7 formalin-inactivated bacteria·mL -1 , for 20 minutes. Vaccinated fish groups presented higher hematocrit, number of erythrocytes and leukocytes than the non-vaccinated group. Serum agglutination titer of intraperitoneally vaccinated fish was higher on both evaluation periods for the three bacteria strains. Only on day 21 post-vaccination fish from the oral and immersion vaccination groups presented higher serum agglutination titer than the non-vaccinated fish for A. hidrophyla and E. durans. Serum antimicrobial activity in vaccinated fish was higher for P. aeroginosa and E. coli than in non-vaccinated fish on both evaluation periods. The different vaccine administration routes stimulated hematological and immunological responses in Nile tilapia 21 days post-vaccination, but intraperitoneal vaccination presented higher total number of leukocytes, lymphocytes and serum agglutination titer.

This

New types of fish feed based on processed animal proteins (PAPs), insect meal, yeast, and microbial biomasses have been used with success in gilthead sea bream. However, some drawback effects on feed conversion and inflammatory systemic markers were reported in different degrees with PAP- and non-PAP-based feed formulations. Here, we focused on the effects of control and two experimental diets on gut mucosal-adherent microbiota, and how it correlated with host transcriptomics at the local (intestine) and systemic (liver and head kidney) levels. The use of tissue-specific PCR-arrays of 93 genes in total rendered 13, 12, and 9 differentially expressed (DE) genes in the intestine, liver, and head kidney, respectively. Illumina sequencing of gut microbiota yielded a mean of 125,350 reads per sample, assigned to 1,281 operational taxonomic unit (OTUs). Bacterial richness and alpha diversity were lower in fish fed with the PAP diet, and discriminant analysis displayed 135 OTUs driving the separation between groups with 43 taxa correlating with 27 DE genes. The highest expression of intestinal pcna and alpi was achieved in PAP fish with intermediate values in non-PAP, being the pro-inflammatory action of alpi associated with the presence of Psychrobacter piscatorii. The intestinal muc13 gene was down-regulated in non-PAP fish, with this gene being negatively correlated with anaerobic (Chloroflexi and Anoxybacillus) and metal-reducing (Pelosinus and Psychrosinus) bacteria. Other inflammatory markers (igm, il8, tnfα) were up-regulated in PAP fish, positively correlating the intestinal igm gene with the inflammasome activator Escherichia/Shigella, whereas the systemic expression of il8 and tnfα was negatively correlated with the Bacilli class in PAP fish and positively correlated with Paracoccus yeei in non-PAP fish. Overall changes in the expression pattern of il10, galectins (lgals1, lgals8), and toll-like receptors (tlr2, tlr5, tlr9) reinforced the anti-inflammatory profile of fish fed with the non-PAP diet, with these gene markers being associated with a wide range of OTUs. A gut microbiota-liver axis was also established, linking the microbial generation of short chain fatty acids with the fueling of scd1- and elovl6-mediated lipogenesis. In summary, by correlating the microbiome with host gene expression, we offer new insights in the evaluation of fish diets promoting gut and metabolism homeostasis, and ultimately, the health of farmed fish.

This study has demonstrated, for the first time, the normal microbiota in the A. gigas intestine during different life stages and the presence of LAB strains. It also demonstrated LAB antibiotic resistance and antagonistic behaviour against pathogens isolated from the same fish.

This study evaluated the efficiency of differently prepared vaccines against Aeromonas hydrophila in the hybrid surubim (Pseudoplatystoma corruscans 9 P. reticulatum). Survival and haematoimmunological parameters were compared between the treatments: non-vaccinated fish (C); bacterin-vaccinated fish (B); bacterin plus oral booster vaccinated fish (B+O); bacterin and toxoid-vaccinated fish (B+T) and bacterin, toxoid and oral booster-vaccinated fish (B+T+O). Fourteendays vaccinated fish from B+O and B+T+O were fed with an oral booster for 4 days. After 1 week, the fish were intraperitoneally challenged with 2 9 10 8 CFU mL À1 of A. hydrophila. Fish from the treatment B+T+O showed the lowest cumulative mortality (11.36%) 96 h after challenge, compared with other treatments (22.72-44.04%), and a relative survival of 74%. Serum immunoglobulin in B+T+O fish was higher than in other treatments. All vaccinated fish showed an increased agglutination titre when compared with non-vaccinated fish, both before and after challenge. Fish fed with oral booster showed an increase in phagocytic percentage before and after challenge. It can be inferred that the oral booster vaccination was efficient in reducing mortality in hybrid surubim by enhancing the response against haemorrhagic septicaemia due to A. hydrophila infection.

