Evaluation of commercial, customized microdilution plates for Ureaplasma parvum, Ureaplasma urealyticum, and Mycoplasma hominis antimicrobial susceptibility testing and determination of antimicrobial resistance prevalence in France.

Antimicrobial susceptibility testing (AST) of human mycoplasmas using microdilution is time-consuming. In this study, we compared the performance of MICRONAUT-S plates (Biocentric-Bruker) designed for AST of Ureaplasma parvum, Ureaplasma urealyticum, and Mycoplasma hominis with the results using the Clinical & Laboratory Standards Institute (CLSI) reference method. Then, we investigated the prevalence and mechanisms of resistance to tetracyclines, fluoroquinolones, and macrolides in France in 2020 and 2021. The two methods were compared using 60 strains. For the resistance prevalence study, U. parvum-, U. urealyticum-, and M. hominis-positive clinical specimens were collected for 1 month each year in 22 French diagnostic laboratories. MICs were determined using the MICRONAUT-S plates. The tet(M) gene was screened using PCR, and fluoroquinolone resistance-associated mutations were screened using PCR and Sanger sequencing. Comparing the methods, 99.5% (679/680) MICs obtained using the MICRONAUT-S plates concurred with those obtained using the CLSI reference method. For 90 M. hominis isolates, the tetracycline, levofloxacin, and moxifloxacin resistance rates were 11.1%, 2.2%, and 2.2%, respectively, with no clindamycin resistance. For 248 U. parvum isolates, the levofloxacin and moxifloxacin resistance rates were 5.2% and 0.8%, respectively; they were 2.9% and 1.5% in 68 U. urealyticum isolates. Tetracycline resistance in U. urealyticum (11.8%) was significantly (P < 0.001) higher than in U. parvum (1.2%). No macrolide resistance was observed. Overall, the customized MICRONAUT-S plates are a reliable, convenient tool for AST of human mycoplasmas. Tetracycline and fluoroquinolone resistance remain limited in France. However, the prevalence of levofloxacin and moxifloxacin resistance has increased significantly in Ureaplasma spp. from 2010 to 2015 and requires monitoring. IMPORTANCE: Antimicrobial susceptibility testing of human urogenital mycoplasmas using the CLSI reference broth microdilution method is time-consuming and requires the laborious preparation of antimicrobial stock solutions. Here, we validated the use of reliable, convenient plates designed for antimicrobial susceptibility testing that allows the simultaneous determination of the MICs of eight antibiotics of interest. We then investigated the prevalence and mechanisms of resistance of each of these bacteria to tetracyclines, fluoroquinolones, and macrolides in France in 2020 and 2021. We showed that the prevalence of levofloxacin and moxifloxacin resistance has increased significantly in Ureaplasma spp. from 2010 to 2015 and requires ongoing monitoring.

Antibiotic resistance among clinical Ureaplasma isolates from Cuban individuals between 2013 and 2018.

Introduction. Acquired resistance against the antibiotics that are active against Ureaplasma species has been described.Hypothesis/Gap Statement. Diagnostics combined with antimicrobial sensitivity testing are required for therapeutic guidance.Aim. To report the prevalence of antimicrobial resistance among Cuban Ureaplasma isolates and the related molecular mechanisms of resistance.Methodology. Traditional broth microdilution assays were used for antimicrobial sensitivity testing in 262 clinical Ureaplasma species isolates from Cuban patients between 2013 and 2018, and a subset of samples were investigated in parallel with the commercial MYCO WELL D-ONE rapid culture diagnostic assay. The underlying molecular mechanisms for resistance were determined by PCR and sequencing for all resistant isolates.Results. Among the tested isolates, the tetracycline and erythromycin resistance rates were 1.9 and 1.5%, respectively, while fluoroquinolone resistance was not found. The tet(M) gene was found in all tetracycline-resistant isolates, but also in two tetracycline-susceptible Ureaplasma clinical isolates. We were unable to determine the underlying mechanism of erythromycin resistance. The MYCO WELL D-ONE kit overestimated tetracycline and erythromycin resistance in Ureaplasma spp. isolates.Conclusions. Although low levels of antibiotic resistance were detected in Cuban patients over a 5-year period, continued surveillance of the antibiotic susceptibility of Ureaplasma is necessary to monitor possible changes in resistance patterns.

Effect of probiotics on vaginal Ureaplasma parvum in women suffering from unexplained infertility.

