ERIC-PCR分子杂交技术分析大熊猫肠道菌群结构

ERIC-PCR based fingerprinting and molecular hybridization to analyze the characteristics of intestinal microflora of giant panda

目的 了解大熊猫肠道微生物区系结构的相似性和稳定性,并找出大熊猫肠道微生物群落结构的变化与健康状况的关系。方法 对上海动物园及上海野生动物园所饲养的3只大熊猫2次采集的粪便样品进行微生物群落总DNA的抽提,并以此为模板获得反映肠道微生物群落结构特征的ERIC- PCR和Southern杂交指纹图谱,比较各DNA样品指纹图谱的相似性指数。结果 除国庆(大熊猫)的第1次采集的样品(当时处于腹泻状态) ,其他各DNA样品的ERIC- PCR及Southern杂交指纹图谱的相似性都达到85 %～10 0 % ;佳斯及川川(大熊猫) 2个个体2次采集的样品之间ERIC指纹图谱的相似性分别为93%和87% ,而国庆腹泻时的样品与健康时的样品之间则为71%。结论 大熊猫不同个体之间肠道微生物群落结构比较相似,而且同一个体在不同时期表现出比较高的稳定性,但当个体的健康出现问题时肠道优势菌菌群结构有一定波动。所采用的DNA提取方法、ERIC- PCR和Southern杂交指纹图谱的高度重复性证明了之一分子生态学技术在大熊猫肠道微生物区系动态监测中的可行性。

Objective To investigate the structural variation and changes in intestinal microbial community of panda and find out whether the structure of intestinal microbial community in panda was related to its health.Methods The total DNAs of microbial community were extracted from three giant pandas' fecal samples,subsequently used as templates in ERIC PCR to generate community fingerprint,then molecular hybridization was carried out.Results Except Guoqing's lax fecal sample GQ1,the similarities of ERIC fingerprints between all samples under healthy condition were more than 85%.The hybridiztion fingerprint also showed the high similarity between different samples.However,the similarity coefficient between Guoqing's lax fecal sample GQ1 and healthy sample GQ2 was 71%.The similarities of ERIC fingerprinting profiles between individual Jiasi and Guoqing which were caged in the same zoo was relatively highter,over 93%,while the similarity between them and individal Chuanchuan which was caged in another zoo were lower,about 85%.Conclusions The above result suggested that the intestinal microflora were very similar between different healthy panda individuals and also very stable when they were under health conditions.And microbial community of individuals under the same feeding condition tends to be more similar.