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基于培养和变性梯度凝胶电泳分析的蚯蚓(Eisenia fetida)肠道细菌群落结构。

Culture-based and denaturing gradient gel electrophoresis analysis of the bacterial community structure from the intestinal tracts of earthworms(Eisenia fetida).

机构信息

Division of Biological Science and Technology, Yonsei University, Wonju 220-710, Korea.

出版信息

J Microbiol Biotechnol. 2011 Sep;21(9):885-92. doi: 10.4014/jmb.1009.09041.

Abstract

The bacterial communities in the intestinal tracts of earthworm were investigated by culture-dependent and - independent approaches. In total, 72 and 55 pure cultures were isolated from the intestinal tracts of earthworms under aerobic and anaerobic conditions, respectively. Aerobic bacteria were classified as Aeromonas (40%), Bacillus (37%), Photobacterium (10%), Pseudomonas (7%), and Shewanella (6%). Anaerobic bacteria were classified as Aeromonas (52%), Bacillus (27%), Shewanella (12%), Paenibacillus (5%), Clostridium (2%), and Cellulosimicrobium (2%). The dominant microorganisms were Aeromonas and Bacillus species under both aerobic and anaerobic conditions. In all, 39 DNA fragments were identified by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis. Aeromonas sp. was the dominant microorganism in feeds, intestinal tracts, and casts of earthworms. The DGGE band intensity of Aeromonas from feeds, intestinal tracts, and casts of earthworms was 12.8%, 14.7%, and 15.1%, respectively. The other strains identified were Bacillus, Clostridium, Enterobacter, Photobacterium, Pseudomonas, Shewanella, Streptomyces, uncultured Chloroflexi bacterium, and uncultured bacterium. These results suggest that PCR-DGGE analysis was more efficient than the culture-dependent approach for the investigation of bacterial diversity and the identification of unculturable microorganisms.

摘要

采用培养依赖和非依赖的方法研究了蚯蚓肠道中的细菌群落。在有氧和厌氧条件下,分别从蚯蚓肠道中分离出 72 株和 55 株纯培养物。好氧细菌可分为气单胞菌(40%)、芽孢杆菌(37%)、发光杆菌(10%)、假单胞菌(7%)和希瓦氏菌(6%)。厌氧菌可分为气单胞菌(52%)、芽孢杆菌(27%)、希瓦氏菌(12%)、类芽孢杆菌(5%)、梭菌(2%)和纤维单胞菌(2%)。在有氧和厌氧条件下,优势微生物均为气单胞菌和芽孢杆菌。通过聚合酶链反应-变性梯度凝胶电泳(PCR-DGGE)分析鉴定了 39 个 DNA 片段。气单胞菌是饲料、肠道和蚯蚓粪便中优势微生物。气单胞菌在饲料、肠道和蚯蚓粪便中的 DGGE 条带强度分别为 12.8%、14.7%和 15.1%。鉴定出的其他菌株有芽孢杆菌、梭菌、肠杆菌、发光杆菌、假单胞菌、希瓦氏菌、链霉菌、未培养的绿弯菌和未培养的细菌。这些结果表明,与培养依赖方法相比,PCR-DGGE 分析更有效,可用于研究细菌多样性和鉴定不可培养微生物。

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