Biology subjects

Lejeune, M.

Publications and source records attributed to Lejeune, M..

15 recordsLinked to original sources

bioRxiv2026

Large-scale exposure of southern seabirds to avian cholera Pasteurella multocida: scavengers as sentinels at different spatial scales

Seabird communities represent spatially structured social networks within which potential sentinels could inform us where multi-host pathogens could be present. Pasteurella multocida, the agent of avian cholera, is responsible for recurrent massive seabird mortality events on remote Amsterdam Island, Southern Indian Ocean, but little is known about its broad spatial occurrence. To explore its distribution and whether avian scavengers and predators could be used as sentinels, we screened 18 southern seabird species from 11 islands for antibodies against P. multocida (2634 plasma samples). We detected widespread but heterogeneous exposure among host populations. High seroprevalences were found in widely roaming predators and scavengers (giant petrels (Macronectes spp), and brown skua (Stercorarius antarcticus)), which could represent efficient sentinels at broad scales. Antibodies were also detected over a series of years in a non-migratory scavenger, the lesser sheathbill (Chionis minor) and an abundant penguin species, the king penguin (Aptenodytes patagonicus), suggesting that the bacterium was circulating locally at some sites. This broad scale serological screening of whole seabird host communities provides critical information to assess disease emergence risks in the context of global change.

ecology↗

bioRxiv2026

Highly persistent antibody levels but limited population immunity in gannets after HPAI outbreak

The recent large-scale circulation of High Pathogenicity Avian Influenza (HP AI) viruses H5Nx of clade 2.3.4.4b has been responsible for massive die-offs in wild species, notably in long-lived seabirds, with unknown implications for the immunity of surviving individuals. In the North Atlantic, northern gannet colonies were heavily affected in 2022, with more than 40% mortality observed among breeding adults and some surviving individuals developing dark irises. Using samples collected in 2023 and 2024 on Rouzic colony (France), we report persistent individual anti-AI antibody levels and seroneutralisation titres, with most of the immune individuals showing dark irises. A modelling approach further stressed the importance of long-lasting immunity in such species by showing that the proportion of individuals which kept their immunity between years strongly limited decreases in population size in case of repeated outbreaks. Overall, our results highlight the existence and importance of long-lasting immunity in long-lived species for population persistence.

ecology↗

bioRxiv2026

Mass mortality of southern elephant seals during multi-species outbreak of HPAI H5N1 on sub-Antarctic Heard Island

High pathogenicity avian influenza (HPAI) has spread across the sub-Antarctic, causing significant wildlife impacts. We report its first detection in an Australian external territory, Heard Island and McDonald Islands, which supports over one million breeding seabirds and seals. Drone and ground surveys (October 2025, January 2026), combined with viral genome analysis, confirmed infection with Influenza A H5N1 clade 2.3.4.4b at Heard Island. Drone surveys revealed mass mortality in southern elephant seals, with 8,573 pups (62%) recorded dead across Heard Island by the final surveys. Mortality increased at an average rate of 5.6% per day in a subset of harems, and the highest observed mortality in a harem was 97%. Based on the average (76%) mortality in the final surveys, total estimated pup mortality at Heard Island was 13,359 (from a total population of 17,364 pups), though this may be an underestimate as mortality was ongoing at this time. HPAI was detected in six of nine species tested and, we suspect, led to elevated mortality in king and gentoo penguins. Phylogenetic analysis indicates the virus was introduced from Crozet Islands, with an estimated arrival around August 2025. These data show the continued easterly spread of HPAI around the sub-Antarctic, with severe but heterogeneous impacts across taxa. Our results demonstrate the value of drones for large-scale monitoring, underscoring the need for continued and enhanced HPAI surveillance across the Southern Ocean.

ecology↗

bioRxiv2026

Structural determinants of endopilus assembly, stability and functional specificity in bacterial type II secretion

