| In the study,we added three different doses of Clostridium butyricum(CBM01)in rabbit basal diets to investigate the effect of Clostridium butyricum on the reproductive performance of female rabbit and the growth performance,digestibility,intestinal immune function and intestinal flora of weaning rex rabbit.60 healthy primiparous estrus female Chuanbai rex rabbit were selected for mating on the same day.Then the breeded female rabbits were randomly divided into 4 groups,including normal control group(NC),low dose group(LD,1.0 × 103 cfu/g CBM01),medium dose group(MD,1.0 x 104 cfu/g CBM01),and high dose group(HD,1.0 × 105 cfu/g CBM01).The female rabbit was fed from the breeding until the young rabbits weaned.After weaning,30 healthy weaning rex rabbits with similar weights were slected in each group.They were fed the same diet with their mother for 28 days.At the end of the feeding period,samples of weaned rabbits were taken for testing.All the results as follows:1.There was no significant difference on the reproductive performance,including litter size,alive litter size,litter weight at brith,alive litter size at 3 week,litter weight at 3 week,litter weight of weaning,litter weight at 5 weeks of age,and body weight at weaning between experimental groups and NC group of female rabbit.2.Growth performance and organ index of weaning rabbit:The average daily gain per week of HD group was significantly higher than that of NC group at the second and third week.The average daily gain of HD group was extremely increased than that of NC and LD groups.Compared with the control group,thymus index of LD group and MD group and vermiform appendix index of HD group were significantly increased.The sacculus rotundus index of HD group was extremely significant higher than that of NC group.3.Activity of digestive enzyme in intestinal contents of weaning rabbit:Compared with NC group,the activity of a-amylase in duodenum and jejunum of experimental groups were evry significantly increased.The activity of a-amylase in ileum of HD group was significantly higher than that of NC group.Chymotrypsin activity in duodenum of HD group was significantly higher than the other groups,and MD group was significantly higher than that of NC group.Chymotrypsin activity in jejunum of HD group was very significantly higher than NC group and LD group.Compared with the NC group,trypsase activity in small intestine contents and cellulase activity in caecum of the experimental groups showed a rising trend,but there was no significant difference.4.Villus and crypts of small intestine of weaning rabbit:Compared with NC group,the villus length of small intestine tissues in experimental groups were significantly increased.The crypt depth of duodenum in the experimental groups were significantly lower than that in NC group,and the crypt depth of jejunum and ileum in MD and HD groups were significantly lower than that in NC group.The ratio of villus and crypt of duodenum,jejunum and ileum in experimental groups was higher than that in NC group.5.Intestinal tissue antioxidant status of weaning rabbit:The activities of SOD,GSH-Px and CAT in duodenum tissue in HD group were significantly higher than those in NC group.The SOD and GSH-Px activity of ileum tissue in HD group were higher than that in NC group.Compared with NC group,there was no significant difference in the activity of SOD and GSH-Px in jejunum.The MDA content of duodenum and jejunum in HD group was obviously lower than in NC group.6.Intestinal mucosa and intestinal barrier of weaning rabbit:Compared with NC group,the SIgA content of duodenal mucosa in MD and HD groups,and of ileal mucosa in MD group significantly increased.The mRNA expression of ZO-1,occludin,and claudin of experimental groups showed an increasing trend in small intestinal tissue.The mRNA expression of ZO-1 and claudin of jejunum tissue in HD group were significantly higher than in NC group.Compared with NC group,the mRNA expression of ZO-1,occlldin and claudin of ileum tissue in HD group were significantly increased,and the expression of ZO-1 and occlldin in HD group were also significantly increased in colon tissue.Compared with NC group,the content of IL-6 in ileum in HD group was significantly reduced.7.Intestinal tissue cytokines of weaning rabbit:The inflammatory cytokines including IL-6,TNF-a and IFN-y showed a decreasing trend in the experimental groups.The TNF-a content of ileum,colon and vermiform appendix tissue in MD and HD groups were significantly lower than those in NC group.And the IFN-y content of ileum and colon in MD and HD groups displayed obviously decreased than in NC group.There was no significant difference in IL-4 and IL-10 levels between the experimental groups and the NC group.8.Intestinal tissue Toll receptor signaling pathway of weaning rabbit:The mRNA relative expression of TLR2/4 in experimental groups displayed a upregulating trend,and the change trend of MyD88 was positively correlated with TLR2/4.Compared with NC group,the mRNA expression of TLR2/4 and MyD88 in duodenum of HD group was significantly rised,as well as the TLR4 and MyD88 mRNA expression levels in LD group of jejunum and in MD group of sacculus rotundus were significantly increased.9.Intestinal flora of weaning rabbit:The PCR-DGGE analysis and PCA diagram of intestinal contents in the duodenum,jejunum,ileum,cecum and colon can be distinguished between the NC group and experimental groups.And PCA diagram of NC group and experimental groups were consistent with changes in cluster analysis.The similarity of cluster analysis from the duodenum to colon were 56%,51%,52%,62%,and 67%,respectively.Compared with the NC group,the Shannon-wiener index,species richness index,and species evenness index of HD group in duodenum as well as the Shannon-wiener index and species evenness index of MD group in ileum were significantly increased.The number of total bacteria in HD group(jejunum,cecum)and MD group(jejunum,ileum)were significantly higher than that in NC group.Compared with NC group,the number of Frimicutes in HD group(duodenum,jejunum,cecum)and MD group(jejunum,ileum and colon)displayed a significant increase trend.Moreover,the number of Bacteroides in MD and HD groups(duodenum,jejunum,ileum and cecum)were evidently higher than that in NC group.Furthermore,the number of cellulolytic bacteria(Clostridium cluster ?,Clostridium cluster ??a,Butyrivibrio fibrisolvens,and Ruminococcus flavefaciens)and beneficial bacteria(Lactobacillus,Bifidobacterium)in experimental groups were higher than those in NC group.And compared with NC group,HD and MD groups showed more significant difference among them.The experimental groups had no significant effect on the number of conditional pathogens(Enterobacteriaceae,Enterococcus spp.).In conclusion,Clostridium butyricum(CBM01)have no significant effect on the reproductive performance of female rabbits.Treatment with CBM01 could promote the growth and development of weaning rex rabbit though regulating the intestinal flora,increasing digestive enzyme activity of intestinal contents,and improving intestinal morphology.Weaning rex rabbit with CBM01 has improved intestinal immunity by adjusting the intestinal tract Toll-like receptor signaling pathway,heightening intestinal mucosal immunity,and enhancing intestinal barrier function for the prevention of intestinal diseases. |
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