is associated with the nestlings' oxidative status, (iii) whether the association between the disease and oxidative status is similar between males and females (iv), and whether cloacal and tracheal swabs might be used to detect herpesvirus. To address our questions, we took advantage of a population of Magnificent frigatebirds (Fregata magnificens) whose nestlings have experienced high mortality rates in recent times. Our work shows that (i) blood lipid oxidative damage is associated with observable clinical signs and survival probabilities of nestling frigatebirds, and (ii) that high glutathione levels in red blood cells are associated with the emergence of visible clinical signs of the disease. Our work provides evidence that differences in the oxidative status of nestlings might underlie individual health and survival.

Trophic ecology drives contaminant concentrations within a tropical seabird community.

To support environmental management programs, there is an urgent need to know about the presence and understand the dynamics of major contaminants in seabird communities of key marine ecosystems. In this study, we investigated the concentrations and trophodynamics of trace elements in six seabird species and persistent organic pollutants (POPs) in three seabird species breeding on Grand Connétable Island (French Guiana), an area where the increase in human population and mining activities has raised concerns in recent years. Red blood cell Hg concentrations in adults were the highest in Magnificent frigatebirds Fregata magnificens (median: 5.6 µg g-1 dw; range: 3.8-7.8 µg g-1 dw) and lowest in Sooty terns Onychoprion fuscatus (median: 0.9 µg g-1 dw; range: 0.6-1.1 µg g-1 dw). Among POPs, dichlorodiphenyldichloroethylene (p,p'-DDE) was the most abundant compound in plasma of Cayenne terns Thalasseus sandvicensis (median: 1100 pg g-1 ww; range: 160 ± 5100 pg g-1 ww), while polychlorinated biphenyls (PCBs) were the most abundant compound class in plasma of Magnificent frigatebirds (median: 640 pg g-1 ww; range 330 ± 2700 pg g-1 ww). While low intensity of POP exposure does not appear to pose a health threat to this seabird community, Hg concentration in several adults Laughing gulls Leucophaeus atricilla and Royal terns Thalasseus maximus, and in all Magnificent frigatebirds was similar or higher than that of high contaminated seabird populations. Furthermore, nestling red blood cells also contained Hg concentrations of concern, and further studies should investigate its potential health impact in this seabird community. Differences in adult trophic ecology of the six species explained interspecific variation in exposure to trace element and POPs, while nestling trophic ecology provides indications about the diverse feeding strategies adopted by the six species, with the consequent variation in exposure to contaminants.

Individual variation of persistent organic pollutants in relation to stable isotope ratios, sex, reproductive phase and oxidative status in Scopoli's shearwaters (Calonectris diomedea) from the Southern Mediterranean.

Little is known about the accumulation of persistent organic pollutants (POPs) and its consequences for seabirds in the Mediterranean basin. We characterised the plasma contaminant profile (polychlorinated biphenyls ΣPCBs; organochlorine pesticides ΣOCPs; polybrominated diphenyl ethers ΣPBDEs) of a population of the seabird Scopoli's shearwater (Calonectris diomedea) that breeds in the southern Mediterranean (Linosa Island) and investigated (i) whether sex, stable isotope ratios (related to diet), reproductive phase (early incubation vs. late breeding season) and body mass explained variation in contaminant burden and (ii) whether they predict health-related variables. The predominant category of POPs was ΣPCBs contributing between 53.0 and 92.4% of the total POPs in each shearwater. The percentage contribution of ΣOCPs to total POPs ranged between 7.6 and 47.0%, while that of ΣPBDEs ranged between <1% and 22.1%. Near the end of the breeding season, concentrations of ΣPCBs, ΣOCPs and ΣPOPs were significantly higher than at the beginning of the incubation period. ΣPBDEs were higher in males than females near the end of the breeding season, while they were higher in females than males at the beginning of the egg incubation period. Carbon- and nitrogen isotope ratios and individual body mass were not significantly associated with any contaminant class. Mates differed in the concentration of POPs, but they had similar stable isotope values. There was little evidence for a connection between contaminants and blood-based markers of oxidative balance. None of the contaminants predicted the probability of a bird being resighted as a breeder the following year. Thus, although POPs were present at high concentrations in some individuals, our study suggests little concern regarding POP exposure for this shearwater population.

