Prevotella Oralis Research Articles

Characterization of bacterial orofacial infections using a new murine model

We devised a new murine orofacial infection model using bacteria from odontogenic infection origins and characterized the experimental infections. In this model, bacteria were injected into the submandible of mice. Streptococcus constellatus and Peptostreptococcus micros produced a single abscess at the injection site and their abscess-forming and lethal abilities were low: the median abscess-forming dose (AF50) of S. constellatus and P. micros were 108.5–10.7and 1010.2–10.6cfu/mouse, and their median lethal dose (LD50) were >11 and 1010.6–11cfu/mouse, respectively. Prevotella oralis and Fusobacterium nucleatum produced multiple abscesses and their abscess-forming and lethal abilities were strong: AF50of P. oralis andF. nucleatum were 106.0–6.4and 107.0–8.7cfu/mouse, and their LD50were 107.0–7.7and 108.3–9.9cfu/mouse, respectively. LD50of P. intermedia andP. gingivalis were 109.4–>11and 108.9–9.1cfu/mouse, respectively. Prevotella intermedia and Porphyromonas gingivalis generated a necrotizing lesion, which progressed rapidly. We conclude that this murine model could reflect human orofacial odontogenic infections and is useful to investigate the pathogenicity of causative bacteria of such infections.

Antibiotic susceptibility of putative periodontal pathogens in advanced periodontitis patients.

In the present study, the antibiotic susceptibility of most prevalent micro-organisms in advanced periodontitis patients was evaluated. In 56 patients, pooled subgingival plaque samples were taken from the deepest site of each quadrant and were cultivated anaerobically. From each patient, the 4 most frequently encountered types of bacterial colonies were subcultured and identified (Rapid ID 32 A). From all bacterial species identified in the 224 subcultures, the 4 most prevalent were used for susceptibility testing to tetracycline, metronidazole and amoxicillin/clavulanate using the E Test. The most prevalent microorganisms were Fusobacterium nucleatum (38/214), Peptostreptococcus micros (33/214), Prevotella oralis (33/214) and Porphyromonas gingivalis (32/214). Regarding antibiotic susceptibility it could be shown that minimal inhibitory concentration (MIC) in all cases was below antibiotic concentrations achievable in gingival crevicular fluid. However, antibiotic resistance was seen in 3 to 29% of the investigated microorganisms.

Evaluation of the Rapid ID 32A system for identification of anaerobic Gram-negative bacilli, excluding the Bacteroides fragilis group

OBJECTIVE: To evaluate the Rapid ID 32A system (bioMérieux, Marcy-l'Etoile, France) for the identification of anaerobic Gram-negative bacilli, excluding the Bacteroides fragilis group. METHODS: Five hundred and twenty-eight identified clinical isolates of non-B. fragilis group anaerobic Gram-negative bacilli were tested in the Rapid ID 32A system, and identifications were compared with those obtained with conventional biochemical tests and gas-liquid chromatography. RESULTS: The Rapid ID 32A system correctly identified 280 (60.9%) of the 460 isolates tested for which taxa were included in the database, without the need for additional testing. A further 97 (21.1%) isolates were correctly identified to species level following the performance of complementary tests recommended by the manufacturer. Fifty-nine (12.8%) isolates were identified at the genus level only, and 21 (4.6%) were misidentified at the species level. Three isolates of Prevotella were not identified by the system. Of the 68 isolates belonging to taxa not included in the database, no identification was obtained for 33 (48.5%), while 35 (51.5%) were misidentified. CONCLUSIONS: The Rapid ID 32A system provided a rapid and reliable method for the identification of non-B. fragilis group, anaerobic Gram-negative bacilli to the genus level, while the success of species-level identification varied with different taxa. There was poor discrimination between Fusobacterium nucleatum and F. necrophorum, between Porphyromonas asaccharolytica and Porphyromonas endodontalis, and between Prevotella buccalis, Prevotella denticola, Prevotella loescheii, Prevotella melaninogenica and Prevotella oralis. The need to perform conventional complementary tests on 149 (32.4%) of the 460 isolates compromised the usefulness of the system for rapid species identification.

