The Effect Of Fecal Microbiota Transplantaion On Growth Performance And Gut Microbiota In Chickens

It has been found that the intestinal microbiota plays an important role in the growth and development of animals.Intervening intestinal microorganisms by changing diets,supplementing probiotics and fecal microbiota transplantation can achieve the goals of regulating metabolism,promoting growth and development and treating diseases.However,there are few studies about the effects of fecal microbiota transplantation on growth and development of chickens.Therefore,in the present study,adult chickens(hybrids of Turpan cockfight and Laihang chicken)with high body weight were used as donors and 1-day-old chickens as recipients for fecal microbiota transplantation.After thirty days of continuous fecal microbiota transplantation,the chicken growth performance and gut microbiota will be tested.The results will provide a theoretical basis for clinical improvement of chicken growth performance and the development of probiotics.The main research contents and research results are as follows:1.The effect of fecal microbiota transplantation on chicken growth performanceIn order to determine the applicable concentration of fecal microbiota,the faeces from adult chickens with high body weight were diluted in 1: 3(high concentration)and 1: 6(low concentration)ratio and transplanted to 1-day-old chickens for 30 days respectively.The results showed that the body weight of the low concentration fecal microbiota transplantation group(group L)was significantly higher than that of the control group from the third day.While the body weight of the high concentration fecal bacteria transplantation group was not significantly different from that of the control group.Further research results showed that the bodyweight,chest muscle weight,leg muscle weight,liver weight and jejunal lenghth of group L was still significantly higher than that of group C.HE staining results showed that the number of muscle fibers in chest and leg muscles in FMT group was significantly higher than that in control group.And the ratio of villus height / crypt depth in duodenum,jejunum,ileum and cecum was significantly higher than that in control group.2.The effect of FMT on chickengut microbiota2.1 Diversity analysis of gut microbiota after FMTIn order to explore the effect of fecal microbiota transplantation on intestinal microbial diversity of chickens,the microbial 16 S r RNA gene sequencing technology was used to detect the abundance and diversity of the contents and mucosal microbiome of the jejunum and cecum.The results of α-diversity showed that the bacterial diversity in cecal of FMT group was higher than that of control group.The results of β diversity analysis showed that there was a significantly difference of the cecal mucosa and contents.The above research results showed that fecal microbiota transplantation can affect the intestinal microbiota diversity of chickens.2.2 Analysis of the abundance of gut microbiota after FMTIn order to explore the effect of fecal microbiota transplantation on the composition of intestinal microbiota,the differences of the microbial composition in jejunum and cecum between the FMT group and the control group were compared and analyzed according to the results of OTU classification and classification status identification.At phylum level,Firmicutes and Proteobacteria were the dominant bacteria of jejunum.The relative abundance of Firmicutes phylum in FMT group was higher than that in control group,while the relative abundance of the Proteobacteria was lower than that in control group.Bacteroidetes and Firmicutes phylum were the dominant bacteria in cecal contents and mucosa.In cecal contents,the relative abundance of Bacteroidetes phylum in FMT group was higher than that in control group,and the relative abundance of Firmicutes bacteria was lower than that in control group.In contact,in the mucosa,the relative abundance of Bacteroides phylum in FMT group was lower than that in control group,and the relative abundance of the Firmicutes phylum was higher than that in control group.At genus level,Lactobacillus and Escherichia-Shigella were the dominant genus in jejunum.The relative abundance of Lactobacillus in FMT group was higher than that in control group,while the relative abundance of Shigella was lower than that in control group.In cecum,Bacteroides was the dominant genus of cecal mucosa and contents,and the relative abundance of it in the cecum mucosa of FMT group was lower than that of control group.The results showed that fecal microbiota transplantation have changed the composition of intestinal microbiota.2.3 Analysis of the differences of gut microbiota after FMTIn order to compare and analyze the differences of intestinal microbiota between FMT group and control group,the Wilcoxon rank-sum test was used to analyze the differential microbiota taxa of FMT group and control group.The results showed that the relative abundance of some SCFA-producers and probiotics such as