Chytrid Research Articles

TEMPERATURE AS A DRIVER OF THE PATHOGENICITY AND VIRULENCE OF AMPHIBIAN CHYTRID FUNGUS BATRACHOCHYTRIUM DENDROBATIDIS: A SYSTEMATIC REVIEW.

Chytridiomycosis, caused by the chytrid fungus Batrachochytrium dendrobatidis (Bd), is a leading cause of global amphibian declines. Severe infections with Bd can lead to cardiac arrest, and mass deaths during epidemics have been reported. Temperature, pH, salinity, and moisture are important determinants of the survival, growth, reproduction, and pathogenicity of Bd, as well as its effect on amphibian populations. Here, we synthesize current knowledge on the role of temperature as a driver of the pathogenicity and virulence of Bd to better understand the effects of temperature on amphibian defense mechanisms against infection. This review advises on research direction and management approaches to benefit amphibian populations affected by Bd. We conclude by offering guidelines for four levels of temperature monitoring in amphibian field studies to improve consistency between studies: regional climate, habitat, microhabitat, and amphibian host.

Enigmatic Near‐Extirpation in a Boreal Toad Metapopulation in Northwestern Montana

ABSTRACTNorth America's protected lands harbor biodiversity and provide habitats where species threatened by a variety of stressors in other environments can thrive. Yet disease, climate change, and other threats are not limited by land management boundaries and can interact with conditions within protected landscapes to affect sensitive populations. We examined the population dynamics of a boreal toad (Anaxyrus boreas boreas) metapopulation at a wildlife refuge in northwestern Montana, USA, over a 16‐year period (2003–2018). We used robust design capture‐recapture models to estimate male population size, recruitment, and apparent survival over time and in relation to the amphibian chytrid fungus (Batrachochytrium dendrobatidis). We estimated female population size in years with sufficient captures. Finally, we examined trends in male and female toad body size and condition. We found no evidence of an effect of disease or time on male toad survival but detected a strong negative trend in recruitment of new males to the population. Estimates of male and female abundance decreased over time. Body size of males and females was inversely related to estimated population size, consistent with reduced recruitment to replace adults, but body condition of adult males was only weakly associated with abundance. Together, these results describe the demography of a near‐extirpation event, and point to dramatic decreases in the recruitment of new individuals to the breeding population as the cause of this decline. We surmise that processes related to the restoration of historical hydrology within the refuge adversely affected amphibian breeding habitat, and that these changes interacted with disease, life history, and other factors to restrict the recruitment of new individuals to the breeding population over time. Our results point to challenges in understanding and predicting factors that influence population change and highlight that current metrics for assessing population status can have limited predictive ability. Published 2021. This article is a U.S. Government work and is in the public domain in the USA.

Rapid population increase of the threatened Australian amphibian Litoria aurea in response to wetlands constructed as a refuge from chytrid-induced disease and introduced fish

Amphibians have declined due to multiple impacts including invasive fish and the disease chytridiomycosis caused by the chytrid fungus Batrachochytrium dendrobatidis (Bd). Wetland restoration can be used to increase amphibian populations. However the design of created wetlands must account for threats such as Bd and introduced fish. There have been no attempts on a landscape level to manage these threats with habitat design. Here we monitored the green and golden bell frog (Litoria aurea) in 2.6 ha of constructed wetlands designed to enhance breeding and increase survival through passive mitigation of Bd and exotic fish. We compared the fecundity, adult population sizes, introduced fish occupancy, Bd prevalence and survival rates of frogs in created wetlands (CW) to three control sites to determine if and why the habitat design was successful. Monitoring involved weekly capture-recapture during the austral spring and summer for three L. aurea breeding seasons. We hypothesised that (1) if the CWs were successful in passively limiting fish colonisation, a larger number of breeding events would be detected compared to control sites which are known to be widely colonised by introduced fish. (2) If the wetlands were successful in passively mitigating Bd, then we would observe an equal or greater survival rate and equal to or lower Bd prevalence compared to control wetlands. We observed a 3.3-fold increase in adult population size in CW from season 1 to 2, and the population increased further in season 3.We found strong support for hypothesis (1) and weak support for (2). Based on these results, we conclude that this design was beneficial shortly after their formation primarily due to fish exclusion, but further study is required to determine if these benefits extend long-term. Future amphibian restoration studies are needed to improve the design of wetlands to enhance suppression of Bd.

