denticola produced large amounts of acetic and lactic acid but no measurable amount of any other VFA (data not shown). Hydrogen sulfide production All isolates and reference species produced Selleck NSC23766 copious amounts of hydrogen sulfide as measured by lead acetate paper suspended above the actively growing culture. Substrate utilization and growth conditions All four of the original Iowa DD isolates shared enzymatic similarity, 16SrRNA gene sequence similarity, and were isolated from the same herd. Consequently, further examination of growth characteristics and nutrient utilization were carried out using isolate 4A. Growth of isolate 4A did not occur in OTI without the

addition of bovine rumen fluid or in the absence of volatile fatty acids in BMV (data not PND-1186 purchase shown). Bovine serum was required for growth in both media types. In contrast to T. vincentii and T. denticola, T. phagedenis and isolate 4A required serum in addition to VFA and complex amino acids for growth [21]. Nutrient utilization was determined for isolate 4A cells grown in BMV medium. Isolate 4A grew in the absence of heart infusion broth but growth was restricted Sotrastaurin in the absence of polypeptone or yeast extract, suggesting an amino acid requirement. Enhanced growth (resulting in an increase in O.D. <0.1 above that seen when isolate 4A was

grown in BMV without carbohydrate) was observed using fructose, glucose, maltose, mannitol, mannose, pectin, ribose and soluble starch as carbohydrate source, whereas

no enhancement of growth was observed for arabinose, cellobiose, galactose, lactose, sucrose, trehalose or xylose. These results are summarized medroxyprogesterone and compared to two other Treponema species (Table 3). Optimal growth temperature for isolate 4A is 40°C with a range of 29-42°C. Cells in OTI exposed to lower temperatures (down to 4°C) do not grow but remain viable for an extended period of time and will resume growth upon incubation in the optimal temperature range (data not shown). Optimal pH for growth of isolate 4A is pH 7.4 with a range of 6.5-8.0. The general description, temperature, pH range and serum requirement for growth of isolate 4A match those given for Treponema phagedenis in Bergey’s Manual of Systematic Bacteriology [18]. Mean generation time in OTI was 4 hours with a maximal density of 109 cells/ml in 96 hours (Additional file 1: Figure S1). Mean generation time in BMV was slightly longer, at 6.8 hours and reaching lower maximal density of 108 cells/ml at 96 hours (Additional file 1: Figure S1). Table 3 Utilization of carbohydrate sources by novel isolate 4A and other known Treponeme species   Strain 4A** T. phagedenis† T. phagedenis (ATCC 27087)** T. denticola (ATCC 35405)** T.