Treponema putidum sp. nov., a medium-sized proteolytic spirochaete isolated from lesions of human periodontitis and acute necrotizing ulcerative gingivitis

Abstract

So far, little phenotypic heterogeneity has been detected in cultured oral treponemes with trypsin-like proteolytic activity, and all have been assigned to the species Treponema denticola. However, comparisons of protein patterns and antigen expression in our collection of proteolytic oral treponemes occasionally identified isolates with a unique phenotype; e.g. strain OMZ 830 (=ATCC 700768), which qualified as a ‘pathogen-related oral spirochaete’ due to the presence of a ∼37 kDa protein reactive with the Treponema pallidum FlaA-specific mAb H9-2. In addition to such single isolates, a homogeneous group of seven independent strains is described that were highly motile, medium-sized, proteolytic but asaccharolytic spirochaetes and were cultured from human gingivitis, periodontitis and acute necrotizing ulcerative gingivitis in medium OMIZ-Pat supplemented with 1 % human serum and antibiotics. Growth of these spirochaetes in OMIZ-Pat was not dependent on, but was stimulated by, human or bovine serum. Carbohydrates were neither required nor stimulatory for growth. The protein and antigen patterns of total cell extracts of these organisms separated by SDS-PAGE were distinct from those of all previously cultured spirochaetes, with highest similarity to T. denticola. The novel spirochaete has a 2 : 4 : 2 arrangement of the periplasmic flagella, similar to T. denticola. However, the flagellin pattern as detected by immunostaining or glycan staining of Western blots readily distinguished the novel group from T. denticola. Also, distinct from reference strains of T. denticola, none of the novel isolates displayed sialidase or dentilisin activities, both of which are expressed by most strains of T. denticola. Trypsin-like activity and other enzymes as detected by API ZYM test were similar to those of T. denticola. The status of a novel species is supported by the 16S rRNA gene sequence, with 98·5 % similarity to its closest cultured relative, T. denticola. The name Treponema putidum sp. nov. is proposed (type strain OMZ 758T=ATCC 700334T=CIP 108088T).

Exilispira thermophila gen. nov., sp. nov., an anaerobic, thermophilic spirochaete isolated from a deep-sea hydrothermal vent chimney

“…Differential characteristics between strain RASEN T and other genera in the phylum Spirochaetes Data for strain RASEN T are from this study. Data for other genera were taken from Holt et al (1994), Paster et al (1991) and the following sources: Treponema (Norris et al, 2006;Smibert, 1984;Wyss et al, 2004;Nordhoff et al, 2005;, Spirochaeta (Leschine et al, 2006;Canale-Parola, 1984a, b;Hoover et al, 2003;Dröge et al, 2006), Borrelia (Caimono, 2006;Kelly, 1984;Güner et al, 2004;Masuzawa et al, 2001;Fukunaga et al, 1996;Richter et al, 2004;Le Fleche et al, 1997;Baranton et al, 1992;Canica et al, 1993;Wang et al, 1997;Karimi et al, 1979), Cristispira (Breznak, 1984), Brevinema (Defosse et al, 1995), Brachyspira (Stanton, 2006;Hampson & La, 2006) and Leptospira/Leptonema (Adler & Faine, 2006;Faine, 1984;Levett et al, 2006;Perolat et al, 1998;Ramadass et al, 1992). +, Positive; 2, negative; V, dependent on the species; ND, no data available.…”

Virulence Factors of the Oral Spirochete Treponema denticola

“…The numbers of both cultivated and uncultivated Treponema phylotypes reported in recent years have rapidly increased, mainly due to the mass of data obtained from efforts to sequence genes encoding 16S rRNA (Choi et al, 1994;Wyss et al, 2004;Demirkan et al, 2006;Molbak et al, 2006;Nordhoff et al, 2008a;Pringle et al, 2008Pringle et al, , 2009Evans et al, 2009;Sayers et al, 2009;Yano et al, 2009). There are currently 49 species of oral Treponema listed on the Human Oral Microbiome Database (Dewhirst et al, 2010), the best-characterized being T. denticola, Treponema amylovorum, T. lecithinolyticum, Treponema maltophilum, Treponema medium, Treponema parvum, …”

Upregulation of Intercellular Adhesion Molecule 1 and Proinflammatory Cytokines by the Major Surface Proteins of Treponema maltophilum and Treponema lecithinolyticum , the Phylogenetic Group IV Oral Spirochetes Associated with Periodontitis and Endodontic Infections

“…These organisms include enormously diverse Treponema species (8,11), and 10 species have been cultivated so far (10,50,54). Epidemiological studies have shown that some species are more often associated with periodontal diseases, and yet uncultivated treponemes have also been detected predominantly at the disease sites (11,32,53).…”