Mobiluncus Curtisii Research Articles

Microbiological Findings in Subjects With Asymptomatic Oral Lichen Planus: A Cross‐Sectional Comparative Study

The bacterial colonization of the oral mucosa was evaluated in patients with asymptomatic oral lichen planus (OLP) and compared to the microbiologic status in mucosally healthy subjects. Bacteria from patients with clinically and histopathologically diagnosed OLP from the Stomatology Service, Department of Oral Surgery and Stomatology, School of Dental Medicine, University of Bern, were collected with a non-invasive swab system. Samples were taken from OLP lesions on the gingiva and from non-affected sites on the contralateral side of the mouth. The control population did not have OLP and was recruited from the student clinic. All samples were processed with the checkerboard DNA-DNA hybridization method using well-defined bacterial species for the analysis. Significantly higher bacterial counts of Bacteroides ureolyticus (P = 0.001), Dialister species (sp.) (P = 0.006), Staphylococcus haemolyticus (P = 0.007), and Streptococcus agalactiae (P = 0.006) were found in samples taken from OLP lesions compared to sites with no clinical evidence of OLP. Significantly higher bacterial counts were found for Capnocytophaga sputigena, Eikenella corrodens, Lactobacillus crispatus, Mobiluncus curtisii, Neisseria mucosa, Prevotella bivia, Prevotella intermedia, and S. agalactiae at sites with lesions in subjects with OLP compared to sites in control subjects (P <0.001). Microbiologic differences were found between sites with OLP and sites in subjects without a diagnosis of OLP. Specifically, higher counts of staphylococci and S. agalactiae were found in OLP lesions.

Molecular Quantification ofGardnerella vaginalisandAtopobium vaginaeLoads to Predict Bacterial Vaginosis

Bacterial vaginosis (BV) is a poorly detected public health problem that is associated with preterm delivery and for which no reliable diagnostic tool exists. Molecular analysis of 231 vaginal samples, classified by Gram stain-based Nugent score, was used to propose molecular criteria for BV; these criteria were prospectively applied to 56 new samples. A quantitative molecular tool targeting 8 BV-related microorganisms and a human gene was developed using a specific real-time polymerase chain reaction assay and serial dilutions of a plasmid suspension. The targeted microorganisms were Gardnerella vaginalis, Lactobacillus species, Mobiluncus curtisii, Mobiluncus mulieris, and Candida albicans (which can be identified by Gram staining), as well as Atopobium vaginae, Mycoplasma hominis, and Ureaplasma urealyticum (which cannot be detected by Gram staining). With use of the Nugent score, 167 samples were classified as normal, 20 were classified as BV, and 44 were classified as intermediate. Except for U. urealyticum, M. mulieris, and Lactobacillus species, DNA of the tested bacteria was detected more frequently in samples demonstrating BV, but the predictive value of such detection was low. The molecular quantification of A. vaginae (DNA level, > or = 10(8) copies/mL) and G. vaginalis (DNA level, > or = 10(9) copies/mL) had the highest predictive value for the diagnosis of BV, with excellent sensitivity (95%), specificity (99%), and positive (95%) and negative (99%) predictive values; 25 (57%) of the samples demonstrating intermediate flora had a BV profile. When applied prospectively, our molecular criteria had total positive and negative predictive values of 96% and 99%, respectively. We report a highly reproducible, quantitative tool to objectively analyze vaginal flora that uses cutoff values for the concentrations of A. vaginae and G. vaginalis to establish the molecular diagnosis of BV.

Quantitative Survival of Aerobic and Anaerobic Microorganisms in Port-A-Cul and Copan Transport Systems

Transport media should preserve the viability and stability of microorganisms in clinical specimens. In this study, the Port-A-Cul transport system and the Copan transport system without charcoal, both designed to preserve anaerobes, were evaluated. Dacron swabs were inoculated with two combinations of facultative and anaerobic organisms typically found in vaginal swab samples. Combination I contained Candida albicans, Escherichia coli, Enterococcus spp., group B streptococci, Lactobacillus crispatus, and Staphylococcus aureus. Combination II contained Lactobacillus iners, Peptoniphilus asaccharolyticus, Mycoplasma hominis, Prevotella bivia, Prevotella corporis, Porphyromonas asaccharolytica, Mobiluncus curtisii, Peptostreptococcus anaerobius, and Gardnerella vaginalis. Duplicate swabs were placed into the two transporters and held for 24, 48, 72, and 96 h at 4 and 24 degrees C. Both transporters maintained the viability of organisms better at 4 degrees C than at 24 degrees C. Prevotella bivia and Prevotella corporis had a loss of viability in both transporters at both temperatures. However, at 24 degrees C, there was a significantly greater loss of viability for Mycoplasma hominis, Prevotella bivia, Prevotella corporis, and Peptoniphilus asaccharolyticus when the organisms were stored in Copan transport medium than when they were stored in Port-A-Cul transport medium for 96 h (P < 0.002). Some organisms proliferated in the transport media, but when transporters were held at 24 degrees C for 96 h, a significantly greater increase in the concentrations of group B streptococci and Candida albicans, Escherichia coli, and Enterococcus spp. organisms in Copan medium than in Port-A-Cul medium was observed (P < 0.002). At room temperature, the Port-A-Cul system is superior to the Copan system with respect to the preservation of fastidious microorganisms and the prevention of the proliferation of facultative organisms.

