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International audience; Peste des petits ruminants (PPR) is a highly contagious viral disease that mainly affects goats and sheep in Asia, Africa and the Middle East, and threatens Europe [R.E.1]. The disease is endemic on the African continent, particularly in West Africa, and is a major factor driving food insecurity in low-income populations. The aim of this research study was to carry out surveillance, genetic characterisation and isolation of recently circulating PPR viruses (PPRV) in sheep and goats from the six agro-ecological zones of Nigeria. A total of 268 post-mortem tissue samples of lung and mesenteric ganglia were collected from clinically suspected sheep and goats in 18 different states, of which five never previously sampled. The presence of PPRV was confirmed using a reverse-transcription coupled with a polymerase chain reaction (RT-PCR) assay. A total of 72 samples, 17 sheep (6%) and 55 goats (21%), were found to be PPR positive. Positive samples were distributed in almost all states, except Kano, where PPR was detected in previous studies. The PPRV-positive samples were further confirmed by sequencing or virus isolation in areas where the infection had never previously been detected. These results confirm the active circulation of PPRV across all six agro-ecological zones of Nigeria, and consequently, the need for introducing strict measures for the control and prevention of the disease in the country.

Peste des petits ruminants (PPR) is a highly contagious and devastating viral disease infecting predominantly sheep and goats. Tracking outbreaks of disease and analysing the movement of the virus often involves sequencing part or all of the genome and comparing the sequence obtained with sequences from other outbreaks, obtained from the public databases. However, there are a very large number (>1800) of PPRV sequences in the databases, a large majority of them relatively short, and not always well-documented. There is also a strong bias in the composition of the dataset, with countries with good sequencing capabilities (e.g. China, India, Turkey) being overrepresented, and most sequences coming from isolates in the last 20 years. In order to facilitate future analyses, we have prepared sets of PPRV sequences, sets which have been filtered for sequencing errors and unnecessary duplicates, and for which date and location information has been obtained, either from the database entry or from other published sources. These sequence datasets are freely available for download, and include smaller datasets which maximise phylogenetic information from the minimum number of sequences, and which will be useful for simple lineage identification. Their utility is illustrated by uploading the data to the MicroReact platform to allow simultaneous viewing of lineage date and geographic information on all the viruses for which we have information. While preparing these datasets, we identified a significant number of public database entries which contain clear errors, and propose guidelines on checking new sequences and completing metadata before submission.

Animal-associated microbiomes are integral to host health, yet key biotic and abiotic factors that shape host-associated microbial communities at the global scale remain poorly understood. We investigated global patterns in amphibian skin bacterial communities, incorporating samples from 2,349 individuals representing 205 amphibian species across a broad biogeographic range. We analysed how biotic and abiotic factors correlate with skin microbial communities using multiple statistical approaches. Global amphibian skin bacterial richness was consistently correlated with temperature-associated factors. We found more diverse skin microbiomes in environments with colder winters and less stable thermal conditions compared with environments with warm winters and less annual temperature variation. We used bioinformatically predicted bacterial growth rates, dormancy genes and antibiotic synthesis genes, as well as inferred bacterial thermal growth optima to propose mechanistic hypotheses that may explain the observed patterns. We conclude that temporal and spatial characteristics of the host’s macro-environment mediate microbial diversity. National Science Foundation/[DEB-1146284]/NSF/Estados Unidos National Science Foundation/[IOS-1121758]/NSF/Estados Unidos National Science Foundation/[DEB-1310036]/NSF/Estados Unidos John Templeton Foundation/[]/JTF/Estados Unidos Deutsche Forschungsgemeinschaft/[]/DFG/Alemania Deutsche Forschungsgemeinschaft/[VE247/9-1]/DFG/Alemania Coordenação de Aperfeiçoamento de Pessoal de Nível Superior/[]/CAPES/Brasil Fundação de Amparo à Pesquisa do Estado de São Paulo/[#2013/50741-7]/FAPESP/Brasil Conselho Nacional de Desenvolvimento Científico e Tecnológico/[]/CNPq/Brasil Simons Foundation/[429440, WTW]//Estados Unidos Deutscher Akademischer Austauschdienst/[]/DAAD/Alemania Universidad de Costa Rica/[801-B2-029]/UCR/Costa Rica Ministerio de Ciencia, Tecnología y Telecomunicaciones/[849-PINN-2015]/MICITT/Costa Rica National Research Foundation of Korea/[2015R1D1A1A01057282]/NRF/Corea del Sur UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Biología Celular y Molecular (CIBCM) UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Estructuras Microscópicas (CIEMIC)

