The Effect Of Combined Use Of Ellagic Acid And Berberine On Diarrhea And Gut Microbiota Of Early Weaned Piglets

Early weaning is an effective means of increasing sow utilization in intensive pig farms and is widely used.Weaning stress induces intestinal inflammatory damage and intestinal flora imbalance in early weaned piglets,which in turn leads to a series of health problems such as diarrhea.Ellagic acid(EA)and berberine(BBR)have the effect of regulating intestinal flora and relieving weaning stress,but the effect of combined use of the two has not been studied.In order to explore the effect and mechanism of adding EA and BBR to the diet of weaned piglets to prevent and treat piglet diarrhea,63 21-day-old Duroc-landrace-yorkshire ternary early weaned piglets were randomly divided into 3 groups(3 pens in each group,each 7 pigs per pen):the control group was fed the basal diet,the EA+BBR group was fed the basal diet supplemented with 20 mg/kg BW of EA and BBR in equal proportions,and the FEA+BBR group was fed the basal diet The fecal flora suspension of the EA+BBR group was transplanted every other day on the basis of 14 days.On the 15th day,7 piglets in each group were randomly selected for slaughter sampling.By measuring jejunum histomorphology,intestinal tight junction protein expression,oxidative stress-related index content,gut microbial 16S rDNA amplicon sequencing,and short chain fatty acids(SCFAs)in jejunum and colon contents and other indicators,to explore the effect of EA combined with BBR on diarrhea and intestinal microbes of early weaned piglets.The main results are as follows:(1)Compared with the Ctrl group,the EA+BBR group and the FEA+BBR group significantly increase the ADG and ADFI of the piglets from the 7th day to the 14th day after weaning(P<0.05);the diarrhea rate statistics show that the EA+BBR group(P<0.01)and the FEA+BBR group(P<0.05)significantly reduce piglet fecal scores on day 14 after weaning.(2)The histomorphological examination of jejunum show that compared with the Ctrl group,the crypt depth in the EA+BBR group and the FEA+BBR group be significantly decrease(P<0.05),and the ratio of villus height to crypt depth be significantly increase(P<0.05).The results of Western blotting show that the EA+BBR group and the FEA+BBR group could significantly restore the expression of the tight junction protein Occludin in the jejunum(P<0.05),but there be no significant difference in the expression of Claudin-1.(3)Compared with the Ctrl group,the EA+BBR group and the FEA+BBR group increase the jejunum antioxidant index of weaned piglets,including T-AOC(P<0.01),CAT(P<0.01)and GSH/GSSG(P<0.01),but significantly decreased the oxidative metabolite MDA in jejunal tissue(P<0.05).(4)The results of 16S rDNA amplicon sequencing show that the gut microbiota composition of the Ctrl group,the EA+BBR group and the FEA+BBR group be significantly different,and the gut microbiota structure of the FEA+BBR group was closer to that of the EA+BBR group.The results of Alpha diversity analysis show that the bacterial community abundance index Chaol and ACE of the EA+BBR group and the Ctrl group be significantly different(P<0.05).At the phylum level,the EA+BBR group and the FEA+BBR group increase the relative abundance of Firmicutes and Bacteroidetes and decrease the relative abundance of Euryarchaeota compared with the Ctrl group(P<0.05).At the genus level,EA+BBR increase the relative abundance of beneficial bacteria such as Lactobacillus(P<0.05)and Ruminococcus.(5)Compared with the Ctrl group,the concentrations of acetate,propionate and butyrate in the intestinal contents of weaned piglets in the EA+BBR and FEA+BBR groups show an increasing trend.The EA+BBR group significantly increase propionate concentrations in colonic and jejunal contents(P<0.05),and increase butyrate concentrations in jejunal contents(P<0.05).The butyrate concentration in the colonic contents of the FEA+BBR group be significantly increase(P<0.05).(6)The correlation analysis between the differential bacteria and the detection indicators show that:Lactobacillus(P<0.01),Prevotella copri(P<0.05),Ruminococcus(P<0.05)be positively correlated with acetate and propionate in the jejunum,and negatively correlate with NO(P<0.01);Prevotella copri and(Eubacterium)be positively correlate with butyric acetate in empty city and colon(P<0.01);Lactobacillus(P<0.01)and Ruminococcus(P<0.05)be positively correlated with colonic acetate,Firmicutes Firmicutes(P<0.05)be positively correlated with colonic propionate.Overall,the differential microbiota be associated with indicators of gut microbial metabolite SCFAs.(7)The correlation analysis between SCFAs and detection indicators show:jejunal acetate and propionate be positively correlated with villus height/crypt depth(P<0.01);colonic acetate and propionate be negatively correlated with crypt depth(P<0.01);Jejunal propionate,jejunal butyrate,and colonic butyrate be positively correlated with total antioxidant capacity(P<0.01);jejunal propionate,jejunal butyrate,and colonic acetate be negatively correlate with MDA(P<0.01).Overall,SCFAs be associated with antioxidant markers,ileal morphology,and other markers.Conclusion:The combined use of ellagic acid and berberine in the diet improves the production performance of weaned piglets,improves the intestinal damage caused by weaning stress,and relieves diarrhea.And by increasing the content of beneficial bacteria in the gut,it promotes the production of SCFAs and maintains intestinal homeostasis.The results of the trial indicate that the gut microbiota mediate beneficial effects on reducing diarrhea and intestinal damage in weaned piglets.