Unravelling the genetic diversity of ruminal bacteria belonging to the CFB phylum

A Ramsak, M Peterka, K Tajima, J C Martin, J Wood, M E A Johnston, R I Aminov, H J Flint, G Avgustin

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Abstract

Molecular biology approaches ware employed to examine the genetic diversity of bacteria from the Cytophaga/Flexibacter/Bacteroides (CFB) phylum in the rumen of cattle. By this means we were able to identify cultured strains that represent some of the larger CFB clusters previously identified only by PCR amplification and sequencing. Complete 16S rDNA sequences were obtained for 16 previously isolated rumen strains, including the type strains of Prevotella ruminicola, P. bryantii, P. brevis and P. albensis to represent a wide range of diversity. Phylogenetic analysis of cultured strains revealed the existence of three clusters of ruminal CFB: (i) a cluster of Prevotella strains, which have been found only in the rumen, including the two type strains, P. brevis GA33(T) and P. ruminicola 23(T), (ii) Prevotella spp. that cluster with prevotellas from other ecological niches such as the oral cavity and which include the type strains, P. bryantii B(1)4(T) and P. albensis M384(T); (iii) two Bacteroides sop. strains clustering with B. forsythus of oral origin. In order to establish whether the cultivated isolates cover the whole range of ruminal CFB genetic diversity, 16S rRNA gene sequences were amplified and cloned from DNA extracted from the same rumen samples (one cow in Slovenia, one in Scotland and three in Japan). Sequencing and phylogenetic analysis of 16S rRNA genes confirmed the existence of two superclusters of ruminal Prevotella, one exclusively ruminal and the other including non-ruminal species. In the case of ruminal Bacteroides spp., however, phylogenetic analysis revealed the existence of three new superclusters, one of which has as yet no cultivable counterpart. Interestingly, these Bacteroides clusters were represented almost exclusively by clone libraries from the Japanese cattle and only three sequences were from the European cattle. This study agrees with previous analyses in showing that rumen Prevotella/Bacteroides strains exhibit a remarkable degree of genetic diversity and suggests that different strain groupings may differ greatly in their recovery by cultural methods. The most important conclusion, however, is that cultured strains can be identified that represent some of the larger clusters previously identified only by PCR amplification and sequencing. (C) 2000 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

Original languageEnglish
Pages (from-to)69-79
Number of pages11
JournalFEMS Microbiology Ecology
Volume33
Issue number1
DOIs
Publication statusPublished - Jul 2000

Keywords

  • ruminal bacterium
  • genetic diversity
  • Cytophaga/Flexibacter/Bacteroides phylum
  • 16S rDNA library
  • sequence-analysis
  • bacteroides-ruminicola
  • phylogenetic analysis
  • rumen
  • DNA
  • identification
  • strains

Cite this

Ramsak, A., Peterka, M., Tajima, K., Martin, J. C., Wood, J., Johnston, M. E. A., Aminov, R. I., Flint, H. J., & Avgustin, G. (2000). Unravelling the genetic diversity of ruminal bacteria belonging to the CFB phylum. FEMS Microbiology Ecology, 33(1), 69-79. https://doi.org/10.1111/j.1574-6941.2000.tb00728.x