Cryptobacterium curtum gen. nov., sp. nov., a new genus of Gram-positive anaerobic rod isolated from human oral cavities

1 Department of Oral Microbiology, School of Dentistry, Niigata University, Niigata, 951-8514, Japan 2 Departments of Oral Biology, School of Dentistry, University of Umeå, Umeå, Sweden 3 Endodontics, School of Dentistry, University of Umeå, Umeå, Sweden Author for correspondence: Etsuro Hoshino. Tel: + 81 25 227 2838. Fax: + 81 25 227 0806. e-mail: hoshino{at}dent.niigata-u.ac.jp ABSTRACT Novel Eubacterium -like isolates, strains 12-3 T and KV43-B, which were isolated from the periodontal pocket of an adult patient with periodontal disease and necrotic dental pulp, respectively, were studied taxonomically and phylogenetically. The morphological and differential biochemical characteristics of these organisms are also described in this paper. These organisms were Gram-positive, anaerobic, non-spore-forming, rod-shaped bacteria that were inert in most of the conventional biochemical tests and closely resembled members of asaccharolytic oral Eubacterium species. On the other hand, protein profiles of whole cells in SDS-PAGE and Western immunoblotting reaction analysis distinguished these isolates from strains of the previously described genus Eubacterium . The G+C content of the DNAs from the novel isolates was 50 and 51 mol%, respectively. The levels of DNA-DNA relatedness to other asaccharolytic oral Eubacterium species, including Eubacterium brachy, Eubacterium lentum, Eubacterium nodatum, Eubacterium timidum, Eubacterium saphenum, Eubacterium minutum and Eubacterium exiguum , was less than 11 %. These organisms also exhibited a very low level of reassociation with the DNA of Eubacterium limosum , the type species of the genus Eubacterium . The results of 16S rDNA sequence comparisons revealed that these organisms represent a novel lineage distinct from all previously described genera of Gram-positive, rod-shaped bacteria. On the basis of our results, it is suggested that strains 12-3 T and KV43-B should be classified in a new genus and species, for which the name Cryptobacterium curtum gen. nov., sp. nov. is proposed. The type strain of Cryptobacterium curtum is 12-3 T (= ATCC 700683 T ). Key Words: Cryptobacterium curtum gen. nov. • sp. nov. • DNA-DNA relatedness • 16S rDNA sequence • phylogenetic analysis The DDBJ/EMBL/GenBank accession number for the 16S rRNA gene sequence of strain 12-3 T is AB019260 .