The Effect Of Compound Orange Peel Powder On Pig Gastrointestinal Microbial Diversity

The purpose of the research was to study the the microbial diversity in gastrointestinal of pig fed with compound orange peel powder and basal diet using metagenomics and metaproteomics methods. Sixty pigs were randomly divided into two groups. The control group was fed with basal diet, while the experimental group was fed with the basal diet supplemented with 0.2% compound orange peel powder, respectively, for 60 days. Fresh fecal samples were collected from the two groups after feeding. Denaturing gradient gel electrophoresis (DGGE) and terminal restriction fragment length polymorphism (T-RFLP) fingerprinting techniques were used to study the effect of the compound orange peel powder on pig gastrointestinal microbial diversity. Two dimensional electrophoresis and mass spectrometry were used to analyze the proteins of the gastrointestinal microbial diversity. Endotoxin detection technology were also used to study endotoxin content of gastrointestinal and analyse the influence of compound orange peel powder on gastrointestinal gram negative bacteria and the endotoxin release.The result shows that during the experimental period, pigs in the experimental group had no diarrhea phenomenon, while the pigs in the control group had frequent diarrhea phenomenon. The DGGE band patterns showed that the pig gastrointestinal microorganisms were abundant, and there were differences between the two groups. 11 bands of DGGE patterns were selected for cloning and sequencing, and the sequences were compared with the existing sequences in the GenBank database with the BLAST tool. Among the two groups the common bands corresponding to the most similar strains in the database are Escherichia coli, new species of uncultured Firmicutes, new species of uncultured bacterium and new species of Streptococcus spp. Specific bands in the control group corresponding to the most similar strains in the database are new species of Treponema brennaborense, new species of uncultured proteobacterium and new species of Clostridium perfringens. Specific bands in the experimental group corresponding to the most similar strains in the database are Lactobacillus rhamnosus, new species of Lactobacillus acidophilus, new species of Faecalibacterium pransnitzii and new species of Weissella paramesenteroides.T-RFLP fingerprint technology were also used, and the results show that there are 27 same T-RFs fragment including 14 genus in the fecal samples of two groups. They arc Bacillus genus, Treponema, tiny bacillus, Clostridium,Lactobacillus, Helicobacter, Eubacterium, Cytophaga, Tissierella, butyrivibrio, Weissella, Pediococcus, Streptococ-cus and nitrospirae genus. Most of them are opportunistic pathogens and gastro-intestinal normal flora members, only a few are pathogen. Specific fragments of control group corresponding to the bacterial species include pathogens, opportunistic bacteria and normal flora members, which account for the majority of harmful bacteria and opportunistic bacteria. Specific fragments of experimental group corresponding to the bacterial species also include pathogens, opportunistic bacteria and normal flora members, but the number of pathogens and opportunistic bacteria are significantly less than the control group.Through comparative proteomics, seven special spots were used to sequence compare protein database to obtain the corresponding bacterial species, which include Lactobacillus rhamnosus, Clostridium Perfringens, Bacillus cereus, Staphylococcus vitulinus, Erysipelothrix rhusiopathiae, Salmonella enterica, Treponema brennaboren-se. Among Lactobacillus rhamnosus is from experimental group and the other six identified proteins are pathogenic or opportunistic pathogen from control group.In the early period of the experiment, the concentration of endotoxin in the two groups had little difference. But in the later period, the endotoxin concentration appeared relatively differences between the two groups. The endotoxin concentra-tion in the control group showed an upward trend, while the experimental group had a slight difference,20 days later, the difference between the experimental group and the control group were significant (P< 0.05).In conclusion, the compound orange peel powder can reduce pig diarrhea and other gastrointestinal diseases. The pharmacodynamic effect of compound orange peel powder improve the gastrointestinal microbial environment, maintain or build a new gastrointestinal flora balance, make the pig gastrointestinal tract flora tend to be more health state. Therefore, it can be concluded that compound orange peel powder pharmacodynamic effect may be relative to change and regulate the animal’s microbial ecological system and then achieve the new microbial ecological balance.