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In this study, we classified the five strains (ChDC F128(T), ChDC F145, ChDC F174, ChDC F206, and ChDC F300) as a novel species of genus Fusobacterium by DNA-DNA hybridization and multi-locus phylogenetic analysis (MLPA), based on a single sequence (24,715 bp) of 22 concatenated housekeeping genes, with morphological and chemotaxonomic characteristics. DNA-DNA hybridization data showed that the values of genomic relatedness between ChDC F128(T) and each of the other novel strains were ranged from 79.0 to 82.6 %, while those of genomic relatedness between ChDC F128(T) and type strain of each of subspecies of F. nucleatum or Fusobacterium periodonticum were ranged from 40.9 to 54.4 %. MLPA revealed that the 5 strains were clustered as one group and clearly discriminated with F. nucleatum and F. periodonticum with 100 % bootstrap value. The DNA G+C content of the five novel strains were ranged from 26.9 to 27.0 mol%. The cellular fatty acid analysis of clinical isolates and type strains revealed C14:0, C16:0, and cis-9 C16:1 as the major fatty acids. The cell wall peptidoglycan of the 5 strains was comprised of meso-lanthionine. These results show that the 5 strains are novel species and belong to the genus Fusobacterium. Strain ChDC F128(T) (=KCOM 1249(T) = KCTC 5108(T) = JCM 30218(T)) is suggested to be the type strain of a novel species of genus Fusobacterium, for which the name Fusobacterium hwasookii sp. nov. is proposed.

Fusobacterium equinum, a gram negative, rod-shaped and an obligate anaerobic bacterium is a newly described species. The organism is associated with necrotic infections of the respiratory tract in horses that include necrotizing pneumonia, pleuritis and paraoral infections. The species is closely related to F. necrophorum that causes liver abscesses in cattle and sheep, calf-diphtheria in cattle, and foot-rot in sheep and cattle. Leukotoxin, an exotoxin, is an important virulence factor in bovine strains of F. necrophorum. Our objective was to examine strains (n=10) of F. equinum for leukotoxin (lktA) gene and its toxic effects on equine leukocytes. Southern hybridization and partial DNA sequencing revealed that all the 10 strains had the lktA gene with greater similarities to F. necrophorum subsp. necrophorum. The secreted leukotoxin was detected in the culture supernatant and its biological activity was determined by viability assays with equine polymorphonuclear cells (PMNs) using flow cytometry. While culture supernatants of four strains (E1, E7, E9, and E10) were highly toxic to equine PMNs; strain E5 was moderately toxic and the remaining strains (E2, E3, E4, E6, and E8) were only mildly toxic. Our data indicated that F. equinum isolates had lktA gene and its product was toxic to equine leukocytes. Therefore, leukotoxin may be an important virulence factor in F. equinum infections.

Background: Bacteria are implicated in certain forms of model chronic colitis but the identity and role of bacteria in human ulcerative colitis (UC) are uncertain., Aims: To isolate pathogenic bacteria from inflamed mucosa of patients with UC, to examine whether the bacteria have a toxin to Vero cells, and to determine whether the toxin induces UC-like lesions in animals., Methods: Bacteria were isolated from UC patients and supernatants from cultures were filtered and tested for cytotoxicity to Vero cells. Bacterial cells producing the cytotoxic supernatants were examined by polymerase chain reaction for verotoxin genes. Culture supernatants of cytotoxic strains were examined by high performance liquid chromatography for organic acid concentrations. Mice were given enemas containing organic acid at the mean concentration in the supernatants of cytotoxic strains to ascertain whether colonic lesions appear in UC., Results: Only supernatants from cultures of Fusobacterium varium killed Vero cells. Bacterial cells lacked verotoxin genes. Bacterial culture supernatants contained high concentrations of n-butyric acid and the mean concentration (32 mmol/l) was cytotoxic to Vero cells. Twenty four hours after mice were given enemas containing either butyric acid or F varium culture supernatants, colonic ulcers with crypt abscesses, inflammatory cell infiltration, and apoptotic changes were observed., Conclusions: Butyric acid in culture supernatants from cultures of F varium caused UC-like lesions in mice. This study indicates that F varium may be one of the elusive pathogenic factors in UC.

Twenty infrequently reported species of gram-negative anaerobic bacilli other than Fusobacterium nucleatum, Fusobacterium necrophorum, and members of the genus Bacteroides were studied with regard to their role in infection and their susceptibility to antimicrobial agents. In addition, the literature regarding the recovery of these organisms from both the normal flora and infections of humans was reviewed. During a six-year period at the Wadsworth Clinical Anaerobic Bacteriology Research Laboratory (Veterans Administration Wadsworth Medical Center, Los Angeles, Calif.), 39 (6%) of 679 specimens obtained from anaerobic infections yielded "other gram-negative anaerobic bacilli" (OGNAB). Fusobacterium naviforme, Fusobacterium gonidiaformans, Fusobacterium varium, Fusobacterium mortiferum, and Fusobacterium russii were the most commonly isolated OGNAB. Most of the OGNAB tested were resistant to erythromycin, and most strains, except for F. varium, were susceptible to beta-lactam antibiotics and clindamycin. Chloramphenicol and metronidazole were active against all strains of OGNAB tested. Certain Fusobacterium species are undoubtedly previously unrecognized members of the normal flora of the oropharynx, upper respiratory tract, or urogenital tract and may be present in infections derived from these floras.

Gas-liquid chromatographic analysis of volatile fatty acid patterns was performed when growth was initially detected in 87 positive blood cultures. Isovaleric acid, butyric acid, or both were found in 39 of 43 (90%) blood cultures that contained anaerobes but were absent in all 44 cultures that contained facultatively anaerobic or aerobic bacteria. The detection of isovaleric acid indicated the presence of Bacteroides fragilis, as this acid was found in 88% of unimicrobial and 75% of polymicrobial bacteremias due to B. fragilis. Butyric acid was detected most often in blood cultures which grew Fusobacterium spp. (three of three), Clostridium spp. (six of seven), or both (three of four).

The demonstration of indole formation is of great value for the identification of pathogenic anaerobes. A reduction of the pH to values below the critical range of 5.1-5.3 in complex media 5.5 in simple testing substrates, respectively, which is caused by the presence of fermentable carbohydrates and will result in false-negative reactions. For this reason, the indol test should be performed with an adequate buffer capacity and an addition of fermentable carbohydrates should be avoided. Strains showing a false-negative reaction in the spot indole test can be reliably accounted for by means of the rapid test in buffered tryptophan solution described here without having to use additional anaerobic culture methods.