Eight strains of anaerobic Gram-negative bacilli isolated from infections of the skin and soft tissues were subjected to a comprehensive range of phenotypic and genotypic tests. 16S rRNA gene sequence analysis revealed the strains to constitute a homogeneous group, distinct from species with validly published names but related to a cluster including Prevotella buccae, Prevotella dentalis and Prevotella baroniae. A novel species, Prevotella bergensis sp. nov., is proposed to accommodate these strains. Prevotella bergensis is saccharolytic and produces acetic and succinic acids as end products of fermentation. The G+C content of the DNA of the type strain is 48 mol%. The type strain of Prevotella bergensis is 94067913(T) (=DSM 17361(T)=CCUG 51224(T)).

Leptotrichia buccalis varies with respect to cellular and colonial morphology, fermentation of sugars and presence of glucosidases. It can be confused with Fusobacterium spp. and gram-positive bacilli related to Lactobacillus . The aim of the present study was to examine the heterogeneity of 60 Leptotrichia isolates, of which 58 had been assigned as L. buccalis and two as ‘ Leptotrichia pseudobuccalis ’ by using enzymatic/biochemical and cellular fatty acid analyses. The rapid ID 32 A kit (API, bioMerieux, Marcy-l’Etoile, France) revealed considerable strain heterogeneity. Most frequently detected were strains producing α -glucosidase, alkaline phosphatase and β -glucosidase. Three other prominent strain clusters generated β -galactosidase-6-phosphate, β -galactosidase and α -galactosidase, respectively. β -Nacetyl- glucosaminidase was also produced by many strains. Mannose and raffinose fermentation was common and all strains were indole negative. Four strains reduced nitrate, and one strain produced urease. The diagnosis L. buccalis was achieved for 36 of the 60 (60%) Leptotrichia isolates. The MIDI (Microbial ID, Inc., Newark, DE, USA) system demonstrated that the most abundant cellular fatty acids were C 16:0 FA (32–59%) and C 18:1 - cis 11/t9/t6 FA or an unidenti? ed compound with an equivalent chain length (ECL) of 17.8 (21–42%). C 12:0 FA (3–16%), C 15:0 DMA or 3-OH C 14:0 FA (4–14%), C 14:0 FA (3–20%), and C 16:1 - cis 9 FA (1–5%) were also detected. Forty-two of the strains (70%) contained C 18:1 - cis 9 FA (1–9%) and 37 (62%) C 18:2 - cis 9,12 FA (1–6%). The MIDI system came up with the diagnosis L. buccalis in 57 of the 60 isolates (95%). There was complete agreement in diagnosis between the two systems in 34 of the 60 isolates (57%). The heterogeneity in enzymatic/biochemical reactions and cellular fatty acid composition may indicate that the genus Leptotrichia contains hitherto unrecognized species. Keywords: enzymatic reactions, biochemical tests, fatty acid analysis, gas chromatography, Leptotrichia .

Results of a polyphasic taxonomic study on an unknown Gram-negative, facultatively anaerobic, coccobacillus-shaped organism isolated from an infected human gall bladder are presented. Phenotypic and molecular taxonomic studies revealed the organism to be close to, but distinct from, organisms designated CDC (Centers for Disease Control and Prevention) group DF-3. The unknown bacterium was readily distinguished from reference strains of Bacteroides, Prevotella, Porphyromonas and related taxa by 16S rRNA gene sequencing, biochemical tests, analysis of cellular long-chain fatty acids and electrophoretic analysis of whole-cell proteins. Based on the results of the present study, it is proposed that the unknown bacterium be classified in a new genus, Dysgonomonas, as Dysgonomonas gadei sp. nov. (type strain CCUG 42882(T) = CIP 106420(T)). In addition, a new species, Dysgonomonas capnocytophagoides sp. nov., is proposed to accommodate strains previously belonging to CDC group DF-3. The type species of the genus Dysgonomonas is Dysgonomonas gadei.