Dietary supplementation (0, 10, 20, 40, 60, 80 g kg−1) of a nucleotide source (NuPro®) was evaluated on growth, nutrient utilization, haemato‐immunological responses and resistance to experimental infection of Nile tilapia. Fish (2.63 ± 0.63 g) were fed experimental diets to satiation three times a day for 75 days. Feed intake increased linearly with increasing levels of NuPro® promoting higher weight gain (P < 0.05). Survival, feed conversion and protein retention were not affected. Following the growth trial, fish were challenged with Aeromonas hydrophila by intraperitoneal injection. Survival and innate immune responses (phagocytosis and lysozyme activity) were not significantly affected. However, among the haematological variables, thrombocyte, leucocyte and monocyte counts increased linearly (P < 0.05). The indirect measurement of digestibility in juvenile Nile tilapia (122.32 ± 11 g) indicated a linear decrease in protein (1.41%), dry matter (5.72%) and energy (4.10%) digestibility coefficients as dietary supplementation increased (P < 0.05). NuPro® showed to be a beneficial additive to be supplemented in diets for juvenile Nile tilapia, providing 32.8% increase in feed intake and 28.8% in weight gain, as well as in thrombocyte, leucocyte and monocyte counts in blood, possibly indicating a more efficient defence response in the highest dietary supplementation levels tested.

This study evaluated the effects of dietary supplementation with two symbiotics in the hybrid surubim (Pseudoplatystoma corruscans 9 P. reticulatum) prior to and after challenge with Aeromonas hydrophila. The fish were divided into unsupplemented fish, supplemented fish with Weissella cibaria and inulin, and supplemented fish with Lactobacillus plantarum and inulin. Twenty days after being fed symbiotics, the fish were sampled for immunological, haematological, and microbiological analysis pre-challenge. After this period, they were challenged with Aeromonas hydrophila, and after 96 h the fish were sampled for blood collection and survival. Fish fed W. cibaria and inulin from the pre-challenge showed the highest values of lysozyme activity. Fish fed L. plantarum and inulin presented a higher number of thrombocytes and granular leukocyte PAS + than the other fish. After challenge, fish fed both symbiotics presented an increased red blood cells number. Fish fed L. plantarum and inulin showed an increased total leucocyte count (WBC), neutrophils, and monocytes number as well as total immunoglobulin. The W. cibaria-and inulin-supplemented fish showed improved haematological and immunological parameters and reduced cumulative mortality after the challenge.

In Europe, turbot aquaculture has a high potential for sustainable production, but the low tolerance to fishmeal replacement in the diet represents a big issue. Therefore, this study investigated the effects of more sustainable feed formulations on growth and feed performance, as well as nutritional status of juvenile turbot in recirculating aquaculture systems. In a 16-week feeding trial with 20 g juvenile turbot, one control diet containing traditional fishmeal, fish oil and soy products and two experimental diets where 20% of the fishmeal was replaced either with processed animal proteins (PAP) or with terrestrial plant proteins (PLANT) were tested. Irrespective of diets, growth performance was similar between groups, whereas the feed performance was significantly reduced in fish of the PAP group compared to the control. Comparing growth, feed utilisation and biochemical parameters, the results indicate that the fish fed on PAP diet had the lowest performance. Fish fed the PLANT diet had similar feed utilisation compared to the control, whereas parameters of the nutritional status, such as condition factor, hepato-somatic index and glycogen content showed reduced levels after 16 weeks. These effects in biochemical parameters are within the physiological range and therefore not the cause of negative performance. Since growth was unaffected, the lower feed performance of fish that were fed the PAP formulation might be balanced by the cost efficient formulation in comparison to the commercial and the PLANT formulations. Present study highlights the suitability of alternative food formulation for farmed fish.