Peste des petits ruminants virus (PPRV) causes disease in domestic and wild ungulates, is the target of a Global Eradication Programme, and threatens biodiversity. Understanding the epidemiology and evolution of PPRV in wildlife is important but hampered by the paucity of wildlife-origin PPRV genomes. In this study, full PPRV genomes were generated from three Mongolian saiga antelope, one Siberian ibex, and one goitered gazelle from the 2016–2017 PPRV outbreak. Phylogenetic analysis showed that for Mongolian and Chinese PPRV since 2013, the wildlife and livestock-origin genomes were closely related and interspersed. There was strong phylogenetic support for a monophyletic group of PPRV from Mongolian wildlife and livestock, belonging to a clade of lineage IV PPRV from livestock and wildlife from China since 2013. Discrete diffusion analysis found strong support for PPRV spread into Mongolia from China, and phylogeographic analysis indicated Xinjiang Province as the most likely origin, although genomic surveillance for PPRV is poor and lack of sampling from other regions could bias this result. Times of most recent common ancestor (TMRCA) were June 2015 (95 per cent highest posterior density (HPD): August 2014 to March 2016) for all Mongolian PPRV genomes and May 2016 (95 per cent HPD: October 2015 to October 2016) for Mongolian wildlife-origin PPRV. This suggests that PPRV was circulating undetected in Mongolia for at least 6 months before the first reported outbreak in August 2016 and that wildlife were likely infected before livestock vaccination began in October 2016. Finally, genetic variation and positively selected sites were identified that might be related to PPRV emergence in Mongolian wildlife. This study is the first to sequence multiple PPRV genomes from a wildlife outbreak, across several host species. Additional full PPRV genomes and associated metadata from the livestock–wildlife interface are needed to enhance the power of molecular epidemiology, support PPRV eradication, and safeguard the health of the whole ungulate community., Lay Summary Recent mass mortality of critically endangered Mongolian saiga antelope due to peste des petits ruminants virus (PPRV) has dramatically highlighted the threat this viral disease represents for biodiversity. The genome of viruses such as PPRV evolves fast, so virus genetic data gathered from infected animals can be used to trace disease spread between livestock and wildlife and to determine if the virus is adapting to infect wildlife more efficiently. Here, we obtained PPRV virus genomes from Mongolian wildlife and compared them with other published PPRV genomes. Using a molecular clock, we estimated that the disease was circulating in Mongolia well before it was first reported. Genetic analyses support the hypothesis of virus spread from livestock to wildlife, with genetic changes potentially helping infection in Asian wild ungulates. However, more PPR virus genomes and epidemiology data are needed from disease outbreaks in areas shared between livestock and wildlife to confirm these results and take efficient actions to safeguard the health of the whole ungulate community.

Peste des petits ruminants (PPR) is a viral disease of goats and sheep that occurs in Africa, the Middle East and Asia with a severe impact on livelihoods and livestock trade. Many wild artiodactyls are susceptible to PPR virus (PPRV) infection, and some outbreaks have threatened endangered wild populations. The role of wild species in PPRV epidemiology is unclear, which is a knowledge gap for the Global Strategy for the Control and Eradication of PPR. These studies aimed to investigate PPRV infection in wild artiodactyls in the Greater Serengeti and Amboseli ecosystems of Kenya and Tanzania. Out of 132 animals purposively sampled in 2015–2016, 19.7% were PPRV seropositive by ID Screen PPR competition enzyme-linked immunosorbent assay (cELISA, IDvet, France) from the following species: African buffalo, wildebeest, topi, kongoni, Grant’s gazelle, impala, Thomson’s gazelle, warthog and gerenuk, while waterbuck and lesser kudu were seronegative. In 2018–2019, a cross-sectional survey of randomly selected African buffalo and Grant’s gazelle herds was conducted. The weighted estimate of PPRV seroprevalence was 12.0% out of 191 African buffalo and 1.1% out of 139 Grant’s gazelles. All ocular and nasal swabs and faeces were negative by PPRV real-time reverse transcription-polymerase chain reaction (RT-qPCR). Investigations of a PPR-like disease in sheep and goats confirmed PPRV circulation in the area by rapid detection test and/or RT-qPCR. These results demonstrated serological evidence of PPRV infection in wild artiodactyl species at the wildlife–livestock interface in this ecosystem where PPRV is endemic in domestic small ruminants. Exposure to PPRV could be via spillover from infected small ruminants or from transmission between wild animals, while the relatively low seroprevalence suggests that sustained transmission is unlikely. Further studies of other major wild artiodactyls in this ecosystem are required, such as impala, Thomson’s gazelle and wildebeest.

