Fungal Pathogen Batrachochytrium Dendrobatidis Research Articles

Differential Host Susceptibility to Batrachochytrium dendrobatidis, an Emerging Amphibian Pathogen

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.

Social Immunity in Amphibians: Evidence for Vertical Transmission of Innate Defenses

Amphibian embryos are at risk of microbial infection. Here we find evidence that innate immune defenses, both antimicrobial skin peptides and mutualistic microbiota, of adult glass frogs, Hyalinobatrachium colymbiphyllum, can be transmitted to embryos deposited on leaves above rain forest streams in Panama and can inhibit the fungal pathogen Batrachochytrium dendrobatidis. Abstract in Spanish is available at http://www.blackwell-synergy.com/loi/btp.

First Record of Batrachochytrium dendrobatidis Infecting Four Frog Families from Peninsular Malaysia

The fungal pathogen Batrachochytrium dendrobatidis (Bd) infects amphibians on every continent where they occur and is linked to the decline of over 200 amphibian species worldwide. At present, only three published Bd surveys exist for mainland Asia, and Bd has been detected in South Korea alone. In this article, we report the first survey for Bd in Peninsular Malaysia. We swabbed 127 individuals from the six amphibian families that occur on Peninsular Malaysia, including two orders, 27 genera, and 47 species. We detected Bd on 10 out of 127 individuals from four of five states and five of 11 localities, placing the 95% confidence interval for overall prevalence at 4-14%. We detected no variation in Bd prevalence among regions, elevations, or taxonomic groups. The infection intensity ranged from 1 to 157,000 genome equivalents. The presence of Bd infections in native species without clinical signs of disease suggests that Bd may be endemic to the region. Alternately, Bd may have been introduced from non-native amphibians because of the substantial amphibian food trade in Peninsular Malaysia. Under both scenarios, management efforts should be implemented to limit the spread of non-native Bd and protect the tremendous amphibian diversity in Peninsular Malaysia.

Phenotypic profiling of Batrachochytrium dendrobatidis, a lethal fungal pathogen of amphibians

The fungal pathogen Batrachochytrium dendrobatidis (Bd) causes the disease chytridiomycosis, which is responsible for many amphibian declines worldwide. The determinants of Bd’s high virulence across a wide range of amphibian species are not well understood but include environmental factors, host responses to infection, and pathogenicity of Bd. Because there are clear differences among Bd isolates, genetic profiling of multiple strains is currently in progress. However, genetic analyses alone may not provide a functional explanation for differential virulence among strains. I aimed to quantify phenotypic traits, such as zoospores densities, that are important for disease development. Among the three isolates tested, zoospore densities were highest in GibboRiver-L.lesueuri-00-LB-P24, an isolate originally collected from a Litoria lesueuri tadpole and highly virulent in infection experiments. Because characterizing differences among isolates is central to understanding Bd pathogenicity, integrating genetic work with the phenotypic profiling methods provided in this study will offer insights into chytridiomycosis and may be important for amphibian conservation.

Stability and permanence in gender- and stage-structured models for the boreal toad

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.

What Drives Chytrid Infections in Newt Populations? Associations with Substrate, Temperature, and Shade

The pathogenic chytrid fungus Batrachochytrium dendrobatidis (Bd) is considered responsible for the population declines and extinctions of hundreds of amphibian species worldwide. The panzootic was likely triggered by human-assisted spread, but once the pathogen becomes established in a given region, its distribution is probably determined by local drivers. To assess the relative importance of potential drivers of infection in red-spotted newts (Notophthalmus viridescens), we measured Bd levels in 16 populations throughout central Pennsylvania. Infected individuals were detected in all but four populations, indicating that Bd is widespread in this region. We quantified local factors hypothesized to influence Bd, and found that infection levels were best predicted by the proportion of the pond substrate consisting of leaf litter or vegetation, along with a significant effect of water temperature. Bd infection in amphibians is temperature-dependent, and one possible explanation of the apparent substrate effect is that tree cover and vegetation provide shade, reducing the availability of shallow, warm-water patches in which newts might reduce or clear Bd infections. Alternatively, leaf litter and emergent vegetation might increase Bd infection more directly, perhaps by providing substrates for environmental growth of the fungus. We also observed a curvilinear relationship between Bd load and snout-vent length (a proxy for age), hinting that newts might develop acquired resistance to Bd infection. Though correlational, these results add to a growing body of evidence suggesting that environmental temperature is an important driver of Bd infection dynamics.