Characteristics and consequences of a disease outbreak in aquatic insects

Abstract Disease can be a powerful driver of population and community dynamics, as well as evolutionary processes. Disease is also emerging at increasing rates, resulting in massive impacts on populations, communities, and ecosystems. However, assessing these impacts requires foundational knowledge of disease agents and hosts, which is often lacking, particularly in aquatic insects. We describe a recent disease outbreak in caddisflies, suggesting potential consequences for population and community dynamics of the host. We use a series of complementary studies to develop a cohesive foundation of information about this disease, including identification using genomic methods, observational prevalence studies, laboratory experiments to establish transmissibility and fitness consequences, and laboratory and field investigations to infer transmission mechanisms. We identified the infection as being caused by the oomycete Saprolegnia—the first time this parasite has been noted in insect eggs. Prevalence surveys found high prevalence (up to 36%) with variation across space, time, and host species. We demonstrated increasing egg mortality with increasing infection within an egg mass (every 10% increase in infection rate doubles odds of mortality), thereby confirming disease. We established transmissibility and show that transmission occurs through both direct contact with infected egg masses and from background sources, which probably interact to create complex patterns of disease. Taken together, our findings show that conditions necessary for population and community consequences are present. Specifically, increased mortality rates almost certainly occurred during the outbreak, yielding lower larval numbers and potentially altering community interactions. Transmission by contact between egg masses combined with observed species‐specific prevalence suggest shifts in the relative performance of different species because of interactions between host and parasite life histories. Outside extreme examples such as chytrid fungus, disease has traditionally received less interest than resource competition or predation in community ecology, although disease ecology is advancing rapidly. One major hurdle is a lack of foundational knowledge characterising disease processes in natural communities, particularly in aquatic insects. Our findings highlight the importance of investigating diseases in insect eggs and provide the foundation for further investigations of how these processes play out at the population and community scale.

The Fungicide Chlorothalonil Changes the Amphibian Skin Microbiome: A Potential Factor Disrupting a Host Disease-Protective Trait

The skin microbiome is an important part of amphibian immune defenses and protects against pathogens such as the chytrid fungus Batrachochytrium dendrobatidis (Bd), which causes the skin disease chytridiomycosis. Alteration of the microbiome by anthropogenic factors, like pesticides, can impact this protective trait, disrupting its functionality. Chlorothalonil is a widely used fungicide that has been recognized as having an impact on amphibians, but so far, no studies have investigated its effects on amphibian microbial communities. In the present study, we used the amphibian Lithobates vibicarius from the montane forest of Costa Rica, which now appears to persist despite ongoing Bd-exposure, as an experimental model organism. We used 16S rRNA amplicon sequencing to investigate the effect of chlorothalonil on tadpoles’ skin microbiome. We found that exposure to chlorothalonil changes bacterial community composition, with more significant changes at a higher concentration. We also found that a larger number of bacteria were reduced on tadpoles’ skin when exposed to the higher concentration of chlorothalonil. We detected four presumed Bd-inhibitory bacteria being suppressed on tadpoles exposed to the fungicide. Our results suggest that exposure to a widely used fungicide could be impacting host-associated bacterial communities, potentially disrupting an amphibian protective trait against pathogens.

Dual Detection of the Chytrid Fungi Batrachochytrium spp. with an Enhanced Environmental DNA Approach.

