Free-living bacteria with spherical cells 0.5-2.5 µm in diameter were isolated from freshwater sediment. 16S rRNA gene sequence analysis placed the new isolates within the phylum Spirochaetes ('spirochaetes'). The isolates never displayed a helical morphology or motility. Growth occurred in the presence of 100 mg ampicillin l(-1) in complex and defined mineral salts medium amended with vitamins, yeast extract and monosaccharides, disaccharides or soluble starch as fermentable substrates. Two distinct isolates, designated Buddy(T) and Grapes(T), exhibited doubling times of 21±2 and 15±1 h in glucose-amended medium and grew at 15-37 and 15-30 °C. Optimum growth was observed between 25 and 30 °C and pH 6.5-7.5, with no growth below pH 5 or above pH 10. Hexose and pentose fermentation yielded ethanol, acetate and formate as major end products. Growth was strictly fermentative and anaerobic, but the isolates tolerated brief oxygen exposure. Nitrate, sulfate, thiosulfate and carbon dioxide were not used as electron acceptors, but soluble Fe(III) was reduced to Fe(II) in glucose-amended medium. The DNA G+C base contents of isolates Buddy(T) and Grapes(T) were 45.5-46.4 and 47.0-49.2 mol%, respectively. Phospholipid fatty acid (PLFA) profiles contained large proportions of C(14:0) and C(16:0) straight-chain saturated fatty acids; C(16:1)ω7c and C(16:1)ω9c dominated the mono-unsaturated PLFAs in isolate Grapes(T), whereas isolate Buddy(T) also possessed C(18:1)ω5c, C(18:1)ω7c and C(18:1)ω9c fatty acids. Branched monoenoic acids accounted for up to 12.4 and 30% of the total PLFA in isolates Grapes(T) and Buddy(T), respectively. Based on their unique morphological features and the phylogenetic distance from their closest relatives, we propose the new genus, Sphaerochaeta gen. nov., to accommodate the new isolates within the novel species Sphaerochaeta globosa sp. nov. (type strain Buddy(T) =DSM 22777(T) =ATCC BAA-1886(T)) and Sphaerochaeta pleomorpha sp. nov. (type strain Grapes(T) =DSM 22778(T) =ATCC BAA-1885(T)). Sphaerochaeta globosa is the type species of the genus.