Batrachochytrium dendrobatidis infection dynamics in the Columbia spotted frog Rana luteiventris in north Idaho, USA

The pathogenic chytrid fungus Batrachochytrium dendrobatidis (Bd) is contributing to amphibian declines worldwide. Temperature plays an important role in both pathogen growth and host immune function, but little is known about seasonal dynamics of Bd infection in north temperate regions. Our objective was to increase understanding of Bd disease ecology by investigating patterns of Bd infection of Columbia spotted frogs Rana luteiventris across seasons, age classes, and sexes in north Idaho, USA. We collected skin swabs from 223 R. luteiventris in spring, summer, and fall 2009 at 7 ponds in the Palouse region and quantified Bd zoospores for each sample using quantitative PCR. Across seasons, Bd prevalence of adults was higher in summer than in spring or fall, suggesting that individuals may be clearing low-level infections over the summer. Among age classes, all but one late stage tadpole (Gosner stage 43-45) tested negative for Bd. Conversely, 100% of metamorphs tested positive for Bd and had the highest Bd loads of all age classes, suggesting they may be the most vulnerable age class. Adult R. luteiventris had high infection prevalence (> 60%) in all seasons, indicating that Bd infection is maintained within populations and that adults likely serve as disease reservoirs across seasons. Among adults, we also found weak evidence for females having higher infection prevalence than males. Further laboratory and field studies are needed to determine whether there are individual and population impacts from Bd on R. luteiventris and other amphibians in north Idaho.

Widespread occurrence of the amphibian chytrid fungus in Kenya

AbstractAmphibians at the global scale are dramatically declining and the pathogenic fungus Batrachochytrium dendrobatidis (Bd) has been suggested to be an important driver in this biodiversity crisis. Increasing evidence points towards the global emergence of Bd being a panzootic caused by pathogen pollution. Africa has been suggested to be the origin of the pathogen but remains one of the least‐studied areas. We have conducted the most comprehensive survey on the continent to date focusing on Kenya for investigating taxonomic and environmental components in the distribution of Bd in tropical Africa. Eleven sites along a 770 km transect from the coast up to the border of Uganda were surveyed. Using quantitative PCR, we screened 861 samples from 23 different species in nine genera. The pathogen was confirmed at all studied sites, with an overall prevalence of 31.5%. No dead or symptomatic specimens were found and no declines have been reported in the region so far. Both prevalence and parasite load ranged from the detection limit to some of the highest ever reported. The parasite load showed a significant taxonomic bias and a strong inverse correlation with temperature. Our findings suggest that Bd may be enzootic in the region. We recommend that further research should focus on comparative experimental studies of susceptibility to Bd in African species. Moreover, we stress the need for improved knowledge on the conservation status of the tropical African amphibian fauna to confirm the enzootic nature of widespread Bd infections.

Toward a General Theory for How Climate Change Will Affect Infectious Disease

Toward a General Theory for How Climate Change Will Affect Infectious Disease

Cutaneous Bacteria of the Redback Salamander Prevent Morbidity Associated with a Lethal Disease

