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Chytridiomycosis, a disease induced by the chytrid fungi Batrachochytrium dendrobatidis (Bd) and B. salamandrivorans (Bsal), has strongly contributed to the ongoing worldwide amphibian conservation crisis. While Bd infection has caused amphibian declines for decades on several continents, Bsal is a novel threat to Central European salamanders and newts, being responsible for the collapse of Fire Salamander populations in the Netherlands, Belgium, and Germany. However, numerous other parasites causing harm to amphibians exist, yet have received much less attention than the chytrid fungi. The goal of the present study was to contribute to the understanding of declines of the Green Toad, Bufotes viridis, at its northwestern distribution border, in the area of Cologne, Germany. We combined the data from four years of Bd monitoring with a metabarcoding approach to detect other, mainly unicellular parasites, from amphibian feces and environmental samples, and also report results from Bsal testing in 2019. Skin swabs of approximately 280 amphibians were tested for Bd and 66 for Bsal, and 150 cloacal swabs and environmental samples from five sites were tested for other pathogens and parasites.We found Bd in all sampled locations with high prevalences and partly high individual infection loads but without clinical signs of chytridiomycosis. None of the samples tested for Bsal was positive for this pathogen. We further detected eight additional potential amphibian pathogens from fecal samples: three metamonads (Tritrichomonas augusta, Trichomitus batrachorum and Hexamita inflata), three ciliates (Balantidium duodeni, Nyctotherus cordiformis and N. hubeiensis), one stramenopile (Blastocystis sp.) and one metazoan (the nematode Rhabdias ranae). In the environmental samples, we detected OTUs of nine organisms potentially harmful for amphibians: Blastocystis sp., Hexamita inflata, Tritrichomonas augusta, Trichomitus batrachorum, two oomycetes (Leptolegnia sp., Saprolegnia sp.), two ichthyosporeans (Amphibiocystidium ranae, Anurofeca sp.) and the myxozoan Myxobolus sp. [ABSTRACT FROM AUTHOR]

Chytridiomycosis, a disease induced by the chytrid fungi Batrachochytrium dendrobatidis (Bd) and B. salamandrivorans (Bsal), has strongly contributed to the ongoing worldwide amphibian conservation crisis. While Bd infection has caused amphibian declines for decades on several continents, Bsal is a novel threat to Central European salamanders and newts, being responsible for the collapse of Fire Salamander populations in the Netherlands, Belgium, and Germany. However, numerous other parasites causing harm to amphibians exist, yet have received much less attention than the chytrid fungi. The goal of the present study was to contribute to the understanding of declines of the Green Toad, Bufotes viridis, at its northwestern distribution border, in the area of Cologne, Germany. We combined the data from four years of Bd monitoring with a metabarcoding approach to detect other, mainly unicellular parasites, from amphibian feces and environmental samples, and also report results from Bsal testing in 2019. Skin swabs of approximately 280 amphibians were tested for Bd and 66 for Bsal, and 150 cloacal swabs and environmental samples from five sites were tested for other pathogens and parasites. We found Bd in all sampled locations with high prevalences and partly high individual infection loads but without clinical signs of chytridiomycosis. None of the samples tested for Bsal was positive for this pathogen. We further detected eight additional potential amphibian pathogens from fecal samples: three metamonads (Tritrichomonas augusta, Trichomitus batrachorum and Hexamita inflata), three ciliates (Balantidium duodeni, Nyctotherus cordiformis and N. hubeiensis), one stramenopile (Blastocystis sp.) and one metazoan (the nematode Rhabdias ranae). In the environmental samples, we detected OTUs of nine organisms potentially harmful for amphibians: Blastocystis sp., Hexamita inflata, Tritrichomonas augusta, Trichomitus batrachorum, two oomycetes (Leptolegnia sp., Saprolegnia sp.), two ichthyosporeans (Amphibiocystidium ranae, Anurofeca sp.) and the myxozoan Myxobolus sp. [ABSTRACT FROM AUTHOR]

The Green Toad (Bufotes viridis) in Germany is mostly confined to secondary habitats and is experiencing severe population declines especially at its northwestern range boundary in the city of Cologne. As a basis for conservation management of this species, viz. to investigate the status of the population structure, we used a library of over 15,000 short tandem repeats to establish a new set of 12 microsatellite markers for this species. We applied these markers to B. viridis individuals from six sampling sites in Cologne, and included outgroup samples from sites in northern and central Germany. The results suggest the sampled Cologne toads form four distinct populations, without an obvious distinction between sites east and west of the river Rhine. Only the green toads of three neighbouring sampling sites clustered as one population, supporting the initial hypothesis that individuals from proximate sampling sites are genetically more similar than those from more isolated sampling sites. However, mitochondrial DNA sequences revealed a haplotype east of the Rhine that was absent west of the river. Toads from the northernmost sampled population in Cologne "Ginsterpfad" were not genetically depauperate, indicating that the severe declines at this site did not lead to a loss of genetic variation and were not related to inbreeding. Some Ginsterpfad individuals were genetically highly distinct from others, which may be explained by human introduction. Despite the encountered population structure, the low number of private alleles in the populations west of the Rhine, and the uniformity in mitochondrial haplotypes with the exception of one deviant allele east of the Rhine, lead us to recommend a pragmatic approach for conservation management. We suggest to consider all Cologne Green toads as belonging to a single management unit, but propose that, wherever possible, populations east and west of the Rhine should not be mixed in reintroduction measures. [ABSTRACT FROM AUTHOR]