Presence Of Bd Research Articles

Analysis of Reproductive Strategies and Immunological Interactions in Batrachochytrium dendrobatidis-Resistant Japanese Tree Frogs

The amphibian chytrid fungus, Batrachochytrium dendrobatidis (Bd), has been implicated as an agent of acute declines in amphibian populations worldwide. East Asian amphibians have been coexisting with Bd for long periods and thus are considered resistant; among the many is the Japanese tree frog, Dryophytes japonicus. Our study focused Bd infection effects on reproductive behaviors and physiological parameters in D. japonicus as a function of better understanding the chronic effect of the disease on long-term population viability. During the peak breeding season, we captured 70 males and quantified the chorus size, calling behaviors, physiological states, innate immunity, and sperm quality of individuals. In a simple comparison, all parameters were not significantly different. However, in the NMDS analysis, we were able to confirm subtle trends in some items according to infection and correlations between several items. Importantly, sperm density and sperm vitality tend to increase with Bd infection load, suggesting increased reproductive effort following infection. Additionally, this analysis indicated that innate immunity was positively related to Bd infection intensities, indicating the activation of immunity upon infection. These findings indicate that Bd-resistant D. japonicus maintains reproductive capabilities and physio-logical stability despite Bd infection, likely due to a co-evolved immune system. The present work offers insight into how amphibian populations may have some endurance in the presence of Bd and points out the importance of studying resistant species as a means to understand long-term ecological effects. Our results suggest that resistance to Bd may not simply prevent cata-strophic declines but actively contribute to the dynamics of Bd prevalence in amphibian communities, and confer implications for conservation strategies.

Influence of Batrachochytrium dendrobatidis isolate and dose on infection outcomes in a critically endangered Australian amphibian

The amphibian chytrid fungus (Batrachochytrium dendrobatidis (Bd)) is a continuing threat globally, causing widespread declines. Corroboree frogs (Pseudophryne corroboree) are critically endangered and no longer have self-sustaining populations in the wild due to the presence of Bd in the environment. We aimed to investigate variation in susceptibility to Bd in juvenile P. corroboree and evaluate the impact of Bd isolate and dose on infection outcomes. We experimentally exposed juvenile P. corroboree to two Bd isolates at five doses, and also characterised the in vitro growth of these isolates. Frogs showed high susceptibility to Bd, with isolate and dose impacting survival time and infection loads. Additionally, differences in the in vitro phenotype of the two Bd isolates were linked to their differential virulence. This study highlights the considerable impact that fungal isolate and dose have in shaping disease outcomes in experimental exposures.

Reintroduction of resistant frogs facilitates landscape-scale recovery in the presence of a lethal fungal disease

Vast alteration of the biosphere by humans is causing a sixth mass extinction, driven in part by an increase in infectious diseases. The emergence of the lethal fungal pathogen Batrachochytrium dendrobatidis (Bd) has devastated global amphibian biodiversity. Given the lack of any broadly applicable methods to reverse these impacts, the future of many amphibians appears grim. The Sierra Nevada yellow-legged frog (Rana sierrae) is highly susceptible to Bd infection and most R. sierrae populations are extirpated following disease outbreaks. However, some populations persist and eventually recover, and frogs in these recovering populations have increased resistance against infection. Here, we conduct a 15-year reintroduction study and show that frogs collected from recovering populations and reintroduced to vacant habitats can reestablish populations despite the presence of Bd. In addition, the likelihood of establishment is influenced by site, cohort, and frog attributes. Results from viability modeling suggest that many reintroduced populations have a low probability of extinction over 50 years. These results provide a rare example of how reintroduction of resistant individuals can allow the landscape-scale recovery of disease-impacted species, and have broad implications for amphibians and other taxa that are threatened with extinction by novel pathogens.

