Stocking density has been identified as one of the main factors affecting fish growth, welfare and behavior. However, few studies have focused on the effects of stocking density on fish health in integrated rice–fish farming systems. Thus, the aim of this study was to evaluate the effects of different stocking densities on the growth performance, physiological parameters, redox status and lipid metabolism of Micropterus salmoides in an integrated rice–fish farming system. The fish were reared at three densities: low density (LD, 40 g/m³), medium density (MD, 80 g/m³) and high density (HD, 120 g/m³) for 90 days. At the end of the experiment, fish reared in the MD and HD groups showed lower growth performance than those from the LD group. The HD treatment significantly altered the physiological parameters, including glucose and lactate. Meanwhile, the HD treatment induced oxidative stress and lipid peroxidation after 90 days of farming. Furthermore, transcriptomic analysis revealed that HD treatment led to abnormal lipid metabolism. Interestingly, we found the suppression of three key pathways related to lipid metabolism, including the PPAR, insulin and adipocytokine signaling pathways, in the HD group. Overall, our data indicated that the HD treatment inhibited growth and caused physiological responses, oxidative stress and abnormal hepatic lipid metabolism in M. salmoides in an integrated rice–fish farming system.

Berberine, a natural alkaloid found abundantly in various medicinal plants, exhibits antioxidative, anti-inflammatory, and lipid metabolism-regulatory properties. Nonetheless, its protective effects and the molecular mechanisms underlying liver injury in fish have not been fully elucidated. The aims of this study were to investigate the antioxidative, anti-inflammatory, and lipid metabolism-regulating effects of berberine against high-fat diet (HFD)-induced liver damage and to clarify the underlying molecular mechanisms. Tilapia were fed diets containing two doses of berberine (50 and 100 mg/kg diet) alongside high fat for 60 days. The results showed that berberine treatments (50 and/or 100 mg/kg) significantly reduced elevated aminotransferases, triglycerides (TG), total cholesterol (TC), and low-density lipoprotein cholesterol (LDL-c) in the plasma. In the liver, berberine treatments significantly increased the expression of peroxisome proliferator-activated receptor α (pparα) and carnitine palmitoyltransferase 1 (cpt-1) genes, leading to a reduction in lipid accumulation. Meanwhile, berberine treatment suppressed lipid peroxidation formation and enhanced antioxidant capacity. Berberine upregulated the mRNA levels of erythroid 2-related factor 2 (nrf2) and its downstream genes including heme oxygenase 1 (ho-1) and glutathione-S-transferase (gstα). Additionally, berberine attenuated the inflammation by inhibiting the expression of toll-like receptor 2 (tlr2), myeloid differential protein-88 (myd88), relb, and inflammatory cytokines such as interleukin-1β (il-1β), tumor necrosis factor-α (tnf-α), and il-8. In summary, this study suggested that berberine offers protection against HFD-induced liver damage in tilapia via regulating lipid metabolism, antioxidant status, and immune response. This protective effect may be attributed to the modulation of the Nrf2, TLR2/MyD88/NF-κB, and PPARα signaling pathways.

Pelteobagrus fulvidraco is a freshwater fish commonly raised in rice fields, yet the optimal stocking density for this species remains unknown. Therefore, this study aimed to investigate the appropriate stocking density of P. fulvidraco in integrated rice–fish farming systems. Three different stocking densities––low density (LD, 125 g/m²), middle density (MD, 187.5 g/m²), and high density (HD, 250 g/m²)––were set up to evaluate P. fulvidraco’s growth performance, stress indices, immune function, antioxidant status, and lipid metabolism after 90 days of farming. The results indicated that HD treatment had a detrimental effect on P. fulvidraco’s growth parameters. HD treatment led to an increase in cortisol (Cor) and lactate (La) levels, but a decrease in glucose (Glu) content in serum. After 90 days of farming, an immune response accompanied by the increase of complement 3 (C3), C4, and immunoglobulin M (IgM) was observed in the HD group. Meanwhile, HD treatment induced oxidative stress and altered antioxidative status evidenced by the levels of catalase (CAT), glutathione peroxidase (Gpx), glutathione (GSH), malondialdehyde (MDA), superoxide dismutase (SOD), and total antioxidant capacity (T-AOC) in serum or liver. Additionally, the lipid metabolism-related genes including lipoprotein lipase (lpl), peroxisome proliferators-activated receptor (pparα), carnitine palmitoyltransferase-1 (cpt-1), and sterol regulatory element binding protein-1 (srebp-1) were markedly downregulated in the HD and/or MD group after 90 days of farming. In conclusion, this study contributes to a better understanding of P. fulvidraco’s response to different stocking densities in integrated rice–fish farming systems. We suggest that the appropriate stocking density for P. fulvidraco in these farming systems should be below 250 g/m², considering both fish growth and physiological responses.

In this study, effects of stocking density on the growth performance and physiological responses of blunt snout bream, Megalobrama amblycephala juveniles were evaluated. The fish (average body weight, 25.76 ± 2.25 g) were randomly stocked at densities of 30F (30 fish/m³), 60F, 90F and 120F in 12 cages (1 m × 1 m × 1 m) in a concrete pond, with three cages for each density, for a period of 6 weeks. The higher stocking densities had a negative effect on individual growth performance. The results indicated that serum cortisol, triglyceride, alanine aminotransferase, aspartate transaminase, alkaline phosphatase and malondialdehyde activities; and Acinetobacter, Aeromonas, Pseudomonas and Vibrio numbers in the intestinal microflora increased significantly as the stocking density increased. In contrast, the viscerosomatic index, hepatosomatic index survival rate; serum glucose, total cholesterol, lipase, protease, glutathione peroxidase and superoxide dismutase activities; and Clostridium, Bacteroides, Lactococcus, Lactobacillus and Bacillus numbers in the intestinal microflora decreased significantly. The 90F and 120F groups showed obvious enlargement of the lamina propria and goblet cell damage, indicating that the gut showed inflammatory responses. The specific growth rate and weight gain rate increased significantly as the stocking density increased from 30 to 60 fish/m³, but decreased significantly when the stocking density was over 60 fish/m³.