Search

Host-associated microbial communities can influence physiological processes of macroorganisms, including contributing to infectious disease resistance. For instance, some bacteria that live on amphibian skin produce antifungal compounds that inhibit two lethal fungal pathogens, Batrachochytrium dendrobatidis (Bd) and Batrachochytrium salamandrivorans (Bsal). Therefore, differences in microbiome composition among host species or populations within a species can contribute to variation in susceptibility to Bd/Bsal. This study applies 16S rRNA sequencing to characterize the skin bacterial microbiomes of three widespread terrestrial salamander genera native to the western United States. Using a metacommunity structure analysis, we identified dispersal barriers for these influential bacteria between salamander families and localities. We also analysed the effects of habitat characteristics such as percent natural cover and temperature seasonality on the microbiome. We found that certain environmental variables may influence the skin microbial communities of some salamander genera more strongly than others. Each salamander family had a somewhat distinct community of putative anti-Bd skin bacteria, suggesting that salamanders may select for a functional assembly of cutaneous symbionts that could differ in its ability to protect these amphibians from disease. Our observations raise the need to consider host identity and environmental heterogeneity during the selection of probiotics to treat wildlife diseases. [ABSTRACT FROM AUTHOR]

Understanding the responses of naïve communities to the invasion of multihost pathogens requires accurate estimates of susceptibility across taxa. In the Americas, the likely emergence of a second amphibian pathogenic fungus (Batrachochytrium salamandrivorans, Bsal) calls for new ways of prioritizing disease mitigation among species due to the high diversity of naïve hosts with prior B. dendrobatidis (Bd) infections. Here, we applied the concept of pathogenic potential to quantify the virulence of chytrid fungi on naïve amphibians and evaluate species for conservation efforts in the event of an outbreak. The benefit of this measure is that it combines and summarizes the variation in disease effects into a single numerical index, allowing for comparisons across species, populations or groups of individuals that may inherently exhibit differences in susceptibility. As a proof of concept, we obtained standardized responses of disease severity by performing experimental infections with Bsal on five plethodontid salamanders from southeastern United States. Four out of five species carried natural infections of Bd at the start of the experiments. We showed that Bsal exhibited its highest value of pathogenic potential in a species that is already declining (Desmognathus auriculatus). We find that this index provides additional information beyond the standard measures of disease prevalence, intensity, and mortality, because it leveraged these disease parameters within each categorical group. Scientists and practitioners could use this measure to justify research, funding, trade, or conservation measures. HIGHLIGHTS: We evaluate the risk of infection by chytrid fungi in five species of salamanders.We apply the concept of pathogenic potential to compare across species.Chytrid invasion could threaten Desmognathus auriculatus and Eurycea cirrigera populations. [ABSTRACT FROM AUTHOR]

Captive breeding programs are a valuable conservation strategy, particularly when integrated with research goals. Panamanian Harlequin frogs (genus Atelopus) serve as a case study for integrating captive breeding and research goals because they have experienced drastic chytridiomycosis-related declines and have large captive populations. Captive breeding efforts in Panama and the United States established secure ex-situ populations of Atelopus certus , A. glyphus , A. limosus , A. varius , and A. zeteki. Atelopus chiriquiensis is presumed to be extinct with no captive populations. The status of one undescribed species, Atelopus aff. limosus , has not been evaluated and no secure captive population has yet been established. Captive breeding efforts that produce a surplus of Atelopus are an important resource for research into collections management, disease mitigation, and adaptive management approaches for Atelopus reintroduction efforts. We reevaluated all Panamanian Atelopus species through the IUCN Redlist and compiled occurrence records for Panamanian Atelopus species to create a historical distribution map. We model Atelopus habitat suitability using Maxent and found annual mean air temperature to be the best predictor of Atelopus occurrence. The model will improve our knowledge of their likely spatial distribution and guide future conservation and reintroduction efforts. The recent proliferation of molecular tools, climate models, bio-banking, and reproductive technologies position us to address multiple applied and basic evolutionary questions such as: What factors cause differential disease outcomes? Do persisting populations have heritable traits associated with improved survivorship? Are there climatic refugia from disease? Ultimately, the answers to these questions will help us develop applied solutions and facilitate the reestablishment of self-sustaining wild populations. • Atelopus species have experienced severe chytridiomycosis-related declines. • This case study documents the wild and captive status of Atelopus in Panama. • Habitat models improve historical distribution maps and guide future efforts. • Captive breeding efforts prevent extinctions and are a valuable research resource. [ABSTRACT FROM AUTHOR]

