title: First Cultivation of Health-Associated Tannerella sp. HOT-286 (BU063).
abstract: Despite significant advances in recent years in culture-independent molecular microbiology methods, the detailed study of individual bacterial species still relies on having pure cultures in the laboratory. Yet, more than a third of the approximately 700 bacterial taxa found in the human oral cavity are as yet uncultivated in vitro. One such taxon, Tannerella sp. HOT-286 (phylotype BU063), is the focus of much interest since it is associated with periodontal health, while Tannerella forsythia, its closest phylogenetic neighbor, is strongly associated with periodontal disease. HOT-286, however, has remained uncultivated despite the efforts of several research groups, spanning over a decade. The aim of this study was to cultivate Tannerella sp. HOT-286. A heavily diluted sample of subgingival plaque was inoculated onto culture plates supplemented with siderophores (pyoverdines-Fe complex or desferricoprogen) or a neat plaque suspension. After 8 d of anaerobic incubation, microcolonies and colonies showing satellitism were passaged onto fresh culture plates cross-streaked with potential helper strains or onto cellulose-acetate membranes placed over lawn cultures of helper strains. Subcultured colonies were identified by 16S rRNA gene sequencing, and purity was confirmed by sequencing 20 clones per library prepared from a single colony. Three colonies of interest (derived from pyoverdines- and plaque-supplemented plates) were identified as Tannerella sp. HOT-286. The isolates were found to be incapable of independent growth, requiring helpers such as Propionibacterium acnes and Prevotella intermedia for stimulation, with best growth on membranes over "helper" lawns. A representative isolate was subjected to phenotypic characterization and found to produce a range of glycosidic and proteolytic enzymes. Further comparison of this novel "periodontal health-associated" taxon with T. forsythia will be valuable in investigating virulence factors of the latter and possible health benefits of the former.
authors.affiliation: Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, UK.
authors.forename: S R
authors.initials: SR
authors.lastname: Vartoukian
authors.affiliation: King's College London Dental Institute, London, UK.
authors.forename: R V
authors.initials: RV
authors.lastname: Moazzez
authors.affiliation: The Forsyth Institute, Cambridge, MA, USA Harvard School of Dental Medicine, Boston, MA, USA.
authors.forename: B J
authors.initials: BJ
authors.lastname: Paster
authors.affiliation: The Forsyth Institute, Cambridge, MA, USA Harvard School of Dental Medicine, Boston, MA, USA.
authors.forename: F E
authors.initials: FE
authors.lastname: Dewhirst
authors.affiliation: Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, UK The Forsyth Institute, Cambridge, MA, USA w.wade@qmul.ac.uk.
authors.forename: W G
authors.initials: WG
authors.lastname: Wade
mesh_terms: D001431:Bacteriological Techniques / Q000379:methods; D003470:Culture Media; D003773:Dental Plaque / Q000382:microbiology; D005260:Female; D006801:Humans; D008875:Middle Aged; D010510:Periodontal Diseases / Q000382:microbiology; D000070436:Tannerella forsythia / Q000254:growth & development* / Q000472:pathogenicity
chemical_list: D003470:Culture Media
keywords: bacteriology; culture; dental plaque; isolation; microbiome; periodontitis