Association of Mobiluncus curtisii With Recurrence of Bacterial Vaginosis

Association of Mobiluncus curtisii With Recurrence of Bacterial Vaginosis

The lower genital tract microbiota in relation to cytokine‐, SLPI‐ and endotoxin levels: application of checkerboard DNA‐DNA hybridization (CDH)

In the present study the lower genital tract microbiota in asymptomatic fertile women (n=34) was identified and quantified by culturing vaginal secretions. Also, vaginal and cervical samples were analyzed by a semiquantitative checkerboard DNA-DNA hybridization technique (CDH) based on genomic probes prepared from 13 bacterial species (Bacteroides ureolyticus, Escherichia coli, Fusobacterium nucleatum, Gardnerella vaginalis, Mobiluncus curtisii ss curtisii, Prevotella bivia, Prevotella disiens, Prevotella melaninogenica, Atopobium vaginae, Lactobacillus iners, Staphylococcus aureus ss aureus, Streptococcus anginosus, and Streptococcus agalactiae). The bacterial species found by either culture or CDH were correlated with proinflammatory cytokines (IL-1 alpha, IL-1 beta, IL-6, IL-8), secretory leukocyte protease inhibitor (SLPI), and endotoxin in the cervicovaginal samples. Grading the women into healthy, intermediate, or bacterial vaginosis (BV) as based on Gram staining of vaginal smears, the viable counts of lactobacilli (L. gasseri) and of streptococci-staphylococci combined were highest in the intermediate group. In BV, particularly the high concentrations of Actinomyces urogenitalis, Atopobium vaginae, and Peptoniphilus harei were noted (>or=10(11) per ml). The total viable counts correlated with both cervical IL-1 alpha and IL-1 beta. A strong negative correlation was observed between L. iners and total viable counts, G. vaginalis, or cervical IL-1 alpha, while it correlated positively with SLPI. Analysis of vaginal and cervical samples from 26 out of the 34 women by CDH showed that anaerobic bacteria were more frequently detected by CDH compared to culture. By this method, A. vaginae correlated with G. vaginalis, and L. iners with S. aureus. With regard to cytokines, B. ureolyticus correlated with both cervical and vaginal IL-1 alpha as well as with cervical IL-8, while F. nucleatum, S. agalactiae, S. anginosus, or S. aureus correlated with vaginal IL-1 alpha. Furthermore, all Gram-negative bacteria taken together, as measured by CDH, correlated with vaginal endotoxin and inversely with vaginal SLPI. The significance of the results is discussed. In summary, mapping of the identity and quantity of vaginal bacterial species and their association with locally produced host innate immune factors will help in defining various types of abnormal vaginal microbiota, developing new ways of assessing the risk of ascending subclinical infections, and in treating them. CDH appears to be a suitable tool for future analyses of large numbers of clinical samples with an extended number of bacterial probes.

Mobiluncus curtisii bacteremia

Mobiluncus curtisii was isolated from the blood of a 35-year-old man with a medical history of ulcerative colitis who was admitted unconscious to the Intensive Care Unit (ICU). A CT scan revealed massive intracerebral hemorrhage in the left hemisphere. Temperature remained constant over 38.5 °C; therefore, two sets of blood cultures were collected. One anaerobic bottle BacT/ALERT SN (bioMerieux, France) was detected as positive after 5 days of incubation and a Gram stain confirmed a gram variable curved-shaped rod. The patient died 18 h after being admitted to the hospital.