Centrifugation is an effective and inexpensive way to determine Batrachochytrium dendrobatidis quantity in water samples with low turbidity

Batrachochytrium dendrobatidis (Bd) is a pathogenic chytrid fungus that is particularly lethal for amphibians. Bd can extirpate amphibian populations within a few weeks and remain in water in the absence of amphibian hosts. Most efforts to determine Bd presence and quantity in the field have focused on sampling hosts, but these data do not give us a direct reflection of the amount of Bd in the water, which are useful for parameterizing disease models, and are not effective when hosts are absent or difficult to sample. Current methods for screening Bd presence and quantity in water are time, resource, and money intensive. Here, we developed a streamlined method for detecting Bd in water with low turbidity (e.g., water samples from laboratory experiments and relatively clear pond water from a natural lentic system). We centrifuged water samples with known amounts of Bd to form a pellet and extracted the DNA from that pellet. This method was highly effective and the resulting concentrations across all tested treatments presented a highly linear relationship with the expected values. While the experimentally derived values were lower than the inoculation doses, the values were highly correlated and a conversion factor allows us to extrapolate the actual Bd concentration. This centrifuge-based method is effective, repeatable, and would greatly expand the domain of tractable questions to be explored in the field of Bd ecology. Importantly, this method increases equity in the field, because it is time- and cost-efficient and requires few resources.

Internalized Stigma in Patients with Bipolar Disorder: A Cross-sectional Study on Its Associations with Sociodemographic, Marital and Clinical Characteristics

IntroductionBipolar disorder (BD) is a chronic and complex affective disorder among top diseases that cause disability worldwide. Internalized stigmatization is a process including the awareness of negative stereotypes adopted by the society, participation in and internalization of these judgements, associated with impaired social functionality. Studies examining internalized stigma and related factors in BD is limited.ObjectivesIn this study, it is aimed to investigate the associations between internalized stigmatization and clinical characteristics, as well as sociodemographic and marital features of patients with BD.MethodsThis observational and cross-sectional study was conducted at a specialized affective disorders clinic in a university hospital between November 2020 and March 2021. During routine follow-up, each consecutive patient with BD was invited and a total of 118 were included in the study. Information about sociodemographic, marital and clinical characteristics of patients was collected through a prepared data form and follow-up documents. Internalized Stigma of Mental Illness Scale (ISMIS) was administered to assess internalized stigma. Statistical analysis of data was conducted by SPSS version 25 and a statistical significance level of p<0.05 was determined.ResultsMean ISMIS total score of the sample was 56.50 ±13.65. Multiple linear regression was used to test the predictors of higher ISMIS scores. Being currently unemployed (p=0.012, Β=0.208), shorter BD duration (p<0.001, Β=0.302) and presence of inter-episode residual symptoms (p=0.004, Β=0.248) predicted higher ISMIS total. Younger age (p=0.002, Β=0.264), being female (p=0.007, Β=0.226) and absence of mania dominance (p=0.019, Β=0.190) predicted higher alienation scores. Presence of inter-episode residual symptoms predicted both stereotype endorsement (p<0.001, Β=0.320) and perceived discrimination (p<0.001, Β=0.358). Younger age (p=0.001, Β=0.281) and total number of depressive episodes (p=0.015, Β=0.212) also predicted perceived discrimination. Shorter BD duration and absence of seasonality predicted higher ISMIS social withdrawal, while history of hospitalization predicted higher ISMIS stigma resistance.ConclusionsOur study demonstrated similar mean ISMIS total scores to the findings previously reported in Türkiye, while roughly lower than results in the international literature. Considering that internalized stigmatization was increased in earlier stages of BD and in younger patients, as well as in patients with inter-episode residual symptoms, it might be important to implement psychosocial interventions for internalized stigmatization and appropriate psychoeducation programs in the earlier periods of BD. Therefore a multidimensional and holistic approach towards internalized stigmatization may positively contribute to the functionality of patients with BD.Disclosure of InterestNone Declared

Gene functions of the Ambystoma altamirani skin microbiome vary across space and time but potential antifungal genes are widespread and prevalent.

Amphibian skin microbiomes can play a critical role in host survival against emerging diseases by protecting their host against pathogens. While a plethora of biotic and abiotic factors have been shown to influence the taxonomic diversity of amphibian skin microbiomes it remains unclear whether functional genomic diversity varies in response to temporal and environmental factors. Here we applied a metagenomic approach to evaluate whether seasonality, distinct elevations/sites, and pathogen presence influenced the functional genomic diversity of the A. altamirani skin microbiome. We obtained a gene catalogue of 92 107 nonredundant annotated genes and a set of 50 unique metagenome assembled genomes (MAGs). Our analysis showed that genes linked to general and potential antifungal traits significantly differed across seasons and sampling locations at different elevations. Moreover, we found that the functional genomic diversity of A. altamirani skin microbiome differed between B. dendrobatidis infected and not infected axolotls only during winter, suggesting an interaction between seasonality and pathogen infection. In addition, we identified the presence of genes and biosynthetic gene clusters (BGCs) linked to potential antifungal functions such as biofilm formation, quorum sensing, secretion systems, secondary metabolite biosynthesis, and chitin degradation. Interestingly genes linked to these potential antifungal traits were mainly identified in Burkholderiales and Chitinophagales MAGs. Overall, our results identified functional traits linked to potential antifungal functions in the A. altamirani skin microbiome regardless of variation in the functional diversity across seasons, elevations/sites, and pathogen presence. Our findings suggest that potential antifungal traits found in Burkholderiales and Chitinophagales taxa could be related to the capacity of A. altamirani to survive in the presence of Bd, although further experimental analyses are required to test this hypothesis.