Fibronectin and laminin binding of urogenital and oral Prevotella species

88 strains of five Prevotella species--P. bivia, P. buccae, P. disiens, P. oralis, and P. oris--were examined for their fibronectin and laminin binding properties with the aid of latex particle agglutination assays. Beside single protein binding activities, all species showed strains that adhered to both fibronectin and laminin. The oral species, P. buccae, P. oralis, and P. oris were found to interact with laminin to a pronouncedly higher extent than with fibronectin. The urogenital species, P. bivia and P. disiens showed comparable activities of binding to fibronectin and laminin, with P. bivia exhibiting higher matrix protein binding rates than P. disiens. Within the oral species group, P. oralis showed a higher percentage of fibronectin and laminin reactive strains than did P. buccae and P. oris. The finding of species-related different binding properties may throw some light on the known differences in clinical relevance and pathogenicity of the urogenital species, P. bivia and P. disiens, but does so only in part concerning the oral species, P. buccae, P. oralis, and P. oris. Moreover, the observed differences in matrix protein binding of Prevotella species may have implications in chemotaxis and opsonization on the one hand and maintenance of colonization activities under antibiotic therapy on the other.

Pathogenicity of Peptostreptococcus micros morphotypes and Prevotella species in pure and mixed culture.

Recently, an atypical rough colony morphotype of Peptostreptococcus micros, a species which is found in ulcerating infections, including periodontitis, was isolated. The virulence of morphotypes alone and in combination with Prevotella intermedia and P. nigrescens was investigated both in vivo and in vitro. All strains tested induced abscesses containing fluid pus in a mouse skin model, and lesions caused by monocultures of the rough morphotype strains of P. micros were statistically significantly larger than those induced by the smooth morphotype strains. Inocula containing both morphotypes produced similar sized abscesses compared to mono-inocula containing the same bacterial load. Both Prevotella species induced small abscesses when inoculated alone, and when Pr. nigrescens was inoculated with one of the other strains, the abscesses were not significantly different from the abscesses induced by the mono-infections of this strain. Synergy, in terms of higher numbers of colony forming units (cfu) in the mixed inocula, was found for all combinations of the rough morphotypes of P. micros and both Prevotella spp. Pus from abscesses caused by combinations of Peptostreptococcus and Prevotella spp. transmitted the infection to other mice, but no abscesses were formed in mice inoculated with pus induced by mono-inocula. These results demonstrated synergic activity between both rough and smooth P. micros strains and oral Prevotella strains. The in-vitro co-culture experiments produced no evidence of growth stimulation. The effect of P. micros strains on the immune system was investigated by testing their ability to initiate luminol-dependent chemiluminescence of polymorphonuclear leucocytes in the presence and absence of human serum. In the latter, the rough morphotype strains initiated higher counts than the smooth morphotype strains. Further work is needed to elucidate the difference in virulence between the smooth and the rough morphotype cells of P. micros and the nature of the interaction with the Prevotella spp.

Role of human intestinal Prevotella oris in hydrolyzing ginseng saponins.

The potential of intestinal bacteria to hydrolyze ginsenoside Rb1 to 20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol (I) was found in 79% of the fecal specimens from 58 human subjects whose age ranged from 1 to 64 years. Following a ginsenoside-Rb1-hydrolyzing activity assay, Prevotella oris strains were then isolated as a major bacterial species possessing the potential. All the intestinal isolates converted ginsenosides Rb1 and Rd to I, ginsenoside Rb2 to 20-O-[alpha-L-arabinopyranosyl(1-->6)-beta-D-glucopyranosyl]-20(S) -protopanaxadiol (II), and ginsenoside Rc to 20-O-[alpha-L-arabinofuranosyl(1-->6)-beta-D-glucopyranosyl]-20(S)- protopanaxadiol (III) like fecal microflora, but did not attack ginsenosides Re or Rg1 (protopanaxatriol-type). The isolates were susceptible to colimycin (MIC, 3.13 micrograms/ml) and then the treatment of specific pathogen free mice with colimycin (20 mg/kg/day) decreased intestinal bacterial Rb1-hydrolyzing potential from 22.1 +/- 1.2% to 4.7 +/- 2.7%, while the decreased potential was restored to 30.7 +/- 3.7% by the inoculation with P. oris isolates. These results suggest that the metabolism of protopanaxadiol saponins to metabolites I-III in the intestines seems most partly due to intestinal P.oris. In addition, the fact that neither intact ginsenoside Rb1 nor its middle metabolic derivatives but only the final metabolite I was detected at 1.0-7.3 micrograms/ml in blood after oral administration of mice with ginsenoside Rb1 (125 mg/kg) allows us to speculate that metabolites I-III are the most likely forms of protopanaxadiol saponins absorbed from the intestines.

