Erysipelothrix amsterdamensis sp. nov., associated with mortalities among endangered seabirds.

Infectious diseases threaten endangered species, particularly in small isolated populations. Seabird populations on the remote Amsterdam Island in the Indian Ocean have been in decline for the past three decades, with avian cholera caused by Pasteurella multocida proposed as the primary driver. However, Erysipelothrix species have also been sporadically detected from albatrosses on Amsterdam Island and may be contributing to some of the observed mortality. In this study, we genomically characterized 16 Erysipelothrix species isolates obtained from three Indian yellow-nosed albatross (Thalassarche carteri) chick carcasses in 2019. Histological analyses suggest that they died of bacterial septicaemia. Two isolates were sequenced using both Illumina short-read and MinION long-read approaches, which - following hybrid assembly - resulted in closed circular genomes. Mapping of Illumina reads from the remaining isolates to one of these new reference genomes revealed that all 16 isolates were closely related, with a maximum of 13 nucleotide differences distinguishing any pair of isolates. The nucleotide diversity of isolates obtained from the same or different carcasses was similar, suggesting all three chicks were likely infected from a common source. These genomes were compared with a global collection of genomes from Erysipelothrix rhusiopathiae and other species from the same genus. The isolates from albatrosses were phylogenetically distinct, sharing a most recent common ancestor with E. rhusiopathiae. Based on phylogenomic analysis and standard thresholds for average nucleotide identity and digital DNA-DNA hybridization, these isolates represent a novel Erysipelothrix species, for which we propose the name Erysipelothrix amsterdamensis sp. nov. The type strain is A18Y020dT (=CIP 112216T=DSM 115297T). The implications of this bacterium for albatross conservation will require further study.

Climate change in the Arctic: Testing the poleward expansion of ticks and tick-borne diseases.

Climate change is most strongly felt in the polar regions of the world, with significant impacts on the species that live there. The arrival of parasites and pathogens from more temperate areas may become a significant problem for these populations, but current observations of parasite presence often lack a historical reference of prior absence. Observations in the high Arctic of the seabird tick Ixodes uriae suggested that this species expanded poleward in the last two decades in relation to climate change. As this tick can have a direct impact on the breeding success of its seabird hosts and vectors several pathogens, including Lyme disease spirochaetes, understanding its invasion dynamics is essential for predicting its impact on polar seabird populations. Here, we use population genetic data and host serology to test the hypothesis that I. uriae recently expanded into Svalbard. Both black-legged kittiwakes (Rissa tridactyla) and thick-billed murres (Uria lomvia) were sampled for ticks and blood in Kongsfjorden, Spitsbergen. Ticks were genotyped using microsatellite markers and population genetic analyses were performed using data from 14 reference populations from across the tick's northern distribution. In contrast to predictions, the Spitsbergen population showed high genetic diversity and significant differentiation from reference populations, suggesting long-term isolation. Host serology also demonstrated a high exposure rate to Lyme disease spirochaetes (Bbsl). Targeted PCR and sequencing confirmed the presence of Borrelia garinii in a Spitsbergen tick, demonstrating the presence of Lyme disease bacteria in the high Arctic for the first time. Taken together, results contradict the notion that I. uriae has recently expanded into the high Arctic. Rather, this tick has likely been present for some time, maintaining relatively high population sizes and an endemic transmission cycle of Bbsl. Close future observations of population infestation/infection rates will now be necessary to relate epidemiological changes to ongoing climate modifications.

Active breeding seabirds prospect alternative breeding colonies.

Compared to other animal movements, prospecting by adult individuals for a future breeding site is commonly overlooked. Prospecting influences the decision of where to breed and has consequences on fitness and lifetime reproductive success. By analysing movements of 31 satellite- and GPS-tracked gull and tern populations belonging to 14 species in Europe and North America, we examined the occurrence and factors explaining prospecting by actively breeding birds. Prospecting in active breeders occurred in 85.7% of studied species, across 61.3% of sampled populations. Prospecting was more common in populations with frequent inter-annual changes of breeding sites and among females. These results contradict theoretical models which predict that prospecting is expected to evolve in relatively predictable and stable environments. More long-term tracking studies are needed to identify factors affecting patterns of prospecting in different environments and understand the consequences of prospecting on fitness at the individual and population level.

Exposure of breeding albatrosses to the agent of avian cholera: dynamics of antibody levels and ecological implications.