Globalized infectious diseases are causing species declines worldwide, but their source often remains elusive. We used whole-genome sequencing to solve the spatiotemporal origins of the most devastating panzootic to date, caused by the fungus Batrachochytrium dendrobatidis, a proximate driver of global amphibian declines. We traced the source of B. dendrobatidis to the Korean peninsula, where one lineage, BdASIA-1, exhibits the genetic hallmarks of an ancestral population that seeded the panzootic. We date the emergence of this pathogen to the early 20th century, coinciding with the global expansion of commercial trade in amphibians, and we show that intercontinental transmission is ongoing. Our findings point to East Asia as a geographic hotspot for B. dendrobatidis biodiversity and the original source of these lineages that now parasitize amphibians worldwide., S.J.O., T.W.J.G., L.Br., A.Lo., A.A.C., D.S.S., E.A.C., C.M., J.B., D.M.A., F.C., and M.C.F. were supported through NERC (standard grant NE/K014455/1). S.J.O. acknowledges a Microsoft Azure for Research Sponsorship (subscription ID: ab7cd695-49cf-4a83-910a-ef71603e708b). T.W.J.G., A.Lo., A.A.C., D.S.S., E.A.C., C.M., J.B., D.M.A., F.C., and M.C.F. were also supported by the EU BiodivERsA scheme (RACE, funded through NERC directed grant NE/G002193/1 and ANR08-Biodiversa-002-03) and NERC (standard grant NE/K012509/1). M.C.F., E.A.C., and C.M. acknowledge the Nouragues Travel Grant Program 2014. R.A.F. was supported by an MIT/Wellcome Trust Fellowship. T.W.J.G. was supported by the People’s Trust for Endangered Species and the Morris Animal Foundation (D12ZO002). J.M.G.S. and M.C.F. were supported by the Leverhulme Trust (RPG-2014-273) and the Morris Animal Foundation (D16ZO-022). F.B. was supported by the ERC (grant ERC 260801–Big_Idea). D.M.A. was funded by Wellcome Trust grant 099202. J.V. was supported by the Hungarian Scientific Research Fund (OTKA K77841) and Bolyai János Research Scholarship, Hungarian Academy of Sciences (BO/00579/14/8). D.J.G. was supported by the Conservation Leadership Programme (grant 0134010) with additional assistance from F. Gebresenbet, R. Kassahun, and S. P. Loader. C.S.-A. was supported by Fondecyt Nº11140902 and 1181758. T.M.D.-B. was supported by the Royal Geographical Society and the Royal Zoological Society of Scotland with assistance from M. Hirschfeld and the Budongo Conservation Field Station. B.W. was supported by the National Research Foundation of Korea (2015R1D1A1A01057282). L.F.T. was supported by FAPESP (#2016/25358-3) and CNPq (#300896/2016-6). L.Be., L.F.S., and R.J.W. were supported by the Australian Research Council (FT100100375, DP120100811). A.A.C. was supported by a Royal Society Wolfson Research Merit award. J.H., A.La., and S.M. were funded by the Swedish Research Council Formas (grant no. 2013- 1389-26445-20). C.W. was funded by the National Research Foundation, South Africa. T.Y.J. and T.S.J. acknowledge NSF grant DEB-1601259. W.E.H. was funded by the NSERC Strategic and Discovery grant programs

Peste des Petits Ruminants (PPR) is a highly contagious viral disease that mainly affects goats and sheep in Asia, Africa and the Middle East. The PPR virus (PPRV) can be classified into four genetically distinct lineages (I, II, III, and IV). All have been historically present in Africa, except the Asian lineage IV that has been spreading across the globe and Africa for the last decades. Previous studies have identified the presence of the lineage IV in Nigeria since 2010. In this study, samples were taken from 268 small ruminants with PPR symptoms across Nigeria in 2017-2018 to provide an update on the distribution and genetic diversity of the lineage IV in the country. Sequence from a portion of the PPRV nucleoprotein (N) gene could be obtained from 72 samples, and all but one belonged to lineage IV. Phylogenetic analysis identified at least 5 lineage IV sub-clusters in Nigeria, re-grouping samples across multiple regions. Our results suggest an extensive endemic circulation of PPRV lineage IV across Nigeria and across borders with neighbouring countries, highlighting the difficulty of the control of this disease in the region.

In the recent past, peste des petits ruminants (PPR) emerged in East Africa causing outbreaks in small livestock across different countries, with evidences of spillover to wildlife. In order to understand better PPR at the wildlife&ndash, livestock interface, we investigated patterns of peste des petits ruminants virus (PPRV) exposure, disease outbreaks, and viral sequences in the northern Albertine Rift. PPRV antibodies indicated a widespread exposure in apparently healthy wildlife from South Sudan (2013) and Uganda (2015, 2017). African buffaloes and Uganda kobs &lt, 1-year-old from Queen Elizabeth National Park (2015) had antibodies against PPRV N-antigen and local serosurvey captured a subsequent spread of PPRV in livestock. Outbreaks with PPR-like syndrome in sheep and goats were recorded around the Greater Virunga Landscape in Kasese (2016), Kisoro and Kabale (2017) from western Uganda, and in North Kivu (2017) from eastern Democratic Republic of the Congo (DRC). This landscape would not be considered typical for PPR persistence as it is a mixed forest&ndash, savannah ecosystem with mostly sedentary livestock. PPRV sequences from DRC (2017) were identical to strains from Burundi (2018) and confirmed a transboundary spread of PPRV. Our results indicate an epidemiological linkage between epizootic cycles in livestock and exposure in wildlife, denoting the importance of PPR surveillance on wild artiodactyls for both conservation and eradication programs.