Inhibition of Prevotella and Capnocytophaga immunoglobulin A1 proteases by human serum.

Oral Prevotella and Capnocytophaga species, regularly isolated from periodontal pockets and associated with extraoral infections, secret specific immunoglobulin A1 (IgA1) proteases cleaving human IgA1 in the hinge region into intact Fab and Fc fragments. To investigate whether these enzymes are subject to inhibition in vivo in humans, we tested 34 sera from periodontally diseased and healthy individuals in an enzyme-linked immunosorbent assay for the presence and titers of inhibition of seven Prevotella and Capnocytophaga proteases. All or nearly all of the sera inhibited the IgA1 protease activity of Prevotella buccae, Prevotella oris, and Prevotella loescheii. A minor proportion of the sera inhibited Prevotella buccalis, Prevotella denticola, and Prevotella melaninogenica IgA1 proteases, while no sera inhibited Capnocytophaga ochracea IgA1 protease. All inhibition titers were low, ranging from 5 to 55, with titer being defined as the reciprocal of the dilution of serum causing 50% inhibition of one defined unit of protease activity. No correlation between periodontal disease status and the presence, absence, or titer of inhibition was observed. The nature of the low titers of inhibition in all sera of the IgA1 proteases of P. buccae, P. oris, and P. loescheii was further examined. In size exclusion chromatography, inhibitory activity corresponded to the peak volume of IgA. Additional inhibition of the P. oris IgA1 protease was found in fractions containing both IgA and IgG. Purification of the IgG fractions of five sera by passage of the sera on a protein G column resulted in recovery of inhibitory IgG antibodies against all three IgA1 proteases, with the highest titer being for the P. oris enzyme. These finding indicate that inhibitory activity is associated with enzyme-neutralizing antibodies.

Characterization of saccharolytic Bacteroides and Prevotella isolates from infected dog and cat bite wounds in humans.

Saccharolytic, nonpigmented, anaerobic gram-negative rods isolated from infected dog and cat bite wounds in humans have been poorly characterized, and most are not included in the databases of kits used for anaerobic identification; thus, they are problematic for clinical laboratories to identify. Fifty strains isolated from such wounds were characterized with commercial kits for preformed-enzyme detection, carbohydrate fermentation, and other biochemical tests. PCR fingerprinting was performed on these strains to further characterize subgroups within these species. Bacteroides tectum is a frequent isolate in bite wounds and resembles Prevotella bivia in colony morphology and saccharolytic activity, except that it grows in 20% bile and hydrolyzes esculin. Profile numbers generated by various kits associate B. tectum with P. bivia, Prevotella oralis group, or Prevotella melaninogenica. PCR fingerprinting identified at least four subgroups and confirmed the heterogeneous nature of this species. Prevotella heparinolytica was also frequently isolated from these bite wounds. It produces indole and generates a profile number in preformed-enzyme kits that is usually associated with Bacteroides uniformis. However, it is bile sensitive and quite distinct from the Bacteroides fragilis group of anaerobes. The PCR fingerprint profiles generated by strains of P. heparinolytica were very similar to that of the type strain and to each other. Prevotella zoogleoformans, occasionally isolated from dog and cat bite wounds in humans, resembles P. heparinolytica except for a negative indole test. Clinical laboratories should be aware of the characteristics of these animal species when identifying isolates from animal bite wounds in humans.

Bacteriologic findings in tonsillitis and pericoronitis.

Bacteriologic samples from 31 young men were cultured quantitatively for aerobes and anaerobes; these samples included 31 specimens of tonsils (16 infected and 15 healthy), 16 specimens from pericoronal pockets of lower third molars (11 infected and 5 symptom-free), and 6 postoperative specimens from lower-third-molar extraction sockets. Anaerobes were isolated more often from infected third molars than from infected tonsils (14.5 isolates vs. 8.4 isolates, respectively; P < .001). Infected tonsil samples contained significantly more anaerobic species if an adjacent partly erupted lower third molar was present rather than absent (10.3 isolates vs. 6.9 isolates, respectively; P < .05). Eubacterium aerofaciens, Clostridium species, Peptostreptococcus micros, and Prevotella oris were frequently isolated. Streptococcus salivarius was found more frequently in tonsillar specimens, whereas Corynebacterium species, Prevotella denticola, Capnocytophaga species, Peptostreptococcus anaerobius, and Lactobacillus species were more common in pericoronal pocket samples. Thus, partial eruption of lower third molars increases the number of anaerobic bacterial species on tonsils and many species can be isolated simultaneously from both tonsils and lower third molars.

