Anti-inflammatory effects of aloe vera on soy meal-induced intestinal inflammation in zebrafish.

Soybean meal is one of the most promising alternatives to replace fishmeal in the aquaculture industry. However, its ingestion triggers an intestinal inflammatory process that compromises fish health and nutrition. Therefore, finding strategies that reduce the deleterious effects of a soy protein-based diet are relevant. In this work we analyzed the effects of an aloe vera (Aloe barbadensis miller, AV) extract on intestinal inflammation and innate immunity of zebrafish by adding it to the water and by supplementing it in a soybean meal-based diet. To search for potential immunomodulatory effects of AV, we tested its effectiveness in two inflammation assays and compared fish fed with either fishmeal or soybean meal-based feed supplemented with AV. Our results show a strong anti-inflammatory effect of AV. Furthermore, while soy-based meal strongly induces the expression of inflammation markers, supplementation with AV reverted this effect. Finally, we show that fish fed with a soy meal diet are highly susceptible to bacterial infection, but that this condition is significantly reduced when the soy meal is supplemented with AV. Our results suggest that AV is a good candidate to be incorporated as an additive in farmed fish diets to facilitate the replacement of fishmeal by soybean meal, maintaining intestinal health.

PCR protocol for detection of Vibrio ordalii by amplification of the vohB (hemolysin) gene.

Vibrio ordalii is the causative agent of atypical vibriosis and has the potential to cause severe losses in salmonid aquaculture. To prevent and control outbreaks, a rapid, reproducible, sensitive, and effective diagnostic method is needed. We evaluated a new conventional polymerase chain reaction (PCR) and real-time PCR (qPCR) protocol using a primer set (VohB_Fw-VohB_Rv) designed to amplify a 112 bp fragment flanking the vohB gene (coding for hemolysin production), against 24 V. ordalii strains isolated from different fish species, the V. ordalii type strain, and 42 representative related and unrelated bacterial species. The primer set was species-specific, recognizing all V. ordalii strains evaluated, with no cross-reaction with the other bacterial species. A sensitivity of 103 copies of the vohB gene was obtained with a standard curve. When the VohB_Fw-VohB_Rv qPCR protocol was applied to Atlantic salmon seeded tissues (kidney, liver, spleen, and muscle), the detection limit ranged from 5.27 × 102 to 4.13 × 103 V. ordalii CFU ml-1, i.e. 62 to 145 copies of the vohB gene, using the previously calculated standard curve. The conventional PCR also detected V. ordalii, but the total reaction time was 1 h longer. When the qPCR protocol was applied to naturally infected cage-cultured Atlantic salmon samples, 5 of 8 fish tested positive for V. ordalii, but only one of them was diagnosed as positive by direct cultivation on agar. We conclude that the PCR protocol evaluated is fast, specific, and sensitive enough to detect V. ordalii in infected tissues and is an important tool for secure diagnosis of atypical vibriosis, and is therefore helpful for the control of the disease through the prompt detection within fish populations.