Lactobacillus,Staphylococcus,Bididobacterium,Brachybacterium genus in jejunum of FMT group were significantly higher than that of control group;the relative abundance of Spirochaetes phylum level and Sphaerochaeta,Alloprevotella,Sutterella,Parasutterella,Eubacterium_brachy_group,Paraprevotella,Slackia,Faecalicoccus genus in the cecum of FMT group were significantly higher than those in control group.The results showed that there was a difference between the FMT group and the control group.2.4 Analysis of the prediction function of gut microbiome after FMTIn order to explore the effect of different intestinal microbiota on the metabolic pathway of chickens,prediction of microbioal function using PICRUST analysis.The results showed that the intestinal microbiota of FMT group in the jejunum and cecum was mainly involved in the glycolipid metabolism and immune pathways.For example,jejunal content microbiota participate in Glycerolipid metabolism,Galactose metabolism,Primary bile acid biosynthesis,Secondary bile acid biosynthesis,Gluconeogenesis etc.In the control group,jejunal intestinal mucosa microbiota were mainly involved in Lipoic acid metablism pathway.The cecal mucosa microbiota of Control group mainly participates in the Glycerolipid metabolism pathways.The results suggest that faecal microbiota transplantation can change the intestinal microbiota and may have an effect on the glucose and lipid metabolism of chickens.3.Functional verification of intestinal microbiota after FMT3.1 The effect of FMT on the concentration of SCFA in chickensThe concentration of SCFA(short-chain fatty acid)in cecum was determined by Gas chromatography.These results suggested that fecal microbiota transplantation can regulate glycolipid metabolism,and affect the production of short chain fatty acids.3.2 The effect of FMT on glycolipid metabolism in chickensIn order to further verify the effect of fecal microbiota transplantation on glycolipid metabolism in chickens,q-PCR and Immunohistochemical staining techniques were used to compare the expression patterns of liver glucose and lipid metablic genes.The results of q-PCR showed that the genes related to fat biosynthesis of Acetyl-Co A carboxylase(ACC),and fatty acid synthase(FAS),and the genes related to fat degration of Carnitine palmitoyltransferase 1(CPT-1)and Peroxisome proliferators-activated receptor α(PPARα)of FMT group were significantly higher than those of control group.The genes related to glycogen biosynthesis of glycogen synthase 2(GYS2)and the genes related to glycogen degration of glycogen phosphorylase L(PYGL)and Isocitrate dehydrogenase 3(IDH3A)of FMT group were significantly higher than those of control group.Immunohistochemical results showed that Sodium-dependent glucose transporters 1(SGLT-1)was mainly expressed on the hepatocyte membrane,and it was significantly higher in FMT group than control group.The results of gas chromatography showed that the concentration of acetic acid,propionic acid,butyric acid,and total short-chain fatty acids in FMT group were significantly higher than those in control group.These results suggested that fecal microbiota transplantation can affect the expression of genes related to glycolipid synthesis and degradation.3.3 The effect of FMT on the expression of genes related growth and developmentIn order to verify the effect of fecal microbiota transplantation on the expression of genes related to growth and development in chickens,q-PCR and immunohistochemical staining technology were used to compare the expression of genes related to growth in chest muscle,leg muscle and liver of FMT group and control group.The q-PCR results showed that the expression level of IGF-1 in the chest muscles,leg muscles liver of FMT group was significantly higher than that of control group.The expression levels of PCNA in chest muscle,leg muscle and liver in FMT group were significantly higher than those in control group,and the expression levels of Myo G in the chest muscle and leg muscles of FMT group were significantly higher than those in control group.Immunohistochemical staining results showed that the IGF-1 protein was mainly distributed in the cytoplasm in the liver;and it was mainly diffusely distributed in the cytoplasm as small particles in muscle.The expression of IGF-1 protein in the chest muscle,leg muscle and liver of FMT group was significantly higher than that of control group.PCNA in the chest muscle,leg muscles and liver of FMT group was significantly higher than that of control group.The results showed that fecal microbiota transplantation could affect the expression of the growth-related genes and affect the growth and development of chickens.In conclusion,fecal microbiota transplantation can change the composition of intestinal microbiota,increase the concentration of probiotics such as Lactobacillus and short chain fatty acids,enhance the metabolism of glucose and lipid,promot the expression of growth related genes such as IGF-1,PCNA and Myo G,and promote the growth and development of chickens.