FIRST RECORD OF BATRACHOCHYTRIUM DENDROBATIDIS IN THE NORTHERN NETHERLANDS.

Batrachochytrium dendrobatidis (Bd) infects amphibians and has been linked to the decline of hundreds of anuran amphibians all over the world. In the province of Groningen in the Netherlands, this fungal pathogen was not detected before this study. To determine whether Groningen was Bd-free, we surveyed 12 locations in this province in 2020 and 2021. Samples were then used to quantify the presence of Bd with a qPCR assay. In total, 2 out of 110 (∼0.02%) collected in 2020 and 11 out of 249 samples collected in 2021 tested positive for Bd. Infected amphibians were found in 4 out of the 12 sites, and the prevalence of Bd was estimated at 4% for both years combined. Our study provides the first record of Bd in Groningen, and we hypothesize that Bd is present throughout the Netherlands in regions currently considered "Bd-free." Furthermore, we warn scientists and policymakers to be apprehensive when calling a site free from Bd when sampling is limited or not recent.

Molecular detection of Batrachochytrium dendrobatidis (Chytridiomycota) and culturable skin bacteria associated with three critically endangered species of Atelopus (Anura: Bufonidae) in Ecuador.

Chytridiomycosis is a fungal disease responsible for massive amphibian die-offs worldwide, caused by the fungus Batrachochytrium dendrobatidis (Bd). Potential symbiotic relationships between frogs and the bacteria residing on their skin-referred to as skin-bacteria-may inhibit Bd growth, aiding in resistance to this lethal disease. This research had three main objectives: (1) to detect the presence of Bd in native populations of Atelopus balios, A. bomolochos, and A. nanay in the central Andes and coastal southern regions of Ecuador; (2) to identify the culturable skin-bacteria; and (3) to analyze differences among the bacterial communities in the three Atelopus species studied. Skin swabs were collected from two populations of A. balios (107-203 m a.s.l.) and one population each of A. bomolochos and A. nanay (3,064-3,800 m a.s.l.). These swabs served two purposes: first, to detect Bd using conventional PCR; and second, to isolate culturable bacteria, which were characterized through DNA sequencing, molecular phylogeny, and community composition similarity analysis (Jaccard index). Results showed that Bd was present in all species, with positive Bd PCR amplification found in 11 of the 12 sampled amphibians. The culturable skin-bacteria were classified into 10 genera: Pseudomonas (31.4%), Stenotrophomonas (14.3%), Acinetobacter (11.4%), Serratia (11.4%), Aeromonas (5.7%), Brucella (5.7%), Klebsiella (5.7%), Microbacterium (5.7%), Rhodococcus (5.7%), and Lelliottia (2.9%). The Jaccard index revealed that bacterial genera were least similar in A. bomolochos and A. balios (J=0.10), while the highest similarity at the genus level was between A. bomolochos and A. nanay (J=0.33). At the clade-species level, only A. bomolochos and A. nanay show common bacteria (J=0.13). Culturable bacterial communities of specimens diagnosed as Bd positive (n=10) or Bd negative (n=1) share a J value of 0.1 at genus and 0.04 at species-clade level. The prevalence of Bd and the composition of cutaneous bacteria could be influenced by Bd reservoirs, Atelopus biology, and intrinsic environmental conditions. This research contributes to understanding the relationship between endangered Andean species and Bd, and explores the potential use of native skin-bacteria as biocontrol agents against Bd.

Batrachochytrium dendrobatidis in the Arid and Thermally Extreme Sonoran Desert.

