Comparison of spore or vegetative Bacillus sp. supplementation on physiological changes and gut microbiota of the olive flounder (Paralichthys olivaceus)

Abstract

The effects of Bacillus sp. supplementation of different types of cells, such as spores and vegetative cells, on fish growth performance, intestinal microbial diversity alteration and immunity in juvenile olive flounder (Paralichthys olivaceus) were investigated. The fish were divided into three groups and provided with either a basal diet (control), Bacillus sp. vegetative cell (VCS) or spore (SS) supplemented diet for 8 weeks. At the end of the experiment, significant differences (P < .05) were found in terms of weight gain, specific growth rate, feed conversion ratio and protein efficiency ratio between the VCS and control groups. Respiratory burst, superoxide dismutase activity, and lysozyme activity, which are innate immune indicators, were observed with positive alterations (P < .05) in SS compared to controls. Intestinal microbial diversity analysis showed that the SS group had an increased richness estimate (Chao 1, ACE, and Jackknife) compared with the control group. Diversity estimates (Shannon and reverse Simpson) revealed that the VCS group was increased compared with the control group. Analyzing the similarity between groups through beta-diversity, Venn diagram, and heatmap analysis, the control and VCS groups were similar, and the SS group was different. IL-1β and TNF-α expression levels in the liver and spleen were increased in the SS group compared with the control group. In growth-related gene expression analysis, only growth hormone increased in the VCS group compared with the control group. In an in vivo challenge experiment with Streptococcus iniae (1 × 108 CFU/mL), the survival rates of the VCS and SS groups were 14.29% and 28.57%, respectively, when the control mortality reached 100%. Therefore, we concluded that supplementation of Bacillus sp. with different physiological processes affects the growth performance and immune activity of flounder. In particular, VCS can induce growth performance and growth-related gene expression, and SS can alter nonspecific immune and immune-related gene expressions.