On the basis of DNA-DNA hybridization data, nine intestinal spirochete strains were grouped into five genospecies. Three of these genospecies were previously recognized species, (type strain, B78), (type strain, B256), and (type strain, P43/6/78; previously “”). The other two genospecies were found to be new species, for which we propose the names sp. nov. (with type strain PWS/A) and sp. nov. (with type strain 56–150). and cells had a typical spirochete ultrastructure with 22 to 28 periplasmic flagella per cell. Various soluble sugars were growth substrates for and . During growth in basal heart infusion broth supplemented with fetal calf serum beneath an O-N (1:99) atmosphere, cells of these new species consumed oxygen and glucose and produced H, CO, acetate, butyrate, and ethanol. The G+C content of the DNA of 56–150 was 27 mol%, and the G+C content of the DNA of PWS/A was 25 mol%. In addition, a restriction fragment length polymorphism-PCR assay for the detection of intestinal spirochetes was developed. The assay was based on generation and restriction endonuclease analysis (with 3A, and II) of a 558-bp amplicon of ribosomal DNA (rDNA) encoding 16S rRNA. The PCR amplification was specific for species and . Four restriction digest patterns were found for the five species. fl. restriction differentiated and from the other species. 3A and I restrictions gave unique fragment patterns for and , respectively. and DNAs gave the same fragment pattern regardless of the enzyme tested. was differentiated from the species by II digestion of the 16S rDNA amplicon.


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