The Anti-RV Effect Of GM3 Regulated By Lactobacillus Amylovorus

Rotavirus(Rotavirus,RV)belongs to the reovirus class,rotavirus genus.It's one of the major pathogens caused young animal viral enteritis,diarrhea.The RV make a great harm to China's pig industry.ganglioside GM3 is One of its main receptors.In order to investigate the Lactobacillus amylovorus(L.A)regulatory mechanism of anti-porcine rotavirus role of GM3,we completed the present study were analyzed ability of anti-RV the L.A;L.A vivo correlation effects and neonatal rat jejunum ganglioside GM3 and other changes of both RV infected mice pups,seeks to clarify food starch L.A anti-RV partial mechanism.This study was divided into two parts: in vitro and in vivo.In vitro experiments were conducted with IPEC-J2 cells as the cell platform,The in vitro anti-RV ability of L.A was analyzed by MTT assay.In vivo,Kunming mice were selected as experimental animals.And the correlation between the changes of jejunum ganglioside GM3 and the above effects were further detected.1?In vitro test part.Including the stability of L.A supernatant anti-RV ability and probiotics co-culture supernatant anti-RV ability to determine two aspects:(1)The stability of anti-RV ability of L.A experiments.L.A was routinely passed 25 times,The anti-RV ability of supernatant was measured every 3 generations,the results showed that: There was no significant difference in the inhibition rate of L.A on RV,which indicated that the strain resistant to L.A strain was more stable.(2)To investigate the anti-RV ability of probiotics co-culture supernatant: Four strains of Enterococcus faecalis,which were different from L.A and anti-RV ability,were selected as experimental materials.The results showed that L.A and different strains of Enterococcus faecium After culture,the difference between anti-RV ability of co-culture supernatant and anti-RV ability of fresh supernatant of fresh starch and supernatant of Enterococcus faecalis alone.The results showed that the supernatant anti-RV ability of z4,z5 and z8 and L.A were mixed between L.A and z4,z5 and z8 supernatant alone.Z7 and L.A were lower than those of the supernatant of Escherichia coli and the supernatant of L.A oleracea alone,and the inhibition rate of RV was significantly lower than that of L.A oleracea supernatant(p<0.05).2?The experimental part of the body.Including the effects of L.A ammodendron on the mice infected with RV and the effect of L.A on the jejunum ganglioside GM3 in neonatal rats.The sucking mouse were divided into four groups: normal group,virus group,Gavaged probiotic before inoculation group(BI group)and Gavaged probiotic after inoculation group(AI group)(106cfu/mL),and the other groups were fed with the same amount of PBS,the virus group,the first group,the first group,the first group After each dose of 10-6.28 TCID50/mL rotavirus 50 ?L was given to each mouse at 4 days after the probiotic group and the first group.(1).The effect of L.A on RV-infected mice: the weight gain of RV mice was increased,the amount of fecal-like virus and the histological changes of jejunum mucosa.1).The results of body weight gain showed that there was no significant difference in body weight gain between 1d and 4d(p>0.05).The weight gain rate of normal group was significantly higher than that of other groups at 4d-11d(p<0.05).The body weight growth rate of BI group and AI group was significantly higher than that of virus group(p<0.05).The body weight of BI group and AI group was significantly higher than that of virus group There was no significant difference in growth rate(p>0.05).2).Fecal viral load measurement results showed that: after infection 2d,3d,4d,5d,and RV content BI group and 7d were significantly lower virus group(p<0.05);AI after infection when 4d RV were significantly lower than Virus group(p<0.05),and there was no significant difference at other time points.3).Histological changes in jejunum mucosa showed: Virus group mild intestinal epithelial cell degeneration,villus internal tissue loss more serious,significant changes in villous epithelial morphogenesis,the BI and AI groups with respect to virus group,significantly reduced the symptoms associated.The length of jejunum villus in BI group was significantly higher than that in virus group(p<0.05)at 1d and 3d after challenge,and the difference was not significant(p>0.05).The length of jejunum villus in AI group was larger than that in virus group,but the difference was not significant(p>0.05).There was no significant difference in villus length between the two groups at 5d and 7d(p>0.05).(2).The effect of Lactobacillus ammodendron on the jejunum ganglioside GM3 in neonatal rats was divided into two aspects:1).The results of quantitative ELISA analysis of jaundice GM3 in neonatal rats showed:Before attacking,2d~4d when BI group and normal group was not significant(P>0.05),After attacking,5d,9d and BI group GM3 ganglioside content was significantly higher than when the group 11 d and virus group(P<0.05);Virus group ganglioside GM3 content and 9d significantly higher than control group(P<0.05);AI was significantly higher than virus group(p<0.05)when the 5d and 11 d,AI Group,in 9d and 11 d significantly lower than BI Group.2).The mammal jejunum tissue GM3 synthase Mus St3gal5 m RNA results showed: Before attacking,BI group and normal group in 2d,3d,4d when the differences were not significant(p>0.05).After attacking,5d,and 9d virus group was significantly lower than the control group(p<0.05);7d,and 9d and 11 d BI group was significantly lower than the viral group(p<0.05);5d,7d when set slightly below the AI virus group(p>0.05).3).Musu GCG m RNA expression in the jejunum GM3 synthase of neonatal rats showed: There was no significant difference between the BI group and the normal group(p>0.05).The level of the virus group was significantly lower than that of the normal group at 7 days and 9 days(p<0.05);7d,9d,10 d BI when the group was significantly lower than the normal group and virus group(p <0.05),11 d when the AI virus group were significantly lower than group(p<0.05).In summary: The L.A has a relatively stable anti-RV ability,and the strain of L.A and 4 strains of Enterococcus faecium co-culture supernatant anti-RV ability no significant change.This L.A can affect the RV receptor ganglioside GM3 in small intestinal epithelial cells,reduce RV invasion of intestinal mucosal cells.Thereby significantly reducing the RV content in the dairy manure,reducing RV on neonatal villi damage,inhibition of small intestine crypt proliferation,improve the growth rate.