Environmental DNA (eDNA) is becoming an indispensable tool in biodiversity monitoring, including the monitoring of invasive species and pathogens. Aquatic chytrid fungi Batrachochytrium dendrobatidis (Bd) and B. salamandrivorans (Bsal) are major threats to amphibians. However, the use of eDNA for detecting these pathogens has not yet become widespread, due to technological and economic obstacles. Using the enhanced eDNA approach (a simple and cheap sampling protocol) and the universally accepted qPCR assay, we confirmed the presence of Bsal and Bd in previously identified sites in Spain, including four sites that were new for Bsal. The new approach was successfully tested in laboratory conditions using manufactured gene fragments (gBlocks) of the targeted DNA sequence. A comparison of storage methods showed that samples kept in ethanol had the best DNA yield. Our results showed that the number of DNA copies in the Internal Transcribed Spacer region was 120 copies per Bsal cell. Eradication of emerging diseases requires quick and cost-effective solutions. We therefore performed cost-efficiency analyses of standard animal swabbing, a previous eDNA approach, and our own approach. The procedure presented here was evaluated as the most cost-efficient. Our findings will help to disseminate information about efforts to prevent the spread of chytrid fungi.

Time in the Laboratory, but Not Exposure to a Chytrid Fungus, Results in Rapid Change in Spring Peeper (Pseudacris crucifer) Skin Bacterial Communities

Interactions between pathogens and the symbiotic microbial communities that reside in hosts can impact disease processes. Symbiotic microbial communities can act to prevent pathogen infection in some instances, while in other instances, pathogens can disrupt these symbiotic communities. We sought to address these interactions between the fungal skin pathogen Batrachochytrium dendrobatidis (Bd) and the skin bacterial communities of adult Spring Peepers (Pseudacris crucifer). In the laboratory, frogs were exposed to Bd zoospores for a 12-hour period and then monitored for five days. Bacterial community composition on the skin was assessed upon initial collection, 24 hours post-exposure, and at the conclusion of the experiment using 16S rRNA gene amplicon sequencing. Little impact on the bacterial community was observed as a result of Bd exposure, suggesting that the timeline was either too short to observe the effects of Bd, or that the skin communities are resilient in the face of such short exposure periods. In contrast, laboratory housing was associated with changes in skin bacterial community composition, in terms of both relative abundances and bacterial taxa that were present. These findings suggest that even a short time away from their natural environment can have strong effects on the composition of amphibian skin communities and that Bd exposure may not always disrupt these skin communities.

Midges not only sucks, but may carry lethal pathogens to wild amphibians

AbstractThe chytrid fungus (Bd) is one of the main causes of recent amphibian population declines and extinctions. However, one possible disease transmission pathway remained unexplored: the insect‐borne transport between hosts. We found Bd DNA on specialized blood‐sucking midges, which could indicate the transport of alive zoospores between amphibian hosts, in this case acting as Bd carriers in the wild.Abstract in Portuguese is available with online material.

Evaluating eDNA-based monitoring of fire salamander larvae under field conditions

Amphibians globally belong to the most threatened animal groups and monitoring their populations is of critical importance for their conservation. The fire salamander (Salamandra salamandra) is one of the European amphibian species which has been experiencing drastic regional population declines due to the spread of the invasive chytrid fungus Batrachochytrium salamandrivorans, making it a key species for monitoring efforts. Here, we evaluated how the sampling and analysis of eDNA can aid the monitoring of larval salamander populations in small streams under field conditions. Nine small streams with known adult and larval fire salamander populations were investigated in Tirol (Austria). Per steam a stretch of 30 m downstream from its source was divided into 10 m sections where salamander larvae were counted. Water samples were taken at the end of each section and filtered on site. The DNA extracted from these filters was tested by a new PCR assay developed for the detection of mitochondrial DNA of S. salamandra. This assay combines endpoint PCR with capillary electrophoresis, allowing to relatively quantify the amount of fire salamander eDNA present in the water samples. In two of the nine streams no eDNA of S. Salamandra could be detected. The outcomes of an analysis of how larval densities, discharge and volume of filtered water affected the detection of salamander eDNA will be presented. Finally, we will conclude on the practical implications of the current findings for eDNA-based monitoring of fire salamander populations.

