动物营养学报2018，Vol.30Issue(5)：1837-1844,8.DOI:10.3969/j.issn.1006-267x.2018.05.026
一种猪胃肠道内容物与粪便微生物梯度离心分离方法
A Gradient Centrifugation Separation Method for Microorganisms in Gastrointestinal Contents and Faeces of Swine
摘要
Abstract
This experiment was conducted to investigate a gradient centrifugation separation method for micro-organisms in gastrointestinal contents and faeces of swine. First, dilute the gastrointestinal contents and faeces with separation buffer to find the fittest dilution ratio. Then gradiently centrifuge and acquire microbial bacteria from gastrointestinal contents and faeces samples. By this method,the best dilution ratio of gastro,jejunum and ileum contents to separation buffer ranged from 8.0% to 12.0%,and the weight of samples ranged from 3.2 to 4.8 g. The best dilution ratio of cecum,colon contents and faeces ranged from 2.0% to 8.0%,and the weight ranged from 0.8 to 1.6 g. We detected the amino acids composition of microorganisms in ileum and faeces as well as the diversity of microorganisms in gastrointestinal content and faeces of weighed at 30 kg pigs fed with three kinds of protein level diets(12.0%,15.0% and 18.0%). The results showed that the most amino acids composition of microbes in ileum contents and faeces were significantly responsible to different protein levels of nutritional model(P<0.05). The bands in polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE)figure were rich and legible with microorganisms separated from gastrointestinal contents and faeces. The study demonstrates that this method has high separation degree and obtains substantial dry cell weight. Meanwhile it owns advantages of less cost, easy operation and good repeatability, which provides a method for studying the physiological function of gastrointestinal microbes deeply.关键词
猪/胃肠道内容物/粪便/微生物/梯度离心Key words
swine/gastrointestinal contents/faeces/microorganism/gradient centrifugation分类
农业科技引用本文复制引用
李金龙,王瑶,马娅君,陈庆菊,卢昌文,唐志如..一种猪胃肠道内容物与粪便微生物梯度离心分离方法[J].动物营养学报,2018,30(5):1837-1844,8.基金项目
国家自然科学基金(31772610) （31772610）
西南大学基本科研业务费(XDJK2017D038) （XDJK2017D038）
重庆市留学人才创新计划重点项目(cx2017024) （cx2017024）
重庆市自然科学基金(cstc2016jcyjA1414) （cstc2016jcyjA1414）
国家留学基金委西部计划(201508505170) （201508505170）
国家973重点基础研究发展计划项目(2013CB127303) （2013CB127303）