International audience; Peste des Petits Ruminants (PPR) is a serious transboundary infectious disease of small ruminants. The causal agent, PPR virus (PPRV), can be separated into four genetically distinct lineages using phylogenetic analysis. In recent decades, lineage IV of PPRV has dramatically extended its geographic distribution from Asia to the Middle East and to Africa, where it has progressively replaced other PPRV lineages. Lineages I and II are historically distributed in West Africa. Currently, lineage II appears to dominate the region, whereas the last recorded occurrence of lineage I dates back to 1994. Recent studies reported the presence of lineage IV in Nigeria, suggesting that this lineage is expanding in West Africa. In Niger, a close neighbour of Nigeria, PPRV has never been genetically characterized, despite reports of PPR incidence. In this study, pathological samples collected from sick goats were collected in 2013 during a suspected PPR outbreak in southern Niger close to the Nigerian border were compared to samples collected in a previous investigation in October 2001 in south-western Niger. These strains were characterized by sequencing and phylogenetic analysis to identify their genetic lineage. Our results show that in 2001, lineages I and II were cocirculating in south-western Niger, whereas the strain that caused the outbreak in 2013 belonged to lineage IV and is closely related to strains identified in Nigeria. These results confirm the progression of lineage IV in West Africa. The process of PPRV lineage replacement and its implications for the epidemiology and the control of the disease in this region are unclear and should be the subject of further studies in the field.

International audience; Peste des Petits Ruminants (PPR) is a severe disease of small ruminants and has high economic impacts in developing countries. Endemic in Africa, the Middle East and Asia, the disease is currently progressing with occurrences reported in North Africa, Turkey and in Georgia, and now threatens Europe. Much remains unknown about the infection dynamics, the virulence of the different strains and species/breed susceptibility. Robust experimental challenge models are needed to explore these fields and to confirm the efficacy of currently sold vaccines. We first assessed virulence of two PPR virus strains (CI89 and MA08) in Saanen goats. Whereas the MA08 strain led to classical severe clinical signs of PPR, the CI89 strain appeared to cause a mild disease in Saanen goats, highlighting the difference in virulence between strains in this animal model. We further demonstrated the importance of the inoculation route in the appearance of clinical signs and that ocular excretion is a better choice than blood for viral detection. After developing a robust challenge model, we assessed the efficacy of a vaccine (PPR-VAC , BVI Botswana) against the MA08 strain and demonstrated that this vaccine blocked viral excretion and significantly reduced clinical signs. These results reinforce the paradigm that a strain from one lineage could protect against strains from other lineages.

Heartwater (or cowdriosis) is a tick-borne disease caused by Ehrlichia ruminantium, an obligatory intracellular bacterium of the order Rickettsiales, transmitted by several ticks of the genus Amblyomma. The organism is genetically highly variable which prevented until now the development of efficient vaccines. The disease is enzootic in sub-Sahelian Africa and in some Caribbean islands. It affects domestic and wild ruminants, the susceptibility to cowdriosis varying greatly between breeds and species: African wildlife shows mainly asymptomatic infections; local cattle breeds are generally protected due to enzootic stability; and introduced cattle breeds and small ruminants, even in enzootic regions, are usually susceptible to heartwater and can suffer high mortality rates. Cowdriosis is characterized by a sudden and acute fever followed by nervous, respiratory, and gastrointestinal symptoms and by hydrothorax and hydropericardium during postmortem examination. In West Africa, the only vector is Amblyomma variegatum, present in areas where pluviometry is higher than 500 mm. Therefore, animals of a high proportion of the Sahelian region are usually not infested by the tick and not infected by the bacterium. They are thus susceptible when introduced in southern parts of the Sahel or in the subhumid neighboring areas of the West African countries, for example during transhumance. Tetracyclines are effective drugs to treat heartwater when administered before occurrence of the nervous symptoms. Various vaccines have been tested, and are still developed, but, up to now, none of them showed enough effectiveness against all the field strains of E. ruminantium to allow its marketing. Prevention is therefore mainly achieved by drastic vector control or, on the contrary, acquisition of enzootic stability following tick infestation combined with tetracycline treatment as soon as hyperthermia occurs.

Parasitic chytrid fungi have emerged as a significant threat to amphibian species worldwide, necessitating the development of techniques to isolate these pathogens into sterile culture for research purposes. However, early methods of isolating chytrids from their hosts relied on killing amphibians. We modified a pre-existing protocol for isolating chytrids from infected animals to use toe clips and biopsies from toe webbing rather than euthanizing hosts, and distributed the protocol to interested researchers worldwide as part of the BiodivERsA project RACE – here called the RML protocol. In tandem, we developed a lethal procedure for isolating chytrids from tadpole mouthparts. Reviewing a database of use a decade after their inception, we find that these methods have been widely applied across at least 5 continents, 23 countries and in 62 amphibian species, and have been successfully used to isolate chytrids in remote field locations. Isolation of chytrids by the non-lethal RML protocol occured in 18% of attempts with 207 fungal isolates and three species of chytrid being recovered. Isolation of chytrids from tadpoles occured in 43% of attempts with 334 fungal isolates of one species (Batrachochytrium dendrobatidis) being recovered. Together, these methods have resulted in a significant reduction and refinement of our use of threatened amphibian species and have improved our ability to work with this important group of emerging fungal pathogens.

