Isolation and Identification of Bacterial Strains from Skin of Terrestrial Amphibians

The microbiota is a vital element of higher organisms, with the cutaneous microbiota significantly contributing to the organism's defense against external agents. There is no standard method to sample Speleomantes (Amphibia: Plethodontidae) skin microbial communities for the microbial culturing approach. To address this, we have developed a practical protocol for isolating bacteria from the skin of individuals and a best practice for handling and storing samples. The steps are straightforward: swabs are stored in a physiological saline solution supplemented with glycerol upon sampling. Once transferred in the laboratory, swabs can be stored at +4 °C for a maximum of 10 days to ensure the integrity of the microbial community. This study offers interesting insights into the cutaneous microbiota investigation of terrestrial amphibian species, such as those belonging to the genus Speleomantes. This approach contributes to implementing, monitoring and conserving measures for endangered species and might improve the knowledge of microbiota's function in the health and ecology of amphibians. This protocol may also have broader significance for microbiome research in many wildlife conservation scenarios.