RESEARCH QUESTION: Does oral probiotic supplementation influence the relative abundance of different vaginal microbiota in women experiencing infertility? DESIGN: A prospective, monocentric randomized controlled trial. To study the influence of probiotics on infertility, 80 patients with primary or secondary infertility were included. Patients were assigned to either a probiotic treatment or a control group. Participants in the treatment group (n = 40) took one sachet (2 g) a day of a defined probiotic supplement limiting Lactobacillus strains. Patients in the control group did not receive any additional probiotic supplements. Vaginal samples were taken on day 20 of the menstrual cycle and 4 weeks later, on day 20, of the consecutive cycle. Subsequently, 16s rRNA gene analysis of the vaginal samples was conducted. RESULTS: After the intervention phase, no effects on alpha diversity resulting from treatment could be observed. The between sample diversity of different women (beta diversity) at baseline had no effects of age, treatment group or body mass index. Primary or secondary sterility, however, had a significant effect on community. Three clusters (Lactobacillus crispatus, Lactobacillus iners and Lactobacillus gasseri) were identified as the leading representatives. Furthermore, patients treated with probiotics showed limited growth of Ureaplasma parvum compared with the control group (P = 0.021). CONCLUSIONS: This study points to a possible protective effect of probiotic supplements on the vaginal microbiota. It is tempting to speculate that this effect assists in containing the growth of non-beneficial bacteria and helps to prevent or cure a dysbiotic vaginal flora.

Molecular exploration for Mycoplasma amphoriforme, Mycoplasma fermentans and Ureaplasma spp. in patient samples previously investigated for Mycoplasma pneumoniae infection.

OBJECTIVES: To determine the presence and genotypic macrolide susceptibility of Mycoplasma amphoriforme, and the presence of Ureaplasma spp. and Mycoplasma fermentans among clinical samples from England previously investigated for Mycoplasma pneumoniae. METHODS: Quantitative and conventional PCR methods were used to retrospectively screen a collection of 160 clinical samples previously submitted to Public Health England (PHE) for the detection of M. pneumoniae between October 2016 and December 2017. Samples which were positive for M. amphoriforme DNA were further investigated for mutations associated with genotypic macrolide resistance by sequencing domain V of the 23s rRNA. RESULTS: M. amphoriforme was detected in 10/160 samples (6.3%), Ureaplasma parvum was detected in 4/160 samples (2.5%), and M. fermentans was not detected in any samples (0/160). Of the nine individuals (two samples were from the same patient) in which M. amphoriforme was detected, eight were male (age range 10-60 years) and one was female (age range 30-40 years). One individual with cystic fibrosis was positive for both M. amphoriforme and U. parvum. All M. amphoriforme DNA was genotypically susceptible to macrolides. CONCLUSIONS: Mycoplasma amphoriforme was found in clinical samples, including lower respiratory tract samples of patients with pneumonia. In the absence of other respiratory pathogens, these data suggest a potential role for this organism in human disease, with no evidence of acquired macrolide resistance. Ureaplasma parvum was detected in cerebrospinal fluid and respiratory tract samples. These data suggest that there is a need to consider these atypical respiratory pathogens in future diagnostic investigations.

Antimicrobial activity of enacyloxin IIa and gladiolin against the urogenital pathogens Neisseria gonorrhoeae and Ureaplasma spp.

AIMS: To determine the antimicrobial activity of enacyloxin IIa and gladiolin against Neisseria gonorrhoeae and Ureaplasma spp. METHODS AND RESULTS: The Burkholderia polyketide antibiotics enacyloxin IIa and gladiolin were tested against 14 N. gonorrhoeae and 10 Ureaplasma spp. isolates including multidrug-resistant N. gonorrhoeae isolates WHO V, WHO X and WHO Z as well as macrolide, tetracycline and ciprofloxacin-resistant ureaplasmas. Susceptibility testing of N. gonorrhoeae was carried out by agar dilution, whereas broth micro-dilution and growth kinetic assays were used for Ureaplasma spp. The MIC range for enacyloxin IIa and gladiolin against N. gonorrhoeae was 0·015-0·06 mg l-1 and 1-2 mg l-1 respectively. The presence of resistance to front line antibiotics had no effect on MIC values. The MIC range for enacyloxin IIa against Ureaplasma spp. was 4-32 mg l-1 with a clear dose-dependent effect when observed using a growth kinetic assay. Gladiolin had no antimicrobial activity on Ureaplasma spp. at 32 mg l-1 and limited impact on growth kinetics. CONCLUSIONS: Enacyloxin IIa and gladiolin antibiotics have antimicrobial activity against a range of antibiotic susceptible and resistant N. gonorrhoeae and Ureaplasma isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights the potential for a new class of antimicrobial against pathogens in which limited antibiotics are available. Development of these compounds warrants further investigation in the face of emerging extensively drug-resistant strains.

