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To investigate the effects of luteolin (LUT) on the growth, antioxidant capacity, non−specific immunity and intestinal microbiota of red swamp crayfish (Procambarus clarkii), 300 individual crayfish (4.17 ± 0.50 g average body weight and 50.19 ± 1.40 mm average body length) were randomly divided into five groups and fed with LUT doses of 0 mg/kg (CG), 50 mg/kg (LG50), 100 mg/kg (LG100), 300 mg/kg (LG300), and 500 mg/kg (LG500) for 8 weeks, respectively. Firstly, the results demonstrated that the survival and growth performance were significantly improved in all LUT added groups compared to the control (CG group), with no significant differences in hepatosomatic index, feed coefficient and muscle components, and the crayfish in LG100 group showed the maximal survival rate (SR) and specific growth rate (SGR) (P < 0.05). Secondly, histological observations displayed that the hepatopancreas and intestine tissues were not impacted in the LUT−supplemented groups, and the length of intestinal folds was significantly increased in LG100 and LG300 groups. Thirdly, LG50, LG100, and LG300 groups produced significantly increased hepatopancreatic antioxidant enzyme activities and hemolymph immune indicators compared with the control. In all experimental groups, malondialdehyde (MDA) contents were significantly reduced, as were aspartate aminotransferase and alanine aminotransferase activities. Lastly, dietary LUT specifically improved the structure and optimized the function of the gut microbiota. Compared to the control, the LG100 group displayed a significant increase in the index of Chao1 and Observed species, an increase in the relative abundance of Proteobacteria and Actinobacteria, a decrease in the ratio of Bacteroidetes to Firmicutes, and an increase in the abundance of Thermomonas , Turicibacter , Bdellovibrio and Enterococcus. PICRUSt analysis demonstrated a significant improvement in the KEGG pathway associated with isoflavone biosynthesis in the LG group. In conclusion, the present study confirms that dietary supplementation with 50−300 mg/kg LUT favors growth, non−specific immunity and antioxidant function in P. clarkii , with optimal additions ranging from 102−165 mg/kg. Furthermore, it optimized gut microbiota function, suggesting that LUT can be used as a potential feed additive for P. clarkii. These findings provide new insights for crayfish's healthy culture. • The optimal level of LUT addition to P. clarkii diet was 102−165 mg/kg. • Dietary addition of LUT improved the survival and growth performance of P. clarkii. • Dietary addition of LUT enhanced the antioxidant and immune capacity of P. clarkii. • Dietary addition of LUT improved intestinal microbiota structure of P. clarkii. [ABSTRACT FROM AUTHOR]

The red swamp crayfish (Procambarus clarkii) is an important farming species in China and there is a high degree of overlap between the main crayfish production areas and areas contaminated with the heavy metal lead (Pb), thus putting crayfish farming at potential risk of Pb contamination. To assess the toxic effects of Pb on crayfish, in this study they were exposed to different concentrations of Pb (0, 0.1, 1, 10, 50 mg/L) for 72 h, and 0.1 mg/L represents the level of Pb in the contaminated water. Histomorphology and activities of antioxidant or immune-related enzymes suggest that the damage of Pb to the hepatopancreas and intestine was dose- and time-dependent, with the intestine being more sensitive to Pb than the hepatopancreas. Notably, after a short period (24 h) of stress at low concentrations (0.1 mg/L) of Pb, the malondialdehyde (MDA) content and antioxidant enzymes such as catalase (CAT) and glutathione peroxidase (GSH-Px) in the intestine of crayfish showed significant changes, indicating that low concentrations of Pb were also highly detrimental to crayfish. High-throughput sequencing of the intestinal microbial community indicated that Pb exposure led to a disturbance in the relative abundance of intestinal bacteria, increasing the abundance of pathogenic bacteria (Bosea, Cloacibacterium, Legionella spp.) and decreasing the abundance of potentially beneficial bacteria (Chitinibacter, Chitinilyticum, Paracoccus, Microbacterium, Demequina , and Acinetobacter spp.). In conclusion, Pb damages the hepatopancreas and intestinal barrier of crayfish, leading to the destruction of their anti-stress ability and immune response, and at the same time disrupts the homeostasis of intestinal microbes, resulting in adverse effects on the gut. This study contributed to the assessment of the ecotoxicity of the heavy metal Pb to the crustacean aquatic animals. [ABSTRACT FROM AUTHOR]

Gastrointestinal probiotics are important members of intestinal microflora in both healthy animals and human beings, and these bacteria may reduce the risk of infection caused by certain opportunistic pathogens through exclusive inhibition, competition, and displacement. The lp_1643 protein on the cell surface of Lactobacillus plantarum WCFSI was assumed to possess a mucus-binding capability. This study aimed to determine if purified His-N2 protein exclusively inhibits pathogen adhesion to porcine mucus. The interaction of the His-N2 protein with porcine mucus was determined by indirect enzyme-linked immunosorbent assay (ELISA), and the adhesion was assessed by a traditional plating method to count the bacteria adhered to the porcine mucus. Indirect ELISA showed that His-N2 protein adhered to porcine mucus, and its interacting molecules existed. The His-N2 protein effectively inhibited the adhesion of Escherichia coli DH5α, Listeria monocytogenes CMCC54004, Salmonella Typhimurium ATCC 14028, and Shigella flexneri CMCC(B)51572 to porcine mucus. Results showed that inhibition of pathogen adhesion to porcine mucus depended on dose and strain. The adhesion of L. monocytogenes CMCC54004, Salmonella Typhimurium ATCC 14028, and S. flexneri CMCC(B)51572 was reduced by 95.7, 97.0, and 95.7%, respectively, by pre-adding 100 µl of 3.92 mg/ml of His-N2 protein, whereas that of E. coli DH5α was only 50.4%. The inhibition of adhesion of some pathogens by His-N2 was different at pH 6.6 and 7.5. The inhibition of E. coli DH5α, L. monocytogenes CMCC54004, and Salmonella Typhimurium ATCC 14028 at pH 6.6 was significantly higher than that at pH 7.5, whereas no statistically significant difference was observed in S. flexneri CMCC(B)51572. These results suggest that various types of inhibition mechanisms of His-N2 were involved in different pathogens. [ABSTRACT FROM AUTHOR]