Associations of endodontic symptoms and signs with particular combinations of specific bacteria

Significant associations have been reported between (a) specific bacterial species isolated from root canals and (b) between individual bacterial species and endodontic symptoms and signs. The prime objective of this study was to determine whether particular combinations of specific bacteria are associated with individual endodontic symptoms and signs. Seventy root canals were investigated microbiologically taking care to maintain the viability of obligate anaerobes, which accounted for 64% of the total species isolated, including Peptostreptococcus micros, Prevotella melaninogenica, Prevotella oralis, Eubacterium aerofaciens, Eubacterium lentum, Fusobacterium nucleatum, Prevotella buccae and Prevotella intermedia. Significant associations were found between individual clinical features and the following pairs of species: (a) pain (37 cases) and Peptostreptococcus spp./Prevotella spp., Peptostreptococcus spp./Prevotella melaninogenica, Pstr. micros/Prev. melaninogenica (all P < 0.01); (b) swelling (23 cases) and Pstr. micros/Prevotella spp. (P < 0.01); (c) 'Wet' canal (57 cases) and Prevotella spp./Eubacterium spp. (P < 0.01), Peptostreptococcus spp./Eubacterium spp. (P < 0.05). Thus data from this investigation suggests that statistically significant associations exist between individual endodontic symptoms and signs and particular combinations of specific bacteria.

Molecular analysis of microflora associated with dentoalveolar abscesses.

The microflora associated with three dentoalveolar abscesses was determined by cultural and molecular methods. 16S rRNA genes were randomly amplified by means of conserved eubacterial primers and cloned. Restriction fragment length polymorphism analysis of the clones and amplified genes encoding 16S rRNA from the cultured bacteria was used to detect putative unculturable bacteria. Clones representative of five predominant groups of uncultured organisms were sequenced. Two were identified as Porphyromonas gingivalis and Prevotella oris, and one was found to be closely related to Peptostreptococcus micros. The remaining two clones did not correspond to known, previously sequenced organisms. One was related to Zoogloea ramigera, a species of aerobic waterborne organisms, while the other was distantly related to the genus Prevotella. This study has demonstrated the possibility of the characterization of microflora associated with human infection by molecular methods without the inherent biases of culture.

In vivo cleavage of immunoglobulin A1 by immunoglobulin A1 proteases from Prevotella and Capnocytophaga species

Immunoglobulin A1 (IgA1) proteases secreted by oral Prevotella and Capnocytophaga species specifically cleave IgA1 at the same peptide bond in the hinge region, leaving intact monomeric Fab and Fc fragments. Assuming that Prevotella- and Capnocytophaga-induced Fab fragments of IgA1 expose a specific immunogenic neoepitope at the cleavage site, we established an enzyme-linked immunosorbent assay to measure human serum antibodies to this neoepitope as indirect evidence of in vivo activity of Prevotella and Capnocytophaga IgA1 proteases. The assay used a monoclonal antibody with specificity for the neoepitope, and the ability to block binding of the monoclonal antibody to the neoepitope was investigated. Absorption of sera with Prevotella melaninogenica-induced Fab fragments of IgA1 resulted in removal of antibodies blocking binding of the monoclonal antibody, whereas absorption with Fab fragments induced by bacterial IgA1 proteases of other cleavage specificities did not remove blocking antibodies. Consequently, we assume that the antibodies detected had been induced by a neoepitope an the Fab fragment of IgA1 exposed exclusively after cleavage with IgA1 proteases from Prevotella and Capnocytophaga, indicating in vivo activity of these IgA1 proteases. Evidence, though indirect, of in vivo activity of Prevotella and Capnocytophaga IgA1 proteases was present in 42 of 92 sera examined and in a significantly higher proportion of sera from adults with periodontal disease compared with control individuals. No correlation with disease was observed for the juvenile periodontitis groups.