Batrachochytrium dendrobatidis (Bd), the causative agent of the devastating global amphibian disease chytridiomycosis, was not projected to threaten amphibians in hot and arid regions due to its sensitivity to heat and desiccation. However, Bd is being detected more frequently than ever in hot and arid regions of Australia and the USA, challenging our current understanding of the environmental tolerances of the pathogen under natural conditions. We surveyed for Bd in an extremely hot and arid portion of the Sonoran Desert, where the pathogen is not projected to occur, and related presence and prevalence of the pathogen to local environmental conditions. We collected eDNA samples from isolated desert water sites including six tinajas and 13 catchments in June and August of 2020 and swabbed a total of 281 anurans of three species (red-spotted toad Anaxyrus punctatus, Couch's spadefoot Scaphiopus couchii, and the Sonoran Desert toad Incillius alvarius) across five catchments and six tinajas from June to September of 2020. Overall, Bd occurred at 68.4% of sites, despite extreme heat and aridity routinely exceeding tolerances established in laboratory studies. Average summer maximum air and water temperatures were 40.7°C and 30.7°C, respectively, and sites received an average of just 16.9mm of precipitation throughout the summer monsoon season. Prevalence was low (5.7%) across species and life stage. Our results demonstrate that Bd is capable of persisting and infecting amphibians beyond its projected range, indicating a need to account for higher thermal tolerances when quantifying risk of Bd presence and infection.

OCCURRENCE OF PATHOGENIC CHYTRID FUNGI BATRACHOCHYTRIUM SALAMANDRIVORANS AND BATRACHOCHYTRIUM DENDROBATIDIS IN THE HONG KONG NEWT (PARAMESOTRITON HONGKONGENSIS) AND OTHER WILD AND IMPORTED AMPHIBIANS IN A SUBTROPICAL ASIAN REGION.

One of the major threats for the massive loss in global amphibian diversity is chytridiomycosis, caused by chytrid fungi Batrachochytrium dendrobatidis (Bd) and B. salamandrivorans (Bsal). Following its discovery in 2013, Bsal has emerged as a severe threat to the global survival of urodelans. In 2018, a study reported a high prevalence of Bsal (65.6%) in the Hong Kong newts (Paramesotriton hongkongensis, Near Threatened) of a southern China population adjacent to Hong Kong (HK). Uncertainty regarding the Bsal infection status of P. hongkongensis inhabiting HK raised deep concern over the risk of introducing Bsal from that population. We screened the skin swabs from wild individuals of P. hongkongensis, 15 sympatric amphibian species, and 16 imported amphibian species in HK for chytrids. We found that both Bsal and Bd occur in low prevalences in P. hongkongensis (Bsal 1.7%, 5/293; Bd 0.34%, 1/293), Hong Kong cascade frog, Amolops hongkongensis, family Ranidae (Bsal only, 5.26%, 1/19), and Asian common toad, Duttaphrynus melanostictus, family Bufonidae (Bsal only, 5.88%, 1/17), populations of HK, with infected individuals being asymptomatic, suggesting a potential role of these species as reservoirs of Bsal. Conversely, Bd, but not Bsal, was present on 13.2% (9/68) of imported amphibians, indicating a high chytrid introduction risk posed by international amphibian trade. Long-term surveillance of the presence of Bd and Bsal in wild and captive amphibians would be advisable, and we recommend that import and export of nonnative chytrid carriers should be prevented, especially to those regions with amphibian populations naïve to Bd and Bsal.

Ecological Barriers for an Amphibian Pathogen: A Narrow Ecological Niche for Batrachochytrium salamandrivorans in an Asian Chytrid Hotspot.