Despite critical implications for disease dynamics and surveillance in wild long-lived species, the immune response after exposure to potentially highly pathogenic bacterial disease agents is still poorly known. Among infectious diseases threatening wild populations, avian cholera, caused by the bacterium Pasteurella multocida, is a major concern. It frequently causes massive mortality events in wild populations, notably affecting nestlings of Indian yellow-nosed albatrosses (Thalassarche carteri) in the Indian Ocean. If adults are able to mount a long-term immune response, this could have important consequences regarding the dynamics of the pathogen in the local host community and the potential interest of vaccinating breeding females to transfer immunity to their offspring. By tracking the dynamics of antibodies against P. multocida during 4 years and implementing a vaccination experiment in a population of yellow-nosed albatrosses, we show that a significant proportion of adults were naturally exposed despite high annual survival for both vaccinated and non-vaccinated individuals. Adult-specific antibody levels were thus maintained long enough to inform about recent exposure. However, only low levels of maternal antibodies could be detected in nestlings the year following a vaccination of their mothers. A modification of the vaccine formulation and the possibility to re-vaccinate females 2 years after the first vaccination revealed that vaccines have the potential to elicit a stronger and more persistent response. Such results highlight the value of long-term observational and experimental studies of host exposure to infectious agents in the wild, where ecological and evolutionary processes are likely critical for driving disease dynamics.

Diversity, prevalence and host specificity of avian parasites in southern Tunisian oases.

Oases are semi-natural woodlots surrounded by an inhospitable desert environment. This insular-like habitat system is known to support a mixture of sedentary and migratory bird species originating from different areas. However, little is known about the interactions between these birds and parasites. In this study, we investigated the diversity, prevalence and host specificity of avian haemosporidian parasites in southern Tunisian oases in two sedentary and common bird species, the laughing dove Spilopelia senegalensis and hybrid sparrow Passer domesticus × hispaniolensis, in six sites that differ regarding vegetation structure and distance to the coast. Two new Haemoproteus lineages, related to other Haemoproteus transmitted by biting midges, were detected in doves. With regard to sparrows, all detected parasites have previously been reported in other sparrow populations, except for one new Haemoproteus lineage. Our results also showed that densely vegetated sites were characterized by the higher prevalence of Plasmodium but a lower prevalence of Haemoproteus compared with less-vegetated sites. This is the first study aiming to explore avian parasites in the oasis habitat. Gathering data on a larger sample of oases with different sizes and isolation levels will be the next step to better understand factors shaping the transmission dynamics of avian parasites in such ecosystems.

Influenza A virus on oceanic islands: host and viral diversity in seabirds in the Western Indian Ocean.

Ducks and seabirds are natural hosts for influenza A viruses (IAV). On oceanic islands, the ecology of IAV could be affected by the relative diversity, abundance and density of seabirds and ducks. Seabirds are the most abundant and widespread avifauna in the Western Indian Ocean and, in this region, oceanic islands represent major breeding sites for a large diversity of potential IAV host species. Based on serological assays, we assessed the host range of IAV and the virus subtype diversity in terns of the islands of the Western Indian Ocean. We further investigated the spatial variation in virus transmission patterns between islands and identified the origin of circulating viruses using a molecular approach. Our findings indicate that terns represent a major host for IAV on oceanic islands, not only for seabird-related virus subtypes such as H16, but also for those commonly isolated in wild and domestic ducks (H3, H6, H9, H12 subtypes). We also identified strong species-associated variation in virus exposure that may be associated to differences in the ecology and behaviour of terns. We discuss the role of tern migrations in the spread of viruses to and between oceanic islands, in particular for the H2 and H9 IAV subtypes.

The role of seabirds of the Iles Eparses as reservoirs and disseminators of parasites and pathogens.