Breakdown of Various Serum Proteins by Periodontal Bacteria

The capacity of subgingival microorgansims to degrade various serum proteins was estimated. To this end, the organisms were incubated in heat-inactivated human serum for 48 h. Porphyromonas endodontalis and Prevotella intermedia NY 365 and NY 655 completely degraded IgG, IgA and IgM. Porphyromonas gingivalis, P. asaccharo-lytica, Prevotella nigrescens and P. intermedia ATCC 25611 also performed complete degradation of IgA and IgM, but partial breakdown of IgG. Cleavage of IgG by the latter four microorganisms, in addition to Prevotella oralis and Capnocytophaga ochracea , yielded Fab and Fc fragments. Degradation of various other serum proteins by P. endodontalis, P. gingivalis, P. intermedia and P. asaccharolytica included the iron-handling proteins albumin, haptoglobin, haemopexin and transferrin. Only P. endodontalis failed to degrade albumin. Extensive breakdown of serum proteins, in particular immunoglobulins and iron-handling proteins, will sustain bacterial growth and hamper local host defences. This suggests an important role for proteolytic microorganisms, such as Porphyromonas and Prevotella species, in the causation of polymicrobial infections. Keywords - Periodontal diseases, Proteolytic activity, Immunoglobulin degradation, Serum proteins, Porphyromonas , Prevotella .

Degradation of immunoglobulin G by periodontal bacteria.

Several subgingival microorganisms were tested for their ability to utilize human immunoglobulin G (IgG) as a substrate for growth. This was done using a protein-free chemically defined medium, supplemented with IgG. Stimulation of growth was observed for Capnocytophaga ochracea, Porphyromonas asaccharolytica, Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella intermedia, Prevotella oralis, Lactobacillus catenaforme and Streptococcus intermedius. Immunoelectrophoresis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and a protein assay demonstrated that P. intermedia and P. endodontalis completely degraded the protein chains of IgG. Partial breakdown of IgG was observed for P. asaccharolytica and C. ochracea, whereas P. oralis cleaved the IgG heavy chain, yielding Fc and Fab fragments. All these bacteria utilized IgG as a substrate for growth. Binding studies using an enzyme-linked immunosorbent assay, revealed complete loss of in vitro antigen-antibody binding capacity after incubation of specific IgG with P. endodontalis and partial loss of binding with P. intermedia, P. gingivalis, C. ochracea or Fusobacterium nucleatum. Degradation or inactivation of IgG by oral bacteria is thought to be important in the causation of polymicrobial infections.

Evidence for the absence of hyaluronidase activity in Porphyromonas gingivalis

The aim of the present study was to evaluate the ability of Porphyromonas gingivalis to degrade hyaluronic acid. No hyaluronidase activity was detected using a turbidimetric method, whereas a standard plate assay showed a positive reaction for P. gingivalis. We postulated that the high proteolytic activity of P. gingivalis may account for this observation. A modified plate assay was designed to avoid false-positive reactions caused by proteolytic bacteria. The new assay, based on the formation of a water-insoluble salt between hyaluronic acid and the polyanion cetylpyridinium chloride, indicated that P. gingivalis does not have hyaluronidase activity. By this modified plate method, it was found that among 24 different oral bacterial species tested, Propionibacterium acnes and Prevotella oris were the only species that possess hyaluronidase activity.

Identification of proteases from periodontopathogenic bacteria as activators of latent human neutrophil and fibroblast-type interstitial collagenases.