The chytrid fungal pathogens Batrachochytrium salamandrivorans (Bsal) and B. dendrobatidis (Bd) are driving amphibian extinctions and population declines worldwide. As their origins are believed to be in East/Southeast Asia, this region is crucial for understanding their ecology. However, Bsal screening is relatively limited in this region, particularly in hotspots where Bd lineage diversity is high. To address this gap, we conducted an extensive Bsal screening involving 1101 individuals from 36 amphibian species, spanning 17 natural locations and four captive facilities in the biodiversity-rich Guangxi Zhuang Autonomous Region (GAR). Our PCR assays yielded unexpected results, revealing the complete absence of Bsal in all tested samples including 51 individuals with Bd presence. To understand the potential distribution of Bsal, we created niche models, utilizing existing occurrence records from both Asia and Europe. These models estimated potential suitable habitats for Bsal largely in the northern and southwestern parts of the GAR. Although Bsal was absent in our samples, the niche models identified 10 study sites as being potentially suitable for this pathogen. Interestingly, out of these 10 sites, Bd was detected at 8. This suggests that Bsal and Bd could possibly co-exist in these habitats, if Bsal were present. Several factors seem to influence the distribution of Bsal in Asia, including variations in temperature, local caudate species diversity, elevation, and human population density. However, it is climate-related factors that hold the greatest significance, accounting for a notable 60% contribution. The models propose that the specific climatic conditions of arid regions, primarily seen in the GAR, play a major role in the distribution of Bsal. Considering the increased pathogenicity of Bsal at stable and cooler temperatures (10-15 °C), species-dependent variations, and the potential for seasonal Bd-Bsal interactions, we emphasize the importance of periodic monitoring for Bsal within its projected range in the GAR. Our study provides deeper insights into Bsal's ecological niche and the knowledge generated will facilitate conservation efforts in amphibian populations devastated by chytrid pathogens across other regions of the world.

Exploring the interaction between 1,4-butanediol and acid phosphatase using spectroscopic techniques and molecular modeling studies

The functional characteristics of enzymes greatly rely on their stability, which is a vital factor to be considered when utilizing these biomolecules in industrial settings. This study investigated the effects of 1,4-butanediol (BD) on the structure and activity of acid phosphatase (AP). To this end, an array of analytical techniques, such as circular dichroism (CD), thermal stability analysis, kinetic studies, fluorescence spectroscopy, molecular docking, and molecular dynamics (MD) simulations, were employed to elucidate the impact of BD on AP. The data gathered indicated that BD has a positive effect on enhancing the stability of AP. BD demonstrated its capability to bind to AP, leading to the quenching of AP's inherent fluorescence through the static quenching mechanism. The binding process was spontaneous, with van der Waals forces and hydrogen bonding serving as the primary interactions between BD and AP. The formation of the complex was also validated via UV–Vis. According to CD studies, the presence of BD induced alternations in the structure of AP. Further, docking studies showed the favored binding site of BD within the structure of AP. Besides, the kinetic parameters showed that BD could act as an activator for AP.

First report of chytridiomycosis in the Southern Yungas Andean forest: a threat to the endangered La Banderita marsupial frog Gastrotheca gracilis

This work reports the first record of Batrachochytrium dendrobatidis (Bd) infection in the endangered La Banderita marsupial frog Gastrotheca gracilis from the Southern Yungas Andean forest. We analysed swab samples from the oral discs of 20 tadpoles and histological sections from two post-metamorphic individuals. We found 60 % of the tadpoles to be infected, and the histological sections revealed the presence of zoosporangia of Bd in different maturation stages. The signs of infection confirm the presence of Bd, which may pose a threat to the endangered La Banderita marsupial frog populations. Keywords: conservation, Batrachochytrium dendrobatidis, chytridfungus infection, Hemiphractidae, tadpoles

Water Pollution Increases the Risk of Chytridiomycosis in Mexican Amphibians

Chytridiomycosis is affecting amphibians worldwide, causing the decline and extinction of several amphibian populations. The disease is caused by the fungus Batrachochytrium dendrobatidis (Bd), a multihost pathogen living in freshwater habitats. While several environmental factors have been associated with the prevalence of Bd and its virulence, the effects of water quality on the pathogen are not clear yet. Some evidence suggests that water pollution may reduce amphibians' immune response and increase prevalence of Bd. To explore this hypothesis, we analyzed the relationship between water quality and the presence of Bd by using spatial data mining of 150 geolocations of Bd in amphibians from 9 families where Bd positive specimens have been previously reported, and water quality in 4,202 lentic and lotic water bodies in Mexico from 2010 to 2021. Our model showed that in the 3 main families where Bd was recorded, its presence is high in locations with low water quality, i.e., water polluted likely contaminated with urban and industrial waste. Using this model, we inferred areas suitable for Bd in Mexico; mainly in poorly studied areas along the gulf and on the pacific slope. We further argue that actions to reduce water pollution should become an integral part of public policies to prevent the spread of Bd and protect amphibians from this deadly pathogen.

