Study On The Effects Of Echinacea Purpurea Extract On Intestinal Flora And Metabonomics In Immunosuppressed Chickens

In this paper,we use metagenomics methods to analyze the structure and function of broiler chicken gut microbiota,to explore the effects of immunosuppression on the diversity of broiler chicken gut microbiota,and to study the intervention effects of Echinacea extract(EE)and Astragalus Polysaccharides(APS)respectively.Metabonomics research is also conducted on the intestinal contents,where the changes of immunosuppressed chicken intestinal metabolites and the intervention effects of EE and APS are detected and analyzed.At the same time combined with serum factors,immune organ index and other detection data,the results of the experiment are expected to demonstrate the changes of the structure,function and metabolites of immunosuppressive broiler chicken gut microflora,and to further explain the mechanism of EE and APS in enhancing immune function.Experiment 1 The effects of immunosuppression on immune organs,serum factors and broiler chicken gut microbiota50 1-day-old broilers including males and females were randomly divided into 2groups and fed for 21 days.One group was blank control group(C group)and the other group was immunosuppressive group(IS group).At the broilers’ age of 4 days,The C group received 0.5 ml normal saline injections into the chest muscles for 3 consecutive days,while the IS group received 100 mg/kg·BW Cyclophosphamide(CY)for 3consecutive days.At the age of 7,14 and 21 days,5 chickens in each group were randomly selected to determine the changes of serum cytokine concentrations.Whole-genome DNA of cecal content was extracted to analyze gut microbiota,structure composition and function condition using metagenomics methods.The results are as follows:Under the state of immunosuppression,the average body weight and bursa coefficient of broilers decreased significantly(P<0.01).The spleen coefficient and thymus coefficient also decreased significantly(P<0.05),and different serum factors changed at different levels.We observed that the diversity of broiler gut microbiota decreased,where Clostridium,Veronococcus,Dehalobacteriaceae,Launospiraceae,Oscillatoria,Molluscules RF39,Mollicutes＿RF39,Anaerobic Corynebacterium,Anaerobic Clostridium and Sutterella of Alcaligenes family were significantly or extremely significantly lower than that of C group,while the composition of Eaglezia genus in IS group was significantly higher than that of C group.Experiment 2 The effects of EE and APS on chicken immunosuppressive model100 1-day-old broilers including males and females were randomly divided into blank control group(C),immunosuppression model group(IS),EE intervention group(EE-IS)and APS intervention group(APS-IS).After feeding to 4 days of age,intramuscular injection of CY was conducted in all the groups except the C group.After the modeling,EE-IS group and APS-IS group were fed with 4 g of EE or 1.2 g of APS per liter of drinking water,respectively.On the 1th,7th and 14 st days after the first administration,serum and cecal contents were collected to test and analyze serum factors and blood biochemical parameters.The structure and function of gut microbiota were analyzed using genomics methods,and the metabolism of serum and intestinal contents were analyzed using metabolomics methods.The results are as follows:EE has an inhibitory effect on anaerobic coryneform bacteria,Holdemaria,Eggerthella,cc＿115,Anaeroplasma and Blautia in gut microbiota of immunosuppressive broiler chickens,while EE has a promoting effect on Enterobacter and Laurespirillum.APS has an inhibitory effect on anaerobic Corynebacterium,Faecoccus and Blautia in gut microbiota of immunosuppressive broiler chickens,while APS has a promoting effect on anaerobic Clostridium,anaerobic Mycoplasma,Ruminococcus and Enterobacter faecalis.EE and APS also play roles in regulating metabolism,which is manifested in metabolic pathways.It has been observed through metabolomics that immunosuppression mainly affects the metabolic pathways of serum alanine,aspartic acid and glutamate and intestinal contents.They also affect metabolic pathways of starch and sucrose and the conversion between pentose and glucuronic acid.EE mainly affects the metabolic pathways of serum alanine,aspartic acid,aminosugar and nucleotide sugar,intestinal TCA cycle in the tract,and the conversion between pentose and glucuronic acid.APS mainly affects the biosynthesis pathway of unsaturated fatty acid in serum,the biosynthesis of aminoacyl t RNA,the metabolism of starch and sucrose in the intestine,and the convention between pentose and glucuronic acid.