Gram-negative bacteria employ the type II secretion system (T2SS) to transport folded protein effectors across the outer membrane. This multiprotein nanomachine assembles the endopilus, a periplasmic helical polymer composed of one major and four minor pilin subunits. Endopili are structurally related to type IV pili but exhibit distinct features including a conserved calcium-binding site stabilizing their major pilin subunits. Endopilus polymerisation is coupled to substrate translocation through a dedicated outer membrane channel. To investigate the structural basis of endopilus assembly, stability and functional specificity, we performed a comparative analysis of the Out T2SS from the plant pathogen Dickeya dadantii and the Pul T2SS from the human pathogen Klebsiella oxytoca. Although their major pilins OutG and PulG share over 77% sequence identity, these bacteria differ markedly in ecological niche and range of secreted effectors. We report here the NMR structure of calcium-bound OutG monomer and cryo-EM structures of OutG and PulG endopili at 3.6 [A] resolution. The integration of structural, mutational, and biophysical analyses with in vivo assays, allowed us to identify the molecular determinants of secretion specificity and endopilus stability. These findings demonstrate how subtle sequence variations in conserved nanomachines have evolved to optimize their function and adapt to their local environments.

microbiology↗

bioRxiv2026

Dispersal, adaptation and persistence of H5N1 in the sub-Antarctic and Antarctica

High pathogenicity avian influenza virus (HPAIV) H5N1 reached the sub-Antarctic and Antarctica in 2023, subsequently spreading to remote locations within this region where it had devastating impacts on seal, penguin and albatross populations. The threat to marine wildlife over this broad area exemplifies the need to understand H5N1 long-distance dispersal and evolution. We obtained 104 novel viral genomic sequences from samples that we collected at South Georgia, Kerguelen, Crozet, Prince Edward, Falklands/Malvinas Islands and the Antarctic Peninsula in a region spanning 8,000 kilometers. Using recent phylogeographic modeling advances we show that H5N1 spread encompassed numerous transmission events between distant locations, accumulating mammalian-adaptive mutations in the process. Seals are the most affected species, but we reveal that the long-distance eastward virus dispersal better aligns with the long-distance movements of large petrels and albatrosses. The risk of H5N1 endemisation, dispersal to other locations and ongoing evolution are highly concerning.

microbiology↗

bioRxiv2026

Drivers of host-infectious agent community associations in seabirds from sub-Antarctic oceanic islands

Understanding the patterns and drivers of infectious disease dynamics at different levels of life organization is a fundamental disease ecology challenge. In this study, we performed a multi-infectious agents (IA) screening of 1,983 individuals belonging to 18 seabird species, sampled between 2017 and 2022 across five sub-Antarctic islands in the Southern Indian and Atlantic Oceans. Our aim was to identify the drivers of IA community in these seabirds by considering intrinsic and extrinsic factors on three dimensions: distance between seabird metapopulations and within-island sites, macro-community (host functional traits based on feeding and breeding features) and intra-host IA community composition. Samples were screened for 24 DNA-based IAs using a high-throughput real-time PCR method, followed by subsequent hierarchical modelling of species communities. Campylobacter lari, Escherichia coli, Pasteurella multocida, Chlamydiaceae and Mycobacterium spp. were found to be present on all islands in some, but not all, species. However, only E. coli was detected in all species. According to our modelling results, the influence of year and within-island site had limited influence on IA presence/absence, accounting respectively for an average of 1.0% and 1.8% of the variation in IA community. Including host species as a response variable produced a better model fit than including host species functional traits, highlighting the difficulty in simplifying systems by focusing on high-level functional categories. Model outputs show how each species influences IA distribution depending on IA traits, but no clear trend has emerged. Interestingly, burrowing species were less frequently infected with directly transmitted IAs, probably due to the low frequency of transmission events associated with their breeding features and limited social contacts. Our findings pave the way for further multi-dimension studies that would combine complementary approaches to disentangle the complex processes at play in the dynamics of hosts and IAs interactions.

ecology↗

bioRxiv2025

Vaccination against H5 HP influenza virus leads to persistent immune response in wild king penguins

