Molecular analysis of bacterial communities and detection of potential pathogens in a recirculating aquaculture system for Scophthalmus maximus and Solea senegalensis

PLoS One. 2013 Nov 21;8(11):e80847. doi: 10.1371/journal.pone.0080847. eCollection 2013.

Abstract

The present study combined a DGGE and barcoded 16S rRNA pyrosequencing approach to assess bacterial composition in the water of a recirculating aquaculture system (RAS) with a shallow raceway system (SRS) for turbot (Scophthalmus maximus) and sole (Solea senegalensis). Barcoded pyrosequencing results were also used to determine the potential pathogen load in the RAS studied. Samples were collected from the water supply pipeline (Sup), fish production tanks (Pro), sedimentation filter (Sed), biofilter tank (Bio), and protein skimmer (Ozo; also used as an ozone reaction chamber) of twin RAS operating in parallel (one for each fish species). Our results revealed pronounced differences in bacterial community composition between turbot and sole RAS, suggesting that in the systems studied there is a strong species-specific effect on water bacterial communities. Proteobacteria was the most abundant phylum in the water supply and all RAS compartments. Other important taxonomic groups included the phylum Bacteriodetes. The saltwater supplied displayed a markedly lower richness and appeared to have very little influence on bacterial composition. The following potentially pathogenic species were detected: Photobacterium damselae in turbot (all compartments), Tenacibaculum discolor in turbot and sole (all compartments), Tenacibaculum soleae in turbot (all compartments) and sole (Pro, Sed and Bio), and Serratia marcescens in turbot (Sup, Sed, Bio and Ozo) and sole (only Sed) RAS. Despite the presence of these pathogens, no symptomatic fish were observed. Although we were able to identify potential pathogens, this approach should be employed with caution when monitoring aquaculture systems, as the required phylogenetic resolution for reliable identification of pathogens may not always be possible to achieve when employing 16S rRNA gene fragments.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aquaculture*
  • Bacteria / classification
  • Bacteria / genetics*
  • Denaturing Gradient Gel Electrophoresis
  • Fish Diseases / microbiology
  • Flatfishes / microbiology*
  • Microbiota / genetics*
  • Phylogeny
  • RNA, Ribosomal, 16S / genetics
  • Sequence Analysis, DNA

Substances

  • RNA, Ribosomal, 16S

Grants and funding

Patricia Martins benefited from a PhD grant (SFRH/BD/73889/2010) provided by the Portuguese FCT (Fundação para a Ciência e Tecnologia). This study has been carried out with the financial support of project AQUASAFE - Development of new technologies to anticipate and diagnose disease outbreaks in aquaculture (PROMAR 31-03-05-FEP-0016) (PROMAR, a Portuguese instrument for the sectors of fisheries and aquaculture funded by the European Fisheries Fund). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.