North American amphibians have recently been impacted by two major emerging pathogens, the fungus Batrachochytrium dendrobatidis (Bd) and iridoviruses in the genus Ranavirus (Rv). Environmental factors and host genetics may play important roles in disease dynamics, but few studies incorporate both of these components into their analyses. Here, we investigated the role of environmental and genetic factors in driving Bd and Rv infection prevalence and severity in a biodiversity hot spot, the southeastern United States. We used quantitative PCR to characterize Bd and Rv dynamics in natural populations of three amphibian species: Notophthalmus perstriatus, Hyla squirella and Pseudacris ornata. We combined pathogen data, genetic diversity metrics generated from neutral markers, and environmental variables into general linear models to evaluate how these factors impact infectious disease dynamics. Occurrence, prevalence and intensity of Bd and Rv varied across species and populations, but only one species, Pseudacris ornata, harbored high Bd intensities in the majority of sampled populations. Genetic diversity and climate variables both predicted Bd prevalence, whereas climatic variables alone predicted infection intensity. We conclude that Bd is more abundant in the southeastern United States than previously thought and that genetic and environmental factors are both important for predicting amphibian pathogen dynamics. Incorporating both genetic and environmental information into conservation plans for amphibians is necessary for the development of more effective management strategies to mitigate the impact of emerging infectious diseases. [ABSTRACT FROM AUTHOR]

Global amphibian declines are linked with the presence of specific, highly virulent genotypes of the emerging fungal disease chytridiomycosis caused by Batrachochytrium dendrobatidis ( Bd) known as the global panzootic lineage ( Bd-GPL). The global trade in amphibians for human consumption is suspected to have facilitated emergence of the disease, but evidence to support this is largely lacking. Here, we investigated the role the Lithobates catesbeianus (North American bullfrog) trade in spreading Bd genotypes by comparing strains associated with L. catesbeianus to a global panel using 36 sequenced loci from multiple chromosomal regions. Most bullfrogs were infected with Bd-GPL genotypes, but we also detected novel, highly divergent Bd genotypes ( Bd-Brazil) from a live bullfrog in a US market and from native Brazilian anurans in the Atlantic Forest where bullfrogs are widely farmed. Sexual reproduction was also detected for the first time in Bd in the form of a hybrid genotype between the Bd-GPL and Bd-Brazil lineages in the Atlantic Forest. Despite the demonstration that ribosomal RNA types in Bd fail to undergo concerted evolution (over 20 sequence types may be found in a single strain), the Bd-GPL and Bd-Brazil lineages form largely separate clusters of related internal transcribed spacer (ITS) RNA sequences. Using ITS sequences, we then demonstrate the presence of Bd-Brazil in Japan, primarily on invasive L. catesbeianus. The finding that Bd is capable of sexual reproduction between panzootic and endemic genotypes emphasizes the risk of international wildlife trade as a source of additional Bd epizootics owing to hybridization. [ABSTRACT FROM AUTHOR]

The amphibian fungal pathogen Batrachochytrium dendrobatidis (Bd) has received considerable attention due to its role in amphibian population declines worldwide. Although many amphibian species appear to be affected by Bd, there is little information on species-specific differences in susceptibility to this pathogen. We used a comparative experimental approach to examine Bd susceptibility in 6 amphibian species from the United States. We exposed postmetamorphic animals to Bd for 30 days and monitored mortality, feeding rates, and infection levels. In all species tested, Bd-exposed animals had higher rates of mortality than unexposed (control) animals. However, we found differences in mortality rates among species even though the amount of Bd detected on the different species' bodies did not differ. Of the species tested, southern toads (Anaxyrus terrestris ) and wood frogs (Lithobates sylvaticus ) had the highest rates of Bd-related mortality. Within species, we detected lower levels of Bd on individuals that survived longer and found that the relationship between body size and infection levels differed among species. Our results indicate that, even under identical conditions, amphibian species differ in susceptibility to Bd. This study represents a step toward identifying and understanding species variation in disease susceptibility, which can be used to optimize conservation strategies. [ABSTRACT FROM AUTHOR]

The boreal toad Bufo boreas boreas, once common in the western USA, is listed as an endangered species in Colorado and New Mexico, and protected in Wyoming. Populations have dramatically declined due to the presence of the fungal pathogen Batrachochytrium dendrobatidis (Bd). A gender- and stage-structured model for the boreal toad is formulated which depends on its life cycle and breeding strategies. In addition, an epizootic model for the spread of Bd is formulated. Analysis of these models provides two thresholds. The first threshold, the basic reproduction number for the population, R0, determines whether the population persists and the second threshold, the basic reproduction number for the fungal disease, RF, determines whether the disease persists. If R0>1 and RF<1, then the population becomes disease-free. However, if both thresholds are greater than one, the population levels are severely reduced by the fungal pathogen. [ABSTRACT FROM AUTHOR]

