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Abstract The bacterial communities of the amphibian skin (i.e., the bacteriome) are critical to the host's innate immune system. However, it is unclear how different drivers can alter this function by modulating the bacteriome's structure. Our aim was to assess the extent to which different host attributes and extrinsic factors influence the structure of the bacterial communities of the skin. Skin bacterial diversity was examined in 148 individuals of the four‐eyed frog (Pleurodema thaul) from 16 localities spanning almost 1800 km in latitude. The richness and beta diversity of bacterial families and the richness and abundance of Bd‐inhibitory bacterial genera were used to describe their structure. Predictors associated with the host (developmental stage, genetic lineage, individual Batrachochytrium dendrobatidis [Bd] infection status) and the landscape (current climate, degree of anthropogenic disturbance) were used in the statistical modeling in an information theoretical approach. Bd infection and host developmental stage were the only predictors affecting bacteriome richness, with Bd+ individuals and postmetamorphic stages (adults and juveniles) having higher richness than Bd− ones and tadpoles. High diversity in Bd+ individuals is not driven by bacterial genera with known anti‐Bd properties. Beta diversity was not affected by Bd infection and was mainly a consequence of bacterial family turnover rather than nestedness. Finally, for those bacterial genera known to have inhibitory effects on chytrid, Bd+ individuals had a slightly higher diversity than Bd− ones. Our study confirms an association between Bd infection and the host developmental stage with the bacterial communities of the skin of P. thaul. Unexpectedly, macroclimate and human impact factors do not seem to play a role in shaping the amphibian skin microbiome. Our study exemplifies that focusing on a single host–parasite system over a large geographic scale can provide essential insights into the factors driving host–parasite–bacteriome interactions.

Abstract Amphibian chytridiomycosis, caused by the fungus Batrachochytrium dendrobatidis (Bd), has caused the greatest known loss of biodiversity due to an infectious disease. We used Bd infection data from quantitative real-time PCR (qPCR) assays of amphibian skin swabs collected across Chile during 2008–2018 to model Bd occurrence with the aim to determine bioclimatic and anthropogenic variables associated with Bd infection. Also, we used Bd presence/absence records to identify geographical Bd high-risk areas and compare Bd prevalence and infection loads between amphibian families, ecoregions, and host ecology. Data comprised 4155 Bd-specific qPCR assays from 162 locations across a latitudinal gradient of 3700 km (18º to 51ºS). Results showed a significant clustering of Bd associated with urban centres and anthropogenically highly disturbed ecosystems in central-south Chile. Both Bd prevalence and Bd infection loads were higher in aquatic than terrestrial amphibian species. Our model indicated positive associations of Bd prevalence with altitude, temperature, precipitation and human-modified landscapes. Also, we found that macroscale drivers, such as land use change and climate, shape the occurrence of Bd at the landscape level. Our study provides with new evidence that can improve the effectiveness of strategies to mitigate biodiversity loss due to amphibian chytridiomycosis.

Abstract Amphibian chytridiomycosis, due to infection with the fungus Batrachochytrium dendrobatidis (Bd), has been associated with the alarming decline and extinction crisis of amphibians worldwide. It is essential for conservation management to identify regions with high or low suitability for Bd. We use a species distribution model to estimate the environmental suitability of Bd in the Chilean Winter Rainfall–Valdivian Forest biodiversity hotspot. Fourteen environmental variables were used as predictors in the statistical modeling (Maxent, generalized linear models, random forest) which also included 56 independent Bd+ localities. High‐risk areas (i.e., suitability above a defined threshold) were validated through prospective field surveys conducted in 2017. As results from Maxent, which only uses presence data, were the only results retained, refugia (i.e., suitability below a defined threshold) were validated with the Bd absences (N = 12) used in the GLM and RF modeling. Our results showed that (1) suitability for Bd increased with human footprint and with shorter distances to urban centers and water bodies and decreased with elevation; (2) climate was not a major factor shaping the current distribution of Bd; and (3) the model predicted high‐risk and refugia areas fairly well. Surveys of 24 new localities in high‐risk areas confirmed that 23 were Bd+; hence, these areas warrant consideration for long‐term Bd surveillance, population monitoring, and disease mitigation. In addition, five localities with apparent Bd absence were found in the predicted high‐risk areas. Our models showed that refugia can exist near high‐risk areas and Bd+ sites. Four localities with apparent Bd absence were located within the refugia predicted by the model. Preventing Bd transmission to such refugia is of paramount importance for persistence of Bd‐susceptible amphibian populations. The identification and validation through prospective field surveys of high‐risk areas and refugia are imperative to develop strategies to prevent further arrival and establishment of Bd and also, by identifying amphibian species or populations of conservation concern in such areas, will help to guide specific actions to reduce the biodiversity loss caused by chytridiomycosis.

