冷冻原肠浸泡解冻池水细菌多样性分析  

Analysis of Bacterial Diversity in Thawing Pool Water of Frozen Intestines

作　　者：仇保丰[1,2] 宋鸿雁[1,3] 董蓉莲[4] 高雪梅[2] 蒋荧梅[3] 顾炳泉[2] 胡顺林[1] 

机构地区：[1]扬州大学江苏省动物预防医学重点实验室,江苏扬州225009 [2]南通出入境检验检疫局,江苏南通226004 [3]南通大学实验动物中心,江苏南通226001 [4]江苏省疾病预防控制中心,江苏南京210009

出　　处：《动物医学进展》2017年第5期43-48,共6页Progress In Veterinary Medicine

基　　金：江苏省动物预防医学重点实验室开放课题项目(K13044);南通市科技项目(BK2012091和MS12016014);江苏检验检疫局科研项目(2010KJ35)

摘　　要：为了研究冷冻原肠浸泡解冻池水的细菌多样性,采集3份水样直接提取细菌基因组总DNA,再分别用PCR技术扩增出细菌16S rDNA并构建基因文库,经PCR鉴定后从3个文库中分别随机挑取50个阳性克隆进行DNA序列测定。分别对3个文库的测序结果进行分析发现,除去24.00%~32.00%为无法鉴定的未培养细菌和未分类细菌以外,其余细菌来自于芽胞杆菌纲、γ-变形菌纲、放线菌纲和黄杆菌纲,分别占各文库克隆总数的38.00%~42.00%、20.00%~26.00%、4.00%~8.00%和0~2.00%。除去44株无法鉴定的细菌,进一步对其他106株细菌进行分析发现,其中2株放线菌目细菌和5株肠杆菌科细菌无法进一步鉴定,其余99个菌株包含了来自19个属的至少30种细菌,且肠球菌属、变形杆菌属和芽胞杆菌属为其中的优势菌属。本研究显示冷冻原肠浸泡解冻池水中细菌多样性较高,其中部分细菌为致病菌和条件致病菌。The aim of this study was to study the bacterial diversity in water of thawing pool for frozen intestines, three water samples were collected and total bacterial genomic DNA was extracted directly.After the 16S rDNA genes were amplified from the total bacterial genomic DNA by PCR, three clone libraries were constructed separately. Positive clones were identified by PCR and then 50 of them in each library were randomly selected for DNA sequencing.Based on the analyses of DNA sequences,it showed that, except 24.00 to 32.00 percent （%） of the unidentified bacteria which including uncultured and unclassified bacteria,the rest bacteria were affiliated with the classes Bacilli, 7-Proteobacteria, Actinobacteria and Fla- vobacteria,which accounting for 38.00%-42.00%, 20.00%-26.00%, 4.00%-8.00% and 0-2.00% of the clones in each library, respectively.Further analysis was conducted on the 106 stains of identified bacteria and no consideration was given to 44 unidentified bacteria,the results showed that, except 2 bacteria belonging to Actinomycetales and 5 bacteria belonging to Enterobacteriaceae which could not be further identified,the remaining 99 bacteria belonged to 19 genera and more than 30 species,and the dominant genera were Enterococcus,Proteus and Bacillus. This study revealed that the bacterial diversity in water of thawing pool for frozen intestines is high, and part of them are pathogenic and opportunistic pathogenic strains.

关 键 词：冷冻原肠 解冻池 细菌多样性 16SrDNA文库 

分 类 号：S852.6[农业科学—基础兽医学]