The role of birds as reservoirs and disseminators of parasites and pathogens has received much attention over the past several years due to their high vagility. Seabirds are particularly interesting hosts in this respect. In addition to incredible long-distance movements during migration, foraging and prospecting, these birds are long-lived, site faithful and breed in dense aggregations in specific colony locations. These different characteristics can favor both the local maintenance and large-scale dissemination of parasites and pathogens. The Iles Eparses provide breeding and feeding grounds for more than 3 million breeding pairs of seabirds including at least 13 species. Breeding colonies on these islands are relatively undisturbed by human activities and represent natural metapopulations in which seabird population dynamics, movement and dispersal can be studied in relation to that of circulating parasites and pathogens. In this review, we summarize previous knowledge and recently-acquired data on the parasites and pathogens found in association with seabirds of the Iles Eparses. These studies have revealed the presence of a rich diversity of infectious agents (viruses, bacteria and parasites) carried by the birds and/or their local ectoparasites (ticks and louse flies). Many of these agents are widespread and found in other ecosystems confirming a role for seabirds in their large scale dissemination and maintenance. The heterogeneous distribution of parasites and infectious agents among islands and seabird species suggests that relatively independent metacommunities of interacting species may exist within the western Indian Ocean. In this context, we discuss how the patterns and determinants of seabird movements may alter parasite and pathogen circulation. We conclude by outlining key aspects for future research given the baseline data now available and current concerns in eco-epidemiology and biodiversity conservation.

Laridae: A neglected reservoir that could play a major role in avian influenza virus epidemiological dynamics.

Avian influenza viruses (AIVs) are of great concern worldwide due to their economic impact and the threat they represent to human health. As wild birds are the natural reservoirs of AIVs, understanding AIV dynamics in different avian taxa is essential for deciphering the epidemiological links between wildlife, poultry and humans. To date, only the Anatidae (ducks, geese and swans) have been widely studied. Here, we aim to shed light on the current state of knowledge on AIVs in Laridae (gulls, terns and kittiwakes) versus that in Anatidae by setting forth four fundamental questions: how, when, where and to which host species are AIVs transmitted? First, we describe ecological differences between Laridae and Anatidae and discuss how they may explain observed contrasts in preferential transmission routes and the evolution of specific AIV subtypes. Second, we highlight the dissimilarities in the temporal patterns of AIV shedding between Laridae and Anatidae and address the role that immunity likely plays in shaping these patterns. Third, we underscore that Laridae may be key in promoting intercontinental exchanges of AIVs. Finally, we emphasize the crucial epidemiological position that Laridae occupy between wildlife, domestic birds and humans.

Long antibody persistence and transgenerational transfer of immunity in a long-lived vertebrate.

Although little studied in natural populations, the persistence of immunoglobulins may dramatically affect the dynamics of immunity and the ecology and evolution of host-pathogen interactions involving vertebrate hosts. By means of a multiple-year vaccination design against Newcastle disease virus, we experimentally addressed whether levels of specific antibodies can persist over several years in females of a long-lived procellariiform seabird-Cory's shearwater-and whether maternal antibodies against that antigen could persist over a long period in offspring several years after the mother was exposed. We found that a single vaccination led to high levels of antibodies for several years and that the females transmitted antibodies to their offspring that persisted for several weeks after hatching even 5 years after a single vaccination. The temporal persistence of maternally transferred antibodies in nestlings was highly dependent on the level at hatching. A second vaccination boosted efficiently the level of antibodies in females and thus their transfer to offspring. Overall, these results stress the need to consider the temporal dynamics of immune responses if we are to understand the evolutionary ecology of host-parasite interactions and trade-offs between immunity and other life-history characteristics, in particular in long-lived species. They also have strong implications for conservation when vaccination may be used in natural populations facing disease threats.

Tracing the colonization and diversification of the worldwide seabird ectoparasite Ixodes uriae.

Historical patterns of dispersal and population isolation are key components shaping contemporary genetic diversity across landscapes and require explicit consideration when examining the relative role of different factors in driving the evolution of host specificity in parasitic organisms. In this study, we investigate the worldwide colonization history of a common ectoparasite of seabirds, the tick Ixodes uriae. This tick has a circumpolar distribution across both hemispheres but has repeatedly formed host-specific races within different regions. By combining mitochondrial and nuclear data, we infer how this species spread to its present-day distribution and how the colonization process may have affected the geographic and host-associated structure of this tick within regions. We demonstrate that I. uriae is highly structured at a global scale and isolates into four genetic groups that correspond to well-defined geographical regions. Molecular dating suggests that the diversification of I. uriae began in the early Miocene (22 Myr) and that this tick colonized most of the southern hemisphere before moving into northern latitudes via two independent routes. However, no relationship between the degree of host race divergence and colonization history was evident, supporting previous hypotheses that host specialization evolves relatively rapidly in this parasite, but does not typically lead to speciation. We discuss the possible historical and contemporary mechanisms of large-scale dispersal for this ectoparasite and how its biological characteristics may condition current patterns of genetic diversity. More generally, our results illustrate how combining broad-scale sampling and modern molecular tools can help disentangle complex patterns of diversification in widespread parasites.

