Effects Of Spo0A Gene On Spore Formation Of Clostridium Tyrobutyricum And Intestinal Microecology In Mice

Clostridium tyrobutyricum is a kind of probiotic that is often used as a feed additive.It has the function of regulating intestinal microflora of animals,repairing intestinal epithelial cells and anti-diarrhea.As an endogenous Clostridium perfringens,it can produce strong resistant spores under certain conditions to prolong the survival time and resist the external environment that is not conducive to growth.Compared with other non-Bacillus species,this feature has unparalleled advantages in commercial production.However,the low rate of sporulation has always been a bottleneck encountered in practical production.At present,most of the researches are mostly focused on the genetic screening of strains and the improvement of the fermentation process.However,no relevant molecular studies have been conducted to increase sporulation rates.In this study,an overexpression vector was constructed to over-express the key regulatory gene spo0 A in the spore-forming pathway in Clostridium tyrobutyricum,and its spore formation rate was determined under common culture conditions with adequate nutrition.The results showed that,after overexpression of spo0 A,the strain showed a higher rate of sporulation.The final spore production was doubled.Subsequently,animal experiments were conducted to explore the effects of Clostridium tyrobutyricum on the morphology of intestinal mucous membranes in mice and the effects on intestinal microflora in mice.The results showed that Clostridium tyrobutyricum can significantly improve the mucous membranes of the intestine in mice,resulting in increased villus height on the ileum,increased V/C value,and thus enhanced the ability of the mice to digest and absorb.It is worth noting that the overexpressed mutant strains have a stronger effect on improving the intestinal mucosa,which may be due to an increase in the rate of sporulation resulting in prolonged survival of the bacteria in the intestine.In addition,Clostridium tyrobutyricum will reduce the species abundance,diversity,and evenness of the intestinal flora in mice.However,it can increase the species abundance of beneficial bacteria such as Lactobacillus and Faecalibaculum,reduce the species abundance of pathogens such as Erysipelotrichaceae,and regulate the intestinal microflora and balance the intestinal microecology.