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Bacterial pathogens destroy the structural integrity of functional organs in fish, leading to severe challenges in the aquaculture industry. Vitamin D3(VD3) prevents bacterial infections and strengthens immune system function via vitamin D receptor (VDR). However, the correlation between VD3/VDR and the structural integrity of functional organs remains unclarified. This study aimed to investigate the influence of VD3supplementation on histological characteristics, apoptosis, and tight junction characteristics in fish intestine during pathogen infection. A total of 540 healthy grass carp (257.24 ± 0.63 g) were fed different levels of VD3(15.2, 364.3, 782.5, 1,167.9, 1,573.8, and 1,980.1 IU/kg) for 70 d. Subsequently, fish were challenged with Aeromonas hydrophila, a pathogen that causes intestinal inflammation. Our present study demonstrated that optimal supplementation with VD3(1) alleviated intestinal structural damage, and inhibited oxidative damage by reducing levels of oxidative stress biomarkers; (2) attenuated excessive apoptosis-related death receptor and mitochondrial pathway processes in relation to p38 mitogen-activated protein kinase signaling (P < 0.05); (3) enhanced tight junction protein expression by inhibiting myosin light chain kinase signaling (P < 0.05); and (4) elevated VDR isoform expression in fish intestine (P < 0.05). Overall, the results demonstrated that VD3alleviates oxidative injury, apoptosis, and the destruction of tight junction protein under pathogenic infection, thereby strengthening pathogen defenses in the intestine. This finding supports the rationale for VD3intervention as an essential practice in sustainable aquaculture.

The present study explored the impacts of Ile on muscle fatty acid and amino acid profiles, lipid metabolism, and autophagy in hybrid catfish. Seven isonitrogenous (387.8 g/kg protein) semi-purified diets were formulated to contain 5.0 (control), 7.5, 10.0, 12.5, 15.0, 17.5, and 20.0 g Ile/kg diet respectively. The fish (initial weight of 33.11 ± 0.09 g) were randomly assigned to 7 groups for a 56-day trial. Each group has 3 replicates with 30 fish per replicate, fed at 08:00 and 18:00 each day. Results showed that muscle protein and lipid, C14:0, C18:0, C22:0, C14:1, C18:1n-9, polyunsaturated fatty acid (PUFA), Arg, Ile, Ala, Cys, Gly, Tyr, essential amino acid (EAA), and total amino acid (TAA) contents and flavor amino acid (FAA)/TAA in muscle had positive linear and/or quadratic responses to dietary Ile levels (P < 0.05). Fatty acid synthase (FAS), stearoyl-CoA desaturase (SCD), acetyl-CoA carboxylase (ACC), and lipoprotein lipase (LPL) activities had positive linear and/or quadratic responses, but carnitine palmitoyl transferase 1 (CPT1) activity had a negative response with increasing dietary Ile levels (P < 0.05). The mRNA expressions of FAS, SCD, ACC, LPL, fatty acid binding protein 4 (FABP4), FATP1, sterol response element-binding protein 1c (SREBP-1c), sequestosome 1 (SQSTM1), and adenosine 5′-monophosphate-activated protein kinase (AMPK) had positive linear and/or quadratic responses to dietary Ile levels (P < 0.05). The mRNA expressions of hormone-sensitive lipase (HSL), CPT1, peroxisome proliferator-activated receptor α (PPARα), PPARγ, uncoordinated 51-like kinase 1 (ULK1), beclin1 (Becn1), autophagy-related protein 9α (Atg9α), Atg4b, Atg7, autophagy marker light chain 3 B (LC3B), and SQSTM1in muscle had negative linear and/or quadratic responses to dietary Ile levels (P < 0.05). The p-AMPK and ULK1 protein levels, and p-AMPK/AMPK were decreased by 12.5 g Ile/kg in the diet (P < 0.05). Finally, SQSTM1 protein level had the opposite effect (P < 0.05). The above results indicate that dietary Ile improves fish muscle fatty acid and amino acid profiles potentially via respectively regulating lipid metabolism and autophagy. The Ile requirement of hybrid catfish (33 to 72 g) were estimated to be 12.63, 13.77, 13.75, 11.45, 10.50, 12.53 and 12.21 g/kg diet based on the regression analysis of protein, lipid, SFA, PUFA, FAA, EAA, and TAA muscle contents, respectively.

