复方陈皮粉对猪胃肠道菌群多样性的影响  

Effect of compound orange peel powder on pig gastrointestinal microbial diversity

作　　者：张文连[1] 黄志坚[2] 殷光文[2] 李清禄[1,2] 

机构地区：[1]福建农林大学生命科学学院 [2]福建农林大学动物科学学院福建省动物药物工程实验室,福建福州350002

出　　处：《福建农林大学学报（自然科学版）》2016年第2期196-202,共7页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：福建省自然科学基金资助项目(2013J01360)

摘　　要：将60头试验猪随机分为2组,每组3个重复,每个重复10头猪;对照组和试验组分别饲喂基础日粮和添加0.2%复方陈皮粉的日粮,共计60 d;饲喂过程中定期采集新鲜粪便,采用变性梯度凝胶电泳(DGGE)和末端限制性内切酶片段长度多态性(T-RFLP)指纹图谱技术研究饲喂复方陈皮粉对猪胃肠道菌群多样性的影响.结果表明,试验期间试验组的猪基本不腹泻,对照组有部分猪经常腹泻.分别对两组粪便样品的胃肠道微生物进行16S rDNA V3区扩增,再通过DGGE技术进行条带分离,DGGE图谱显示,猪胃肠道菌群物种丰富,两组样品之间存在明显的条带差异.对两组粪便样品胃肠道微生物16S rDNA全长进行扩增,用MspⅠ和HaeⅢ对扩增产物进行限制性酶切,再运用T-RFLP技术检测.通过RDP数据库查询比对,在两组粪便样品中,相同的T-RFs片段有27个,相对于试验组,对照组样品中差异部分片段对应的菌种多为致病菌.研究结果表明,临床上使用复方陈皮粉能有效预防猪腹泻等胃肠道疾病,这可能与药物能改善胃肠道微生态坏境,调节菌群平衡有关.Denaturing gradient gel electrophoresis（ DGGE） and terminal-restriction fragment length polymorphism（ T-RFLP） fingerprinting technique were used to study the effect of compound orange peel powder on gastrointestinal microbial diversity of swine.Sixty pigs were randomized into two groups,with the control group being fed with basal diet only while the experimental group being fed with basal diet supplemented with 0. 2% compound orange peel powder for 60 days,respectively. Fresh fecal samples were collected after feeding,and V3 region of 16 Sr DNA of intestinal microorganism from the fecal samples were amplified by PCR,and the diversity of intestinal microbial was studied by DGGE. During the raising period,diarrhea frequently occurred in pigs in the control group but much less in the experimental group. Band patterns from DGGE showed that gastrointestinal microorganism was abundant in swine,and varied significantly between two groups. Furthermore,full length of 16 Sr DNA of intestinal microorganism was amplified,with PCR products being digested by MspⅠ and HaeⅢ and followed by digested fragments being detected by T-RFLP techniques. Referring to recombination detection programme RDP database,27 T-RFs fragments were found in fecal samples in both group. Most of fragments that existed in the control but not the experimental group belonged to pathogenic bacteria. To summarize,compound orange peel powder can effectively prevent pig from diarrhea and other gastrointestinal diseases clinically. This may be attributed to medicine that improved gastrointestinal micro-ecological environment and balanced bacteria population.

关 键 词：复方陈皮粉 DGGE和T-RFLP指纹图谱 胃肠道微生物多样性 胃肠道菌群平衡 

分 类 号：S859.3[农业科学—临床兽医学]