Use of DNA hybridization to detect vaginal pathogens associated with bacterial vaginosis among asymptomatic pregnant women

The purpose of this study was to determine whether microbial DNA hybridization is a useful method to study bacterial vaginosis in asymptomatic pregnant women. Vaginal specimens were collected at <26 weeks' gestation from 230 women, and analyzed for bacterial vaginosis by both Gram stain using Nugent criteria and DNA hybridization for Gardnerella vaginalis, Prevotella bivia, Bacteroides ureolyticus, and Mobiluncus curtisii. Results were analyzed using McNemar's paired test and chi-square test for trend, with significance set at P < .05. By Gram stain, 60 (26.1%) of 230 were positive for bacterial vaginosis, and 134 (58.3%) were negative. By DNA hybridization, 99 (43%) were positive for at least 1 pathogen, and DNA results were significantly associated with Gram-stain results (P < .01). As the Nugent score progressed from normal to abnormal flora, the proportion with >1 pathogen detected by DNA hybridization increased significantly (P < .001). DNA hybridization may be a useful method to study shifts in vaginal flora during pregnancy.

The use of Sorbarod biofilms to investigate some organisms involved in bacterial Vaginosis

Sorbarod biofilms were investigated for their suitability in establishing a continuous growth for the study of bacterial vaginosis. Important organisms in the condition, Gadnerella vaginalis, Mobiluncus curtisii and Lactobacillus acidophilus were studied. In contrast to growth in broth culture, L. acidophilus and G. vaginalis were maintained for at least 96 hours in steady state on the biofilms. The strict anaerobe M. curtisii was maintained in these biofilms for at least 48 hours an indication that anaerobic conditions do exist in these biofilms. The haemolytic activity of G. vaginalis was consistently maintained in the biofilms in contrast to the short-term activity in the broth culture, which matched the bacterial titre. Gram stains of thin sections of paraffin wax embedded Sorbarods showed the micro-colonies as distinct irregular aggregates of organisms adherent to the cellulose matrix for G. vaginalis while L. acidophilus appeared as large latticed networks that formed between the cellulose fibre matrix. The simple Sorbarod system appears to be suitable for studying the growth of bacteria in continuous culture and has a potential for investigating interactions between micro- organisms. Journal of Tropical Microbiology Vol.1(1) 2002: 36-40

Transfer of Members of the Genus Falcivibrio to the Genus Mobiluncus, and Emended Description of the Genus Mobiluncus

It has long been thought that the genera Mobiluncus and Falcivibrio contain the same organisms. Using a polyphasic approach, it was found that Mobiluncus curtisii and Mobiluncus mulieris were the same as Falcivibrio vaginalis and Falcivibrio grandis, respectively. As the genus name Mobiluncus takes precedence, it is proposed that F. vaginalis and F. grandis be transferred to the genus Mobiluncus. In agreement with previous studies, results from phenotypic tests did not support the separation of M. curtisii strains into its two subspecies, M. curtisii subsp. curtisii and M. curtisii subsp. holmesii. Phenotypic complexity within M. curtisii dictates that the species should be treated as a complex until more in-depth analyses of the species have been performed. Phylogenetic analyses, based on 16S rRNA gene sequences, demonstrated that the genus Mobiluncus was associated with Varibaculum cambriense and the two subspecies of Actinomyces neuii, and that A. neuii is only distantly related to Actinomyces sensu stricto.

A multiplex polymerase chain reaction-based diagnostic method for bacterial vaginosis

OBJECTIVE:To develop a polymerase chain reaction (PCR)-based diagnostic method for bacterial vaginosis using bacterial vaginosis-associated anaerobes.METHODS:A multiple PCR assay was developed using primers specific to 16S ribosomal deoxyribonucleic acid (DNA) (Mobiluncus mulieris and Mobiluncus curtisii), nanH (Bacteroides fragilis), and an internal spacer region of ribosomal DNA (Gardnerella vaginalis). The vaginal swabs from pregnant and nonpregnant women were examined by Gram stain-based Nugent scoring system. One hundred seventy-two samples of 853 Gram stain-interpretable samples were randomly selected and subjected to multiplex PCR assay.RESULTS:The sensitivity of the PCR assay ranged from 103 to 104 colony-forming units per vaginal swab. The prevalence of the bacterial vaginosis, intermediate, and normal categories was found by Nugent scoring system to be 21.6% (184/853), 26.0% (222/853), and 52.4% (447/853), respectively. By the multiplex PCR-based diagnostic method, 20.3% (35/172) of the samples were identified as bacterial vaginosis. The diagnostic sensitivity, specificity, positive predictive value, and negative predictive value of multiplex PCR in comparison with Gram stain examination were 78.4% (95% confidence interval [CI] 65.1%, 91.6%), 95.6% (95% CI 92.1%, 99.0%), 82.9% (95% CI 70.4%, 95.4%), and 94.2% (95% CI 90.3%, 98.1%), respectively.CONCLUSION:This multiplex PCR can be used as a diagnostic or screening test for bacterial vaginosis.