Many declines of amphibian populations have been associated with chytridiomycosis, a disease caused by the aquatic fungus Batracjiochytrium dendrobatidis (Bd). Despite the relatively high prevalence of chytridiomycosis in stream amphibians globally, most surveys in North America have focused primarily on wetland-associated species, which are frequently infected. To better understand the distribution and prevalence of Bd in headwater amphibian communities, we sampled 452 tailed frogs (Ascaphus truei and Aseaphus inontanus) and 304 stream salamanders (seven species in the Dicamptodonti- dae and Plethodontidae) for Bd in 38, first- to third-order streams in five montane areas across the United States. We tested for presence of Bd by using PCR on skin swabs from salamanders and metamorphosed tailed frogs or the oral disc of frog larvae. We detected Bd on only seven individuals (0.93%) in four streams. Based on our study and results from five other studies that have sampled headwater- or seep-associated amphibians in the United States, Bd has been detected on only 3% of 1,322 individuals from 21 species. These results differ strongly from surveys in Central America and Australia, where Bd is more prevalent on stream-breeding species, as well as results from wetland-associated anurans in the same regions of the United States that we sampled. Differences in the prevalence of Bd between stream- and wetland-associated amphibians in the United States may be related to species-specific variation in susceptibility to chytridiomycosis or habitat differences. [ABSTRACT FROM AUTHOR]

The article discusses the survey conducted between June-October of 2012-2013 regarding the amphibian fungal pathogen Batrachochytrium dendrobatidis in the North Cascades National Park Service Complex, Washington, which is part of the study focused on the Coastal Tailed Frog, Ascaphus truei. It mentions that the hand-capturing and dipnetting were used in collecting amphibians.

Amphibians are globally threatened by anthropogenic habitat loss, the wildlife trade and emerging diseases. Previous authors have hypothesized that the spread of the amphibian disease chytridiomycosis (Batrachochytrium dendrobatidis) and amphibian ranaviruses are associated with the international trade in live amphibians. The North American bullfrog (Rana catesbeiana) is thought to be a carrier of these pathogens, is globally traded as a live commodity, and is sold live in US markets. We obtained importation data for all live amphibians, and parts thereof, into three major US ports of entry (Los Angeles, San Francisco and New York) from 2000 to 2005. Importation of live amphibians into these ports totaled almost 28 million individuals over this 6-year period. We collected samples from freshly-imported market frogs and found infection with both pathogens in all three cities and all seasons, with an overall infection prevalence of 62% (306/493) and 8.5% (50/588) for B. dendrobatidis and ranaviruses, respectively, by PCR. This study definitively identifies these two important pathogens in recently imported live market frogs and suggests that the amphibian trade can contribute to introductions of these pathogens into new regions. It provides support for the recent listing of B. dendrobatidis and ranaviral diseases by the OIE, and provides evidence for measures to be taken to eradicate these pathogens from the trade. [Copyright &y& Elsevier]

Amphibian populations continue to be imperiled by the chytrid fungus (Batrachochytrium dendrobatidis). Understanding where B. dendrobatidis (Bd) occurs and how it may be limited by environmental factors is critical to our ability to effectively conserve the amphibians affected by Bd. We sampled 1247 amphibians (boreal toads and surrogates) at 261 boreal toad (Bufo boreas) breeding sites (97 clusters) along an 11° latitudinal gradient in the Rocky Mountains to determine the distribution of B. dendrobatidis and examine environmental factors, such as temperature and elevation, that might affect its distribution. The fungus was detected at 64% of all clusters and occurred across a range of elevations (1030–3550m) and latitudes (37.6–48.6°) but we detected it in only 42% of clusters in the south (site elevations higher), compared to 84% of clusters in the north (site elevations lower). Maximum ambient temperature (daily high) explained much of the variation in Bd occurrence in boreal toad populations and thus perhaps limits the occurrence of the pathogen in the Rocky Mountains to areas where climatic conditions facilitate optimal growth of the fungus. This information has implications in global climate change scenarios where warming temperatures may facilitate the spread of disease into previously un- or little-affected areas (i.e., higher elevations). This study provides the first regional-level, field-based effort to examine the relationship of environmental and geographic factors to the distribution of B. dendrobatidis in North America and will assist managers to focus on at-risk populations as determined by the local temperature regimes, latitude and elevation. [Copyright &y& Elsevier]

