No abstract

A new species, Bacteroides salivosus sp. nov., is proposed for black-pigmented, asaccharolytic Bucteroides strains from cats, isolated from subcutaneous abscesses and empyemas, as well as from gingival margins of normal mouths. The bacterium is an obligately ahaerobic, gram-negative, brown-or black-pigmented, asaccharolytic, nonmotile, nonsporeforming rod that does not grow in 20% bile and has a guanine-pluscytosine content of 42 to 44 mol% . It has 12 % deoxyribonucleic acid homology with the human type strain of Bucteroides gingivalis (ATCC 33277T) and 1 % deoxyribonucleic acid homology with the human type strain of Bacteroides asacchurolyticus (ATCC 25260T). Strain VPB 157 (NCTC 11632T) is the type strain. Unlike B. gingivalis, B . salivosus produces catalase, and the colonies of the type strain fluoresce at 24 and 48 h, although some other strains do not fluoresce. It does not agglutinate sheep erythrocytes. Unlike either B. asaccharolyticus or Bacteroides endodontulis, it has trypsinlike activity and produces large quantities of phenylacetic acid.During our studies of anaerobic bacteria from soft tissue infections and of normal flora of the oral cavity of cats, we isolated a number of strains of pigmented, asaccharolytic Bacteroides spp. resembling Bacteroides gingivalis, which is isolated from humans. The strains have been described previously (4), but as no deoxyribonucleic acid (DNA) homology data were available at the time, it was not possible to propose a new species. In the present paper, we report the results of DNA homology studies of the strains from cats. We also propose a new species, Bacteroides salivosus. MATERIALS AND METHODSBacterial strains. All 18 strains studied were isolated from cats with either a solitary closed subcutaneous "fight" wound abscess or a pyothorax, or from the gingival margins in normal cats. These organisms were compared in all reactions with the type strains (obtained from the American Type Culture Collection [ATCC]) Bacteroides. ATCC 33277T and Bacteroides asaccharolyticus ATGC 25260T.Methods and characterization. The general methods of growth and biochemical characterization have been described previously (4). DNA isolation. The organisms were grown in 800-ml amounts of prereduced brain heart infusion broth (Oxoid Ltd.) supplemented with yeast extract and vitamin K and heme (PRBHIB) (1). Bottles containing 800 ml of medium were inoculated with 18 ml of an overnight culture grown in PRBHIB and then incubated for 24 h at 37°C. The harvested cells were suspended in a 0.15 M NaCI-0.01 M ethylenediaminetetraacetic acid-salt solution (pH 8.0). The cells were lysed by adding sodium dodecyl sulfate to a final concentration of 1%. DNA preparations for hybridization experiments and for DNA guanine-plus-cytosine (G + C) determinations were isolated by a hydroxyapatite procedure (2). The DNA preparations were stored in 0 . 1~ SSC (SSC is 0.15 M NaCl and 0.015 M sodium citrate, pH 7.0) at -20°C.Gi-C content of DNA. Thermal melting points were used to determine the G+C contents of t...

This study used a previously described multiplex PCR-based reverse line blot (mPCR/RLB) assay to assess the prevalence and distribution of 14 urogenital pathogens or putative pathogens, namely Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium, Mycoplasma hominis, Trichomonas vaginalis, Gardnerella vaginalis, Ureaplasma parvum, Ureaplasma urealyticum, Neisseria meningitidis, Streptococcus pneumoniae, Haemophilus influenzae, herpes simplex virus types 1 and 2, and human adenovirus. First-voided urine specimens and endocervical and self-collected vaginal swabs from each of 216 women attending three sexual health clinics in Sydney, Australia, were tested and the results were compared with those of reference methods for each organism. One hundred and sixty-eight women (77.7 %) had at least one and 105 (48.6 %) had more than one target organism, most commonly G. vaginalis and Ureaplasma spp. The prevalence of each of the four known sexually transmissible pathogens was ,5 %. Of the 216 women, 111 (51.4 %) reported at least one symptom consistent with genital or urethral infection, including discharge, pain or discomfort. Only G. vaginalis was detected more frequently in women with symptoms (P50.05). The specificity of the mPCR/RLB assay compared with that of the reference methods for each organism and for all specimen types was 100 %. The mean sensitivities of the mPCR/RLB assay compared with those of the reference methods for self-collected vaginal swabs, cervical swabs and first-voided urine specimens for all organisms were 99.3, 98.1 and 84.6 %, respectively; however, these differences were not significant. There were no differences in sensitivities between specimen types for C. trachomatis, N. gonorrhoeae, T. vaginalis and H. influenzae, although all were found infrequently. Overall, the mPCR/RLB platform was found to be an accurate testing platform in a sexual health clinic setting.

Sixty strains of obligately anaerobic gram-negative nonsporeforming nonpigmented Bacteroides species were isolated from subcutaneous abscesses and pyothoraxes of cats and dogs. All of these strains grew well in bile and produced acetic, propionic, and succinic acids as major products of fatty acid metabolism. Phenotypic and deoxyribonucleic acid (DNA) homology data divided the strains into five groups. Only three strains were members of Bacteroides fragilis, as determined by biochemical and DNA homology analyses. All of the other strains showed negligible levels of DNA homology with human strains of B. fragilis, Bacteroides distasonis, Bacteroides vulgatus, Bacteroides thetaiotaomicron, and Bacteroides ovatus, although they did have substantial ribosomal ribonucleic acid homology with these species. One group comprising 52 strains is proposed as a new species, Bacteruides tectum, having three distinct homology clusters. Strains of B. tectum are obligately anaerobic, gram-negative, nonmotile, nonpigmented, nonsporeforming rods that grow well in bile, do not produce indole, and only weakly ferment carbohydrates. They have a DNA guanine-plus-cytosine ratio of 46 mol% but do not exhibit DNA homology with phenotypically similar human strains of Bacteroides.During the past several years we have been investigating strains of Bacteroides isolated from the oral floras, subcutaneous abscesses, and pyothoraxes of cats and dogs. Many of these isolates are phenotypically similar, but not identical, to some of the saccharolytic Bacteroides species that are capable of growth in bile-containing media (13). The purpose of this study was to extend our phenotypic characterization of these organisms and to determine their taxonomic position by using deoxyribonucleic acid (DNA) and ribosomal ribonucleic acid (rRNA) homology techniques. Culture media and methods. The organisms were maintained and subcultured by using previously described techniques (13). The basal media used for biochemical tests were those of Holdeman et al. (3), and the range of biochemical tests performed included the tests considered discriminatory by Cat0 and Johnson (1) for characterization of Bacteroides species. Fermentation of carbohydrates (Table 1) Fatty acids were extracted and analyzed by gas-liquid chromatography with a Hewlett-Packard model 5830A gas chromatograph as described previously (11,12). Gram stain reactions and morphology were determined from 1-day-old cultures in cooked meat-carbohydrate medium (11) and peptone-yeast extract-glucose medium (3) and from 2-dayold blood agar plates (3). The broth disk test of Wilkins and Thiel (19) was used to determine susceptibilities to antimicrobial agents. MATERIALS AND METHODSDNA isolation. The organisms were grown in medium that was prepared as described previously (2); this medium contained mineral salts, 1% pepticase, 0.5% yeast extract, 1% dehydrated brain heart infusion broth (Difco Laboratories, Detroit, Mich.), 1% glucose, 0.03% cysteine, 0.03% sodium formaldehyde sulfoxalate, 0.01% heme, and 0.05...