Drug resistance-associated mutations in Mycoplasma genitalium in female sex workers, Japan.

Mycoplasma genitalium was detected in 21 (14.1%) of 149 vaginal swab samples and in 1 (0.7%) of 149 throat washing samples from female sex workers during 2013-2014 in Japan. Prevalences of M. genitalium with macrolide resistance-associated 23S rRNA mutations and fluoroquinolone resistance-associated parC alterations were 47.1% and 36.8%, respectively.

Bacterial loads of Ureaplasma parvum contribute to the development of inflammatory responses in the male urethra.

Ureaplasma parvum, which has been recognised as a coloniser in the male urethra, is detected in some men with non-gonococcal urethritis. In this study, we quantified the 16 S rRNA genes of U. parvum by a real-time polymerase chain reaction-based assay in first-voided urine from 15 symptomatic and 38 asymptomatic men who were positive only for U. parvum. We also determined the leukocyte counts by automated quantitative urine particle analysis in their first-voided urine. Positive correlations were observed between copies of the 16 S rRNA genes of U. parvum/ml and the leukocyte counts/µl in first-voided urine (p = 0.0019). The loads of ≥10(4) copies of the 16 S rRNA gene/ml, corresponding to ≥5 × 10(3) cells of U. parvum/ml, were significantly associated with the presence of ≥12.5 leukocytes/µl in first-voided urine that might document the presence of inflammatory responses in the urethra. However, a large portion of the subjects (83.0%) had bacterial loads of <5 × 10(3) cells of U. parvum/ml, and 79.5% of them showed <12.5 leukocytes/µl. The ambiguity of the pathogenic role of U. parvum in non-gonococcal urethritis could, in part, be due to its low bacterial loads, which might not give rise to inflammatory responses in the male urethra.

Prediction of the persistence of Mycoplasma genitalium after antimicrobial chemotherapy by quantification of leukocytes in first-void urine from patients with non-gonococcal urethritis.

Mycoplasma genitalium is regarded as another pathogen of male non-gonococcal urethritis (NGU). Failure to eradicate this mycoplasma is associated with persistent or recurrent NGU, but this mycoplasma is not routinely examined in clinical practice. In cases of M. genitalium-positive NGU, therefore, some criteria are needed to assess the success or failure of antimicrobial chemotherapy other than microbiological outcomes. We enrolled 49 men with M. genitalium-positive non-chlamydial NGU. At successive visits after treatment, we inquired about their symptoms, observed their urethral meatus for urethral discharge, and examined their first-void urine (FVU) for quantification of leukocytes and for the persistence of M. genitalium. M. genitalium was eradicated in 34 patients after treatment, whereas the mycoplasma persisted in 15. Urethritis symptoms and urethral discharges were not found to be predictors of the persistence of M. genitalium up to the 25th day after the start of treatment. Leukocyte counts in FVU from the patients with persistence of M. genitalium were significantly higher than those from the patients with eradication of the mycoplasma. Leukocyte counts of 10 leukocytes/µl or more between the 18th and 24th day after the start of treatment were most significantly associated with the persistence of M. genitalium. Quantification of leukocytes in FVU would appear to be crucial to judge the outcome of treatment in patients with non-chlamydial NGU and could be helpful to predict the persistence of M. genitalium after treatment when M. genitalium is not routinely examined in clinical specimens in clinical practice.

The role of Mycoplasma genitalium and Ureaplasma urealyticum biovar 2 in postgonococcal urethritis.

BACKGROUND: There are few studies on coinfection with genital mycoplasmas and ureaplasmas in men with gonococcal urethritis (GU). The role of these species in postgonococcal urethritis (PGU) is poorly understood. Thus, we conducted a study to determine the prevalence of coinfection with genital mycoplasmas and ureaplasmas among men with GU and to assess the role of these pathogens in PGU. METHODS: Three hundred ninety men infected with culture-confirmed Neisseria gonorrhoeae participated in the study. Chlamydia trachomatis, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum biovar 1, and Ureaplasma urealyticum biovar 2 in first-voided urine samples were detected by polymerase chain reaction-based assay at the patients' initial visits. PGU was judged to be present if the urethral smear was positive for polymorphonuclear leucocytes 7-14 days after treatment for gonorrhea. The association between each microorganism and PGU, measured by the odds ratio, was estimated by multivariate logistic regression analysis. RESULTS: C. trachomatis, M. genitalium, M. hominis, U. parvum biovar 1, and U. urealyticum biovar 2 were detected in 85 (21.8%), 16 (4.1%), 8 (2.1%), and 33 men (8.5%), respectively. In patients with chlamydia-negative GU, coinfection with M. genitalium was associated with a 14.54-fold greater risk of PGU (95% confidence interval, 2.91-72.74), and coinfection with U. urealyticum biovar 2 was associated with a 3.64-fold greater risk of PGU (95% confidence interval, 1.24-10.63). CONCLUSIONS: Coinfection with M. genitalium or U. ureaplasma biovar 2 in men with GU was significantly associated with PGU, independent of C. trachomatis. Men with GU should be treated presumptively with antibiotics that are active against C. trachomatis, M. genitalium, and U. urealyticum biovar 2.

Emergence and spread of Neisseria gonorrhoeae clinical isolates harboring mosaic-like structure of penicillin-binding protein 2 in Central Japan.

Of 150 clinical isolates of Neisseria gonorrhoeae recovered in 2001, we examined 55 clinical isolates of N. gonorrhoeae for which cefixime MICs were > or=0.125 microg/ml and randomly selected 15 isolates for which cefixime MICs were < or =0.06 microg/ml for analysis of alterations in the penicillin-binding protein 2 (PBP 2) gene. We found insertion of an extra codon (Asp-345a) in the transpeptidase domain of PBP 2, and this insertion occurred alone or in conjunction with other amino acid substitutions. We also found a mosaic PBP 2 that was composed of fragments of the PBP 2 proteins from Neisseria cinera and Neisseria perflava. This mosaic PBP 2 was significantly associated with decreased susceptibilities to penicillin and cephalosporins, especially oral cephalosporins. For most of the isolates with a mosaic PBP 2, the cefixime MICs were > or =0.5 microg/ml and the cefdinir MICs were > or =1 microg/ml. Analysis of chromosomal DNA restriction patterns by pulsed-field gel electrophoresis revealed that most isolates with the mosaic PBP 2 were genetically similar. The recombination events that generated the mosaic PBP 2 would likely have contributed to the decreased sensitivities to cephalosporins. Isolates with the mosaic PBP 2 appear to threaten the efficacy of the currently recommended regimen with cefixime. The emergence of such strains may be the result of the in vivo generation of clones in which interspecies recombination occurred between the penA genes of N. gonorrhoeae and commensal Neisseria species.