Darwin's frogs (Rhinoderma darwinii and R. rufum) are two species of mouth brooding frogs from Chile and Argentina that have experienced marked population declines. Rhinoderma rufum has not been found in the wild since 1980. We investigated historical and current evidence of Batrachochytrium dendrobatidis (Bd) infection in Rhinoderma spp. to determine whether chytridiomycosis is implicated in the population declines of these species. Archived and live specimens of Rhinoderma spp., sympatric amphibians and amphibians at sites where Rhinoderma sp. had recently gone extinct were examined for Bd infection using quantitative real-time PCR. Six (0.9%) of 662 archived anurans tested positive for Bd (4/289 R. darwinii; 1/266 R. rufum and 1/107 other anurans), all of which had been collected between 1970 and 1978. An overall Bd-infection prevalence of 12.5% was obtained from 797 swabs taken from 369 extant individuals of R. darwinii and 428 individuals representing 18 other species of anurans found at sites with current and recent presence of the two Rhinoderma species. In extant R. darwinii, Bd-infection prevalence (1.9%) was significantly lower than that found in other anurans (7.3%). The prevalence of infection (30%) in other amphibian species was significantly higher in sites where either Rhinoderma spp. had become extinct or was experiencing severe population declines than in sites where there had been no apparent decline (3.0%; x(2) = 106.407, P

Darwin's frogs (Rhinoderma darwinii and R. rufum) are two species of mouth-brooding frogs from Chile and Argentina. Here, we present evidence on the extent of declines, current distribution and conservation status of Rhinoderma spp.; including information on abundance, habitat and threats to extant Darwin's frog populations. All known archived Rhinoderma specimens were examined in museums in North America, Europe and South America. Extensive surveys were carried out throughout the historical ranges of R. rufum and R. darwinii from 2008 to 2012. Literature review and location data of 2,244 archived specimens were used to develop historical distribution maps for Rhinoderma spp. Based on records of sightings, optimal linear estimation was used to estimate whether R. rufum can be considered extinct. No extant R. rufum was found and our modelling inferred that this species became extinct in 1982 (95% CI, 1980-2000). Rhinoderma darwinii was found in 36 sites. All populations were within native forest and abundance was highest in Chiloé Island, when compared with Coast, Andes and South populations. Estimated population size and density (five populations) averaged 33.2 frogs/population (range, 10.2-56.3) and 14.9 frogs/100 m(2) (range, 5.3-74.1), respectively. Our results provide further evidence that R. rufum is extinct and indicate that R. darwinii has declined to a much greater degree than previously recognised. Although this species can still be found across a large part of its historical range, remaining populations are small and severely fragmented. Conservation efforts for R. darwinii should be stepped up and the species re-classified as Endangered.

Wildlife conservation on private land is an important approach that is increasingly utilized to protect biodiversity and can help contribute to the 30 by 30 target of the Global Biodiversity Framework. In 2018, a Chilean non-profit conservation organization launched a land conservation programme aiming to build long-term partnerships with private landowners to protect critical habitat for threatened amphibians in Chilean Patagonia. Here we describe a new locality record of the micro-endemic, Endangered Barrio's frog Insuetophrynus acarpicus found at a site that joined the programme in 2020. Barrio's frog is ranked 11th in the list of Evolutionarily Distinct and Globally Endangered amphibians. Our systematic literature search showed that most aspects of the natural history and ecology of this species are unknown, limiting our ability to provide actionable science to inform its conservation. The newly described Barrio's frog population is the eighth known locality of this species and one of only three occurring within a protected area. Habitat quality assessments indicated optimal conditions for most of the measured habitat parameters in the high-gradient stream where the species occurs. This case study illustrates that long-term partnerships between private landowners and conservationists can be used as an effective tool to protect the habitat of highly threatened amphibians.

Emerging infectious diseases in wildlife are increasingly associated with animal mortality and species declines, but their source and genetic characterization often remains elusive. Amphibian chytridiomycosis, caused by the fungus Batrachochytrium dendrobatidis (Bd), has been associated with catastrophic and well-documented amphibian population declines and extinctions at the global scale. We used histology and whole-genome sequencing to describe the lesions caused by, and the genetic variability of, two Bd isolates obtained from a mass mortality event in a captive population of the threatened Chilean giant frog (Calyptocephalella gayi). This was the first time an association between Bd and high mortality had been detected in this charismatic and declining frog species. Pathological examinations revealed that 30 dead metamorphosed frogs presented agnathia or brachygnathia, a condition that is reported for the first time in association with chytridiomycosis. Phylogenomic analyses revealed that Bd isolates (PA1 and PA2) from captive C. gayi group with other Bd isolates (AVS2, AVS4, and AVS7) forming a single highly supported Chilean Bd clade within the global panzootic lineage of Bd (BdGPL). These findings are important to inform the strengthening of biosecurity measures to prevent the impacts of chytridiomycosis in captive breeding programs elsewhere.