笼养海兰褐蛋雏鸡肠道菌群的聚合酶链式反应-变性梯度凝胶电泳分析

Analysis of Intestinal Bacterial Flora of Hylan Brown Laying Chicken Fed in Cage by Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis

本试验旨在采用聚合酶链式反应(PCR)-变性梯度凝胶电泳(DGGE)法分析笼养海兰褐蛋雏鸡肠道菌群。随机取5只8周龄笼养鸡剖杀,取全部小肠,将5只鸡的十二指肠、空肠、回肠和盲肠内容物分别混匀,提取基因组DNA,以细菌通用引物扩增16S rDNA高变区序列,对PCR产物进行DGGE分析。结果表明:雏鸡不同肠段中细菌菌群组成差别很大,十二指肠、空肠、回肠和盲肠分别分离出8、12、8和9株菌,共20个菌株,其中有乳酸菌属7株,包括2株鸟乳酸菌(Lactobacillus aviaries)、2株棉籽糖乳球菌(Lactococcus raffinolactis)、1株敏捷乳杆菌(Lactobacillus agilis)、1株陪伴粪球菌(Coprococcus comes)和1株同代乳杆菌(Lactobacillus equigenerosi);且各肠段有共同菌3株,为鲁梅利杆菌(Rummeliibacillus stabekisii)、Lactobacillus equigenerosi和阴道加德菌(Gardnerella vaginalis)。结果提示,同龄笼养海兰褐蛋雏鸡肠道细菌菌群结构与肠道的不同部位有关,各肠段中细菌菌群的组成差异很大。

This study aimed to analyze the intestinal bacterial flora of Hylan brown laying chicken fed in cage by polymerase chain reaction （ PCR）-denaturing gradient gel electrophoresis （ DGGE） .Five 8-week-old chick-ens fed in cage were randomly selected and slaughtered to collect whole intestinal samples.Intestinal contents in duodenum, jejunum, ileum and cecum were individually mixed, and genome DNA of them were extracted, then sequences in hypervariable region of 16S rDNA were amplified with bacterial common primers, finally, the PCR products were used in DGGE analysis.The results showed as follows：there was obvious difference in bacterial flora composition among different intestinal segments.Eight, twelve, eight and nine strains of bacteri-a were isolated from duodenum, jejunum, ileum and cecum, respectively, twenty strains in all, and seven strains of Lactobacillus among those （ including two strains of Lactobacillus aviaries, two strains of Lactococ-cus raffinolactis, one strain of Lactobacillus agilis, one strain of Coprococcus comes and one strain of Lactoba-cillus equigenerosi.There were 3 common strains of bacteria in all intestinal segments, which were one strain of Rummeliibacillus stabekisii, one strain of Lactobacillus equigenerosi and one strain of Gardnerella vaginalis. The results indicate that bacterial flora structure is relative with intestinal segment of Hylan brown laying chick-en fed in cage, and there are differences in intestinal bacterial flora composition among different intestinal seg-ments.