Study On The Mechanism Of Microbiota Transplantation Regulating Ileum And Cecum Microbiome Based On 16S RRNA Sequencing Technology In Broilers

The aim of this study was to determine the effects of microbiota transplantation on growth performance,the microfloras of the ileum and the cecum in broiler chickens,and the feasibility of substituting antibiotics by microbiota transplantation was discussed.We randomly assigned 480 1-d old Arbor Acre(AA)male broilers to 4 groups(n=120;10 broilers per replicate).There were 4 treatments by a 2(with or without Chlortetracycline)×2(with or without microbiota transplantation)factorial design.All the groups were fed a basal diet based on corn and soybean meal.The antibiotic group was added 100 mg/kg Chlortetracycline to the basal diet.The microbiota transplantation group was inoculated with the microbiota from the ceca of healthy adult AA male broilers on d 1 and d 2,respectively,over a trial period of 42 d.The results were as follows:Effects of microbiota transplantation on growth performance in broiler chickens The birds were weighed on d 21 and 42,respectively,and the average body weight(ABW),average daily gain(ADG),average daily feed intake(ADFI),and feed:gain(F:G)were calculated.The results showed that the mortality and elimination rate was 4.58%,and the birds kept healthy;There were no differences in mortality and elimination rate(P>0.05),ABW,ADG,ADFI or F:G(P>0.05)in the early and late periods among groups.Effects of microbiota transplantation on the microfloras of the ileum and the cecum in broiler chickensIllumina MiSeq was used to analyze the effects of microbiota transplantation on operational taxonomic unit partition and classification,alpha and beta diversity,taxonomic composition analysis and the microflora comparative analysis and the key species screening.An average of 9 phyla,16 classes,18 orders,27 families,53 genera and 92 species per broiler were identified in the cecum of the control group,and an average of 3 phyla,6 classes,7 orders,9 families,10 genera and 25 species per broiler were identified in the ileum of the control group.There was an average of 8 phyla,15 classes,16 orders,25 families,49 genera,and 87 species per broiler were found in the cecum of the microbiota transplantation group,and an average of 2 phyla,4 classes,5 orders,6 families,7 genera and 19 species per broiler were identified in the ileum of the microbiota transplantation group.The Chao1,ACE,Simpson and Shannon indices for the control group in cecum were 295.64,328.79,0.95,and 5.70,respectively,whereas in ileum were 60.01,93.09,5.32,3.28.In the microbiota transplantation group,the chao1,ACE,Simpson and Shannon indices in cecum were 274.36,288.80,0.92 and 5.42,respectively,whereas in ileum were 79.45,84.54,0.77,3.02.It was seen that Aestuariispira,Christensenella,Fervidicella,Gracilibacter,Haloferula,Mycoplasma,Novispirillum and Pantoea were significantly more abundant(P<0.05)in the microbiota transplantation group than in the control group,while Acidaminococcus,Alkalimonas,Anaerovorax,Brassicibacter,Caldlcoprobacter,Celluloslbacter,Cltrobacter,Cyanoblum,Desulfonispora,Desulfotomaculum,Desulfotomaculum,Fusicatenibacter and Paraprevotella were significantly less abundant in the microbiota transplantation group than in the control group.Our hierarchical GraPhlAn tree indicated that Fimicutes,Bacilli,Bacteroidetes,Clostridia,Clostridiales,Lactobacillales,and Lactobacillus were the groups most affected by microbiota transplantation.These might be useful targets for future studies of microbiota transplantation.Our heatmap results indicated that microbiota transplantation changed the gut microbiota composition substantially when compared with the control group:the relative abundances of species of Olsenella,Coprobacter,Sutterella,Parabacteroides,Sporobacter,Vampirovibrio,Lutispora,and Pelomonas increased,while those of Faeceliibacterium,Campylobacter,Acetivibrio,Paraprevotella,Odoribacter,Eisenbergiella and Ruminiclostridium decreased.After microbiota transplantation,most metabolic pathways were more prevalent than in the control group,particularly carbohydrate metabolism.These results indicated that microbiota transplantation influenced the metabolic activities of the gut microbiota to maintain homeostasis.In conclusion,this is the first study to apply 16S rRNA sequencing to evaluate microbial effects after microbiota transplantation in broilers.Microbiota transplantation has significantly effects on phylum,class,order,family,genus and species.Microbiota transplantation can rebuild microflora rapidly,and rebuild the dominant status of Actinobacteria,Firmicutes and Bacteroides.After microbiota transplantation,the abundance of most metabolic pathways was higher than that of the control group,especially for the metabolism of carbohydrates.Our results provide insights into the impact of transplantation treatment on microbial communities in the gut and lay a foundation for further investigations into substitution of microbiota transplantation for antibiotics.