An experimental test of host specialization in a ubiquitous polar ectoparasite: a role for adaptation?

The evolution of host specificity is considered to be an essential mechanism driving parasite diversity. It may be governed by adaptive constraints that lead to host-dependent fitness trade-offs. Alternatively, specificity may arise via transmission constraints that isolate parasite populations, without necessarily involving adaptation per se. Here, we ask whether the repeated observation of host-associated genetic races across the worldwide distribution of the seabird ectoparasite Ixodes uriae is associated with host adaptation. We conducted a field-based experiment to test for adaptive specialisation in host races of I. uriae. We transferred unengorged ticks of two life stages (nymphs and adults) originating from three host species (black-legged kittiwake, common guillemot and Atlantic puffin) onto young kittiwake nestlings and followed attraction and attachment rates, engorgement times and feeding success of the transplanted ticks. All ticks were also typed genetically to match exploitation patterns with genetic differences among races. Ticks from atypical hosts were significantly less attracted to nestlings than ticks from the typical host, and showed lower feeding success and higher mortality. The degree of host specificity matched patterns of neutral genetic variation among races, with puffin ticks being more specific than guillemot ticks. Differences in specificity were also apparent among tick life stages, suggesting that nymphal ticks may be less discriminating of host type than adult ticks. Our results indicate that the genetic divergence previously observed among sympatric I. uriae host races is at least partially linked to adaptive specialisation to the host species and not simply to host-mediated transmission. They also suggest that the adaptation process may evolve differently in different life stages based on trade-offs with physiological constraints. The identification of the selective forces acting in host specialization will now be necessary to better characterize these patterns and to understand how transmission interacts with the adaptation process to generate parasite biodiversity.

Egg sampling as a possible alternative to blood sampling when monitoring the exposure of yellow-legged gulls (Larus michahellis) to avian influenza viruses.

We explored whether antibody detection in egg yolks could serve as an alternative to antibody detection in plasma samples when monitoring yellow-legged gulls (Larus michahellis) for exposure to avian influenza viruses (AIVs). We tested female plasma and eggs for anti-AIV antibodies and used the data we obtained to check whether the two sample types yielded the same antibody status (positive or negative) and to compare the antibody prevalence estimated from the blood data with that estimated from the yolk data. Our results showed that sampling one egg per clutch, regardless of that egg's position in the laying sequence, is sufficient to provide an unbiased estimate of antibody prevalence across clutches. The results also showed that almost 25% of the clutches laid by positive females contained only antibody-negative eggs, which suggests that yolk samples might underestimate female antibody prevalence. However, this result may stem from differences in the methods used to assess plasma versus yolk antibody status. Further research is needed to clarify this issue; while the number of false negatives could be reduced by adapting antibody detection techniques, it may be that they are an unavoidable consequence of natural avian maternal transfer dynamics.

Rapid loss of maternal immunity and increase in environmentally mediated antibody generation in urban gulls.

Monitoring pathogen circulation in wildlife sentinel populations can help to understand and predict the spread of disease at the wildlife-livestock-human interface. Immobile young provide a useful target population for disease surveillance, since they can be easily captured for sampling and their levels of antibodies against infectious agents can provide an index of localized circulation. However, early-life immune responses include both maternally-derived antibodies and antibodies resulting from exposure to pathogens, and disentangling these two processes requires understanding their individual dynamics. We conducted an egg-swapping experiment in an urban-nesting sentinel seabird, the yellow-legged gull, and measured antibody levels against three pathogens of interest (avian influenza virus AIV, Toxoplasma gondii TOX, and infectious bronchitis virus IBV) across various life stages, throughout chick growth, and between nestlings raised by biological or non-biological parents. We found that levels of background circulation differed among pathogens, with AIV antibodies widely present across all life stages, TOX antibodies rarer, and IBV antibodies absent. Antibody titers declined steadily from adult through egg, nestling, and chick stages. For the two circulating pathogens, maternal antibodies declined exponentially after hatching at similar rates, but the rate of linear increase due to environmental exposure was significantly higher in the more prevalent pathogen (AIV). Differences in nestling antibody levels due to parental effects also persisted longer for AIV (25 days, vs. 14 days for TOX). Our results suggest that yellow-legged gulls can be a useful sentinel population of locally transmitted infectious agents, provided that chicks are sampled at ages when environmental exposure outweighs maternal effects.

