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Organic acids (OA) and nature-identical compounds (NIC) such as monoterpenes and aldehydes are well-known growth and health promoters in terrestrial livestock while their application for fish production is recent and their mechanisms of action require further study. Hence, this study tested the increasing dietary level (D0, D250, D500, D1000; 0, 250, 500 and 1000 mg kg feed−1 respectively) of a microencapsulated blend containing citric and sorbic acid, thymol and vanillin over 82 days on rainbow trout to assess the effects on growth, feed utilization, intestine cytokine gene expression and gut microbiota (GM). Furthermore, the effects on intestinal cytokine gene expression and GM were also explored after one week at high water temperature (23 °C). OA and NIC improved specific growth rate (SGR) and feed conversion rate (FCR) during the second half (day 40–82) of the feeding trial, while at the end of the trial protein (PER) and lipid efficiency (LER) increased with increasing dietary level. GM diversity and composition and cytokine gene expression analysis showed no significant differences in fish fed with increasing doses of OA and NIC (82 days) demonstrating the absence of inflammatory activity in the intestinal mucosa. Although there were no statistical differences, GM structure showed a tendency in clustering D0 group separately from the other dietary groups and a trend towards reduction of Streptococcus spp. was observed in the D250 and D1000 groups. After exposure to high water temperature, lower GM diversity and increased gene expression of inflammatory intestinal cytokines were observed for both inclusions (D0 vs. D1000) compared to groups in standard condition. However, the gene up-regulation involved a limited number of cytokines showing the absence of a substantial inflammation process able to compromise the functional activity of the intestine. Despite further study should be conducted to fully clarify this mechanism, cytokines up-regulation seems to be concomitant to the reduction of the GM diversity and, particularly, to the reduction of specific lactic acid bacteria such as Leuconostoc. The application of the microencapsulate blend tested can be a useful strategy to improve growth and feed utilization in rainbow trout under normal temperature conditions. According to the results organic acids and nature-identical compounds did not revert the effects triggered by the increased temperature of water.

To provide practical feeding management guidelines preceding a stressful episode during farming practices, European sea bass juveniles (initial weight: 72.3 g) were fed for 60-days different fish meal (FM) and fish oil (FO) dietary levels [high (30% FM, 15% FO, FM30/FO15), intermediate (20% FM, 7% FO, FM20/FO7), and low (10% FM, 3% FO, FM10/FO3)] in triplicate conditions. Fish were then fasted for 36 h and exposed to a 2-h acute crowding (80 kg m–3 biomass). Plasma biochemistry, skin mucus parameters and gene expression of stress and immune-related genes were performed before, at 2 and 24 h after crowding. At the end of the trial, the FM10/FO3 group showed lower final body weight, weight gain, and specific growth rate compared to the other treatments. Most of the plasma parameters were mainly affected by crowding condition rather than diet; however, after stress, lactate was higher in the FM30/FO15 group compared to the other treatments. Similarly, protease, antiprotease, peroxidase and lysozyme in skin mucus were mostly affected by crowding conditions, while fish fed FM10/FO3 displayed higher skin mucosal IgM and bactericidal activity against Vibrio anguillarum and V. harveyi. Most of the stress-related genes considered (hsp70 and gr-1 in the brain; hsp70, gr-1 and gr-2 in the head kidney), showed an overall expression pattern that increased over time after stress, in addition, hsp70 in the head kidney was also up-regulated in fish fed FM30/FO15 after stress. Higher plasmatic lactate together with the up-regulation of some stress-related transcripts suggest a higher reactivity to acute crowding of the stress-response mechanism in fish fed high FM and FO dietary levels. Otherwise, the higher skin mucosal IgM and bactericidal activity observed in fish fed FM10/FO3 dietary levels seems to indicate that acute crowding was able to activate a higher pro-inflammatory response in this treatment. Overall, the results of the present study seem to indicate that 10% FM and 3% FO dietary levels might affect stress and immune responses.

A 121-day feeding trial was undertaken to test the effects of two dietary lipid levels (16 and 21% L16, L21) in triplicated gilthead sea bream groups (initial weight: 67.5 g) reared at two different water temperatures (high, H 23°C and low, L 17°C) in the same recirculation system but exposed to a switch in temperature after 58 days. Fish kept at H were transferred to L (HL transition, autumn shift), and the fish kept at L were exposed to H (LH transition, summer shift), while continuing to receive the same diet to apparent satiation in each group. At the end of the trial, no significant diet effect on specific growth rate (SGR), feed intake (FI), and feed conversion rate (FCR) were detected in fish exposed to HL transition compared with those exposed to LH transition, while gross lipid efficiency (GLE) and lipid efficiency ratio (LER) were higher in L16. After temperature changes, L16 displayed higher SGR, FI, GLE, and LER, while mesenteric fat index was reduced. After temperature changes, the combined effects of low lipid diet and low temperature conditions resulted in higher pepsin activity, while trypsin, chymotrypsin, and lipase activities were generally higher at high lipid content. The combined effect of diet and temperature did not alter the metabolic plasma profile, except for the observed final higher aspartate aminotransferase (AST) and alkaline phosphatase (ALP) values when combining high dietary lipid (L21) and temperature changes. Different diets showed a significantly different gut microbiome layout, only at high temperature with L16 diet resulting in a higher load of Lactobacillus. On the contrary, no dietary impact on ecosystem diversity was observed, independently from the temperature. In addition, L16 diet in the HL transition favored an increase in Weissella and Bradyrhizobium genera in the gut microbiome, while in the final condition of LH transition, L21 diet favored a significant increase in Streptococcus and Bacillus. According to the results, the utilization of 16% dietary lipid levels in gilthead sea bream should be preferred during seasonal temperature changes in order to optimize feed utilization and gut health., This research was undertaken under the MedAID (Mediterranean Aquaculture Integrated Development) project, which has received funding from the European Union’s Horizon 2020 Research and Innovation Programme, Call H2020-SFS-23-2016, Grant agreement no. 727315 (http://www.medaid-h2020.eu/). MY and NG received support from the Spanish Ministry of Science, Innovation and Universities (MCIU), the State Research Agency (AEI), the European Regional Development Fund (FEDER/ERDF), and project Thermodigest (RTI2018-096134-B-I00) granted to MY.

