Open AccessDevelopment and use of competitive PCR assays for the rumen cellulolytic bacteria: Fibrobacter succinogenes , Ruminococcus albus and Ruminococcus flavefaciens
Author(s) -
Koike Satoshi,
Kobayashi Yasuo
Publication year - 2001
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2001.tb10911.x
Subject(s) - fibrobacter succinogenes , ruminococcus , rumen , biology , caecum , microbiology and biotechnology , streptococcus bovis , population , bacteria , food science , feces , fermentation , medicine , genetics , demography , sociology
Competitive PCR assays were developed for the enumeration of the rumen cellulolytic bacterial species: Fibrobacter succinogenes , Ruminococcus albus and Ruminococcus flavefaciens . The assays, targeting species‐specific regions of 16S rDNA, were evaluated using DNA from pure culture and rumen digesta spiked with the relevant cellulolytic species. Minimum detection levels for F. succinogenes , R. albus and R. flavefaciens were 1–10 cells in pure culture and 10 3–4 cells per ml in mixed culture. The assays were reproducible and 11–13% inter‐ and intra‐assay variations were observed. Enumeration of the cellulolytic species in the rumen and alimentary tract of sheep found F. succinogenes dominant (10 7 per ml of rumen digesta) compared to the Ruminococcus spp. (10 4–6 per ml). The population size of the three species did not change after the proportion of dietary alfalfa hay was increased. All three species were detected in the rumen, omasum, caecum, colon and rectum. Numbers of the cellulolytic species at these sites varied within and between animals.