Severe sepsis caused by Mobiluncus curtisii subsp. curtisii in a previously healthy female: Case report and review

We present the case of a 54-year-old female with life-threatening septicaemia due to Mobiluncus curtisii subsp. curtisii. Her admission was complicated by septic shock, renal failure, disseminated intravascular coagulation, the adult respiratory distress syndrome and spontaneous splenic rupture. The patient survived with full intensive care support and intravenous ceftriaxone. Extra-genital infection with Mobiluncus species is rarely diagnosed and has been confined to breast abscesses and non-life-threatening bacteraemia. A review of extra-genital infections with Mobiluncus species is presented.

Humoral antibody to Mobiluncus curtisii, a potential serological marker for bacterial vaginosis.

While bacterial vaginosis (BV) is a polymicrobial syndrome, Mobiluncus spp. are the organisms most highly associated with this condition. It is possible that serum antibody to Mobiluncus spp. could be used as a serological marker for BV. Using immunofluorescence techniques, we studied the prevalence of antibody to M. curtisii among three cohorts-pregnant women, pediatric patients, and sexually inexperienced women. The prevalence of antibody in each of these three groups was 75, 6, and 0%, respectively. Of the three pediatric patients with antibody to Mobiluncus curtisii, two were neonates, and the only class of antibody detected was immunoglobulin G. Among the cohort of pregnant women, the presence of antibody could not be correlated with a clinical history of BV. Serum antibody to M. curtisii could be a useful serological marker for BV. The lack of correlation of antibody positivity to historical information regarding BV suggests that unrecognized or undiagnosed episodes of BV may be common.

Broad-range PCR amplification and DNA sequence analysis reveals variable motifs in 16S rRNA genes of Mobiluncus species.

Using DNA primers based on highly conserved regions of bacterial 16S ribosomal RNA genes, a technique was established for detection of Mobiluncus species by polymerase chain reaction (PCR) and hybridization analysis. Part of the 16S rRNA genes of Mobiluncus mulieris, Mobiluncus curtisii and uncharacterized Mobiluncus strains were analyzed by broad-range PCR amplification and direct DNA sequencing analysis. Sequence comparison of the partial 16S rRNA genes of Mobiluncus curtisii, Mobiluncus mulieris and atypical Mobiluncus strains studied indicated genus and species-specific motifs within the variable regions V3, V4 and V9 of 16S ribosomal DNAs. A Mobiluncus curtisii-specific primer, located within the variable region V3 of the 16S rRNA gene, was designed for Southern blot hybridization analysis of broad-range PCR products. Broad-range amplification combined with a M. curtisii-specific hybridization probe, Mob V3, distinguished between Mobiluncus curtisii, Mobiluncus mulieris, and atypical Mobiluncus strains.

Mobiluncus curtisii Bacteremia Following Septic Abortion

Mobiluncus curtisii Bacteremia Following Septic Abortion

Bacterial vaginosis and vaginal microorganisms in idiopathic premature labor and association with pregnancy outcome.

The vaginal microflora of 49 women in idiopathic preterm labor was compared with that of 38 term controls to determine whether the presence of bacterial vaginosis (BV) and/or specific microorganisms would influence the rate of preterm delivery. Demographic factors, pregnancy outcome, and reproductive history were also studied. BV, as defined by the presence of clue cells in a vaginal wet mount and characteristic microbial findings in a stained vaginal smear and vaginal culture, was more common in women with preterm labor and delivery than in controls (P < 0.01). The condition, diagnosed in 41% of women who had both preterm labor and delivery (n = 22) and in 11% each of women who had preterm labor but term delivery (n = 27) and controls, was associated with a 2.1-fold risk (95% confidence intervals, 1.2 to 3.7) for preterm birth prior to 37 weeks of gestation. BV was associated with low birth weight. Of 49 women with preterm labor, 67% (8 of 12) of women with BV were delivered of low-birth-weight neonates (< 2,500 g) compared with 22% (8 of 37) of women without the condition (P < 0.0005). The presence of hydrogen peroxide-producing facultative Lactobacillus spp. was strongly negatively associated with both preterm delivery and BV. BV-associated microorganisms, i.e., Mobiluncus, Prevotella, and Peptostreptococcus species, Porphyromonas asaccharolytica, Fusobacterium nucleatum, Mycoplasma hominis, and high numbers of Gardnerella vaginalis were significantly associated with preterm delivery; all species also strongly associated with BV (P = 0.0001 for each comparison). Mobiluncus curtisii and Fusobacterium nucleatum were recovered exclusively from women with preterm delivery. Our study clearly indicates that BV and its associated organisms are correlated with idiopathic premature delivery.