Establishement of a national monitoring program based on environmental DNA for amphibians and the chytrid fungus Batrachochytrium dendrobatidis

National monitoring programs provide the basis for evaluating the integrity of ecosystems, their responses to disturbances, and the success of actions taken to conserve or recover biodiversity. In this study, we successfully established a national program for the invasive chytrid fungus Batrachochytrium dendrobatidis (Bd) based on dual TaqMan assays. Amphibian diversity based on metabarcoding of the mitochondrial 12S rRNA gene was also performed. Assays were optimized for sensitive detection of target species from a wide range of amphibian ponds with variable potential of inhibitions for eDNA based detection. An amphibian mock community of 5 species was used to validate the metabarcoding approach while internal standards were used in the case of the dual TaqMan assays. First sampling of over 170 ponds in Norway resulted in Bd detection in 12 environmental samples and one swab sample taken over multiple years indicating the establishment of Bd in Norway. Five amphibian species Bufo bufo, Lissotriton vulgaris, Triturus cristatus, Rana arvalis and Rana temporaria as predicted from data in long-term citizen science reporting systems were widely detected in the collected eDNA samples. Our large scale-monitoring program indicates a low risk of a Bd outbreak and amphibian decline caused by chytridiomycosis but continued monitoring is recommended in the future. These findings indicate that eDNA is an effective method to detect invasive species, and to monitor endangered amphibian species. Still, several shortcomings (such as PCR inhibitors and sample volume) were identified that need to be addressed to improve eDNA-based monitoring at the national level.

Niche Contraction of an Endangered Frog Driven by the Amphibian Chytrid Fungus.

Introduced pathogens can alter the geographic distribution of susceptible host species. For example, Batrachochytrium dendrobatidis (Bd) is a fungal pathogen that has been linked to the global decline and extinction of numerous amphibian species during the last four decades. A growing number of studies have described the distribution of Bd and susceptible hosts across the globe; however, knowledge on how Bd may shape the climatic niche of susceptible species is still missing. We estimated the effect of Bd on the geographic distribution and niche dynamics of the critically endangered lowland robber frog (Craugastor ranoides) in Costa Rica. We found a reduction of 98% in the geographic range of this species by 1995, following the epizootic outbreaks of Bd that affected Costa Rica in the 1980 and early 1990s. We also quantified niche contraction and found that the species is currently restricted to dry and warm environments that have been considered unsuitable for Bd. Our results contribute to the understanding of how emerging pathogens shape the climatic niches and geographic distribution of susceptible species.

Amphibian chytridiomycosis, a lethal pandemic disease caused by the killer fungus Batrachochytrium dendrobatidis: New approaches to host defense mechanisms and techniques for detection and monitoring

Chytridiomycosis is a catastrophic disease currently decimating worldwide amphibian populations, caused by the panzootic chytrid fungus Batrachochytrium dendrobatidis. Massive species decline to extinction catalyzes radical changes in ecosystems globally, including the largest continuous rainforest ecosystem on Earth, the Amazon rainforest. Innovative research that aims to propose feasible mechanisms of mitigation and the origins of the disease is vital, including studies addressing climatic effects on the expansion of chytridiomycosis. Thus, this publication aims to provide a comprehensive review of: i) the current technologies used for B. dendrobatidis detection and monitoring, and ii) the known Neotropical amphibian's skin microbiota with anti-fungal properties against B. dendrobatidis. Several immunologic and DNA-based methods are discussed to understand the emerging fungal pathogens and their effects on the biosphere, which can help to mitigate the devastating ecological impacts of mass amphibian morbidity. The establishment of rapid and highly accurate B. dendrobatidis detection techniques and methods for monitoring amphibian's cutaneous microbiome is crucial in the fight against chytridiomycosis.

Bad neighbours: amphibian chytrid fungus Batrachochytrium dendrobatidis infection dynamics in three co-occurring frog species of southern Sydney, Australia.