Pathogenic fungus causes density‐ and trait‐mediated trophic cascades in an aquatic community

AbstractPathogens can alter species composition and ecosystem function by causing direct and indirect effects on communities. Zoospores of the chytrid fungus Batrachochytrium dendrobatidis (hereafter, Bd), a pathogen implicated in worldwide amphibian declines, can be consumed by filter‐feeding zooplankton and can damage mouthparts of infected amphibian larvae. Consequently, we hypothesized that this pathogen would affect the abundance of zooplankton and survival and feeding abilities of larval amphibian hosts. In turn, this could affect the algal food resources of zooplankton and tadpoles, which can include phytoplankton and periphyton. We tested these hypotheses by manipulating the presence of western toad (Anaxyrus boreas) larvae and Bd in outdoor mesocosms and quantifying the densities of amphibians, zooplankton, phytoplankton, and periphyton. Bd infections reduced amphibian larval densities, but this only weakly benefitted periphyton (a density‐mediated indirect effect). After controlling for larval densities, mesocosms with Bd‐exposed larvae had significantly more periphyton biomass than mesocosms with larvae but no Bd, consistent with infection‐induced mouthpart damage reducing larval feeding rates on attached algae (a trait‐mediated indirect effect). In fact, the estimated trait‐mediated effect of Bd on periphyton biomass was larger than the density‐mediated effect. However, we did not find evidence that Bd exposure triggered a switch to filter‐feeding phytoplankton. The presence of Bd was associated with increased copepod abundance, consistent with zooplankton consuming chytrid zoospores. These results suggest that Bd has the potential to cause both trait‐ and density‐mediated indirect effects that can alter community composition and perhaps the primary productivity of freshwater ecosystems.

A Successful Reintroduction of Columbia Spotted Frog (Rana luteiventris) through Repatriation of Recently Hatched Larvae

The Columbia Spotted Frog (Rana luteiventris; CSF) is widely distributed across northwestern North America; however, declines in the southernmost populations, including those in Utah, have resulted in the consideration of this species for protection under the Endangered Species Act. In 1998, a conservation agreement and strategy for Utah's populations of CSF was developed and identified needed conservation actions, including range expansion. We repatriated CSF larvae during 2008–2010 from an extant population in the Provo River into Beaver Creek, a beaver-dominated stream, in the Weber River, a watershed where contemporary surveys indicate CSF have likely been extirpated. In 2011, CSF breeding was first observed in the repatriated population when 11 egg masses were documented in four depositional areas. After ten years of monitoring egg mass numbers, we consider the repatriation a success as this population continues to grow and expand with a high of 54 egg masses (2019) within ten depositional areas (2020). High quality habitat and the large number of CSF larvae transplanted likely attributed to the success of this repatriation. We used 1–2 day old CSF larvae (Gosner life stage 20–21) as the repatriation life stage that we believed would be the most likely to prevent the spread of amphibian chytrid fungus (Batrachochytrium dendrobatidis, Bd) from the known chytrid fungus positive donor site in the Provo River. Genetic testing of a robust sample (n = 59) of CSF tadpoles across the repatriation site in 2020 did not detect the presence of Bd, potentially indicating that we did not move Bd through the repatriation. Additional replicates would be required beyond this single experiment, however, to better determine if our transplant techniques are effective at preventing the spread of Bd.

Infectious disease threats to amphibians in Greece: new localities positive for Batrachochytrium dendrobatidis.

In the early 2000s, numerous cases of European amphibian population declines and mass die-offs started to emerge. Investigating those events led to the discovery that wild European amphibians were confronted with grave disease threats caused by introduced pathogens, namely the amphibian and the salamander chytrid fungi Batrachochytrium dendrobatidis (Bd) and B. salamandrivorans (Bsal) and ranaviruses. In Greece, Bd was previously documented among wild amphibian populations in 2 different locations and 3 different species. However, no disease-related mass declines or mortality events have been reported. In this work, we build upon previous findings with new, subsequently obtained data, resulting in a 225-sample dataset of 14 species from 17 different locations throughout Greece, in order to examine the occurrence status of all 3 pathogens responsible for emerging infectious diseases in European amphibians. No positive samples for Bsal or ranavirus were recorded in any location. We confirmed the presence of Bd in 4 more localities and in 4 more species, including 1 urodelan (Macedonian crested newt Triturus macedonicus) and 1 introduced anuran (American bullfrog Lithobates catesbeianus). All insular localities were negative for Bd, except for Crete, where Bd was identified in 2 different locations. Again, no mass declines or die-offs were recorded in any Bd-positive area or elsewhere. However, given the persistence of Bd across Greece over the past ~20 yr, monitoring efforts should continue, and ideally be further expanded.

