| Chinese herbal medicine has dual roles in nutrition and medicinal, besides low drug residues,so the research of Chinese herbal medicine in ruminant diet nutrition regulation has important theoretical and practical significance.This project chose prepared by Chinese medicine compound treatment groups "Zhuangniulingl,2,3" "Yufeisan 1,2,3 "and anti-heat stress, monensin as reference,blank control group as contrast,Chose 36 goats with been fitted with permanent rumen fistula,unit group average by random grouped,combined with the traditional methods and molecular analysis methods researched traditional Chinese medicine compound on the impacts of rumen fermentation and bacterial diversity,to explore the effect of Chinese medicine compounds for rumen regulation and controlling.Part I the influence of Chinese medicine compound for goat rumen fermentationFeeding goats after 20 days, gathering rumen fluid at the seven time points 0,2,4, 6,8,10,12h for consecutive 3 days, researched the Chinese medicine compound to the influence for goat rumen pH, the concentration of MCP,NH3-N and VFA.Results showed:"Yufeisan 2" TVFA was significantly higher than monensin (P<0.05),and other treatment groups TVFA content were reduced,no significant difference (P>0.05);"Yufeisan 2" had a significantly higher than monensin on the acetate content (P<0.05),"Zhuangniuling 1,2,3"," Yufeisan 3",heat stress resist treatment group in the rumen acetate had a tendency to be reduced, the content of the "Zhuangniuling 1" treatment group had the greater influence;In addition to the "Yufeisan 2", monensin propionate was significantly higher than other groups (P<0.05), treatment group "Yufeisan 2" propionic acid content increased;"Yufeisan 2",heat stress resist and "Zuangniuling 3" treatment group butyrate content were increased significantly(P<0.05),monensin butyrate content was significantly lower than other groups (P<0.05);monensin’s acetate-propionate ratio decreased significantly (P<0.05)," Yufeisan 1" and heat stress been increased significantly (P <0.05).Conclusion:Chinese medicine compound could maintain goat rumen acid-base balance,the compound Chinese medicine "Yufeisan 2"promoted the growth of rumen microbes and increased VFA concentration,other groups reduced VFA concentration.Part II the influence of compound Chinese medicine for goat rumen bacteria communityFeeding goats after 20 days, gathering rumen fluid at rumen ventral before early feeding 0 h, extracting bacterial DNA, using bacterial 16 SrDNA area V4 variable region sequence as a primer for PCR amplification, based on Illumina Miseq sequencing platform, built small fragment library carrying on the double end sequencing, analyzed the annotation and the abundance of species, further analyzed the Alpha Diversity and Beta Diversity analysis. Results show:the 36 samples received 1445331 tags, about 81% of the original reads passed all quality control and filtered the total number of tags used for splicing sequence,then built OTUs and accessed to classified information in the number of tags, At the phylum classification level,monensin got OTUs adundance more,except "Zhuangniling 2",increased significantly (p<0.05), "Zhuangniuling 3" adundance decreased significantly(P<0.05 Based on the number of OTU, Chaol value and Shannon index results, "Yufeisan 2", "Yufeisan 1" and blank obtained numbers were more, moninsen treatment group obtained the number of at least at the three dilution curves,the same results as the rank abundance. Based on the phylum classification level,all sequences were identified as 11 phylums, Bacteroidetes,Proteobacteria and Firmicutes were all samples’advantage bacterium phylums. "Zhuangniuling 1 ","Zhuangniuling 3", heat stress resist and monensin significantly reduced Firmicutes’ relative abundance(P<0.05),monensin significantly improved Proteobacterias’relative abundance(P<0.05).According to the relative abundance of OTUs in the top 10 Genus species annotation.Prevotella was all samples’advantage bacterium genus,Succinivibrio was at high rate in the monensin and heat stress resist,TG5 was at high rate in the "Zhuangniuling 1". In the analysis of the PCoA, included in this study 36 goats rumen bacteria present continuous distribution, the characteristics of all the samples there is no obvious clustering phenomenon. "Zhuangniuling 1","Zhuangniuling 2","Yufeisan 3",heat stress resist and monensin had been clustered trend, "Zhuangniuling 1"and"Zhuangniuling 2"had obvious overlap phenomenon. Conclusion:except "Yufeisan 1" and "Yufeisan 2",other chinese medicine compound let the rumen bacterial community diversity been decreased significantly, "Zhuangniuling 3" let the rumen bacterial community abundance decreased significantly, "Yufeisan 2" had big individual difference, Bacteroidetes,Proteobacteria and Firmicutes were all samples’advantage bacterium phylums, "Zhuangniuling 1 ","Zhuangniuling 3" and heat stress resist significantly reduced Firmicutes’relative abundance, Prevotella was all samples’advantage bacterium genus.Part Ⅲ the influence of compound Chinese medicine for goat rumen cellulose degradation bacterium amountGathering rumen fluid at rumen ventral before early feeding 0 h from 20 to 22 days, extracting bacterial DNA, Real Time PCR method was applied to detect the number of Fibrobacter succinogenes, Ruminococcus flavefaciens,Ruminococcus albus and all bacterias.Results show:except "Zhuangniuling 1" and "Yufeisan 2",other treatment groups reduced the number of Ruminococcus albus significantly(P<0.05);Except moncnsin, "Yufeisan 2" increased the number of Ruminococcus flavefaciens significantly (P<0.05);Monensin increased the number of Fibrobacter succinogenes significantly,chinese medicine compound had not significant difference (P>0.05).Conclusion:except "Zhuangniuling 1" and "Yufeisan 2",other treatment groups reduced the number of Ruminococcus albus significantly.besides "Yufeisan 2" increased the number of Ruminococcus flavefaciens significantly. |
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