Tick-borne zoonotic flaviviruses and Borrelia infections in wildlife hosts: What have field studies contributed?

Tick-borne flaviviruses and Borrelia spp. are globally spread pathogens of zoonotic potential that are maintained by a transmission cycle at the interface between ticks and vertebrate hosts, mainly wild animals. Aside data on pathogen burden in ticks, information on the status of various hosts relative to infection is important to acquire. We reviewed how those infections have been studied in wildlife host species in the field to discuss how collected data provided relevant epidemiological information and to identify needs for further studies. The literature was screened for observational studies on pathogen or antibody detection for tick-borne Borrelia spp. and flaviviruses in wildlife host animals. Overall, Borrelia spp. were more studied (73% of case studies, representing 297 host species) than flaviviruses (27% of case studies, representing 114 host species). Studies on both Borrelia spp. and flaviviruses focused mainly on the same species, namely bank vole and yellow-necked mouse. Most studies were order-specific and cross-sectional, reporting prevalence at various locations, but with little insight into the underlying epidemiological dynamics. Host species with potential to act as reservoir hosts of these pathogens were neglected, notably birds. We highlight the necessity of collecting both demographics and infection data in wildlife studies, and to consider communities of species, to better estimate zoonotic risk potential in the One Health context.

Evolution of the temporal persistence of immune protection.

The evolution of resistance to parasites has been the focus of numerous theoretical studies and several mechanisms, ranging from innate to acquired immune responses, have been considered. Life-history theory predicts that long-lived species should invest more resources into maintenance and immunity than short-lived species. Here, we provide further theoretical and empirical support for this hypothesis. First, an analysis of the evolution of the persistence of immune protection in a theoretical framework accounting for maternal transfer of immunity reveals that longer-lived hosts are expected to invest in more persistent intragenerational and transgenerational immune responses. Controlling for phylogenetic structure and for the confounding effect of catabolic activity, we further showed that immunoglobulin half-life and longevity are positively correlated in mammal species. Our study confirms that persistence of immunity has evolved as part of elaborate anti-parasitic defence strategies.

Female blue tits with brighter yellow chests transfer more carotenoids to their eggs after an immune challenge.

Female ornaments are present in many species, and it is more and more accepted that sexual or social selection may lead to their evolution. By contrast, the information conveyed by female ornaments is less well understood. Here, we investigated the links between female ornaments and maternal effects. In birds, an important maternal effect is the transmission of resources, such as carotenoids, into egg yolk. Carotenoids are pigments with antioxidant and immunomodulatory properties that are crucial for females and developing offspring. In blue tits, we evaluated whether ultraviolet (UV)/blue and yellow feather colouration signals a female's capacity to allocate carotenoids to egg yolk. Because mounting an immune response is costly and trade-offs are more detectable under harsh conditions, we challenged the immune system of females before laying and examined the carotenoid level of their eggs afterward. A positive association between feather carotenoid chroma and egg carotenoid level would be expected if yellow colouration signals basal immunity. Alternatively, if female colouration more generally reflects maternal capacity to invest in reproduction under challenging conditions, then other components of colouration (i.e. yellow brightness and UV/blue colouration) could be linked to maternal capacity to invest in eggs. No association between egg carotenoid levels and UV/blue crown colouration or female yellow chest chroma was found; the latter result suggests that yellow colouration does not signal immune capacity at laying in this species. By contrast, we found that, among females that mounted a detectable response to the vaccine, those with brighter yellow chests transmitted more carotenoids into their eggs. This result suggests yellow brightness signals maternal capacity to invest in reproduction under challenging conditions, and that male blue tits may benefit directly from choosing brighter yellow females.

Avian influenza spread and seabird movements between colonies.

Seabirds have recently been experiencing high rates of mortality across wide scales due to highly pathogenic avian influenza (HPAI). During breeding, seabird populations are highly spatially structured, while over their lifetimes they spend much time at sea. This makes them unique systems in which to document how movement and interspecies interactions affect eco-epidemiological dynamics.

Occurrence of ESBL- and AmpC-Producing E. coli in French Griffon Vultures Feeding on Extensive Livestock Carcasses.

