title: PCR detection of uncultured rumen bacteria.
abstract: 16S rRNA sequences of ruminal uncultured bacterial clones from public databases were phylogenetically examined. The sequences were found to form two unique clusters not affiliated with any known bacterial species: cluster of unidentified sequences of free floating rumen fluid uncultured bacteria (FUB) and cluster of unidentified sequences of bacteria associated with rumen epithelium (AUB). A set of PCR primers targeting 16S rRNA of ruminal free uncultured bacteria and rumen epithelium adhering uncultured bacteria was designed based on these sequences. FUB primers were used for relative quantification of uncultured bacteria in ovine rumen samples. The effort to increase the population size of FUB group has been successful in sulfate reducing broth and culture media supplied with cellulose.
authors.affiliation: Institute of Animal Physiology and Genetics, Vídeňská 1083, 142 20, Prague 4, Czech Republic.
authors.forename: Jaime A
authors.initials: JA
authors.lastname: Rosero
authors.forename: Lenka
authors.initials: L
authors.lastname: Strosová
authors.forename: Jakub
authors.initials: J
authors.lastname: Mrázek
authors.forename: Kateřina
authors.initials: K
authors.lastname: Fliegerová
authors.forename: Jan
authors.initials: J
authors.lastname: Kopečný
mesh_terms: D000818:Animals; D001419:Bacteria / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D002417:Cattle; D003470:Culture Media / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D006041:Goats; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D012417:Rumen / Q000382:microbiology*; D012756:Sheep
chemical_list: D003470:Culture Media; D017931:DNA Primers