Chytridiomycosis, caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd), is an infectious disease that causes population declines of many amphibians. Cutaneous bacteria isolated from redback salamanders, Plethodon cinereus, and mountain yellow-legged frogs, Rana muscosa, inhibit the growth of Bd in vitro. In this study, the bacterial community present on the skin of P. cinereus individuals was investigated to determine if it provides protection to salamanders from the lethal and sub-lethal effects of chytridiomycosis. When the cutaneous bacterial community was reduced prior to Bd exposure, salamanders experienced a significantly greater decrease in body mass, which is a symptom of the disease, when compared to infected individuals with a normal bacterial community. In addition, a greater proportion of infected individuals with a reduced bacterial community experienced limb-lifting, a behavior seen only in infected individuals. Overall, these results demonstrate that the cutaneous bacterial community of P. cinereus provides protection to the salamander from Bd and that alteration of this community can change disease resistance. Therefore, symbiotic microbes associated with this species appear to be an important component of its innate skin defenses.

Batrachochytrium dendrobatidis: requirement for further isolate collection and archiving

The fungal pathogen Batrachochytrium dendrobatidis (Bd) causes the disease chytridiomycosis, which is lethal to many species of amphibians worldwide. Many studies have investigated the epidemiology of chytridiomycosis in amphibian populations, but few have considered possible host-pathogen coevolution. More specifically, investigations focused on the evolution of Bd, and the link with Bd virulence, are needed. Such studies, which may be important for conservation management of amphibians, depend on access to Bd isolates. Here we provide a summary of known Bd isolates that have been collected and archived in various locations around the world. Of 257 Bd isolates, we found that 53% originate from ranids in the United States. In many cases, detailed information on isolate origin is unavailable, and it is unknown how many isolates are cryo-archived. We suggest the creation of a centralized database of isolate information, and we urge researchers and managers to isolate and archive Bd to facilitate future research on chytridiomycosis.

Dynamics of an emerging disease drive large-scale amphibian population extinctions

Epidemiological theory generally suggests that pathogens will not cause host extinctions because the pathogen should fade out when the host population is driven below some threshold density. An emerging infectious disease, chytridiomycosis, caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd) is directly linked to the recent extinction or serious decline of hundreds of amphibian species. Despite continued spread of this pathogen into uninfected areas, the dynamics of the host-pathogen interaction remain unknown. We use fine-scale spatiotemporal data to describe (i) the invasion and spread of Bd through three lake basins, each containing multiple populations of the mountain yellow-legged frog, and (ii) the accompanying host-pathogen dynamics. Despite intensive sampling, Bd was not detected on frogs in study basins until just before epidemics began. Following Bd arrival in a basin, the disease spread to neighboring populations at approximately 700 m/yr in a wave-like pattern until all populations were infected. Within a population, infection prevalence rapidly reached 100% and infection intensity on individual frogs increased in parallel. Frog mass mortality began only when infection intensity reached a critical threshold and repeatedly led to extinction of populations. Our results indicate that the high growth rate and virulence of Bd allow the near-simultaneous infection and buildup of high infection intensities in all host individuals; subsequent host population crashes therefore occur before Bd is limited by density-dependent factors. Preventing infection intensities in host populations from reaching this threshold could provide an effective strategy to avoid the extinction of susceptible amphibian species in the wild.

IMMUNIZATION IS INEFFECTIVE AT PREVENTING INFECTION AND MORTALITY DUE TO THE AMPHIBIAN CHYTRID FUNGUS BATRACHOCHYTRIUM DENDROBATIDIS

The fungal pathogen Batrachochytrium dendrobatidis (Bd), the causative agent of chytridiomycosis, has been implicated in amphibian declines worldwide. It has been hypothesized that low inherent immunogenicity in Bd may be related to the high rates of morbidity and mortality that are associated with Bd-infected anuran populations. To test this idea, juvenile Rana muscosa (mountain yellow-legged frogs) were immunized with adjuvants in combination with a formalin-killed Bd culture to determine if it is possible to stimulate a protective immune response when challenged with a live inoculum of B. dendrobatidis. Three groups of juvenile R. muscosa (6 mo postmetamorphosis) were immunized with saline, Freunds Complete (FCA) and Incomplete Adjuvant (FIA), or the adjuvants in combination with a formalin-killed culture of B. dendrobatidis. The effects of immunization were modeled using survival analysis and a proportional hazards model. No significant differences were found between the groups in overall mortality, time to infection, infection prevalence, or intensity. While this study suggests that immunizing anurans against chytridiomycosis will not alter rates of infection or mortality among individuals, it does raise several questions regarding the attenuation and efficacy of anuran adaptive immune responses and whether they may be protective against this disease.