In order to protect the unique fauna of the Galapagos Islands, it is critical that we understand how and when parasites spread throughout the archipelago. There are several key components of host-parasite dynamics that influence parasite spread, including the basic reproductive rate of the parasite, host density, transmission mode, and host movement, among other factors. Host movement could be especially important in determining parasite spread in island systems like the Galapagos, because parasites are not always able to move larger distances unaided. Because some hosts (and vectors) show population genetic structure within the Galapagos Islands, we can make inferences about potential parasite spread from knowledge of host population connectivity. In this chapter, we review patterns of population connectivity in Galapagos vertebrates (hosts), arthropod vectors, and parasites, focusing on population genetic studies. Hosts with little to no population genetic structure and high rates of inferred movement (e.g., Galapagos fur seal, Galapagos penguin, great frigatebird, Galapagos dove, small ground finch, small tree finch, large tree finch) are the most likely to spread parasites. More research is needed on parasite spread, particularly studies that simultaneously estimate population connectivity of both host (or multiple hosts, including vectors) and parasite.

International audience; In organisms with complex life histories, dramatic changes in microbial community structure may occur with host development and immune system maturation. Amphibian host susceptibility to diseases such as chytridiomycosis may be affected by the reorganization of skin microbial community structure that occurs during metamorphosis. We tracked changes in the bacterial communities inhabiting skin of Korean fire-bellied toads (Bombina orientalis) that we infected as tadpoles with different strains of Batrachochytrium dendrobatidis (Bd), the pathogenic fungus that causes chytridiomycosis. We found that B. orientalis undergoes a major change in skin bacterial community composition between 5 and 15 days following metamorphosis. Richness indices and phylogenetic diversity measures began to diverge earlier, between aquatic and terrestrial stages. Our results further reveal differences in skin bacterial community composition among infection groups, suggesting that the effect of Bd infection on skin microbiome composition may differ by Bd strain. Additional studies are needed to further investigate the structural and temporal dynamics of microbiome shifts during metamorphosis in wild and captive amphibian populations. Analyses of the ontogeny of microbiome shifts may contribute to an understanding of why amphibians vary in their susceptibility to chytridiomycosis.

Colonization comprises the physical arrival of a species in a new area, but also its successful establishment within the local community. Oceanic islands, like the Hawaiian and the Galapagos archipelagos, represent excellent systems to study the mechanisms of colonization because of their historical isolation. In this chapter, we first review some of the major mechanisms by which parasites and vectors could arrive to an oceanic island, both naturally or due to human activities, and the factors that may influence their successful establishment in the insular host community. We then explore examples of natural and anthropogenic colonization of the Galapagos Islands by parasites and vectors, focusing on one or more case studies that best represent the diversity of colonization mechanisms that has shaped parasite distribution in the archipelago. Finally, we discuss future directions for research on parasite and vector colonization in Galapagos Islands.

Population declines and extinctions of amphibians have been attributed to the chytrid fungus Batrachochytrium dendrobatidis (Bd), especially one globally emerging recombinant lineage ('Bd-GPL'). We used PCR assays that target the ribosomal internal transcribed spacer region (ITS) of Bd to determine the prevalence and genetic diversity of Bd in South Korea, where Bd is widely distributed but is not known to cause morbidity or mortality in wild populations. We isolated Korean Bd strains from native amphibians with low infection loads and compared them to known worldwide Bd strains using 19 polymorphic SNP and microsatellite loci. Bd prevalence ranged between 12.5 and 48.0%, in 11 of 17 native Korean species, and 24.7% in the introduced bullfrog Lithobates catesbeianus. Based on ITS sequence variation, 47 of the 50 identified Korean haplotypes formed a group closely associated with a native Brazilian Bd lineage, separated from the Bd-GPL lineage. However, multilocus genotyping of three Korean Bd isolates revealed strong divergence from both Bd-GPL and the native Brazilian Bd lineages. Thus, the ITS region resolves genotypes that diverge from Bd-GPL but otherwise generates ambiguous phylogenies. Our results point to the presence of highly diversified endemic strains of Bd across Asian amphibian species. The rarity of Bd-GPL-associated haplotypes suggests that either this lineage was introduced into Korea only recently or Bd-GPL has been outcompeted by native Bd strains. Our results highlight the need to consider possible complex interactions among native Bd lineages, Bd-GPL and their associated amphibian hosts when assessing the spread and impact of Bd-GPL on worldwide amphibian populations.

Pathogen-driven selection can favour major histocompatibility complex (MHC) alleles that confer immunological resistance to specific diseases. However, strong directional selection should deplete genetic variation necessary for robust immune function in the absence of balancing selection or challenges presented by other pathogens. We examined selection dynamics at one MHC class II (MHC-II) locus across Panamanian populations of the túngara frog, Physalaemus pustulosus , infected by the amphibian chytrid fungus Batrachochytrium dendrobatidis (Bd). We compared MHC-II diversity in highland túngara frog populations, where amphibian communities have experienced declines owing to Bd, with those in the lowland region that have shown no evidence of decline. Highland region frogs had MHC variants that confer resistance to Bd. Variant fixation appeared to occur by directional selection rather than inbreeding, as overall genetic variation persisted in populations. In Bd-infected lowland sites, however, selective advantage may accrue to individuals with only one Bd-resistance allele, which were more frequent. Environmental conditions in lowlands should be less favourable for Bd infection, which may reduce selection for specific Bd resistance in hosts. Our results suggest that MHC selection dynamics fluctuate in túngara frog populations as a function of the favourability of habitat to pathogen spread and the vulnerability of hosts to infection.

