Study On Effect Of Danggui Buxue Tang Fermented On Growth Performance In Broilers And Diversity Of Intestinal Microflora

Danggui Buxue Tang(DBT)is one of the classic prescriptions of traditional Chinese medicine,it has been found to have certain effects such as supplementing qi and blood and enhancing immunity.However,DBT has less clinical application in veterinary medicine,and its' specific effect is not clear,especially the research on DBT fermentation product as feed additives has not been studied.This research intends to study the technological process of DBT fermented by Bacillus subtilis(B.subtilis),use High Performance Liquid Chromatography(HPLC)to determine the changes on content of relevant active ingredients of DBT before and after fermentation,and clarify the effects of DBT fermentation product on the growth,immunity,antioxidant capacity and morphology of intestinal villi in broilers and the regulation of intestinal microbiota diversity in SD rats,which aims to provide a theoretical basis for DBT fermentation product as a new feed additive.1.Optimization of process parameters for DBT fermented by B.subtilis.Single factor and BoxBehnken Central combined test design were used to investigate the effects of fermentation medium components,bacterial solution inoculation volume and fermentation time on polysaccharide extraction rate.The best process conditions under the experimental condition 0.95% of glucose addition,1.52% of peptone addition,2.92% of inoculum,and 35.8 h of fermentation time.Under this condition,the polysaccharide extraction rate reached 9.23±0.067%,which was higher than that of the unfermented group(4.62±0.057%).2.Changes on the content of relevant active ingredients of DBT before and after fermentation was determined by HPLC.The HPLC-DAD method was used to determine the content of ferulic acid and calycosin-7-O-?-D-glucoside.The separation of the ingredients was achieved by a Agilent ZORBAX Eclipse XDB-C18 column at 30 ? and mobile phase composed of 0.2% formic acid(A)and acetonitrile(B)with gradient elution(0~10 min,82%A;10~20 min,82~90%A)at a flow rate of 0.8 mL/min and 10 ?L as sample size.It was found that compared with unfermented DBT,the content of the ferulic acid and calycosin-7-O-?-D-glucoside in the DBT fermentation extract increased significantly by 43.94 ± 0.92% and 14.10 ± 2.38%,respectively(P <0.01).3.The effects of DBT fermentation product on growth,immune,antioxidant capacity and duodenal villus morphology in broilers.In this study,a total of 210 healthy white feather broiler with similar initial body weight of 1 day old were randomly divided into 5 groups: the control group fed the basal diet,unfermented group with 1.0% DBT powder added to the diet,and the fermentation group with 0.5%,1.0% and 2.0% DBT fermentation products added to the diet,respectively.Compared with the control and unfermented groups,1.0% DBT fermentation product significantly reduced the average daily feed intake(AFDI)(P<0.05)and feed conversion ratio(FCR)(P<0.05),and increased average daily gain(ADG)(P<0.05).At the same time,ND-HI Immune Globulin,IL-2,IL-6,TNF-?,INF-?,SOD,GSH-Px and T-AOC contents in serum and bursa index in 1.0% fermentation group were significantly increased(P<0.05).What's more,the ratio of villus height to crypt depth in 1.0% fermentation group was significantly higher than that in control group and unfermented group(P<0.05).4.Regulation of DBT fermentation product on diversity of intestinal microbiota in SD rat.SD rats were randomly divided into the following four groups with six rats in each group: the control group,B.subtilis group,DBT non-fermentation group and DBT fermentation group.The 16 s r RNA of faecal microflora from SD rats was analyzed by high-throughput Illumina sequencing technology.In total,3483 operational taxonomical units(OTUs)were identified,and 1236 OTUs were shared among all samples.The most abundant phyla identified in this study were Bacteroidetes(29.65~38.19%)and Firmicutes(48.30~67.04%).Moreover,compared with the other three test groups,DBT fermentation product can significantly increase the abundance of Lactobacillus and downregulate the abundance of pathogenic bacteria such as Alistipes and Parabacteroides.In summary,DBT fermented with B.subtilis could significantly improve the content of polysaccharides,ferulic acid and calycosin-7-O-?-D-glucoside in DBT.In addition,adding 1.0% of DBT fermentation product to the diet could significantly improve growth performance in broilers and regulate actively intestinal microbiota structure in SD rats,promote the growth of beneficial bacteria and inhibit the reproduction of pathogenic bacteria.Which providing a theoretical basis for the fermentation product of DBT as a new feed additive and technical support.