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鸵鸟盲肠微生物多样性的 16S 核糖体 RNA 基因克隆文库分析及新型纤维杆菌属的检测

Microbial diversity in ostrich ceca as revealed by 16S ribosomal RNA gene clone library and detection of novel Fibrobacter species.

机构信息

Graduate School of Bioresources, Mie University, 1577 Kurimamachiya-cho, Tsu, Mie 514-8507, Japan.

出版信息

Anaerobe. 2010 Apr;16(2):83-93. doi: 10.1016/j.anaerobe.2009.07.005. Epub 2009 Jul 24.

Abstract

The ostrich (Struthio camelus) is a herbivorous bird and although the hindgut is known as the site for fiber digestion, little is known about the microbial diversity in the ostrich hindgut. Our aim was to analyze the microbial diversity in ostrich ceca using a 16S ribosomal RNA gene (rDNA) clone library approach. A total of 310 clones were sequenced and phylogenetically analyzed and were classified into 110 operational taxonomy units (OTUs) based on a 98% similarity criterion. The similarity of the sequences ranged from 86 to 99% and 95 OTUs had less than 98% similarity to the sequences in the public databases. Coverage and the Shannon-Wiener index (H') of the library were 83.9% and 4.29, respectively. The sequences were assigned to the following 6 phyla: Firmicutes (50.9% of the total number of sequences), Bacteroidetes (39.4%), Fibrobacteres (6.5%), Euryarchaeota (1.9%), Spirochaetes (1.0%), and Verrucomicrobia (0.3%); approximately 90% of the sequences were affiliated with Firmicutes and Bacteroidetes. The only OTU of Fibrobacteres (OTU 107), had 93 and 90% similarity to Fibrobacter succinogenes and F. intestinalis, respectively, suggesting a new species of Fibrobacter in ostrich ceca. Clostridium coccoides and C. leptum formed major groups within the Firmicutes. There was no OTU with high similarity (> or =98%) to the 16S rDNA of cultivated fibrolytic bacteria in our library. Although two OTUs were affiliated with Euryarchaeota, no sequence was affiliated with methanogenic Archaea. This study presents the very complex ostrich cecal microbial community, in which the majority of the bacterial species have not yet been cultivated.

摘要

鸵鸟(Struthio camelus)是一种草食性鸟类,虽然后肠被认为是纤维消化的部位,但对鸵鸟后肠的微生物多样性知之甚少。我们的目的是使用 16S 核糖体 RNA 基因(rDNA)克隆文库方法分析鸵鸟盲肠中的微生物多样性。共测序并分析了 310 个克隆,并根据 98%相似性标准将其分类为 110 个操作分类单元(OTU)。序列的相似性范围为 86%至 99%,95 个 OTU 与公共数据库中的序列相似度小于 98%。文库的覆盖率和 Shannon-Wiener 指数(H')分别为 83.9%和 4.29。序列被分配到以下 6 个门:厚壁菌门(总序列数的 50.9%)、拟杆菌门(39.4%)、纤维杆菌门(6.5%)、广古菌门(1.9%)、螺旋体门(1.0%)和疣微菌门(0.3%);约 90%的序列与厚壁菌门和拟杆菌门有关。唯一属于纤维杆菌门的 OTU(OTU107)与纤维形丁酸弧菌和 F. intestinalis 的相似性分别为 93%和 90%,提示在鸵鸟盲肠中存在一种新的纤维形丁酸弧菌类群。梭菌属 coccoides 和 C. leptum 是厚壁菌门中的主要群组。我们的文库中没有与培养的纤维分解菌 16S rDNA 具有高相似性(≥98%)的 OTU。虽然有两个 OTU 与广古菌门有关,但没有序列与产甲烷古菌有关。本研究展示了非常复杂的鸵鸟盲肠微生物群落,其中大多数细菌尚未被培养。

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