Microbiome network analysis in skin and gills of Sparus aurata fed with Nannochloropsis gaditana microalgae

1416 and 694 ASVs in gills and skin respectively were preserved in the taxonomic analysis after filtering. The predominant phylum in gills was Proteobacteria (~50%) in the control and hydrolyzed groups and reached 70.58% in the raw diet. The Bacteroidota phylum was the most represented and Firmicutes and Actinobacteria were close to 3% in all treatments in this tissue. However, in all three different treatments, the phyla abundance was similar in skin samples, ~90% of which corresponded to Proteobacteria. Bateroidota (5.5%–5.9%), Firmicutes (1.8%–2%) and Actinobacteria (<1%) constitute the other phyla. In gills, 13 significantly higher ASVs were obtained in the control (such as Achromobacter, Acidobacter) versus 4 whose abundance was higher in the hydrolyzed diet. The number of ASVs that differed significantly between the gill microbiota in the control vs. raw group amounted to 70, most of them being ASVs corresponding to the genus Shewanella (43) higher in the raw diet. Nevertheless, in the skin samples, Control group showed a significant increase of abundance related to Acinetobacter, Achromobacter, Pseudomonas, Shewanella, Vibrio and Sphingomonas among others. In the hydrolyzed group, the most significant abundance was associated with the genus Pseudomonas, Vibrio, Pseudoalteromonas, Ralstonia o Cutibacterium. In the case of the skin raw samples, there were ASVs corresponding to the genus Acinetobacter, Streptococcus or Pseudoalteromonas that were significantly different respect of control. Taking ASV abundance matrix for each diet and tissue, 6 co-occurrence networks were constructed. In all treatments in gills, Acinetobacter was a central genus in the network, and exhibited a negative correlation with Polaribacter. Besides, in the raw group, Polaribacter also showed negative correlation with Aeromonas, Pseudomonas and Francisellacea. This work shows that Acinetobacter has a key role in the balance of mucosa microbiota and was in co-exclusion with Polaribacter.This work was funded by research projects for young researchers, CEIMAR 2019 (Evaluation of hydrolysates of Nannochloropsis gaditana for use in high value-added finishing feed for farmed gilthead sea bream (Sparus aurata) (NAN2BREAM))

Microbiota composition and intestinal integrity remain unaltered after the inclusion of hydrolysed Nannochloropsis gaditana in Sparus aurata diet

The use of lysed microalgae in the diet of carnivorous fish can increase the bioavailability of proteins and bioactive compounds, such as unsaturated fatty acids or vitamins in the digestive tract. These are essential molecules for the proper physiological development of fish in aquaculture. However, some antinutritional components and other undesirable molecules can be released from an excess of microalgae supplied, compromising the integrity of the intestine. The inclusion of small amounts of hydrolized microalgae in the fish diet can be a good strategy to avoid negative effects, improving the availability of beneficial compounds. Nannochloropsis gaditana is an interesting microalgae as it contains nutraceuticals. Previous studies reported beneficial effects after its inclusion in the diet of Sparus aurata, a widely cultured species in Europe and in all Mediterranean countries. However, administration of raw microalgae can produce intestinal inflammation, increased intestinal permeability, bacterial translocation and disturbance of digestion and absorption processes. The aim of this study was to evaluate changes in the intestinal microbiota and barrier stability of S. aurata fed with low inclusion (5%) hydrolysed N. gaditana. Intestinal microbiota was analyzed using Illumina MiSeq technology and libraries were constructed using variable regions V3-V4 of 16S rDNA molecules. Analysis were based in the identification, quantification and comparison of sequences. The predictive intestinal microbial functionality was analyzed with PICRUSt software. The results determined that the intestinal microbiota bacterial composition and the predictive intestinal microbiota functionality did not change statistically after the inclusion of N. gaditana on the diet. The study of gene expression showed that genes involved in intestinal permeability and integrity were not altered in fish treated with the experimental diet. The potential functionality and bacterial taxonomic composition of the intestinal microbiota, and the expression of integrity and permeability genes in the intestine of the carnivorous fish S. aurata were not affected by the inclusion of hydrolysed 5% N. gaditana microalgae