Vitamin D3(VD3), an essential nutrient for animals, has been demonstrated to stimulate the uptake of certain amino acids. However, the role of VD3in the intestine, the primary site for digestion and absorption of nutrients, remains poorly characterized. Here, the grass carp (Ctenopharyngodon idella) was studied to assess the influence of different doses of VD3(15.2, 364.3, 782.5, 1,167.9, 1,573.8, and 1,980.1 IU/kg) on growth performance, intestinal morphology, digestive absorption, amino acid transport, and potential signaling molecule levels in a feeding experiment. As a result, dietary VD3improved growth performance, intestinal structure, and digestive and brush border enzyme activities. Additionally, most intestinal free amino acids and their transporters were upregulated after VD3intake, except for Ala, Lys, Asp, Leu, solute carrier (SLC) 7A7, SLC1A5, and SLC1A3mRNA in different segments, Leu and SLC6A14 mRNA in the proximal intestine, and SLC7A5mRNA in the mid and distal intestine. In the crucial target of rapamycin (TOR) signal pathway of amino acid transport, the gene and protein expression of TOR, S6 kinase 1, and activating transcription factor 4 were elevated, whereas 4E-binding protein 1 was decreased, further suggesting an advanced amino acid absorption capacity in the fish due to VD3supplementation. Based on percentage weight gain, feed efficiency, and trypsin activity, the VD3requirements of on-growing grass carp were estimated to be 968.33, 1,005.00, and 1,166.67 IU/kg, respectively. Our findings provide novel recommendations for VD3supplementation to promote digestion and absorption capacities of fish, contributing to the overall productivity of aquaculture.

The present study explored effects of L-glutamate (Glu) levels on growth, digestive and absorptive capability, and intestinal physical barrier functions of Jian carp (Cyprinus carpio). A total of 600 Jian carp (126.40 ± 0.21 g) were randomly distributed into 5 groups with 3 replicates each, fed diets containing graded levels of Glu (53.4 [control], 57.2, 60.6, 68.4, and 83.4 g/kg) for 63 d. Results showed compared with control diet, feed intake and percent weight gain (PWG) in fish fed 83.4 g of Glu/kg diet were increased and feed conversion ratio in fish fed 68.4 g of Glu/kg diet was decreased (P < 0.05). Similarly, body crude protein and lipid contents in fish fed 68.4 g of Glu/kg diet were higher (P < 0.05). The activities of trypsin and chymotrypsin in the hepatopancreas and intestine, and amylase, alkaline phosphatase (AKP), Na+, K+-ATPase (NKA), and creatine kinase (CK) in intestine were higher in fish fed 68.4 g of Glu/kg diet (P < 0.05). Dietary Glu (57.2 to 83.4 g/kg diet) decreased malondialdehyde (MDA) and protein carbonyl (PCO) contents in the intestine (P < 0.05). The activities of catalase (CAT), glutathione peroxidase (GPx), and glutathione S-transferase (GST) in the hepatopancreas and intestine were higher in fish fed 60.6 and 68.4 g of Glu/kg diets (P < 0.05). Intestinal the glutathione reductase (GR) activity and glutathione (GSH) content in fish fed 60.6, 68.4, and 83.4 g of Glu/kg diet were increased (P < 0.05). The GPx1a, GST, and nuclear factor erythroid 2-related factor 2 (Nrf2) mRNA expressions in the intestine were up-regulated in fish fed 60.6 and 68.4 g of Glu/kg diet (P < 0.05). The zonula occludens protein-1 (ZO-1), occludin1, and claudin3 mRNA expressions were also up-regulated in fish fed 83.4 g of Glu/kg diet (P < 0.05). Fish fed 68.4 g of Glu/kg diet had higher levels of claudin 2, claudin7, and protein kinase C (PKC) mRNA (P < 0.05). These results indicated that Glu improved fish growth, digestive and absorptive ability, and intestinal physical barrier functions. Based on the quadratic regression analysis of PWG, and MDA of the hepatopancreas and intestine, the optimal dietary Glu levels were estimated to be 81.97, 71.06, and 71.36 g/kg diet, respectively.