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Berberine (BBR) is known for its strong antioxidant, anti-inflammatory, and capacity to preserve intestinal microbiota balance in fish. This study aimed to investigate the protective effects of berberine against copper-induced toxicity in the intestine of freshwater grouper Acrossocheilus fasciatus. The experiment involved four groups: a control group, a Cu group exposed to 0.02 mg/L Cu 2+ , and two BBR groups fed with 100 or 400 mg/kg of berberine diets and exposed to the same Cu 2+ concentration. Three replicates of healthy fish (initial weight 1.56 ± 0.10 g) were subjected to their respective treatments for 30 days. Results showed that none of the treatments significantly affected the survival rate, final weight, weight gain, and feed intake (P > 0.05). However, supplementation with 100 and 400 mg/kg of BBR significantly lowered the antioxidant activities, and glutathione peroxidase (gpx) and superoxide dismutase (sod) expression levels, as well as reduced malondialdehyde (MDA) content caused by Cu 2+ exposure (P < 0.05). Berberine inclusion significantly downregulated proinflammatory factors NLR family pyrin domain containing 3 (nlrp3), interleukin 1 beta (il1β), interleukin 6 cytokine family signal transducer (il6st) but upregulated transforming growth factor beta 1 (tgfβ1) and heat shock 70 kDa protein (hsp70) expression. Moreover, berberine at both levels maintained the intestinal structural integrity and significantly improved gap junction gamma-1 (gjc1) mRNA level compared to the Cu group (P < 0.05). Based on 16S rDNA sequencing, the richness and diversity of intestinal microbiota in different groups were not significantly influenced. Berberine reduced the Firmicutes/Bacteroidota ratio and stifled the growth of some specific pathogenic bacteria such as Pseudomonas, Citrobacter, and Acinetobacter, while boosting the richness of potential probiotic bacteria, including Roseomonas and Reyranella compared with the Cu group. In conclusion, berberine showed significant protective effects against Cu 2+ -induced intestinal oxidative stress, inflammation response, and microbiota disturbance in freshwater grouper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Context: Leucine (Leu) is one of the branched-chain amino acids that is necessary for muscle protein synthesis. Basically, poultry feed ingredients contain abundant amounts of Leu. But little is known about dietary Leu deficiency in performance and health-attributing parameters of female broiler chickens, which may occur in crude protein-restricted diets. Aims: This study evaluates the effect of Leu deficiency in diets on the growth performance, body composition, immunity, and caecum microbiota of female broilers. Methods: Female broiler chicks (Cobb 500; 8 days old; n = 540) were randomly divided into six groups with six replicates (15 birds in each replicate) in a completely randomised design. The experimental diets included six concentrations of Leu (80%, 84%, 88%, 92%, 96%, and 100%) fed for 14 days. A basal Leu-deficient diet was formulated at the 80% recommended level, and the experimental diets were created by gradual replacing of crystalline L-glutamic acid with crystalline L-Leu. Average daily intake (ADI), average daily gain (ADG), feed efficiency (gain:feed), body composition, cutaneous basophil hypersensitivity (CBH) test to phytohaemagglutinin-P (PHA-P), antibody response to sheep red blood cell (SRBC) and caecum microbiota were investigated. Key results: The birds receiving the lowest Leu application level had the lowest ADI (P < 0.05) and a 20% higher fat content (P < 0.01) than did the birds fed 96% Leu. The population of Escherichia coli and coliform bacteria count was increased (P < 0.001; by 36% and 10.3% compared with the birds fed 100% Leu in the diet respectively), and Lactobacillus (but not Enterococcus) count was decreased (P < 0.001; by 26.4% compared with the birds fed 100% Leu in the diet) with decreasing concentrations of dietary Leu. Average daily gain, feed efficiency, carcass dry matter, protein and ash percentages, and immunity were not affected markedly by dietary treatments with different Leu concentrations. Conclusions: The Leu deficiency influenced the broiler performance, body composition, and caecum microbiota, but not the immune response. Implications: This study points to the importance of Leu supplementation in the diet of broilers to improve their growth performance, fat content, and intestine health. Poultry diets contain abundant amounts of leucine, a necessary feed ingredient for higher meat production. Leucine is one of the essential amino acids, whose deficiency impairs the broilers' performance, body composition, and caecum microbiota. This study points to the importance of leucine supplementation in the diet of broilers to improve their growth performance, fat content, and intestine health. [ABSTRACT FROM AUTHOR]

The present study aimed to investigate the effects of different inclusion levels of full-fat Hermetia illucens larvae (HI) meal as a protein source on the growth performance, intestinal morphology, health, and microbiota of largemouth bass. Five isonitrogenous and isolipidic diets were formulated by adding HI to replace 0 % (HI0, control), 10 % (HI10), 20 % (HI20), 30 % (HI30) and 40 % (HI40) of fish meal (FM) in diets. Each diet was randomly assigned to triplicate groups of 30 fish per aquarium. Fish were fed two times daily to apparent satiation for 80 days. The results indicated that weight gain rate, specific growth rate and protein efficiency ratio were not appreciably reduced until more than 30 % of the FM protein was replaced with HI. Fish fed HI40 diet had the lowest fatty acid (C18:3, C20:5, C22:6) contents (P < 0.05). Moreover, the villi height was decreased, villi width and goblet cell number in hindgut was increased in the HI40 group compared with the other groups, respectively. Meanwhile, serum diamine oxidase activity, D-lactate and endotoxin contents increased markedly as the proportion of HI increased (P < 0.05). Notably, the increased dietary HI levels down-regulated the mRNA expression levels of cldnd1b and ocel1 in intestine. Higher malondialdehyde content and down-regulated expression of sod and gpx1a in the intestine were also observed in the HI40 group. Accordingly, the increased dietary HI levels upregulated the expression of intestinal pro-inflammatory cytokines (tumor necrosis factor-α and interleukin-1β) and down-regulated anti-inflammatory cytokines (transcriptional growth factor-β1 and interleukin-10) (P < 0.05). In addition, the abundance of both intestinal Lactobacillus and Streptococcus increased significantly, while the abundance of Cetobacterium decreased in the HI20 group (P < 0.05). Broken-line model analysis based on weight gain rate against dietary HI replacement level indicated that the optimum replacement level was 26.79 %. However, including too high proportion of HI could induce intestinal dysbiosis, impair the intestine physical barrier, and finally reduced growth of Micropterus salmoides. • The dietary inclusion of 16.78 % full-fat H. illucens meal improved growth of largemouth bass. • The 25.06 % full-fat H. illucens meal inhibited the growth and damaged the gut health. • Full-fat H. illucens meal inclusion in diets affected the intestinal barrier function of fish. • The H. illucens meal addition modulated the gut microbiota. [ABSTRACT FROM AUTHOR]