In-vitro activity of lipoic acid against Ureaplasma urealyticum and Ureaplasma parvum isolated from women with infections of the urogenital tract. A pilot study.

Several species of Ureaplasma bacteria are known to be present in the urogenital tract of humans, in both healthy individuals and symptomatic patients. These pathogens are associated with urogenital tract infections, infertility problems and spontaneous abortion in humans. The present study involved 77 strains of Ureaplasma species (Ureaplasma spp.), including 21 Ureaplasma urealyticum (U. urealyticum) strains and 56 Ureaplasma parvum (U. parvum) strains. Lipoic acid (LA) and its reduced form dihydrolipoic acid (DHLA) are synthesized in all prokaryotic and eukaryotic cells. Research of recent years increasingly points to therapeutic properties of exogenously supplemented LA. In our study, we examined for the first time the effect of LA on the bacteria multiplication and its bactericidal activity against U. urealyticum and U. parvum. The LA concentrations used were: 1200 µg/ml, 120 µg/ml, and 12 µg/ml. The titer for each strain of Ureaplasma spp. was estimated using the color changing units (CCU) assay. For CCU measurements, a series of 10-fold dilutions of each cell culture in 0.9% NaCl (titration) was prepared and 1 CCU/ml was defined as the highest dilution of cells at which color change was detected. The strongest bacteriostatic and bactericidal effect of LA was observed at a concentration of 1200 µg/ml. In contrast, at lower LA concentrations, stimulation of the bacteria multiplication was noted for 14% of the total number of strains tested. Taken together, the current data provide novel findings about potential beneficial antimicrobial effects of LA.

Rare case of Ureaplasma parvum septic arthritis in an immunocompetent patient.

Mycoplasmatacea family comprises two genera: Mycoplasma and Ureaplasma Ureaplasma parvum (previously known as U. urealyticum biovar 1) commonly colonises the urogenital tract in humans. Although Ureaplasma species have well-established pathogenicity in urogenital infections, its involvement in septic arthritis has been limited to prosthetic joint infections and immunocompromised individuals. We present a rare case of native right knee infection due to U. parvum identified using next-generation sequencing of microbial cell-free DNA testing and confirmed with PCR assays. This rare case of Ureaplasma septic arthritis was diagnosed using newer next-generation DNA sequencing diagnostic modalities and a literature review of prior cases, antibiotic coverage and antimicrobial resistance is incorporated as part of the discussion.

Antimicrobial resistance, genetic characterization, and molecular epidemiology of Ureaplasma species in males with infertility.

This study aimed to study the antimicrobial resistance, genetic characterization, and molecular epidemiology of Ureaplasma species in order to provide clinicians sufficient data to select optimal strategies of treatment for genitourinary tract infections of infertile male patients. Firstly, a total of 817 clinical semen specimens were detected for Ureaplasma species by molecular detection. Secondly, culture and identification of Ureaplasma species were achieved by using Mycoplasma ICS Test, and the antimicrobial susceptibility tests were determined by using broth microdilution assay. Then, the tetracycline resistance genetic determinants in Ureaplasma species were identified by PCR, and the fluoroquinolone and macrolide resistance genetic determinants were identified by DNA sequencing. Finally, the molecular epidemiology of Ureaplasma species was studied by both multilocus sequence typing (MLST) and expanded MLST (eMLST) schemes. Among the 817 semen specimens, 320 (39.17%) specimens were positive for Ureaplasma species. The percentages of resistance in 320 isolates against LEV, MXF, TET, and ERY were 47.5%, 39.38%, 19.69%, and 3.75%, respectively. The tet(M) and int-Tn genes were detected positive in all the tetracycline-resistant isolates. One macrolide-resistant UU isolate had a novel amino acid alteration (R66T) in L4 ribosomal protein and another UU isolate harbored a novel alteration (S109T) in L22. In fluoroquinolone-resistant isolates, S83L substitution in the ParC was predominant. In this area, ST22 and eST16 were the most prevalent ST and eST, respectively. One ST and 3 eSTs were newly identified in this study. This study has demonstrated that ERY can be first-line therapy for Ureaplasma species infections.

MYCO WELL D-ONE detection of Ureaplasma spp. and Mycoplasma hominis in sexual health patients in Wales.

