蜜蜂成虫工蜂肠道可培养细菌群落结构分析

Diversity and community structure of intestinal cultivable bacteria in adult worker-bee

【目的】研究2种蜜蜂(健康意大利蜜蜂和健康中华蜜蜂)成虫工蜂肠道可培养细菌的群落结构组成。【方法】利用16S r RNA基因的聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)分析技术,结合菌落形态观察和生理生化特征鉴定细菌种类。【结果】从2种蜜蜂成虫工蜂肠道200株可培养细菌得到18种不同细菌遗传型,分属于肠杆菌科(Enterobacteriaceae)、弧菌科(Vibrionaceae)和肠球菌科(Enterococcaceae)3个科。其中肠杆菌科是肠道可培养细菌最优势的细菌种类。同样以序列相似性大于97%的菌株归为相同细菌种类为标准,找到了2种蜜蜂可培养细菌的共有菌种,结合菌落形态观察和生理生化特征鉴定,确定肠道可培养细菌为肠杆菌属8株,克雷伯氏菌属1株,肠球菌属2株,以及气单胞菌属1株。【结论】通过研究健康意大利蜜蜂和中华蜜蜂成虫工蜂肠道可培养细菌群落结构组成,可为开展蜜蜂的微生态研究提供基础性资料。

[Objective] In order to identify the diversity and community structure of the cultivable bacteria inhabiting the intestinal tract in worker adults of honeybee from herlthy Apis mellifera ligustica and Apis cerana cerana Fabricius populations. [Methods] 16 S r RNA gene PCR-DGGE（denaturing gradient gel electrophoresis） molecular methods was employed to study the bacterial diversity. In addition, the cultivable bacteria were identified by colony morphology, physiological and biochemical characteristics tests. [Results] A total of 200 culturable bacteria from aboved two populations were classified into 18 unique phylotypes. All sequenced bacteria strains were grouped into three families： Enterobacteriaceae, Vibrionaceae and Enterococcaceae. The Enterobacteriaceae was dominated in all the populations. The closely related sequences（97% sequence similarity） which had been retrieved from culturable bacteria of two populations were grouped as one common species. We identified the cultivable bacteria： eight strains of bacteria as Enterobacter, one strain of Klebsiella, two strains of Enterococcus, and one strain of Aeromonas. [Conclusion] The present study significantly contributes to the available information on bacterial isolates from worker adults of honeybee.