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人类粪便微生物群中“阿托波菌属菌群”的动态变化与多样性,以及“阿托波菌属菌群”分离株的表型特征

Dynamics and diversity of the 'Atopobium cluster' in the human faecal microbiota, and phenotypic characterization of 'Atopobium cluster' isolates.

作者信息

Thorasin Thanikan, Hoyles Lesley, McCartney Anne L

机构信息

Microbial Ecology & Health Group, Department of Food and Nutritional Sciences, School of Chemistry, Food and Pharmacy, University of Reading, Whiteknights, PO Box 226, Reading RG6 6AP, UK.

Department of Biomedical Sciences, Faculty of Science and Technology, University of Westminster, 115 New Cavendish Street, London W1W 6UW, UK.

出版信息

Microbiology (Reading). 2015 Mar;161(Pt 3):565-79. doi: 10.1099/mic.0.000016. Epub 2014 Dec 22.

Abstract

This study monitored the dynamics and diversity of the human faecal 'Atopobium cluster' over a 3-month period using a polyphasic approach. Fresh faecal samples were collected fortnightly from 13 healthy donors (six males and seven females) aged between 26 and 61 years. FISH was used to enumerate total (EUB338mix) and 'Atopobium cluster' (ATO291) bacteria, with counts ranging between 1.12×10(11) and 9.95×10(11), and 1.03×10(9) and 1.16×10(11) cells (g dry weight faeces)(-1), respectively. The 'Atopobium cluster' population represented 0.2-22 % of the total bacteria, with proportions donor-dependent. Denaturing gradient gel electrophoresis (DGGE) using 'Atopobium cluster'-specific primers demonstrated faecal populations of these bacteria were relatively stable, with bands identified as Collinsella aerofaciens, Collinsella intestinalis/Collinsella stercoris, Collinsella tanakaei, Coriobacteriaceae sp. PEAV3-3, Eggerthella lenta, Gordonibacter pamelaeae, Olsenella profusa, Olsenella uli and Paraeggerthella hongkongensis in the DGGE profiles of individuals. Colony PCR was used to identify 'Atopobium cluster' bacteria isolated from faeces (n = 224 isolates). 16S rRNA gene sequence analysis of isolates demonstrated Collinsella aerofaciens represented the predominant (88 % of isolates) member of the 'Atopobium cluster' found in human faeces, being found in nine individuals. Eggerthella lenta was identified in three individuals (3.6 % of isolates). Isolates of Collinsella tanakaei, an 'Enorma' sp. and representatives of novel species belonging to the 'Atopobium cluster' were also identified in the study. Phenotypic characterization of the isolates demonstrated their highly saccharolytic nature and heterogeneous phenotypic profiles, and 97 % of the isolates displayed lipase activity.

摘要

本研究采用多相方法,在3个月的时间内监测了人类粪便中“阿托波菌属菌群”的动态变化和多样性。每两周从13名年龄在26至61岁之间的健康供体(6名男性和7名女性)中采集新鲜粪便样本。荧光原位杂交(FISH)用于计数总细菌(EUB338mix)和“阿托波菌属菌群”(ATO291)细菌,计数范围分别为1.12×10¹¹至9.95×10¹¹以及1.03×10⁹至1.16×10¹¹个细胞/(克干重粪便)⁻¹。“阿托波菌属菌群”数量占总细菌的0.2 - 22%,比例因供体而异。使用“阿托波菌属菌群”特异性引物的变性梯度凝胶电泳(DGGE)显示,这些细菌的粪便菌群相对稳定,在个体的DGGE图谱中鉴定出的条带对应的细菌有产气柯林斯菌、肠道柯林斯菌/粪便柯林斯菌、田形柯林斯菌、科里杆菌科PEAV3 - 3菌属、迟缓埃格特菌、帕氏戈登菌、丰富奥尔森菌、尤氏奥尔森菌和香港副埃格特菌。菌落PCR用于鉴定从粪便中分离出的“阿托波菌属菌群”细菌(n = 224株分离株)。对分离株的16S rRNA基因序列分析表明,产气柯林斯菌是人类粪便中“阿托波菌属菌群”的主要成员(占分离株的88%),在9名个体中被发现。在3名个体中鉴定出迟缓埃格特菌(占分离株的3.6%)。在该研究中还鉴定出了田形柯林斯菌、一种“Enorma”菌属以及属于“阿托波菌属菌群”的新物种代表的分离株。对分离株的表型特征分析表明它们具有高度的糖分解特性和异质的表型特征,并且97%的分离株具有脂肪酶活性。

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