The microbiota is an essential component of higher organisms, and the cutaneous microbiota plays a crucial role in defending the organism from external agents. Within the scope of the project FFORIC24.35 we aim to develop the first study characterizing the microbiota of European plethodontid salamanders (genus Speleomantes), a group of eight strictly protected species endemic or sub-endemic to Italy. Given the lack of universally applied protocols in the literature, the primary objective of our project is to develop a best practice for sampling and preserving cutaneous bacterial samples. We used as models 12 individuals of the Italian cave salamander (S. italicus) from an epigean population located in Abruzzo. Samples were collected using swabs moistened with saline solution (0.9% NaCl), and immediately stored in saline solution with and without glycerol supplementation. Upon transfer to the laboratory, sample preservation was tested under various conditions: uncontrolled room temperature, +4°C, and –20°C for 10 days. Cultivable bacterial communities were compared using culture-based methods on generic and semi-selective media (LB, TSYE, TSA, NA). Culture-based analyses showed significant differences in microbial densities (CFU/mL) across media types and preservation conditions. Results revealed that swabs collected with physiological saline solution supplemented with glycerol and preserved at +4°C represented the protocol that allowed the maintenance of the highest integrity of microbial community. This study provides critical insights into the development of protocols for studying cutaneous microbiota in endangered amphibian species like Speleomantes. The established protocol can aid in ongoing monitoring efforts, contribute to conservation strategies, and enhance our understanding of the role of microbiota in amphibian health and ecology. Additionally, the findings may have broader implications for microbiota research in other wildlife conservation contexts.
Development of a sampling and preservation protocol for cutaneous bacterial communities of Speleomantes
Beatrice Farda;Luca Coppari;Enrico Lunghi;Marika Pellegrini
2024-01-01
Abstract
The microbiota is an essential component of higher organisms, and the cutaneous microbiota plays a crucial role in defending the organism from external agents. Within the scope of the project FFORIC24.35 we aim to develop the first study characterizing the microbiota of European plethodontid salamanders (genus Speleomantes), a group of eight strictly protected species endemic or sub-endemic to Italy. Given the lack of universally applied protocols in the literature, the primary objective of our project is to develop a best practice for sampling and preserving cutaneous bacterial samples. We used as models 12 individuals of the Italian cave salamander (S. italicus) from an epigean population located in Abruzzo. Samples were collected using swabs moistened with saline solution (0.9% NaCl), and immediately stored in saline solution with and without glycerol supplementation. Upon transfer to the laboratory, sample preservation was tested under various conditions: uncontrolled room temperature, +4°C, and –20°C for 10 days. Cultivable bacterial communities were compared using culture-based methods on generic and semi-selective media (LB, TSYE, TSA, NA). Culture-based analyses showed significant differences in microbial densities (CFU/mL) across media types and preservation conditions. Results revealed that swabs collected with physiological saline solution supplemented with glycerol and preserved at +4°C represented the protocol that allowed the maintenance of the highest integrity of microbial community. This study provides critical insights into the development of protocols for studying cutaneous microbiota in endangered amphibian species like Speleomantes. The established protocol can aid in ongoing monitoring efforts, contribute to conservation strategies, and enhance our understanding of the role of microbiota in amphibian health and ecology. Additionally, the findings may have broader implications for microbiota research in other wildlife conservation contexts.Pubblicazioni consigliate
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