The black salt-marsh mosquito (Aedes taeniorhynchus) is the only native mosquito in the Galapagos Islands and potentially a major disease vector for Galapagos wildlife. Little is known about its population structure, or how its dynamics may be influenced by human presence in the archipelago. We used microsatellite data to assess the structure and patterns of A. taeniorhynchus gene flow among and within islands, to identify potential barriers to mosquito dispersal, and to investigate human-aided transport of mosquitoes across the archipelago. Our results show that inter-island migration of A. taeniorhynchus occurs frequently on an isolation by distance basis. High levels of inter-island migration were detected amongst the major ports of the archipelago, strongly suggesting the occurrence of human-aided transport of mosquitoes among islands, underlining the need for strict control measures to avoid the transport of disease vectors between islands. The prevalence of filarial nematode infection in Galapagos flightless cormorants is correlated with the population structure and migration patterns of A. taeniorhynchus, suggesting that A. taeniorhynchus is an important vector of this arthropod-borne parasite in the Galapagos Islands. Therefore mosquito population structure in Galapagos may have the potential to influence mosquito-borne parasite population dynamics, and the subsequent impacts of such pathogens on their host species in the islands.

Characterization of the fine-scale population dynamics of the mosquito Aedes taeniorhynchus is needed to improve our understanding of its role as a disease vector in the Galapagos Islands. We used microsatellite data to assess the genetic structure of coastal and highland mosquito populations and patterns of gene flow between the two habitats through time on Santa Cruz Island. In addition, we assessed possible associations of mosquito abundance and genetic diversity with environmental variables. The coastal and highland mosquito populations were highly differentiated from each other all year round, with some gene flow detected only during periods of increased precipitation. The results support the hypothesis that selection arising from ecological differences between habitats is driving adaptation and divergence in A. eniorhynchus, and maintaining long-term genetic differentiation of the populations against gene flow. The highland and lowland populations may constitute an example of incipient speciation in progress. Highland populations were characterized by lower observed heterozygosity and allelic richness, suggesting a founder effect and/or lower breeding site availability in the highlands. A lack of reduction in genetic diversity over time in highland populations suggests that they survive dry periods as dormant eggs. Association between mosquito abundance and precipitation was strong in the highlands, whereas tide height was the main factor affecting mosquito abundance on the coast. Our findings suggests differences in the infection dynamics of mosquito-borne parasites in the highlands compared to the coast, and a higher risk of mosquito-driven disease spread across these habitats during periods of increased precipitation. (Resume d'auteur)

This study assesses the ecological factors associated with microfilariae prevalence in wild populations of endangered flightless cormorants (Phalacrocorax harrisi) and Galápagos penguins (Spheniscus mendiculus). Prevalence values were tested for correlation with a large number of environmental variables, as modeled from weather station data and measured by satellite-borne sensors. Predictions were made based on the expected effects of climatic and landscape variables on sustained populations of arthropod vectors required for transmission of microfilariae. In general, findings were consistent with predictions in both cormorants and penguins; prevalence correlated positively with temperature, precipitation, and vegetation density, and negatively with measures of environmental variability. Resulting correlates were used to derive predictive distributions of prevalence values in cormorants throughout the archipelago. Evidence is presented implicating the mosquito Aedes taeniorhynchus as a likely vector. Knowledge of environmental variables that predict risk of disease transmission by arthropod vectors may be useful in control measures should novel pathogens be introduced to the ecosystem.

Cutaneous bacteria may play an important role in the resistance of amphibians to the pathogenic fungus Batrachochytrium dendrobatidis (Bd). Microbial communities resident on hosts' skin show topographical diversity mapping to skin features, as demonstrated by studies of the human microbiome. We examined skin microbiomes of wild and captive fire-bellied toads (Bombina orientalis) for differences across their body surface. We found that bacterial communities differed between ventral and dorsal skin. Wild toads showed slightly higher bacterial richness and diversity in the dorsal compared to the ventral region. On the other hand, captive toads hosted a higher richness and diversity of bacteria on their ventral than their dorsal skin. Microbial community composition and relative abundance of major bacterial taxonomic groups also differed between ventral and dorsal skin in all populations. Furthermore, microbiome diversity patterns varied as a function of their Bd infection status in wild toads. Bacterial richness and diversity was greater, and microbial community structure more complex, in wild than captive toads. The results suggest that bacterial community structure is influenced by microhabitats associated with skin regions. These local communities may be differentially modified when interacting with environmental bacteria and Bd. A better understanding of microbiome variation across skin regions will be needed to assess how the skin microbiota affects the abilities of amphibian hosts to resist Bd infection, especially in captive breeding programs.

The pathogenic fungus Batrachochytrium dendrobatidis (Bd) is a major conservation concern because of its role in decimating amphibian populations worldwide. We used quantitative PCR to screen 244 museum specimens from the Korean Peninsula, collected between 1911 and 2004, for the presence of Bd to gain insight into its history in Asia. Three specimens of Rugosa emeljanovi (previously Rana or Glandirana rugosa), collected in 1911 from Wonsan, North Korea, tested positive for Bd. Histology of these positive specimens revealed mild hyperkeratosis – a non-specific host response commonly found in Bd-infected frogs – but no Bd zoospores or zoosporangia. Our results indicate that Bd was present in Korea more than 100 years ago, consistent with hypotheses suggesting that Korean amphibians may be infected by endemic Asian Bd strains.