The genital mycoplasmas are a unique group of inherently antibiotic-resistant sexually transmitted bacteria, often associated with non-gonococcal urethritis and bacterial vaginosis. The MYCO WELL D-ONE is a culture-based assay that aims to detect these organisms whilst concurrently screening them for antibiotic resistance. Urine and/or swabs from 856 informed and consented participants attending Welsh sexual health clinics were subjected to MYCO WELL D-ONE analysis, alongside qPCR and culture titration methodologies to determine sensitivity, specificity, PPV, NPV and accuracy. Resistance was confirmed by CLSI-compliant susceptibility testing and genetic mechanisms determined. The MYCO WELL D-ONE displayed a sensitivity and specificity of 91.98% and 96.44% for the detection of Ureaplasma spp., with sensitivity and specificity values of 78.23% and 98.84% for Mycoplasma hominis, compared with qPCR. Swabs harboured significantly greater bacterial loads than urine samples for both Ureaplasma spp. and M. hominis. Levofloxacin resistance rates, mediated by Ser83Leu mutation in ParC, for Ureaplasma spp. were 0.54%. Tetracycline resistance rates, mediated by tet(M), were 0.54% and 2% for Ureaplasma spp. and M. hominis, respectively; sequence analysis of tet(M)-positive Ureaplasma spp. and M. hominis strains isolated from a single individual confirmed separate resistance gene origins. The MYCO WELL D-ONE is a sensitive and specific assay for the detection of Ureaplasma spp. and M. hominis in genitourinary medicine samples, facilitating the accurate detection of these organisms within low-technology environments. While good for antibiotic resistance screening, accurate confirmation by MIC determination or molecular methods are required, and more optimally performed on urine samples.

Hyperammonemia From Ureaplasma Infection in an Immunocompromised Child.

Idiopathic hyperammonemia is a rare, poorly understood, and often lethal condition that has been described in immunocompromised patients. This report describes an immunocompromised patient with acute myelogenous leukemia who developed persistent hyperammonemia up to 705 µmol/L (normal, 0 to 47 µmol/L) refractory to multiple different therapies. However, after beginning azithromycin and then doxycycline therapy for Ureaplasma species infection, the patient showed immediate and sustained clinical improvement and resolution of ammonia levels. Recognizing disseminated Ureaplasma species infection as a potential cause of idiopathic hyperammonemia, an unexplained, often fatal condition in immunocompromised patients, and empirically treating for this infection could potentially be lifesaving.

Current understanding and treatment of intra-amniotic infection with Ureaplasma spp.

Considerable evidence has shown that intra-amniotic infection with Ureaplasma spp. increases the risk of chorioamnionitis and preterm labor. Ureaplasma spp. are among the smallest organisms, and their isolation is uncommon in routine clinical practice because of their size and high auxotrophy. Although Ureaplasma spp. have been reported as causative agents of preterm birth, they also have a high incidence in vaginal swabs collected from healthy reproductive-age women; this has led to questions on the virulence of Ureaplasma spp. and to them being considered as harmless commensal bacteria. Therefore, many efforts have been made to clarify the pathogenicity of Ureaplasma spp. at the molecular level. Ureaplasma spp. are surrounded by lipoproteins, including multiple-banded antigen. Both multiple-banded antigen and its derivative, that is, the synthetic lipopeptide, UPM-1, induce an inflammatory response in a preterm mice model, which was adequate to cause preterm birth or stillbirth. In this review, we present an overview of the virulence mechanisms of Ureaplasma spp. and treatment of ureaplasma infection during pregnancy to prevent possible serious sequelae in infants. In addition, relevant mechanisms underlying antibiotic resistance in Ureaplasma spp. are discussed. Ureaplasma spp. are naturally resistant against ß-lactam antibiotics because of the lack of a cell wall. Azithromycin is one of the effective agents that can control intrauterine ureaplasma infection. In fact, macrolide- and fluoroquinolone-resistant isolates of Ureaplasma spp. have already been observed in perinatal practice in Japan.

Antibiotic susceptibilities and genetic variations in macrolide resistance genes of Ureaplasma spp. isolated in China.

Macrolides are widely used for the treatment of Ureaplasma spp. infection. The aim of this study was to investigate possible genetic resistance determinants in Ureaplasma spp. isolates. A total of eleven macrolide- resistant Ureaplasma spp. isolates, recovered from urogenital specimens, were investigated for genetic mechanisms of macrolide resistance. The 23S rRNA operons, as well as L4 and L22 ribosomal protein genes, were amplified and sequenced. Our study identified that the mutation A2066G in the 23S rRNA and four mutations (G361T, A406G, C422T and G196A) in the L22 ribosomal protein, may be responsible for the resistance of Ureaplasma spp. to macrolides, in China.