Wildlife disease is a major cause of global biodiversity loss. Amongst the most devastating is the disease chytridiomycosis, caused by the amphibian chytrid fungus Batrachochytrium dendrobatidis (Bd). This disease has contributed to declines and extinctions in hundreds of amphibian species, but not all species are affected equally. Some amphibian hosts are capable of carrying high levels of Bd infection without population declines, acting as reservoir species for the pathogen and driving population declines in sympatric species. In Australia, several species have been proposed as reservoir species; however, our understanding of Bd is derived from studies that are highly geographically and taxonomically biased, and our ability to extrapolate from these systems is unknown. We examined the prevalence and intensity of Bd infection in 3 frog species in a previously unstudied host-pathogen system in temperate eastern Australia: the Blue Mountains tree frog Litoria citropa, a poorly-known species predicted to be susceptible to Bd infection; and the common eastern froglet Crinia signifera and the stony creek frog L. lesueuri, which have both been identified as reservoir species in other regions. We found that L. citropa and L. lesueuri were infected with Bd at a high prevalence and often high intensity, while the reverse was true for C. signifera. All species were detected at moderate abundance and there was no evidence of morbidity and mortality. Our findings do not support C. signifera and L. lesueuri being reservoir species in this system, highlighting the importance of region-specific studies to inform conservation management.

The actin networks of chytrid fungi reveal evolutionary loss of cytoskeletal complexity in the fungal kingdom

Cells from across the eukaryotic tree use actin polymer networks for a wide variety of functions, including endocytosis, cytokinesis, and cell migration. Despite this functional conservation, the actin cytoskeleton has undergone significant diversification, highlighted by the differences in the actin networks of mammalian cells and yeast. Chytrid fungi diverged before the emergence of the Dikarya (multicellular fungi and yeast) and therefore provide a unique opportunity to study actin cytoskeletal evolution. Chytrids have two life stages: zoospore cells that can swim with a flagellum and sessile sporangial cells that, like multicellular fungi, are encased in a chitinous cell wall. Here, we show that zoospores of the amphibian-killing chytrid Batrachochytrium dendrobatidis (Bd) build dynamic actin structures resembling those of animal cells, including an actin cortex, pseudopods, and filopodia-like spikes. In contrast, Bd sporangia assemble perinuclear actin shells and actin patches similar to those of yeast. The use of specific small-molecule inhibitors indicate that nearly all of Bd's actin structures are dynamic and use distinct nucleators: although pseudopods and actin patches are Arp2/3 dependent, the actin cortex appears formin dependent and actin spikes require both nucleators. Our analysis of multiple chytrid genomes reveals actin regulators and myosin motors found in animals, but not dikaryotic fungi, as well as fungal-specific components. The presence of animal- and yeast-like actin cytoskeletal components in the genome combined with the intermediate actin phenotypes in Bd suggests that the simplicity of the yeast cytoskeleton may be due to evolutionary loss.

Disease influences male advertisement and mating outcomes in a critically endangered amphibian

The sublethal effects of infectious disease on reproductive behaviour and mating success are not well understood. Here, we investigated predictors of male mating success in one of Australia's most critically endangered vertebrates: the northern corroboree frog, Pseudophryne pengilleyi. Using a genomic approach to assign parentage, we explored whether infection with the amphibian chytrid fungus Batrachochytrium dendrobatidis (Bd), a pathogen responsible for amphibian declines globally, influenced male calling behaviour and mating success. We also explored whether male mating success was predicted by phenotypic traits (age, body size, coloration, call characters) that potentially signal genetic quality, and the soil moisture (water potential) of male-constructed terrestrial nests, which may directly impact offspring survival. We found that Bd significantly influenced male advertisement; Bd-infected males produced calls with significantly higher pulse repetition rates than uninfected males. Older males had a higher probability of mating; however, variation in the number of eggs in a nest was most strongly explained by an interaction between male Bd infection status and call pulse repetition rate. We propose that these relationships may result from either pathogen-mediated changes to host behaviour or host-mediated changes to behaviour (e.g. terminal investment). Regardless of the mechanism, this is the first evidence that male mating success in an amphibian can be influenced by male Bd infection status, highlighting a novel mechanism through which this virulent pathogen can affect amphibian fitness. More broadly, these findings add to a growing body of evidence that pathogens can alter the reproductive biology of their hosts. From a conservation perspective, increased consideration of how sexual selection operates in altered environments has the potential to assist with the management of threatened amphibians worldwide.