The pathogenic chytrid fungusBatrachochytrium dendrobatidis(Bd) can cause precipitous population declines in its amphibian hosts. Responses of individuals to infection vary greatly with the capacity of their immune system to respond to the pathogen. We used a combination of comparative and experimental approaches to identify major histocompatibility complex class II (MHC-II) alleles encoding molecules that foster the survival of Bd-infected amphibians. We found that Bd-resistant amphibians across four continents share common amino acids in three binding pockets of the MHC-II antigen-binding groove. Moreover, strong signals of selection acting on these specific sites were evident among all species co-existing with the pathogen. In the laboratory, we experimentally inoculated Australian tree frogs with Bd to test how each binding pocket conformation influences disease resistance. Only the conformation of MHC-II pocket 9 of surviving subjects matched those of Bd-resistant species. This MHC-II conformation thus may determine amphibian resistance to Bd, although other MHC-II binding pockets also may contribute to resistance. Rescuing amphibian biodiversity will depend on our understanding of amphibian immune defence mechanisms against Bd. The identification of adaptive genetic markers for Bd resistance represents an important step forward towards that goal.

We present temperature measurements recorded at a 15 min time step between October 2002 and April 2003 inside the La Peyrere cave (Baget karstic system, Central Pyrenees) on six 0.01 °C sensitivity sensors distributed on the whole recognized cave, down to a depth of 57 m. Rainy events are associated to a steep signal of a few hundredth to a few tenth of degrees in amplitude starting 1 h to one day after rain, and with a sign depending mostly of the outside temperature. The temperature signal does not correlate well with the rainfall, while the water level does with a maximum water rising rate 2.5 h after rain. The spectral density of the temperature signal displays one peak at a period of 3 days, which is also present in the rain signal and which is likely to be related to climatic parameters, and a broad smooth zone of spectral energy close to a 1 day period. By comparing a ‘superficial’ and a ‘deepest’ group of sensors, we note that the flood signal is more pronounced on the deepest sensor group whatever the outside temperature, and that the annual temperature variation is also larger on this sensor group. This could indicate that rain water flows in the cave through its base and explain the development in depth of the cave. Due to the arrival of water with different temperatures and mineralizations at different levels, double diffusive convection is likely to be triggered in the cave. We suggest that the onset of convection could be associated to the oscillations observed on the temperature signal at the onset of the flood.

The pathogenic chytrid fungus, Batrachochytrium dendrobatidis (denoted Bd), causes large-scale epizootics in naive amphibian populations. Intervention strategies to rapidly respond to Bd incursions require sensitive and accurate diagnostic methods. Chytridiomycosis usually is assessed by quantitative polymerase chain reaction (qPCR) amplification of amphibian skin swabs. Results based on this method, however, sometimes yield inconsistent results on infection status and inaccurate scores of infection intensity. In Asia and other regions where amphibians typically bear low Bd loads, swab results are least reliable. We developed a Bd-sampling method that collects zoospores released by infected subjects into an aquatic medium. Bd DNA is extracted by filters and amplified by nested PCR. Using laboratory colonies and field populations of Bombina orientalis, we compare results with those obtained on the same subjects by qPCR of DNA extracted from swabs. Many subjects, despite being diagnosed as Bd-negative by conventional methods, released Bd zoospores into collection containers and thus must be considered infected. Infection loads determined from filtered water were at least 1000 times higher than those estimated from swabs. Subjects significantly varied in infection load, as they intermittently released zoospores, over a 5-day period. Thus, the method might be used to compare the infectivity of individuals and study the periodicity of zoospore release. Sampling methods based on water filtration can dramatically increase the capacity to accurately diagnose chytridiomycosis and contribute to a better understanding of the interactions between Bd and its hosts.

Host selection in blood-sucking arthropods has important evolutionary and ecological implications for the transmission dynamics, distribution and host-specificity of the parasites they transmit. The black salt-marsh mosquito (Aedes taeniorhynchus Wiedemann) is distributed throughout tropical to temperate coastal zones in the Americas, and continental populations are primarily mammalphilic. It is the only indigenous mosquito in the Galapagos Islands, having colonised the archipelago around 200,000 years ago, potentially adapting its host selection, and in the process, altering the dynamics of vector mediated pathogen interactions in the archipelago. Here, we use blood–meal analysis and PCR-based parasite screening approach to determine the blood-feeding patterns of A. taeniorhynchus in the Galapagos Islands and identify potential parasite transmission with which this mosquito could be involved. Our results show that A. taeniorhynchus feeds equally on mammals and reptiles, and only one avian sample was observed in 190 successful PCR amplifications from blood meals. However, we detected endemic filarial worms and Haemoproteus parasites known to infect various Galapagos bird species in mosquito thoraces, suggesting that feeding on birds must occur at low frequency, and that A. taeniorhynchus may play a role in maintaining some avian vector-borne pathogens, although more work is needed to explore this possibility. We also isolated three different DNA sequences corresponding to hemogregarine parasites of the genus Hepatozoon from mosquito and iguana blood samples, suggesting that more than one species of Hepatozoon parasites are present in Galapagos. Phylogenetic analysis of Hepatozoon 18sRNA sequences indicates that A. taeniorhynchus may have facilitated a recent breakdown in host–species association of formerly isolated Hepatozoon spp. infecting the reptile populations in the Galapagos Islands.