Diversity of Mobiluncus Strains as Demonstrated by their Electrophoretic Protein Patterns

The analysis of electrophoretic protein profiles of 21 Mobiluncus curtisii, 11 Mobiluncus mulieris and 3 reference strains (Mobiluncus curtisii subsp. curtisii ATCC 35241, Mobiluncus curtisii subsp. holmesii ATCC 35242 and Mobiluncus mulieris BV 64-5) demonstrated species-related patterns. A highly variable region appeared at 70-85 kDa for Mobiluncus curtisii and at 75-95 kDa for Mobiluncus mulieris, which was likely to correspond to cell surface located proteins. When performed under standardized conditions, PAGE-protein analysis allowed to define intraspecies clusters, from which some strains appeared identical. Thus, the method seemed to provide a useful additive to identify a strain at the species level and might be of epidemiological interest.

Characterization of unusual tetracycline-resistant gram-positive bacteria

Tetracycline-resistant Tet M-negative isolates of Actinomyces viscosus, Eubacterium lentum, Mobiluncus curtisii, and Mobiluncus mulieris were screened with the Tet K, Tet L, and Tet O DNA probes. Ten (71%) of the resistant Mobiluncus strains hybridized with the Tet O probe, two of the three E. lentum strains hybridized with the Tet K probe, and the A. viscosus isolate hybridized with the Tet L probe.

Normal vaginal aerobic and anaerobic bacterial flora of the rhesus macaque (Macacca mulatta)

The most common bacterial species isolated from the vaginas of 37 healthy rhesus macaques were Streptococcus viridans, coagulase negative Staphylococcus, Mobiluncus curtisii ss. curtisii, Corynebacterium renale-like organisms, Peptostreptococcus anaerobius, Gardnerella-like organisms, and other Corynebacterium species. The vaginal flora of the rhesus macaque differs from that previously reported for five other primate species. A two-year retrospective review of clinical cases of vaginitis and metritis found Escherichia coli and coagulase positive Staphylococci to be the most common pathogens isolated.

Reservoir of four organisms associated with bacterial vaginosis suggests lack of sexual transmission

This study consisted of a search for the possible reservoir and mode of spread of the four bacterial vaginosis-associated organisms Mobiluncus mulieris, Mobiluncus curtisii, Mycoplasma hominis, and Gardnerella vaginalis. Their occurrence in rectal, oral, and pharyngeal specimens from women with and without bacterial vaginosis, their male sexual consorts, four homosexual men, and children (altogether, 374 people) was studied. Genital samples were also obtained from all adults. All four organisms were isolated from the rectums of 45 to 62% of women with bacterial vaginosis and 10 to 14% of women without bacterial vaginosis. They also occurred in the rectums of males and children. M. hominis was recovered from the oropharynxes of 12 adults whose sexual consorts had genital occurrences of the organism. Mobiluncus spp. occurred only in the vaginas of women with bacterial vaginosis (97%). The organisms were only infrequently recovered from genital samples from 135 males. Organisms were recovered from the urethras and/or coronal sulci of 10 of 44 male consorts of women with bacterial vaginosis. However, after 2 weeks of condom use during sexual intercourse, only M. hominis remained in the urethra of one man. These findings suggest that the organisms associated with bacterial vaginosis are not spread sexually but colonize the vagina from an endogenous intestinal tract site. The pathophysiological mechanisms leading to bacterial vaginosis in a subpopulation of all women are still unknown.

Demonstration of heterogeneity among the antigenic proteins of Mobiluncus species

The protein and antigenic profiles of the American Type Culture Collection type strains of Mobiluncus species and those of 114 clinical isolates were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and immunoblotting with homologous polyvalent antisera. The majority of isolates (82%) possessed characteristic protein profiles and could be identified to the species level by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. The major protein bands were also antigenic, and some antigenic cross-reactivity was noted between the two Mobiluncus species. All of the isolates were examined for reactivity with a panel of 12 monoclonal antibodies previously prepared against the type strains. While 56 of 60 clinical isolates of Mobiluncus curtisii (93%) reacted with one or more of the monoclonal antibodies, only 23 of 54 clinical isolates which were identified as Mobiluncus mulieris by biochemical methods (48%) reacted with one or more of the monoclonal antibodies. One of the 4 M. curtisii isolates (25%) and 11 of the 31 M. mulieris isolates (35%) which did not react with the monoclonal antibodies also had atypical protein profiles. These results demonstrate a high degree of heterogeneity in the protein and antigenic profiles of Mobiluncus isolates and suggest that further taxonomic division may be appropriate.