Genotyping and differential bacterial inhibition of Batrachochytrium dendrobatidis in threatened amphibians in Costa Rica.

Amphibians have declined around the world in recent years, in parallel with the emergence of an epidermal disease called chytridiomycosis, caused by the chytrid fungus Batrachochytrium dendrobatidis (Bd). This disease has been associated with mass mortality in amphibians worldwide, including in Costa Rica, and Bd is considered an important contributor to the disappearance of this group of vertebrates. While many species are susceptible to the disease, others show tolerance and manage to survive infection with the pathogen. We evaluated the pathogen Bd circulating in Costa Rica and the capacity of amphibian skin bacteria to inhibit the growth of the pathogen in vitro. We isolated and characterized - genetically and morphologically - several Bd isolates from areas with declining populations of amphibians. We determined that the circulating chytrid fungus in Costa Rica belongs to the virulent strain Bd-GPL-2, which has been related to massive amphibian deaths worldwide; however, the isolates obtained showed genetic and morphological variation. Furthermore, we isolated epidermal bacteria from 12 amphibian species of surviving populations, some in danger of extinction, and evaluated their inhibitory activity against the collection of chytrid isolates. Through bioassays we confirmed the presence of chytrid-inhibitory bacterial genera in Costa Rican amphibians. However, we observed that the inhibition varied between different isolates of the same bacterial genus, and each bacterial isolation inhibited fungal isolation differently. In total, 14 bacterial isolates belonging to the genera Stenotrophomonas, Streptomyces, Enterobacter, Pseudomonas and Klebsiella showed inhibitory activity against all Bd isolates. Given the observed variation both in the pathogen and in the bacterial inhibition capacity, it is highly relevant to include local isolates and to consider the origin of the microorganisms when performing in vivo infection tests aimed at developing and implementing mitigation strategies for chytridiomycosis.

Post-epizootic microbiome associations across communities of neotropical amphibians.

Microbiome-pathogen interactions are increasingly recognized as an important element of host immunity. While these host-level interactions will have consequences for community disease dynamics, the factors which influence host microbiomes at larger scales are poorly understood. We here describe landscape-scale pathogen-microbiome associations within the context of post-epizootic amphibian chytridiomycosis, a disease caused by the panzootic chytrid fungus Batrachochytrium dendrobatidis. We undertook a survey of Neotropical amphibians across altitudinal gradients in Ecuador ~30years following the observed amphibian declines and collected skin swab-samples which were metabarcoded using both fungal (ITS-2) and bacterial (r16S) amplicons. The data revealed marked variation in patterns of both B.dendrobatidis infection and microbiome structure that are associated with host life history. Stream breeding amphibians were most likely to be infected with B.dendrobatidis. This increased probability of infection was further associated with increased abundance and diversity of non-Batrachochytrium chytrid fungi in the skin and environmental microbiome. We also show that increased alpha diversity and the relative abundance of fungi are lower in the skin microbiome of adult stream amphibians compared to adult pond-breeding amphibians, an association not seen for bacteria. Finally, stream tadpoles exhibit lower proportions of predicted protective microbial taxa than pond tadpoles, suggesting reduced biotic resistance. Our analyses show that host breeding ecology strongly shapes pathogen-microbiome associations at a landscape scale, a trait that may influence resilience in the face of emerging infectious diseases.