Fatal chytridiomycosis in the Moroccan midwife toad Alytes maurus and potential distribution of Batrachochytrium dendrobatidis across Morocco

ABSTRACT Multiple threats, including emerging infectious diseases, are contributing to the extinction of amphibians worldwide. One of the most devastating diseases is the fatal amphibian skin disease chytridiomycosis caused by the fungus Batrachochytrium dendrobatidis (Bd). The presence of Bd in North Africa was described in 2011 and this included the distribution range of the endemic Moroccan midwife toad (Alytes maurus). Here we report new Bd positive occurrences across several distant regions of Morocco, augment the known number of infected species, and describe the first evidence of lethal chytridiomycosis in A. maurus. Although population declines in this species were not recorded, the family Alytidae has been identified as the most susceptible taxonomic group to chytridiomycosis of the Palearctic. An environmental niche model, taking into account new records of Bd in the country, confirms the Mediterranean coast and the Rif and Middle Atlas Mountains as very favourable areas for Bd. Our results suggest that the real impact of chytridiomycosis in North Africa is poorly understood, and that this continent cannot be identified as a region less impacted by chytridiomycosis, as was previously proposed.

Emerging infectious diseases of amphibians in Poland: distribution and environmental drivers.

Emerging infectious diseases are a threat to biodiversity and have taken a large toll on amphibian populations worldwide. The chytrid fungi Batrachochytrium dendrobatidis (Bd) and B. salamandrivorans (Bsal), and the iridovirus Ranavirus (Rv), are of concern as all have contributed to amphibian declines. In central and eastern Europe, their geographical and host distributions and main environmental drivers determining prevalence are poorly known. We screened over 1000 amphibians from natural and captive populations in Poland for the presence of Bd, Bsal and Rv. In wild amphibian populations, we found that Bd is widespread, present in 46 out of 115 sampled localities as well as 2 captive colonies, and relatively common with overall prevalence at 14.4% in 9 species. We found lower prevalence of Rv at 2.4%, present in 11 out of 92 sampling sites, with a taxonomic breadth of 8 different amphibian species. Bsal infection was not detected in any individuals. In natural populations, Pelophylax esculentus and Bombina variegata accounted for 75% of all Bd infections, suggesting a major role for these 2 species as pathogen reservoirs in Central European freshwater habitats. General linear models showed that climatic as well as landscape features are associated with Bd infection in Poland. We found that higher average annual temperature constrains Bd infection, while landscapes with numerous water bodies or artificial elements (a surrogate for urbanization) increase the chances of infection. Our results show that a combination of climatic and landscape variables may drive regional and local pathogen emergence.

INFECTION DYNAMICS OF BATRACHOCHYTRIUM DENDROBATIDIS IN TWO FROG SPECIES INHABITING QUITO'S METROPOLITAN GUANGÜILTAGUA PARK, ECUADOR

Batrachochytrium dendrobatidis (Bd) infection is one of the principal causes of amphibian declines worldwide. The presence of Bd has been determined in Gastrotheca riobambae tadpoles that inhabit ponds in Quito's Metropolitan Guangüiltagua Park, Ecuador. This study sought to determine whether these tadpoles are infected and to determine the presence of chytridiomycosis in another frog species, Pristimantis unistrigatus, which also inhabits the park and has different reproductive biology and distinct behavioral habits. We used end-point and real-time PCR techniques to detect and quantify Bd infection. At 1 yr, samples were taken from the skin of P. unistrigatus using swabs and were also taken from the mouthparts of G. riobambae tadpoles. It was found that the two species were infected with a Bd prevalence of 39% (53/135) in G. riobambae tadpoles and 15% (57/382) in P. unistrigatus frogs. The two types of samples (tissue and swabs) from mouthparts showed differences in the zoospores per microliter loads (x̄=1,376.7±3,450.2 vs. x̄=285.0±652.3). Moreover, a correlation (r2=0.621) was discovered between the monthly mean maximum temperature of the pond with disease prevalence in G. riobambae tadpoles. Infection levels in the P. unistrigatus population varied significantly over time, and distance to the pond was a determinant factor for infection intensity.