Activation of latent human fibroblast-type and neutrophil interstitial procollagenases as well as degradation of native type I collagen by supra- and subgingival dental plaque extracts, an 80-kDa trypsinlike protease from Porphyromas gingivalis (ATCC 33277), a 95-kDa chymotrypsinlike protease from Treponema denticola (ATCC 29522), and selected bacterial species commonly isolated in periodontitis was studied. The bacteria included were Prevotella intermedia (ATCC 25261), Prevotella buccae (ES 57), Prevotella oris (ATCC 33573), Porphyromonas endodontalis (ES 54b), Actinobacillus actinomycetemcomitans (ATCC 295222), Fusobacterium nucleatum (ATCC 10953), Mitsuokella dentalis (DSM 3688), and Streptococcus mitis (ATCC 15909). None of the bacteria activated latent procollagenases; however, both sub- and supragingival dental plaque extracts (neutral salt extraction) and proteases isolated from cell extracts from potentially periodontopathogenic bacteria P. gingivalis and T. denticola were found to activate latent human fibroblast-type and neutrophil interstitial procollagenases. The fibroblast-type interstitial collagenase was more efficiently activated by bacterial proteases than the neutrophil counterpart, which instead preferred nonproteolytic activation by the oxidative agent hypochlorous acid. The proteases were not able to convert collagenase tissue inhibitor of metalloproteinase (TIMP-1) complexes into active form or to change the ability of TIMP-1 to inhibit interstitial collagenase. None of the studied bacteria, proteases from P. gingivalis and T. denticola, or extracts of supra- and subgingival dental plaque showed any significant collagenolytic activity. However, the proteases degraded native and denatured collagen fragments after cleavage by interstitial collagenase and gelatinase. Our results indicate that proteases from periodontopathogenic bacteria can act as direct proteolytic activators of human procollagenases and degrade collagen fragments. Thus, in concert with host enzymes the bacterial proteases may participate in periodontal tissue destruction.

Comparative in vitro activity of cefpodoxime against anaerobes other than bacteroides fragilis

To assess the in vitro activity of cefpodoxime against anaerobic respiratory tract and oropharyngeal pathogens 77 strains belonging to 18 gram-negative and 7 gram-positive species were studied by means of agar dilution tests. For comparison cefuroxime, amoxicillin, amoxicillin + clavulanic acid and clindamycin were also tested. Cefpodoxime was found to be active at concentrations of less than or equal to 0.125 mg/l against Prevotella oralis, Prevotella buccalis, Prevotella bivia, Porphyromonas asaccharolytica, Bacteroides corporis, Bacteroides gracilis, Fusobacterium necrophorum, Fusobacterium naviforme and Propionibacterium acnes. Prevotella oris, Prevotella buccae, Fusobacterium nucleatum, Peptostreptococcus asaccharolyticus, and Ruminococcus bromii were inhibited at concentrations of less than or equal to 1 mg/l and Prevotella denticola, Prevotella melaninogenica, Prevotella intermedia, Porphyromonas gingivalis, Bacteroides pneumosintes, and Peptostreptococcus micros at concentrations of less than or equal to 4 mg/l. Strains of Veillonella parvula were inhibited by cefpodoxime at 0.25-8 mg/l, and single strains of Peptostreptococcus anaerobius and Peptostreptococcus magnus showed MICs of 32 and 64 mg/l, respectively. The results obtained warrant the use of cefpodoxime in therapy of anaerobic and mixed aerobic-anaerobic infections of the upper and lower respiratory tract and similar infections not involving Bacteroides fragilis.

Multivariate analyses of cellular fatty acids in Bacteroides, Prevotella, Porphyromonas, Wolinella, and Campylobacter spp

The genera Bacteroides, Wolinella, and Campylobacter contain several similar species that require taxonomic revision. Fatty acid profiles of whole bacterial cells have proven useful for taxonomy. In this study, cellular fatty acids from Bacteroides, Prevotella, Porphyromonas, Wolinella, and Campylobacter spp. were identified and quantitated by gas chromatography and gas chromatography-mass spectrometry, and the data were subjected to principal component analyses. Bacteroides fragilis, the type species of the genus Bacteroides, was distinct from the other organisms. While Bacteroides gracilis, Wolinella succinogenes, Wolinella curva, Wolinella recta, and Campylobacter fetus subsp. venerealis were close to each other, Prevotella (Bacteroides) buccae, Prevotella oralis, Prevotella oris, Prevotella disiens, Prevotella veroralis, Prevotella heparinolyticus, Porphyromonas (Bacteroides) endodontalis, and Bacteroides ureolyticus could be distinguished. B. fragilis was characterized by the presence of C3OH-i-1-, Ca-15, and Ci-15 and the absence of C12:0 and unsaturated fatty acids. For comparison, B. gracilis, B. ureolyticus, W. succinogenes, W. curva, W. recta, and Campylobacter fetus subsp. venerealis contained C12:0, C16:1, C18:1, and C3-OH-14 acids but lacked branched hydroxy and branched nonhydroxy acids. B. gracilis and B. ureolyticus are not "true" bacteroides.