Since 2021, the panzootic of high pathogenicity avian influenza (HPAI) represents an increasing threat to wild vertebrate populations. In this context, recent vaccines developed for poultry could provide tools for the conservation of wild endangered birds populations. The king penguin (Aptenodytes patagonicus), a long-lived seabird breeding in dense colonies with an extended chick-rearing period, was identified as a possible surrogate species for a vaccination trial in a sub-Antarctic natural setting. We investigate here the immune response of king penguin chicks to a self-amplifying mRNA vaccine against a H5 HPAI clade 2.3.4.4b protein. Thirty chicks were vaccinated (primo- and boost-injections), 20 were kept as controls. Along 250 days of monitoring, vaccinated chicks showed a high and persistent immune response, granting a strong seroneutralisation capacity against the virus, up to fledging. No adverse effects were observed. The screening for antibodies against unspecific avian influenza viruses suggested that no natural infection occurred over the entire trial. The emergence of HPAI in the Southern Indian Ocean in October 2024 highlights the timeliness of such experimental tests. Our results show the vaccine could provide a powerful tool for mitigation and emphasises the need for studies considering ethical and practical issues of vaccination for wildlife conservation.

ecology↗

bioRxiv2025

Early-life adversity modulates growth trajectories and red blood cell mitochondrial metabolism in king penguin chicks

In many avian species, variation in breeding phenology is known to affect reproductive success. In king penguins, the breeding cycle lasts more than a year, and the start of egg-laying extends over 3 months, with early-breeders laying eggs around December and late-breeders around February. Consequently, late-born chicks have less time to grow and build up their energy reserves before the Austral winter. It is however not known whether late-born chicks display alternative physiological strategies to catch up before winter. Variation in metabolic rate is one important pathway driving differences in growth patterns, as it is directly involved both in energy allocation processes and fitness. The conversion of resources into energy occurs in mitochondria, and the efficiency of this conversion is likely to play a fundamental role in explaining individual heterogeneity in growth and survival. The purposes of this study were to investigate the differences in red blood cell mitochondrial metabolism between early- and late-born king penguin chicks and to assess whether morphometric phenotypes could be explained by differences in mitochondrial metabolism. Late-chicks expressed higher mitochondrial metabolism compared to early-chicks at 100 days post-hatching, probably linked to the stress related to winter environmental conditions and a decrease in parental feeding rates. We did not find a clear association between chick mitochondrial metabolism and growth patterns, suggesting that mostly environmental conditions contributed in explaining different metabolic phenotypes. As king penguin populations in Crozet may face changing breeding conditions (changes in feeding area and foraging trips duration) in relation to global changes, studying the physiological traits and adaptations underlying the chick growth and survival may help understanding the response of king penguins facing challenging conditions.

physiology↗

bioRxiv2025

Mass mortality events in the sub-Antarctic Indian Ocean caused by long-distance circumpolar spread of highly pathogenic avian influenza H5N1 clade 2.3.4.4b

Since 2020, the outbreak of highly pathogenic avian influenza (HPAI) virus clade 2.3.4.4b has turned into the largest documented panzootic to date, reaching the sub-Antarctic region and Antarctica via the tip of South America in 2023. Here, we describe its recent arrival into the Indian Ocean sub-Antarctic archipelagos of Crozet and Kerguelen, where we first detected the virus in October 2024 in dead southern elephant seals, king penguins, gentoo penguins, brown skuas and kelp gulls. While the panzootic is ongoing, it has already caused unprecedented and alarming mortalities of southern elephant seals. We collected brain swabs from various seal and bird carcasses, subsequently isolated the virus and obtained 25 novel HPAI H5N1 clade 2.3.4.4b sequences. Our phylogenetic and phylogeographic analyses show that there have been independent introductions of the virus to Crozet and Kerguelen, from the distant South Georgia Islands in the Southern Atlantic, and not from the more nearby coasts of South Africa. Our results point to a year-long gap in genomic surveillance in the south polar region, obscuring how HPAI H5N1 clade 2.3.4.4b is spreading in the sub-Antarctic and illustrating the difficulties in tracking pathogen dispersal in the region. Locally, our phylogenetic analyses show that the virus is transmitted between different species. Moreover, our serological analyses show that some southern elephant seal pups had mounted an anti-H5 antibody response. With the spread to Crozet and Kerguelen, HPAI H5N1 2.3.4.4b is moving ever closer to Australia and New Zealand, which currently remain free from infections with this strain, and represents a major threat to the sub-Antarctic wildlife. Our results provide key elements to enable stakeholders to anticipate the arrival and spread of the virus in remote areas of critical wildlife conservation concerns.