Prevalence of the pathogenic chytrid fungus, Batrachochytrium dendrobatidis, in an endangered population of northern leopard frogs, Rana pipiens

BackgroundEmerging infectious diseases threaten naïve host populations with extinction. Chytridiomycosis, an emerging infectious disease of amphibians, is caused by the pathogenic fungus Batrachochytrium dendrobatidis (Bd) and has been linked to global declines in amphibians.ResultsWe monitored the prevalence of Bd for four years in the Northern leopard frog, Rana pipiens, which is critically imperiled in British Columbia (BC), Canada. The prevalence of Bd initially increased and then remained constant over the last three years of the study. Young of the year emerging from breeding ponds in summer were rarely infected with Bd. Some individuals cleared their Bd infections and the return rate between infected and uninfected individuals was not significantly different.ConclusionsThe BC population of R. pipiens appears to have evolved a level of resistance that allows it to co-exist with Bd. However, this small population of R. pipiens remains vulnerable to extinction.

An Aquatic Disease on a Terrestrial Salamander: Individual and Population Level Effects of the Amphibian Chytrid Fungus, Batrachochytrium dendrobatidis, on Batrachoseps attenuatus (Plethodontidae)

The pathogenic chytrid fungus Batrachochytrium dendrobatidis, typically associated with anuran amphibians, is present in natural populations of the terrestrial salamander, Batrachoseps attenuatus, from California, USA, and four congeners from California and Oregon, USA. I demonstrate that the chytrid has been present in wild populations of B. attenuatus since at least 1973, and while infected salamanders collected in the wild exhibited 100% mortality in the laboratory, wild populations appear to have remained stable with seasonally variable infection rates. Laboratory experiments showed that inoculated salamanders housed in dry microhabitats, mimicking summer aestivation conditions, are able to shed the chytrid infection. Combining these data with the decrease in prevalence from spring to fall suggests that environmental conditions in the natural range of B. attenuatus mediate the effects of this potentially highly lethal pathogen, stabilizing this host–pathogen relationship. While B. attenuatus continues to be an abundant salamander, other amphibians are experiencing marked declines. An understanding of the relationship between the amphibian chytrid fungus and species of Batrachoseps may be applicable to patterns of declines and persistence in other species of plethodontid salamanders, and amphibians in general.

Silent Springs: Why Are All the Frogs “Croaking”?

Silent Springs: Why Are All the Frogs “Croaking”?

Batrachochytriumdendrobatidisinfection patterns among Panamanian amphibian species, habitats and elevations during epizootic and enzootic stages

The pathogenic fungus Batrachochytrium dendrobatidis (Bd) has caused declines of many amphibian populations, yet the full course of the epizootic has rarely been observed in wild populations. We determined effects of elevation, habitat, and aquatic index (AI) on prevalence of infection among Panamanian amphibians sampled along 2 elevational transects. Amphibian populations on the Santa Fé transect (SFT) had declined in 2002, while those on the El Copé transect (ECT) were healthy until September 2004. In 2004 we sampled Bd along both transects, surveying the SFT 2 yr after decline, and surveying the ECT 4 mo prior to the arrival of Bd, during the epizootic, and 2 mo later. Overall prevalence of Bd along the ECT increased from 0.0 (95% CI 0.00-0.0003) to 0.51 (95% CI 0.48-0.55) over a 3 mo period, accompanied by significant decreases in amphibian abundance and species richness in all habitats. Prevalence of infection on the ECT was highest along riparian transects and at higher elevations, but not among levels of AI. Prevalence of infection on the SFT was highest in pool transects, and at higher elevations, but not among levels of AI. Riparian amphibian abundance and species richness also declined at SFT following detection of Bd in 2002. Variation among species, microenvironmental conditions, and the length of coexistence with Bd may contribute to observed differences in prevalence of Bd and in population response.