Lipopolysaccharide of Fusobacterium nucleatum strain Fevl was split by acid hydrolysis. The split products, i.e. lipid A and degraded polysaccharide were mitogenic for murine spleen cells as measured by uptake of [3H]thymidine. The uptake of [3H]thymidine was dose-dependent. Incubation of spleen cells with stimulants for 3 days resulted in a polyclonal activation of immunoglobulin synthesis. Higher mitogenic response and immunoglobulin production were found in spleen cells of athymic mice compared to those of thymic mice. The activity of lipid A in stimulating immunoglobulin synthesis was comparable with the parent lipopolysaccharide-Fevl, the degraded polysaccharide being the less potent stimulator.

A clone of serogroup A Neisseria meningitidis recognized by multilocus enzyme electrophoresis and designated as clone III-1, has caused major epidemics of meningococcal disease in various parts of the world since the 1970s. In Norway, serogroup B meningococci have been responsible for an epidemic since the mid-1970s. We have studied a sample of 53 meningococci isolated from patients in western Norway prior to the serogroup B epidemic. 22/35 meningococcal isolates collected 1969-73 represented clone III-1, whereas this clone was not found in the 18 isolates from 1962-68. It has been speculated that the epidemic of meningococcal disease in Finland caused by clone III-1 in 1973-75 had spread from an epidemic in USSR that began in 1969. Our findings demonstrate, however, that the clone III-1 was present in Scandinavia in 1969.

The immunochemical reactions of rabbit polyclonal antibodies directed to different preparations of Fusobacterium nucleatum i.e, whole cells, peptidoglycan associated proteins, a peptidoglycan-protein complex and a purified 40 kiloDalton (kDa) protein, were investigated on outer membrane preparations of Fusobacterium species and a restricted number of Leptotrichia buccalis after their separation on sodium dodecyl sulphate polyacrylamide gels and electrotransfer to nitrocellulose. All F. nucleatum strains had identical reaction patterns with the immune sera tested. Surface exposed parts of a restricted number of proteins with apparent molecular weights at 70 kDa (a doublet band), 60 kDa, 55 kDa and 40 kDa seemed to be major immunogens. Antigenic related proteins either of identical or slightly deviating electrophoretic mobilities to the 40-kDa protein were observed with the other members of Bacteroidaceae tested. The characteristic 70-kDa protein doublet seemed to be restricted to F. nucleatum although single protein bands of near identical molecular weights belonging to the other species tested also reacted. The data also indicate that the 60-kDa and 55-kDa polypeptides might be present in other species of Fusobacterium.

The adhesion of Campylobacter jejuni and C. coli to isolated porcine intestinal brush border membranes was studied by phase‐contrast and electron microscopy. Approximately 45% of the cell population adhered to the brush borders, possibly in a specific manner. Pretreatment of the brush borders with trypsin or pronase, and competitive inhibition with L‐rhamnose caused a slight reduction of the adhesion. Different forms of pretreatment of the bacterial cells reduced their ability to adhere, but also their motility.

(1985). Pathogenesis and Treatment of Anaerobic Infections: A Symposium Held in Bergen, Norway June 21, 1984. Scandinavian Journal of Infectious Diseases: Vol. 17, No. sup46, pp. 1-114.

Reference strains and clinical isolates of Bacteroides and Fusobacterium species were examined by the Anaerobe-Tek System (A/T-system). Of 104 strain, only 57 (54.8%) were identified correctly to species level. 38 strains (36.5%) were incorrectly identified and for 9 strains (8.7%) there were no codes in the manufacturers' data base manual. The results indicate that, in our hands, the A/T-system in its present form, is not suitable for the identification of clinical isolates of Bacteroides and Fusobacterium species.

