Effect of Clostridium butyricum in different forms on growth performance, disease resistance, expression of genes involved in immune responses and mTOR signaling pathway of Litopenaeus vannamai.

A 42-day feeding trial was conducted to evaluate the effects of diet supplemented with various additives from Clostridium butyricum (fermentation supernatant, FS; live cells, LC; cell-free extract, CE; spray-dried spores, DS; mixture of live cells and supernatant, LCS) on the growth, intestinal morphology, disease resistance, immune gene expression and mTOR signaling-related gene expression in Litopenaeus vannamai. The feeding trial showed that the final weight and specific growth rate of the shrimp were improved significantly while the feed conversion ratio were reduced significantly in LC, CE, DS and LCS groups compared to the control. The villus height and intestinal wall thickness of shrimp's mid-intestine in LC, DS and LCS group increased significantly. After challenge test to Vibrio parahaemolyticus, the cumulative mortalities of the shrimp in LC, CE, DS and LCS groups were significantly lower than that of the control. As compared to the control, the relative expression levels of superoxide dismutase, lysozyme, prophenoloxidase (proPO), Toll, Immune deficiency (Imd), Relish, TOR, 4E-BP, eIF4E1α and eIF4E2 genes in the shrimp of DS and LCS groups enhanced significantly, whereas the relative expression levels of proPO, SOD, Toll, Imd, Relish, elF4E1α and elF4E2 genes were statistically the same between FS group and the control. These results suggested that the spray-dried spores and mixture of live cells and supernatant of C. butyricum exerted better probiotic benefits in modulating immune responses of shrimp. In addition, single supernatant could not be helpful to shrimp while mixture of live cells and supernatant could better improve the immune responses of shrimp in comparison to single live cells. The integration of C. butyricum and their metabolites supplemented into feed could significantly improve growth performance, intestinal morphology, immunity capacity and resistance against V. parahaemolyticus of L. vannamei.