The Analysis Of Microorganism Diversity In The Intestinal Tract Of Mink

Mink is akind of precious fur animal. In the long-term evolution,the mink has builded a unique digestive method. Mink’s gut is short, straight and the structuresimple, digestive enzyme is mainly protease and lipase. Microbes live in intestinal tract, they constraeted a complieated microbial eommunity. Gut microbes in auxiliary digest food in the large intestine, these microorganisms including bacteria, fungi and parasites, etc., an overwhelming majority of bacteria.These microorganisms and their metabolic products areplays animportant roleto keep the health of the host,absorption of nutrition, metabolism and regulation of immune responses. The purpose of this study is studiedenteric bacterial species, analysised predominant bacteria, explored the structure of intestinal florain the gut of mink,byusing PCR-DGGE and high-throughput sequencing technologies.This study selected 10 healthy male mink, similar weight,feed in the Chinese Academy of Agricultural Sciences Institute of specialty of experimentalanimal base. The diet was traditional without antibiotics. All animals feed from July to December. After slaughter, the intestinal contents were removed immediately, and putthem in liquid nitrogen tube, and stored in the refrigerator for-80℃.1. The first experiment, three different methods of extracting DNA was used, and the extraction was analyzed.Theresults showed that:(1) Three kinds of methods could be extracted microbial genomic DNA from intestinalmink in contents, pure genomic DNA used for molecular analysis of following.(2) Comparing of efficiency and DNA access rate,FastDNA SPIN Kit for fecesmethod obtained the highest rate and relatively speaking, relatively fast, So the extraction of mink genomic DNA using this method, for following analysis.2 The second experiment used PCR-DGGE technology studied microbial diversity of duodenum、jejunum、ileum、colon, and the results showed that:Denaturing gradient gel electrophoresis lanes number and brightness was differentin the duodenum、jejunum、ileum、colon microbiota genomic DNA samples, andmicrobial number was gradually increasingwithgut segment extending back. The microbes werethe most diverse in colonthan other parts.3 The third experiment analyzed the gut microbial diversity and advantage bacterium group of 10 minks by high-throughput sequencing technology, the results showed that:(1) 10 mink gut microbe standardized to 146287 sequences, 2936 OTUs, representing 17 phyla, 168 genera.(2) The most microbe was classified five Bacterial phyla in mink distal gut flora, respectivelyFirmicutes、Bacteroidetes、Fusobacteria、ProteobacteriaandActinobacteria. Firmicuteswas the biggest one, about 59.99%, and Bacteroideteswas the second one, about 16.20%.(3) Mink distal intestinal bacteria was classified at genus level, Streptococcus was the largest proportion, accounted for the total bacterial genus 13.32%, Prevotella and Lactobacillus genus also occupy8.77% and 5.92%, respectively.