Despite the fact that the selective pressure of antibiotics on wild birds is supposed to be very weak, they are considered potential vectors of antimicrobial resistance (AMR). Obligate scavengers such as vultures can present high proportions of resistance to extended-spectrum cephalosporins (ESC) and multi-drug-resistant (MDR) bacteria, partially due to feeding stations that are provisioned with livestock carcasses from intensive farming. Here we investigated whether griffon vultures (Gyps fulvus) from two populations located in the French Alps, which feed on livestock carcasses from extensive farms, may carry such resistant bacteria. Phenotypic and genotypic characterization showed an 11.8% proportion of ESC-resistant bacteria, including five extended-spectrum beta-lactamase (ESBL)-producing and one AmpC-producing E. coli. The five ESBL-positive E. coli were clonal and all came from the same vulture population, proving their spread between animals. The ESBL phenotype was due to a blaCTX-M-15 gene located on the chromosome. Both ESBL- and AmpC-positive E. coli belonged to minor STs (ST212 and ST3274, respectively); interestingly, ST212 has already been identified in wild birds around the world, including vultures. These results suggest that actions are needed to mitigate the spread of MDR bacteria through wild birds, particularly in commensal species.

Temporal dynamics of antibody level against Lyme disease bacteria in roe deer: Tale of a sentinel?

Changes in the risk of exposure to infectious disease agents can be tracked through variations in antibody prevalence in vertebrate host populations. However, information on the temporal dynamics of the immune status of individuals is critical. If antibody levels persist a long time after exposure to an infectious agent, they could enable the efficient detection of the past circulation of the agent; if they persist only a short time, they could provide snap shots of recent exposure of sampled hosts. Here, we explored the temporal dynamics of seropositivity against Lyme disease agent Borrelia burgdorferi sensu lato (Bbsl) in individuals of a widespread medium-sized mammal species, the roe deer (Capreolus capreolus), in France. Using a modified commercially available immunoassay we tested 1554 blood samples obtained in two wild deer populations monitored from 2010 to 2020. Using multi-event capture-mark-recapture models, we estimated yearly population-, age-, and sex-specific rates of seroconversion and seroreversion after accounting for imperfect detection. The yearly seroconversion rates indicated a higher level of exposure in early (2010-2013) than in late years (2014-2019) to infected tick bites in both populations, without any detectable influence of sex or age. The relatively high rates of seroreversion indicated a short-term persistence of antibody levels against Bbsl in roe deer. This was confirmed by the analysis of samples collected on a set of captive individuals that were resampled several times a few weeks apart. Our findings show the potential usefulness of deer as a sentinel for tracking the risk of exposure to Lyme disease Bbsl, although further investigation on the details of the antibody response to Bbsl in this incompetent host would be useful. Our study also highlights the value of combining long-term capture-mark-recapture sampling and short-time analyses of serological data for wildlife populations exposed to infectious agents of relevance to wildlife epidemiology and human health.

Highly pathogenic avian influenza affects vultures' movements and breeding output.

An exceptional highly pathogenic avian influenza (HPAI) outbreak due to H5N1 virus genotypes belonging to clade 2.3.4.4.b has been affecting birds worldwide since autumn 2021.1,2,3 Mortality caused by viral infection has been well documented in poultry and more recently in wild birds, especially in seabird-breeding colonies.4,5,6 However, there is a critical lack of knowledge about how terrestrial birds deal with HPAI virus infections in terms of behavior and space use, especially during the breeding season.7,8,9 Understanding how birds move when they are infected could help evaluate the risk of spreading the virus at a distance among other populations of wild or domestic birds, this latter risk being especially important for commensal bird species. Through long-term GPS tracking, we described the changes in daily movement patterns of 31 adult griffon vultures Gyps fulvus in two French sites in 2022 compared with 3 previous years. In spring 2022, 21 vultures at both sites showed periods of immobility at the nest, during 5.6 days on average. Positive serological status of 2 individuals confirmed that they had been infected by HPAI viruses. Death was recorded for 3 of the 31 tracked individuals, whereas all others recovered and returned quickly to their foraging routine, although at least 9 birds failed breeding. Such immobility patterns and death rates were never observed in previous years and were not related to weather conditions. The high immobility behavior of infected birds could reduce the risks of transmission. The observed vulnerability to HPAI viruses questions the resistance of endangered vulture species worldwide if infected.