Fusobacierium nucleatum ATCC 10953 LPS was split by hydrolysis with 1 per cent acetic acid into acid-soluble polysaccharide and lipid A. Gel filtration of the acid-soluble polysaccharide on Bio-Gel P-60 gave a high-molecular-weight fraction eluted with the void volume (V 0 ), and a fraction eluted at 2.4 × V o . The high-molecular-weight fraction was serologically active, and contained glucose as the only sugar. The other fraction, which was serologically inactive, contained L- glycero -D- manno -hepioss, galactose, glucosamine, keto-deoxy-octonate, and phosphorus. The fraction eluted at 2.4 × V o is thought to constitute the polysaccharide core region, whereas the high-molecular-weight fraction may represent O-antigenic side chains.

Fusobacterium nucleatum Fev1 lipopolysaccharide was split by hydrolysis with 1% acetic acid into acid-soluble polysaccharide and lipid A. Gel filtration of the polysaccharide on Bio-Gel P-60 gave a high-molecular-weight fraction eluted with the void volume, and a fraction eluted at 2.4 x Vo. The high-molecular-weight fraction contained L-glycero-D-manno-heptose in relatively large amounts, glucose, glucosamine, an unknown amino compound and small amounts of (or no) D-glycero-D-manno-heptose. Phosphorus and 3-deoxy-D-manno-octulosonic acid were not detected. The other fraction contained L- and D-glycero-D-manno-heptose, glucose, glucosamine, 3-deoxy-d-manno-octulosonic acid and phosphorus. Further fractionation experiments and serological investigations indicated that the high-molecular-weight fraction carried the O-antigenic side chains, whereas the material eluted from Bio-Gel P-60 at 2.4 x Vo represented the core oligosaccharide.

SUMMARY: Fatty acids of five strains of Leptotrichia buccalis were examined by gas-liquid chromatography. The strains showed identical patterns, characterized by the presence of n-hexadecanoate. octadecenoate and 3-hydroxytetradecanoate as major acids. The general acid pattern showed a distinct similarity to that of Fusobacterium species.

In line with our on-going efforts to create a multivalent anti-Campylobacter jejuni vaccine based on its capsule polysaccharides (CPSs), we report here the chemical structure and the genetic locus of the CPS produced by C. jejuni strain CG8486, which belongs to the serotype HS:4 CPS complex. C. jejuni CG8486 CPS was observed to be composed of approximately 17 disaccharide repeating blocks of 4-substituted N-acetyl-β-d-glucopyranosamine and 3-substituted 6-deoxy-β-d-ido-heptopyranose. A small number of 6-deoxy-β-d-ido-heptopyranose units were observed to carry O-methyl phosphoramidate moieties at the O-2 or O-7 position. The gene content and organization of the CPS locus of C. jejuni CG8486 were comparable to those of C. jejuni strains NCTC 11168 and 81-176, but several CG8486 CPS genes were observed to be more divergent from those present in the CPS loci of NCTC 11168 and 81-176 CPS, which indicated that there are genetic characteristics specific to the C. jejuni HS:4 CPS complex. The efficacy of a glycoconjugate vaccine based on C. jejuni CG8486 CPS is presently being tested in an animal model, the results of which will be presented in future communications.

The API ZYM patterns of 97Bacteroides and 25Fusobacterium strains were examined. The system discriminated betweenB. asaccharolyticus, B. melaninogenicus ss.melaninogenicus, andB. melaninogenicus ss.melaninogenicus, but not between otherBacteroides species or between species ofFusobacterium. The results provided new information on the enzymatic activity of theses gorups of bacteria.

A serologically highly active polysaccharide antigen (PS 02/725) consisting of equimolar amounts ofd-glycero-d-galacto-heptose and a 6-deoxyheptose was isolated fromEubacterium saburreum, strain 02/725, by trypsin digestion and subsequent gel filtration and ion-exchange chromatography. Whole bacterial cells injected intravenously stimulated rabbits to produce precipitins and complement-binding antibodies of the IgG class specific for PS 02/725.