BackgroundThe constant increase of aquaculture production and wealthy seafood consumption has forced the industry to explore alternative and more sustainable raw aquafeed materials, and plant ingredients have been used to replace marine feedstuffs in many farmed fish. The objective of the present study was to assess whether plant-based diets can induce changes in the intestinal mucus proteome, gut autochthonous microbiota and disease susceptibility of fish, and whether these changes could be reversed by the addition of sodium butyrate to the diets. Three different trials were performed using the teleostean gilthead sea bream (Sparus aurata) as model. In a first preliminary short-term trial, fish were fed with the additive (0.8%) supplementing a basal diet with low vegetable inclusion (D1) and then challenged with a bacteria to detect possible effects on survival. In a second trial, fish were fed with diets with greater vegetable inclusion levels (D2, D3) and the long-term effect of sodium butyrate at a lower dose (0.4%) added to D3 (D4 diet) was tested on the intestinal proteome and microbiome. In a third trial, the long-term effectiveness of sodium butyrate (D4) to prevent disease outcome after an intestinal parasite (Enteromyxum leei) challenge was tested.ResultsThe results showed that opposed forces were driven by dietary plant ingredients and sodium butyrate supplementation in fish diet. On the one hand, vegetable diets induced high parasite infection levels that provoked drops in growth performance, decreased intestinal microbiota diversity, induced the dominance of the Photobacterium genus, as well as altered the gut mucosal proteome suggesting detrimental effects on intestinal function. On the other hand, butyrate addition slightly decreased cumulative mortality after bacterial challenge, avoided growth retardation in parasitized fish, increased intestinal microbiota diversity with a higher representation of butyrate-producing bacteria and reversed most vegetable diet-induced changes in the gut proteome.ConclusionsThis integrative work gives insights on the pleiotropic effects of a dietary additive on the restoration of intestinal homeostasis and disease resilience, using a multifaceted approach.Electronic supplementary materialThe online version of this article (10.1186/s40168-017-0390-3) contains supplementary material, which is available to authorized users.

Characterization of Vibrio damsela strains isolated from turbot Scophthalmus maximus in Spain Belen ~o u z ' , Jens L. ~a r s e n~, Bent ~i e l s e n~, Juan L. ~arja', Alicia E. ~o r a n z o '

Strains of Vibrio vuln~flcus biotype 2 were recovered from diseased European eels Angujlla anguilla during 2 epizootics These stralns were biochemically and serologically identical to the ATCC 33149 strain, which was lsolated from the Japanese eel Angujlla japonica. The isolates of biotype 2 from European eels were more virulent for elvers (LD50 < 10' cfu) than the Japanese strain used as control (LDS0 = 104 cfu). All strains of biotype 2 were able to adhere to human and fish cell lines and were also strongly cytotoxic. The extracellu1a1-products (ECPs) of the Japanese strain exhibited lipolytic, phospholipolytic, proteolytic, and haernolyt~c activities, whereas the ECPs of the Spanish isolates displayed only proteolytic activity. Strains of biotype 1 were recovered from tank water and healthy eels during the second outbreak. These isolates were biochemically similar to the type strain ATCC 27562, and serologically distinct from the isolates of biotype 2. Strains of biotype 1 produced proteases, haemolysins, and cytotoxins and showed specific adhesion to human cells, but they were non-pathogenic for eels. The strains of biotype 2 were more virulent for mice (LDS" = 10' to 106cfu) than the strains of biotype 1 (LDS(, = 10%fu), suggesting that this eel pathogen may also represent a health hazard problem for fish farmers.

The routine use of chemotherapy to control bacterial diseases in aquatic populations has resulted in the development and spread of antibiotic resistance. The inclusion of immunostimulants in fish diets (functional diets) is one of the main strategies to solve this threat. This study aimed to analyse the intestinal microbiota of cultured European sea bass (Dicentrarchus labrax) fed two functional diets applying pyrosequencing of PCR-amplified 16S rRNA gene. Quality-filtered reads were assigned to family and genus taxonomic levels using the Ribosomal Database Project classifier. The autochthonous intestinal microbiota of sea bass consisted of two dominant bacterial genera: Dysgonomonas (Bacteroidetes) and Ralstonia (Betaproteobacteria), but effects of diet on this dominance were observed. In fact, the genus Dysgonomonas significantly decreased in samples from fish fed functional diets, recovering control levels at the end of the study. However, Ralstonia proportion significantly raised in samples from fish fed diet C and maintained this high level along the study period. The developed protocol could be used to study the composition of bacterial communities in the fish intestine under different nutritional and environmental conditions and its impact on infection, immune system and general fitness of fish.

Vibrio vulnificus biotype 2 is classically considered an obligate eel pathogen. However, it has recently been associated with one human septicemic case. In this paper, the opportunistic behavior of this pathogen is discussed. The bacterium can survive alone in brackish water or attached to eel surfaces for at least 14 days. It is able to spread through water and infect healthy eels by using skin as a portal of entry. These results suggest that water and infected eels may act as reservoirs of infection. A capsule seems to be essential for waterborne infectivity, which would explain why cells recovered from naturally diseased eels give rise to pure cultures of opaque colonies. The spread of the disease is dependent on temperature and water salinity, thus suggesting a method to reduce the risk of epizootics and that of infection for humans.