microbiology↗

bioRxiv2024

From birds to mammals: spillover of highly pathogenic avian influenza H5N1 virus to dairy cattle led to efficient intra- and interspecies transmission

Infections with the highly pathogenic avian influenza (HPAI) H5N1 clade 2.3.4.4b virus have resulted in the death of millions of domestic birds and thousands of wild birds in the U.S. since January, 20221-4 Throughout this outbreak, spillovers of the virus to mammals have been frequently documented5-12. Here, we report the detection of HPAI H5N1 virus in dairy cattle herds across several states in the U.S. The affected cows displayed clinical signs encompassing decreased feed intake, altered fecal consistency, respiratory distress, and decreased milk production with abnormal milk. Infectious virus and RNA were consistently detected in milk collected from affected cows. Viral staining in tissues revealed a distinct tropism of the virus for the epithelial cells lining the alveoli of the mammary gland in cows. Analysis of whole genome sequences obtained from dairy cows, birds, domestic cats, and a racoon from affected farms indicated multidirectional interspecies transmissions. Epidemiologic and genomic data revealed efficient cow-to-cow transmission after healthy cows from an affected farm were transported to a premise in a different state. These results demonstrate the transmission of HPAI H5N1 clade 2.3.4.4b virus at a non-traditional interface and to a new and highly relevant livestock species, underscoring the ability of the virus to cross species barriers.

microbiology↗

bioRxiv2023

No evidence for highly pathogenic avian influenza virus H5N1 (clade 2.3.4.4b) in the Antarctic region during the austral summer 2022/23

The current highly pathogenic avian influenza H5N1 panzootic has substantial impacts on wild birds and marine mammals. Although major outbreaks occurred in South America, incursion to Antarctica emerged late in the breeding season of 2023/2024 and was confined the wider region of the Antarctic Peninsula. To infer potential underlying processes, we compiled H5N1 surveillance from Antarctica and Sub-Antarctic Islands prior to the first confirmed cases.

microbiology↗

bioRxiv2023

Ton Motor Conformational Switch and Peptidoglycan Role in Bacterial Nutrient Uptake

Active nutrient uptake is fundamental for survival and pathogenicity of Gram-negative bacteria, which operate a multi-protein Ton system to transport essential nutrients like metals and vitamins. This system harnesses the proton motive force at the inner membrane to energize the import through the outer membrane, but the mechanism of energy transfer remains enigmatic. Here, we study the periplasmic domain of ExbD, a crucial component of the proton channel of the Ton system. We show that this domain is a dynamic dimer switching between two conformations representing the proton channels open and closed states. By in vivo phenotypic assays we demonstrate that this conformational switch is essential for the nutrient uptake by bacteria. The open state of ExbD triggers a disorder to order transition of TonB, enabling TonB to supply energy to the nutrient transporter. We also reveal the anchoring role of the peptidoglycan layer in this mechanism. Herein, we propose a mechanistic model for the Ton system, emphasizing ExbD duality and the pivotal catalytic role of peptidoglycan. Sequence analysis suggests that this mechanism is conserved in other systems energizing gliding motility and membrane integrity. Our study fills important gaps in understanding bacterial motor mechanism and proposes novel antibacterial strategies.

microbiology↗

bioRxiv2023

Bridges instead of boats? The Mla system of diderm Firmicute Veillonella parvula reveals an ancestral transenvelope core of phospholipid trafficking

