Skin Microbiota of Salmonids: A Procedure to Examine Active Bacterial Populations Using an RNA-Based Approach

Fish microbiota studies have mostly addressed intestinal bacterial communities because of their role in fish physiology; however, the skin microbiota has been studied less despite its role as the first pathogen barrier in the host. DNA-based molecular techniques have contributed to improving our understanding of the skin microbiota, but this approach faces challenges, such as the low count of bacteria in healthy fish skin. To overcome this limitation, an RNA-based approach was developed to study the skin microbiota in salmonids, including tissue sampling, RNA extraction, and downstream procedures to obtain PCR amplicons for next-generation sequencing. The protocol originated in this work overcomes the limitations of low bacterial counts and is useful for describing active microbiota in fish skin. The application of the protocol to salmonids reared in both an experimental setting and on a farm revealed that the trout skin microbiota was dominated by bacteria from the phylum Proteobacteria (>65%). At the genus level, Piscirickettsia (46%) was highlighted as the most abundant in the experimental unit samples; in contrast, Pseudoalteromonas (26%), followed by Escherichia_Shigella (~25%), was the most abundant in farmed trout.