Genomic Correlates of Virulence Attenuation in the Deadly Amphibian Chytrid Fungus, Batrachochytrium Dendrobatidis

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Description

Emerging infectious diseasespose a significant threat to global health, but predicting disease outcomes for particular species can be complicated when pathogen virulence varies across space, time, or hosts. The pathogenic chytrid fungus Batrachochytrium dendrobatidis (Bd) has caused worldwide declines in

Emerging infectious diseasespose a significant threat to global health, but predicting disease outcomes for particular species can be complicated when pathogen virulence varies across space, time, or hosts. The pathogenic chytrid fungus Batrachochytrium dendrobatidis (Bd) has caused worldwide declines in frog populations. Not only do Bd isolates from wild populations vary in virulence, but virulence shifts can occur over short timescales when Bd is maintained in the laboratory. We leveraged changes in Bd virulence over multiple generations of passage to better understand mechanisms of pathogen virulence. We conducted whole-genome resequencing of two samples of the same Bd isolate, differing only in passage history, to identify genomic processes associated with virulence attenuation. The isolate with shorter passage history (and greater virulence) had greater chromosome copy numbers than the isolate maintained in culture for longer, suggesting that virulence attenuation may be associated with loss of chromosome copies. Our results suggest that genomic processes proposed as mechanisms for rapid evolution in Bd are correlated with virulence attenuation in laboratory culture within a single lineage of Bd. Moreover, these genomic processes can occur over extremely short timescales. On a practical level, our results underscore the importance of immediately cryo-archiving new Bd isolates and using fresh isolates, rather than samples cultured in the laboratory for long periods, for laboratory infection experiments. Finally, when attempting to predict disease outcomes for this ecologically important pathogen, it is critical to consider existing variation in virulence among isolates and the potential for shifts in virulence over short timescales.

Date Created
2015-09-01
Agent

A Fungal Pathogen of Amphibians, Batrachochytrium Dendrobatidis, Attenuates in Pathogenicity With In Vitro Passages

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Description

Laboratory investigations into the amphibian chytrid fungus, Batrachochytrium dendrobatidis (Bd), have accelerated recently, given the pathogen’s role in causing the global decline and extinction of amphibians. Studies in which host animals were exposed to Bd have largely assumed that lab-maintained

Laboratory investigations into the amphibian chytrid fungus, Batrachochytrium dendrobatidis (Bd), have accelerated recently, given the pathogen’s role in causing the global decline and extinction of amphibians. Studies in which host animals were exposed to Bd have largely assumed that lab-maintained pathogen cultures retained the infective and pathogenic properties of wild isolates. Attenuated pathogenicity is common in artificially maintained cultures of other pathogenic fungi, but to date, it is unknown whether, and to what degree, Bd might change in culture. We compared zoospore production over time in two samples of a single Bd isolate having different passage histories: one maintained in artificial media for more than six years (JEL427-P39), and one recently thawed from cryopreserved stock (JEL427-P9). In a common garden experiment, we then exposed two different amphibian species, Eleutherodactylus coqui and Atelopus zeteki, to both cultures to test whether Bd attenuates in pathogenicity with in vitro passages. The culture with the shorter passage history, JEL427-P9, had significantly greater zoospore densities over time compared to JEL427-P39. This difference in zoospore production was associated with a difference in pathogenicity for a susceptible amphibian species, indicating that fecundity may be an important virulence factor for Bd. In the 130-day experiment, Atelopus zeteki frogs exposed to the JEL427-P9 culture experienced higher average infection intensity and 100% mortality, compared with 60% mortality for frogs exposed to JEL427-P39. This effect was not observed with Eleutherodactylus coqui, which was able to clear infection. We hypothesize that the differences in phenotypic performance observed with Atelopus zeteki are rooted in changes of the Bd genome. Future investigations enabled by this study will focus on the underlying mechanisms of Bd pathogenicity.

Date Created
2013-10-10
Agent

Susceptibility to the Amphibian Chytrid Fungus Varies With Ontogeny in the Direct-Developing Frog, Eleutherodactylus Coqui

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Description

Age-related differences in susceptibility to infectious disease are known from a wide variety of plant and animal taxonomic groups. For example, the immature immune systems of young vertebrates, along with limited prior exposure to pathogens and behavioral factors, can place

Age-related differences in susceptibility to infectious disease are known from a wide variety of plant and animal taxonomic groups. For example, the immature immune systems of young vertebrates, along with limited prior exposure to pathogens and behavioral factors, can place juveniles at greater risk of acquiring and succumbing to a pathogen. We studied the ontogenetic susceptibility of terrestrial direct-developing frogs (Eleutherodactylus coqui) to the fungal pathogen, Batrachochytrium dendrobatidis (Bd), which is responsible for the decline of amphibian species worldwide. By exposing juvenile and adult frogs to the same dose and strain of Bd, we uncovered ontogenetic differences in susceptibility. Froglets exposed to the pathogen had significantly lower survival rates compared with control froglets, while adult frogs largely cleared infection and had survival rates indistinguishable from control frogs, even when exposed to a much higher dose of Bd. The high disease-induced mortality rate of juveniles may explain ongoing population declines in eastern Puerto Rico, where Bd is endemic and juveniles experience higher prevalence and infection intensity compared to adults. Our results have important implications for understanding and modeling the decline, possibly to extinction, of amphibian populations and species.

Date Created
2014-07-01
Agent

Chytridiomycosis in the direct-developing frogs of Puerto Rico

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Description
Epidemiological theory normally does not predict host extinction from infectious disease because of a host density threshold below which pathogens cannot persist. However, host extinction can occur when a biotic or abiotic pathogen reservoir allows for density-independent transmission. Amphibians are

Epidemiological theory normally does not predict host extinction from infectious disease because of a host density threshold below which pathogens cannot persist. However, host extinction can occur when a biotic or abiotic pathogen reservoir allows for density-independent transmission. Amphibians are facing global population decline and extinction from the emerging infectious disease chytridiomycosis, caused by the fungus Batrachochytrium dentrobatidis (Bd). I use the model species Eleutherodactylus coqui to assess the impact of Bd on terrestrial direct-developing frog species, a common life history in the tropics. I tested the importance of two key factors that might influence this impact and then used laboratory experiments and published field data to model population-level impacts of Bd on E. coqui. First, I assessed the ontogenetic susceptibility of E. coqui by exposing juvenile and adult frogs to the same pathogen strain and dose. Juveniles exposed to Bd had significantly lower survival rates compared with control juveniles, while adult frogs often cleared infection. Second, I conducted experiments to determine whether E. coqui can become infected with Bd indirectly from contact with zoospores shed onto vegetation by an infected frog and from direct exposure to an infected frog. Both types of transmission were observed, making this the first demonstration that amphibians can become infected indirectly in non-aquatic habitats. Third, I tested the hypothesis that artificially-maintained cultures of Bd attenuate in pathogenicity, an effect known for other fungal pathogens. Comparing two cultures of the same Bd strain with different passage histories revealed reduced zoospore production and disease-induced mortality rates for a susceptible frog species (Atelopus zeteki) but not for the less-susceptible E. coqui. Finally, I used a mathematical model to project the population-level impacts of chytridiomycosis on E. coqui. Model analysis showed that indirect transmission, combined with either a high rate of zoospore production or low rate of zoospore mortality, is required for Bd to drive E. coqui populations below an extinction threshold. High rates of transmission plus frequent re-infection could lead to poor recruitment of infected juveniles and population decline. My research adds further insight into how emerging infectious disease is contributing to the loss of amphibian biodiversity.
Date Created
2013
Agent