A 71-day study was conducted to explore the effect of increasing dietary levels (0, 250, 500, 1000 mg kg feed−1; D0, D250, D500 and D1000, respectively) of a blend of microencapsulated organic acids (OA, specifically citric and sorbic acid) and nature identical compounds (NIC, specifically thymol and vanillin), on growth, intestinal immune parameters and gut microbiota (GM) of European sea bass juveniles reared under normal and subsequently suboptimal environmental conditions (high temperature, 30.0 ± 0.4 °C and low oxygen, 4.6 ± 0.6 mg L−1). OA and NIC did not promote growth, feed utilisation and feed intake at the inclusion tested but induced a significantly upregulation of IL-8, IL-10 and TGFβ. GM analyzed by next-generation sequencing showed that OA and NIC were able to exert prebiotic properties stimulating the development of beneficial bacteria taxa such as Lactobacillus, Leuconostoc, and Bacillus sp. Picrust analyses displayed a significant potential functional reconfiguration of GM promoting a decrease in inflammation-promoting and homeostatic functions at increasing OA and NIC administration. For the first time on this species the exposure to suboptimal rearing conditions was able to modify GM structure reducing LAB and increasing Proteobacteria, findings which were consistent with the inflammatory process observed at mRNA level.

A 9-week feeding trial was performed to evaluate the effects of fishmeal (FM) replacement by a mixture of plant proteins (PP), consisted of wheat gluten, soybean meal and soy protein concentrate, on the growth performance and welfare of turbot juveniles (initial weight 9.7 ± 0.2 g). Four isonitrogenous and isolipidic diets contained FM at 500 g kg−1 (FM50), 350 g kg−1 (FM35), 200 g kg−1 (FM20) and 50 g kg−1 (FM5). A decreased feed intake and an increased feed conversion rate was observed in FM5 group. Specific growth rate was significantly reduced in FM20 and FM5 groups, whereas protein and lipid utilization and proximate whole body composition were significantly different in FM5 group. Serum cortisol significantly increased in FM20 and FM5 groups whereas cholesterol, triglycerides, NEFA, total protein and urea concentrations significantly decreased. Serum lysozyme and blood phagocytes increased in FM20 and FM5 groups. FM35 ensured growth close to FM50, without significant effects on health and welfare of animals. FM20 and FM5 groups displayed reduced growth, metabolic stress and an immune response with effects on health and welfare. Results highlighted the consistency between growth performance and welfare status, suggesting the usefulness of their combined assessment for evaluating the suitability of PP and to improve dietary formulation for turbot.

The present study was undertaken in order to determine the effect of a dietary incorporation of soy bean meal (SBM, trypsin inhibitor activity 1.6 mg/g) on growth performance, nutrient utilization and gut histology of Egyptian sole Solea aegyptiaca. This species, which is genetically and morphologically close to Solea solea, is currently present in the Mediterranean Sea and reared in some Italian farms, using the same techniques for Solea solea production and it could represent a promising new species for Mediterranean aquaculture. Three isoproteic and isolipidic extruded diets (crude protein, 47%, crude fat, 20%) were formulated containing 0 (diet 1), 18 (diet 2) and 30% SBM (diet 3). 540 juveniles (initial body weight of 6.9±0.2g) were equally distributed into nine 150 litres squared tanks (bottom surface: 520 cm2) connected with a recirculation system (temperature 20±1 °C and dissolved oxygen above 5 ppm). Animals have been hand-fed twice a day (at 9.00 a.m. and 5.00 p.m.) at a daily ratio of 1.5% body weight-1. Diets were tested in triplicate for 87 days. The fish were weighed at days 28, 57 and 87, and they were sampled at days 0, 57 and 87 for nutrient retention determination. One-way ANOVA with Newmann – Keuls’ post test were used to analyze data (P≤0.05). No differences in terms of palatability were observed and any diet has been consumed completely all through the experiment. At day 57, fish fed diet 2 (18%SBM) showed a statistically higher growth and nutrient utilization and a lower feed conversion rate than fish fed the diet 1 (fish meal based diet) and the diet 3 (30%SBM). At day 87, fish fed diet 1, 2 and 3 reached weights of 25.8±0.8g, 27.5±0.9g and 26.1±1.5g, respectively and no differences in performances or nutrient retention were found. Histopathological gut examinations have revealed no noticeable differences in the appearances of the intestines between any of diet groups. Intestinal mucosal cells presented well vacuolated upper, with well defined microvillar brush borders. On the basis of the results of this trial, SBM seems to be a good protein source for Egyptian sole and can be added in the diet up to 30% without any reduction in growth rate and minimal effect on gut histology.