An emerging disease of amphibians caused by the chytrid fungus Batrachochytrium dendrobatidis has been associated with morbidity, mortality, and extinction of species. Typically, researchers have detected B. dendrobatidis only when examining amphibians for causes of mortalities; few data exist on infection rates where mortalities are lacking. During May-September 2000-2002 we obtained amphibian specimens killed by vehicles and others collected at remote off-road sites throughout Maine, USA, and from federal lands in 5 states in the Northeast. We detected infected specimens, mostly green frogs (Rana clamitans), at 5 of 7 national wildlife refuges, a federal waterfowl production area, and Acadia National Park. Seven of 9 species, including all Ranidae species, were infected throughout Maine; rates ranged from 14.6% in American toads (Bufo americanus) to 25.7% in northern leopard frogs (Rana pipiens). We did not detect any infections in 50 eastern gray tree frogs (Hyla versicolor) or 21 spring peepers (Pseudacris crucifer). Species that hibernate in terrestrial habitats seem to have lower rates of infection than species that hibernate in aquatic habitats. Infections peaked in spring and autumn and were associated with air temperatures optimal for B. dendrobatidis growth. The relatively high infection rates among species without documented die-offs suggest that either losses have occurred undetected, that the fungus is endemic and species have attained a level of resistance to infections becoming lethal, or that climatic conditions of the Northeast have a role in preventing infections from being lethal. Data on prevalence and distribution of this chytrid fungus in the Northeast may be useful in modeling its origins and predicting long-term ecosystem effects involving anurans. [ABSTRACT FROM AUTHOR]

Symbiotic bacteria can produce secondary metabolites and volatile compounds that contribute to amphibian skin defense. Some of these symbionts have been used as probiotics to treat or prevent the emerging disease chytridiomycosis. We examined 20 amphibian cutaneous bacteria for the production of prodigiosin or violacein, brightly colored defense compounds that pigment the bacteria and have characteristic spectroscopic properties making them readily detectable, and evaluated the antifungal activity of these compounds. We detected violacein from all six isolates of Janthinobacterium lividum on frogs from the USA, Switzerland, and on captive frogs originally from Panama. We detected prodigiosin from five isolates of Serratia plymuthica or S. marcescens, but not from four isolates of S. fonticola or S. liquefaciens. All J. lividum isolates produced violacein when visibly purple, while prodigiosin was only detected on visibly red Serratia isolates. When applied to cultures of chytrid fungi Batrachochytrium dendrobatidis (Bd) and B. salamandrivorans (Bsal), prodigiosin caused significant growth inhibition, with minimal inhibitory concentrations (MIC) of 10 and 50 μM, respectively. Violacein showed a MIC of 15 μM against both fungi and was slightly more active against Bsal than Bd at lower concentrations. Although neither violacein nor prodigiosin showed aerosol activity and is not considered a volatile organic compound (VOC), J. lividum and several Serratia isolates did produce antifungal VOCs. White Serratia isolates with undetectable prodigiosin levels could still inhibit Bd growth indicating additional antifungal compounds in their chemical arsenals. Similarly, J. lividum can produce antifungal compounds such as indole-3-carboxaldehyde in addition to violacein, and isolates are not always purple, or turn purple under certain growth conditions. When Serratia isolates were grown in the presence of cell-free supernatant (CFS) from the fungi, CFS from Bd inhibited growth of the prodigiosin-producing isolates, perhaps indicative of an evolutionary arms race; Bsal CFS did not inhibit bacterial growth. In contrast, growth of one J. lividum isolate was facilitated by CFS from both fungi. Isolates that grow and continue to produce antifungal compounds in the presence of pathogens may represent promising probiotics for amphibians infected or at risk of chytridiomycosis. In a global analysis, 89% of tested Serratia isolates and 82% of J. lividum isolates were capable of inhibiting Bd and these have been reported from anurans and caudates from five continents, indicating their widespread distribution and potential for host benefit.

The emerging amphibian disease chytridiomycosis varies in severity depending on host species. Within species, disease susceptibility can also be influenced by pathogen variation and environmental factors. Here, we report on experimental exposures of American bullfrogs (Lithobates catesbeianus) to three different isolates of Batrachochytrium dendrobatidis (Bd), including one implicated in causing mass mortality of wild American bullfrogs. Exposed frogs showed low infection prevalence, relatively low infection load, and lack of clinical disease. Our results suggest that environmental cofactors are likely important contributors to Bd-associated American bullfrog mortality and that this species both resists and tolerates Bd infection.