Corticosterone, inflammation, immune status and telomere length in frigatebird nestlings facing a severe herpesvirus infection.

Herpesvirus outbreaks are common in natural animal populations, but little is known about factors that favour the infection and its consequences for the organism. In this study, we examined the pathophysiological consequences of a disease probably attributable to herpesvirus infection for several markers of immune function, corticosterone, telomere length and inflammation. In addition, we assessed whether any markers used in this study might be associated with the occurrence of visible clinical signs of the disease and its impact on short-term survival perspectives. To address our questions, in spring 2015, we collected blood samples from nestlings of the magnificent frigatebird (Fregata magnificens) that were free of any clinical signs or showed visible signs of the disease. We found that the plasma concentration of haptoglobin was strongly associated with the infection status and could predict probabilities of survival. We also found that nestlings with clinical signs had lower baseline corticosterone concentrations and similar telomere length compared with healthy nestlings, whereas we did not find any association of the infection status with innate immune defenses or with nitric oxide concentration. Overall, our results suggest that the plasma concentration of haptoglobin might be a valuable tool to assess survival probabilities of frigatebird nestlings facing a herpesvirus outbreak.

High levels of mercury and low levels of persistent organic pollutants in a tropical seabird in French Guiana, the Magnificent frigatebird, Fregata magnificens.

In the present study, trace elements and persistent organic pollutants (POPs) were quantified from Magnificent frigatebirds (Fregata magnificens) breeding at a southern Atlantic island. Stable isotope ratio of carbon (δ(13)C) and nitrogen (δ(15)N) were also measured to infer the role of foraging habitat on the contamination. For another group from the same colony, GPS tracks were recorded to identify potential foraging areas where the birds may get contaminated. Fourteen trace elements were targeted as well as a total of 40 individual POPs, including organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs). The concentration of Hg in the blood was up to 6 times higher in adults (5.81 ± 1.27 µg g(-1) dw.) than in nestlings (0.99 ± 0.23 µg g(-1) dw.). A similar pattern was found for POPs. ∑PCBs was the prevalent group both in adults (median 673, range 336-2801 pg g(-1) ww.) and nestlings (median 41, range 19-232 pg g(-1) ww.), followed by the sum of dichlorodiphenyltrichloroethanes and metabolites (∑DDTs), showing a median value of 220 (range 75-2342 pg g(-1) ww.) in adults and 25 (range 13-206 pg g(-1) ww.) in nestlings. The isotope data suggested that the accumulation of trace elements and POPs between adults and nestlings could be due to parental foraging in two different areas during incubation and chick rearing, respectively, or due to a shift in the feeding strategies along the breeding season. In conclusion, our work showed high Hg concentration in frigatebirds compared to non-contaminated seabird populations, while other trace elements showed lower values within the expected range in other seabird species. Finally, POP exposure was found generally lower than that previously measured in other seabird species.

Oxidative stress favours herpes virus infection in vertebrates: a meta-analysis.

Herpes viruses are responsible for a variety of pathological effects in humans and in both wild and domestic animals. One mechanism that has been proposed to facilitate replication and activity of herpes viruses is oxidative stress (OS). We used meta-analytical techniques to test the hypotheses that (1) herpes virus infection causes OS and (2) supplementation of antioxidants reduces virus load, indicating that replication is favoured by a state of OS. Results based on studies on mammals, including humans, and birds show that (1) OS is indeed increased by herpes virus infection across multiple tissues and species, (2) biomarkers of OS may change differently between tissues, and (3) the effect size does not differ among different virus strains. In addition, the increase of oxidative damage in blood (tissue commonly available in ecological studies) was similar to that in the tissues most sensitive to the herpes virus. Our results also show that administration of antioxidants reduces virus yield, indicating that a condition of OS is favorable for the viral replication. In addition, some antioxidants may be more efficient than others in reducing herpes virus yield. Our results point to a potential mechanism linking herpes virus infection to individual health status.