Chytridiomycosis and Amphibian Population Declines Continue to Spread Eastward in Panama

Chytridiomycosis is a globally emerging disease of amphibians and the leading cause of population declines and extirpations at species-diverse montane sites in Central America. We continued long-term monitoring efforts for the presence of the fungal pathogen Batrachochytrium dendrobatidis (Bd) and for amphibian populations at two sites in western Panama, and we began monitoring at three new sites to the east. Population declines associated with chytridiomycosis emergence were detected at Altos de Campana National Park. We also detected Bd in three species east of the Panama Canal at Soberanía National Park, and prevalence data suggests that Bd may be enzootic in the lowlands of the park. However, no infected frogs were found further east at Tortí (prevalence <7.5% with 95% confidence). Our results suggest that Panama's diverse and not fully described amphibian communities east of the canal are at risk. Precise predictions of future disease emergence events are not possible until factors underlying disease emergence, such as dispersal, are understood. However, if the fungal pathogen spreads in a pattern consistent with previous disease events in Panama, then detection of Bd at Tortí and other areas east of the Panama Canal is imminent. Therefore, development of new management strategies and increased precautions for tourism, recreation, and biology are urgently needed.

Effect of Temperature on Host Response to Batrachochytrium dendrobatidis Infection in the Mountain Yellow-legged Frog (Rana muscosa)

The pathogenic chytrid fungus Batrachochytrium dendrobatidis, which causes the disease chytridiomycosis, has been implicated in declines of amphibian populations throughout the world, including declines and extinctions of local populations of mountain yellow-legged frogs, Rana muscosa, in the California Sierra Nevada. Previous studies have shown B. dendrobatidis achieves its maximum growth rate in culture in the temperature range of 17-25 C, and exposure to very high temperatures can clear frogs of B. dendrobatidis infection. Here we present the results of a laboratory experiment in which experimentally infected R. muscosa tadpoles were followed through metamorphosis at temperatures of 17 and 22 C. All infected animals developed clinical disease within a similar time frame. However, frogs housed at 22 C exhibited a significantly lower mortality than those housed at 17 C. Within 35 days after metamorphosis, 50% of the frogs housed at 22 C died, while 95% of the frogs housed at 17 C died. Clinical signs subsided in the surviving frogs at 22 C, despite persistent infection. Because both temperatures are within the optimal thermal range for growth of B. dendrobatidis, we propose that the difference in outcome indicates the effect of temperature on the host's resistance to chytridiomycosis, rather than an effect on the fungus alone.

Decline and extirpation of an endangered Panamanian stream frog population ( Craugastor punctariolus) due to an outbreak of chytridiomycosis

We conducted a mark-recapture study of three subpopulations of Craugastor punctariolus at a mid-elevation site in central Panama between 1999 and 2005. The study spans a period over which the pathogenic fungus Batrachochytrium dendrobatidis ( Bd) was absent from the site, invaded the site and decimated all amphibian populations, and now persists. We quantified natural demographic parameters prior to and during an event of mass mortality due to chytridiomycosis caused by Bd. Prior to the event of mass mortality, all three subpopulations of C. punctariolus were large (19–68 animals/200 m), showed a stable age-size distribution, and had high survival. All age-size classes of this species co-occurred on boulder clumps along streams, and adults showed high site fidelity and were likely territorial. Following detection of Bd at this site in late September 2004, four dead C. punctariolus were found infected with Bd and all three subpopulations completely disappeared from this site within 2 months. The association of all age-size classes with microhabitats appropriate for survival and growth of Bd likely contributed to the rapid and severe degree of decline of this species at this site. These data provide insight into the patterns and mechanisms of decline within a species due to Bd.