Effect of Batrachochytrium Dendrobatidis on Anura Populations Batrachochytrium

Chytrium (Batrachochytrium Dendrobatidis) is a chytrium fungus that causes amphibian chytriomycosis. They were first discovered in 1998 and have been responsible for a large number of amphibian deaths over the next decade. Chytrid fungus grows on amphibian skin and produces free aquatic spores. They are widely distributed in deserts and lowland forests to cold hilltops. They can cause disease in upland (low temperature) amphibians, causing significant mortality. Chytrid infection has been associated with large numbers of amphibian deaths in north, south and central America, Europe and Australia. In this paper, the author summarized several factors influencing the transmission of chytrid by analyzing the existing data and data, including the transmission route, the worldwide distribution range and harm degree of chytrid, and the existing research and discussion on the elimination and prevention methods of chytrid. After collecting and analyzing data and literature, the author comes to the following conclusions: Firstly, the fungus originated in Africa and spread around the world through trade in animals such as xenopus africanus, and was endemic in Africa for many years before spreading around the world. Secondly, chytridiomycosis is one of the important factors contributing to the extinction of some amphibians in many places, including Africa and Australia. Thirdly, the factors that will affect the chytrid strain of chytrid fungus, the active degree and spread range of factors are temperature, human activities, amphibian’s own immunity, chemicals in pesticides, etc.

Non-random patterns of chytrid infections on phytoplankton host cells: mathematical and chemical ecology approaches

Host-parasite interactions between phytoplankton and fungi (chytrids) are key processes in aquatic ecosystems. However, individual-level heterogeneity in these interactions remains unexplored, although its importance in predicting the spread of diseases has been demonstrated in epidemiology. In this study, we experimentally tested whether individual-level heterogeneity could be a good indicator of phytoplankton-chytrid interactions, using a freshwater green algaStaurastrumsp., the diatomsUlnariasp. andFragilaria crotonensis, and chytrid fungi. The number of attached fungi per host cell showed a non-random clumped parasite distribution onUlnariasp. andF. crotonensis, but a random Poisson distribution onStaurastrumsp. To explore the potential mechanisms of these patterns, we developed a mathematical model describing sequential encounters between chytrid zoospores and host cells. The statistical fits of the model explained the parasite distributions forUlnariasp. andF. crotonensiswell, indicating that the clumped parasite distributions may result from an infection rate, increasing with the number of infections that already occurred on each host cell. Simultaneous analysis of volatile organic compounds (VOCs) from uninfected and infected host populations revealed that, among 13 VOCs detected, 6 components characterized the differences in VOC compositions between species and infection status. In particular, the level of beta-ionone, potentially acting against fungal activities, was significantly reduced in the presence of chytrid infection ofStaurastrumsp. These VOCs are targets for future studies, which potentially act as chemical signals influencing chytrid zoospores’ behaviors. The combination of mathematical and chemical analyses represents a promising approach to better understand the individual-level processes of phytoplankton-chytrid interactions.

Translocation does not influence amphibian chytrid fungus prevalence among wild eastern hellbenders Cryptobranchus alleganiensis.

Disease monitoring is an essential step in translocation projects, specifically in amphibians where emerging pathogens such as the chytrid fungus Batrachochytrium dendrobatidis (Bd) are linked to population declines. The eastern hellbender Cryptobranchus alleganiensis is a large, fully aquatic salamander experiencing precipitous range-wide population declines; however, the role Bd plays in these declines is unclear. To augment declining hellbender populations and determine effects of translocation on Bd prevalence, we conducted a translocation study of wild adult hellbenders from 2 source streams with abundant hellbender populations to 2 streams with declining populations in east Tennessee, USA. In 2018, we implanted radio transmitters into 30 hellbenders and sampled them periodically for Bd until 17 of the 30 hellbenders were translocated in 2019. We attempted to recapture translocated hellbenders approximately every 45 d for 3 mo to determine Bd prevalence post-release. We used qPCR to detect Bd and quantify zoospore loads on positive samples. Hellbenders had a pre-translocation Bd prevalence of 50% (15 of 30), which decreased to 10% (1 of 10) post-translocation. The average zoospore load for positive samples was 73.63 ± 30.82, and no hellbenders showed signs of chytridiomycosis throughout the study. Although we detected no significant effect of translocation on Bd prevalence, we observed a reduction in Bd prevalence post-release. Our results indicate that translocation did not lead to an increase in pathogen prevalence in translocated wild adult hellbenders, suggesting that chytrid did not impact the success of short-term translocations of eastern hellbenders in the Blue Ridge ecoregion.