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Cultivation of a Synergistetes strain representing a previously uncultivated lineage.
Environ Microbiol. 2010 Apr; 12(4):916-28.EM

Abstract

Subgingival plaque samples obtained from human subjects with periodontitis, shown to include previously uncultivable members of the phylum Synergistetes, were used to inoculate Cooked Meat Medium (CMM). The presence of Cluster A (uncultivable) Synergistetes was monitored by fluorescent in situ hybridization (FISH) and quantitative PCR (Q-PCR). Cluster A Synergistetes were found to grow in CMM in co-culture with other plaque bacteria and growth was stimulated by the addition of mucin and serum. Plaque samples were also used to inoculate Blood Agar (BA) plates and growth of Cluster A Synergistetes was revealed after anaerobic incubation, by colony hybridization with specific probes. Surface growth on the plates in regions identified by colony hybridization was harvested and used to inoculate fresh plates, thus enriching for Cluster A Synergistetes. Cross-streaks of other plaque bacteria were also used to stimulate Synergistetes growth. In the early passages, no discrete Synergistetes colonies were seen, but after eight passages and the use of cross-streaks of other bacteria present in the enriched community, colonies arose, which consisted solely of Cluster A Synergistetes cells, as determined by 16S rRNA gene PCR and cloning. This is the first report of the successful culture of a member of the uncultivable branch of this phylum.

Authors+Show Affiliations

King's College London Dental Institute, Infection Research Group, Guy's Campus, London SE1 9RT, UK.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20074237

Citation

Vartoukian, S R., et al. "Cultivation of a Synergistetes Strain Representing a Previously Uncultivated Lineage." Environmental Microbiology, vol. 12, no. 4, 2010, pp. 916-28.
Vartoukian SR, Palmer RM, Wade WG. Cultivation of a Synergistetes strain representing a previously uncultivated lineage. Environ Microbiol. 2010;12(4):916-28.
Vartoukian, S. R., Palmer, R. M., & Wade, W. G. (2010). Cultivation of a Synergistetes strain representing a previously uncultivated lineage. Environmental Microbiology, 12(4), 916-28. https://doi.org/10.1111/j.1462-2920.2009.02135.x
Vartoukian SR, Palmer RM, Wade WG. Cultivation of a Synergistetes Strain Representing a Previously Uncultivated Lineage. Environ Microbiol. 2010;12(4):916-28. PubMed PMID: 20074237.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Cultivation of a Synergistetes strain representing a previously uncultivated lineage. AU - Vartoukian,S R, AU - Palmer,R M, AU - Wade,W G, Y1 - 2010/01/13/ PY - 2010/1/16/entrez PY - 2010/1/16/pubmed PY - 2010/7/9/medline SP - 916 EP - 28 JF - Environmental microbiology JO - Environ Microbiol VL - 12 IS - 4 N2 - Subgingival plaque samples obtained from human subjects with periodontitis, shown to include previously uncultivable members of the phylum Synergistetes, were used to inoculate Cooked Meat Medium (CMM). The presence of Cluster A (uncultivable) Synergistetes was monitored by fluorescent in situ hybridization (FISH) and quantitative PCR (Q-PCR). Cluster A Synergistetes were found to grow in CMM in co-culture with other plaque bacteria and growth was stimulated by the addition of mucin and serum. Plaque samples were also used to inoculate Blood Agar (BA) plates and growth of Cluster A Synergistetes was revealed after anaerobic incubation, by colony hybridization with specific probes. Surface growth on the plates in regions identified by colony hybridization was harvested and used to inoculate fresh plates, thus enriching for Cluster A Synergistetes. Cross-streaks of other plaque bacteria were also used to stimulate Synergistetes growth. In the early passages, no discrete Synergistetes colonies were seen, but after eight passages and the use of cross-streaks of other bacteria present in the enriched community, colonies arose, which consisted solely of Cluster A Synergistetes cells, as determined by 16S rRNA gene PCR and cloning. This is the first report of the successful culture of a member of the uncultivable branch of this phylum. SN - 1462-2920 UR - https://www.unboundmedicine.com/medline/citation/20074237/Cultivation_of_a_Synergistetes_strain_representing_a_previously_uncultivated_lineage_ L2 - https://doi.org/10.1111/j.1462-2920.2009.02135.x DB - PRIME DP - Unbound Medicine ER -