Characterization of the fine-scale population dynamics of the mosquito Aedes taeniorhynchus is needed to improve our understanding of its role as a disease vector in the Galapagos Islands. We used microsatellite data to assess the genetic structure of coastal and highland mosquito populations and patterns of gene flow between the two habitats through time on Santa Cruz Island. In addition, we assessed possible associations of mosquito abundance and genetic diversity with environmental variables. The coastal and highland mosquito populations were highly differentiated from each other all year round, with some gene flow detected only during periods of increased precipitation. The results support the hypothesis that selection arising from ecological differences between habitats is driving adaptation and divergence in A. taeniorhynchus, and maintaining long-term genetic differentiation of the populations against gene flow. The highland and lowland populations may constitute an example of incipient speciation in progress. Highland populations were characterized by lower observed heterozygosity and allelic richness, suggesting a founder effect and/or lower breeding site availability in the highlands. A lack of reduction in genetic diversity over time in highland populations suggests that they survive dry periods as dormant eggs. Association between mosquito abundance and precipitation was strong in the highlands, whereas tide height was the main factor affecting mosquito abundance on the coast. Our findings suggests differences in the infection dynamics of mosquito-borne parasites in the highlands compared to the coast, and a higher risk of mosquito-driven disease spread across these habitats during periods of increased precipitation.

Emerging infectious diseases of wildlife have been recognized as a major threat to global biodiversity. Endemic species on isolated oceanic islands, such as the Galápagos, are particularly at risk in the face of introduced pathogens and disease vectors. The black salt-marsh mosquito ( Aedes taeniorhynchus ) is the only mosquito widely distributed across the Galápagos Archipelago. Here we show that this mosquito naturally colonized the Galápagos before the arrival of man, and since then it has evolved to represent a distinct evolutionary unit and has adapted to habitats unusual for its coastal progenitor. We also present evidence that A. taeniorhynchus feeds on reptiles in Galápagos in addition to previously reported mammal and bird hosts, highlighting the important role this mosquito might play as a bridge-vector in the transmission and spread of extant and newly introduced diseases in the Galápagos Islands. These findings are particularly pertinent for West Nile virus, which can cause significant morbidity and mortality in mammals (including humans), birds, and reptiles, and which recently has spread from an introductory focus in New York to much of the North and South American mainland and could soon reach the Galápagos Islands. Unlike Hawaii, there are likely to be no highland refugia free from invading mosquito-borne diseases in Galápagos, suggesting bleak outcomes to possible future pathogen introduction events.

Wildlife on isolated oceanic islands is highly susceptible to the introduction of pathogens. The recent establishment in the Galápagos Islands of the mosquitoCulex quinquefasciatus, a vector for diseases such as avian malaria and West Nile fever, is considered a serious risk factor for the archipelago's endemic fauna. Here we present evidence from the monitoring of aeroplanes and genetic analysis thatC. quinquefasciatusis regularly introduced via aircraft into the Galápagos Archipelago. Genetic population structure and admixture analysis demonstrates that these mosquitoes breed with, and integrate successfully into, already-established populations ofC. quinquefasciatusin the Galápagos, and that there is ongoing movement of mosquitoes between islands. Tourist cruise boats and inter-island boat services are the most likely mechanism for transportingCulexmosquitoes between islands. Such anthropogenic mosquito movements increase the risk of the introduction of mosquito-borne diseases novel to Galápagos and their subsequent widespread dissemination across the archipelago. Failure to implement and maintain measures to prevent the human-assisted transport of mosquitoes to and among the islands could have catastrophic consequences for the endemic wildlife of Galápagos.

International audience; The aim of the hydraulic stimulations in the Soultz-sous-Forêts, France, Enhanced Geothermal System (EGS) project was to create, in crystalline rocks, a fractured reservoir 750 m high, 750 m long and 35 m thick interconnecting the injection and production wells. Increasing the permeability in a zone with a high geothermal gradient will trigger free convection, which will interact with the forced flow driven by pumping. A systematic numerical study of the coupling between forced and free convective flows has been performed by considering a large range of injection rates and Rayleigh numbers. The simulations showed that if there is weak or no free convection in an EGS reservoir, economic exploitation of the system will rapidly end because of a decrease in produced fluid temperature. The maximum injection rate preventing such a temperature drop increases with the Rayleigh number and the height of the stimulated domain. The model establishes constraints on the conditions for achieving optimal heat extraction at the Soultz-sous-Forêts EGS site. It was also shown that, although mineral precipitation may partially close or heal some open fissures, it does not lead to a major decrease of the hydraulic conductivity in the stimulated reservoir.