Despite extensive characterisation of envelope biogenesis systems in diderm bacteria, glycerophospholipid (GPL) trafficking remains poorly understood, and has only been studied in a handful of model species. Within the Proteobacteria, the maintenance of lipid asymmetry (Mla) system facilitates retrograde GPL trafficking via six proteins, MlaA-F. GPLs are extracted from the outer leaflet of the outer membrane by the lipoprotein MlaA which associates with porin trimers, then shipped through the periplasmic space by the chaperone MlaC, which finally delivers GPLs to the inner membrane complex formed by MlaBDEF. Here, we investigate GPL trafficking in Veillonella parvula, a diderm member of the Firmicutes which encodes an Mla system devoid of MlaA and MlaC. V. parvula {Delta}mla mutants display phenotypes characteristic of disrupted lipid asymmetry such as hypervesiculation and detergent hypersensitivity, and lipid content analysis from outer membrane vesicles reveals an enrichment for the major lipid component phosphatidylethanolamine. Interestingly, suppressor analysis identifies mutations in tamB that rescue detergent hypersensitivity and hypervesiculation of {Delta}mla strains, supporting the involvement of these two systems in antagonistic GPL trafficking functions across diverse bacterial lineages. A combination of structural modeling and subcellular localisation assays shows that MlaDVp is longer than in classical diderm models and forms a transenvelope bridge, encoding both an inner membrane-localised MCE domain and an outer membrane {beta}-barrel. These results strongly suggest that V. parvula possesses a minimal Mla system for GPL trafficking, replacing the need for chaperones and outer membrane lipoproteins by directly connecting the two membranes. Finally, phylogenomic analysis indicates that this MlaEFD self-contained architecture is widely distributed in diderm bacteria and most likely represents the ancestral functional core of the Mla system, which subsequently increased in complexity in Proteobacteria and closely related phyla following the emergence of MlaABC. Our work broadens the diversity of current models of GPL trafficking in diderm bacteria, challenging the paradigm set by classical models and shedding light on the evolution of a crucial system in the biogenesis and maintenance of the bacterial outer membrane.

microbiology↗

bioRxiv2021

Autoinhibition and regulation by phosphoinositides of ATP8B1, a human lipid flippase associated with intrahepatic cholestatic disorders

P4-ATPases flip lipids from the exoplasmic to the cytosolic leaflet, thus maintaining lipid asymmetry in eukaryotic cell membranes. Mutations in several human P4-ATPase genes are associated with severe diseases, e.g. in ATP8B1 causing progressive familial intrahepatic cholestasis, a rare inherited disorder progressing toward liver failure. ATP8B1 forms a binary complex with CDC50A and displays a broad specificity to glycerophospholipids, but regulatory mechanisms are unknown. Here, we report functional studies and the cryo-EM structure of the human lipid flippase ATP8B1-CDC50A at 3.1 [A] resolution. We find that ATP8B1 is autoinhibited by its N- and C-terminal tails, which form extensive interactions with the catalytic sites and flexible domain interfaces. Consistently, ATP hydrolysis is unleashed by truncation of the C-terminus, but also requires phosphoinositides, most markedly phosphatidylinositol-3,4,5-phosphate (PI(3,4,5)P3), and removal of both N- and C- termini results in full activation. Restored inhibition of ATP8B1 truncation constructs with a synthetic peptide mimicking the C-terminal segment further suggests molecular communication between N- and C-termini in the autoinhibition and demonstrates that the regulatory mechanism can be interfered with by exogenous compounds. A recurring (G/A)(Y/F)AFS motif of the C-terminal segment suggests that this mechanism is employed widely across P4-ATPase lipid flippases in plasma membrane and endomembranes.

biochemistry↗

bioRxiv2021

Interaction of the TonB dependent transporter HasR with its cognate TonB-like protein HasB in a membrane environment

The envelope of Gram-negative bacteria is composed of a double membrane separated by the periplasmic space. This organization imposes geometrical and distance constraints that are key for the mechanism of action of multicomponent systems spanning the envelope. However, consideration of all three compartments by experimental approaches is still elusive. Here we used the state-of-the-art molecular dynamics simulation in an Escherichia coli envelope model to obtain a dynamic view of molecular interactions between the outer membrane heme transporter HasR and the inner membrane TonB-like protein HasB. Their interaction allows the transfer of the inner membrane proton motive force derived energy to the transporter for heme internalization. The simulations which incorporate both membranes show the key role of periplasmic domains of both proteins, and their dynamics in the complex formation and stability. They revealed a previously unidentified atomic network of interactions, as well as the sequences of the interactions and their variations with the presence of external substrates. Experimental validation (mutations, phenotypic and in vitro assays) confirms the robustness of our approach and provides verification of the simulation-predicted interactions. Based on structural and sequence conservation, the network of interaction revealed in this study is expected to occur in other nutrient import systems. The integrative approach presented here is highly useful to study the dynamic interplay between components of any other bacterial transmembrane systems.

biophysics↗