Effect of antenatal azithromycin for Ureaplasma spp. on neonatal outcome at ≤30 weeks' gestational age.

BACKGROUND: Ureaplasma spp. in the maternal genitourinary tract has come to attention as a cause of preterm labor, spontaneous abortion, chorioamnionitis and adverse outcomes. A few controversies, however, still remain, namely, whether it should be treated aggressively or not. The aim of this study was to evaluate the effect of maternal azithromycin (AZ) treatment for Ureaplasma colonization on neonatal morbidities including bronchopulmonary dysplasia (BPD). METHODS: A retrospective case-control study of preterm babies delivered at ≤30 weeks of gestational age (GA) from 2012 to 2016 was conducted. Infants whose mothers had confirmed Ureaplasma colonization and treatment with AZ (m-AZ, cases) were matched by GA to control subjects whose mothers did not have Ureaplasma colonization. A subgroup analysis (nUU(+), infants with neonatal respiratory Ureaplasma colonization; nUU(-), infants without colonization) was also performed. RESULTS: Fifty-five control subjects were matched to 110 m-AZ subjects. The incidence of preterm premature rupture of membranes (P = 0.003) and of moderate-severe BPD (P = 0.010) was significantly higher in the m-AZ group. On subgroup analysis with post-hoc analysis (m-AZ + nUU(+) [I, n = 55] vs m-AZ + nUU(-) [II, n = 55] vs controls [n = 55]), the incidence of moderate-severe BPD was significantly different: 26% (I) vs 22% (II) vs 7% (controls), P = 0.033. CONCLUSIONS: Maternal Ureaplasma colonization was associated with moderate-severe BPD despite the use of AZ treatment. In addition, if the neonatal respiratory tract was colonized, then moderate-severe BPD developed even with maternal AZ treatment. Hence, selective antenatal and postnatal treatment of Ureaplasma colonization would be needed to control BPD development.

Antibiotics for amniotic-fluid colonization by Ureaplasma and/or Mycoplasma spp. to prevent preterm birth: A randomized trial.

OBJECTIVE: To assess whether antibiotics used for treatment in asymptomatic second-trimester women positive for Mycoplasma or Ureaplasma spp. detected by amniotic-fluid PCR prevents preterm delivery. DESIGN: A randomized, double-blind, placebo-controlled trial. SETTING: 10 maternal fetal medicine centers in France. POPULATION: Women with a singleton pregnancy who underwent amniocentesis between 16 and 20 weeks' gestation (weeks) for Down syndrome screening. A sample of 238 women with PCR-positive findings per treatment group was needed to show a 50% reduction in the preterm delivery rate. METHODS: Amniotic fluid was tested. Women with positive findings on real-time PCR of amniotic fluid for Mycoplasma hominis, Mycoplasma genitalium, Ureaplasma urealyticum and Ureaplasma parvum were randomized to receive josamycin or placebo. Amniotic fluid was also tested for 16S PCR. MAIN OUTCOME MEASURES: The primary outcome was delivery before 37 weeks. RESULTS: In total, 1043 women underwent amniotic-fluid screening with specific PCR detection between July 2008 and July 2011: PCR detection failed in 27 (2.6%), and 20 (1.9%) underwent termination of pregnancy. Among the 1016 women with PCR results, 980 had available data for the primary outcome (delivery before 37 weeks) and 29 (3.0%) were positive for Ureaplasma and/or Mycoplasma spp. Because of the low rate of women with PCR-positive findings, the trial was stopped prematurely. In total, 19 women were randomized to receive placebo (n = 8) or josamycin (n = 11) and their characteristics were comparable, as was the rate of preterm delivery and secondary outcomes. In comparing all PCR-positive and -negative women regardless of treatment, PCR positivity for Ureaplasma and/or Mycoplasma spp. was not associated with any adverse pregnancy or neonatal outcome. Amniotic-fluid screening by 16S PCR showed no other bacterial colonization associated with preterm birth. CONCLUSIONS: Because of a low amniotic fluid colonization rate, the trial was interrupted. Maternal amniotic-fluid colonization by Mycoplasma and/or Ureaplasma spp. at 16-20 weeks in asymptomatic women is rare and not associated with adverse pregnancy outcomes. TRIAL REGISTRATION: ClinicalTrials.gov NCT00718705.

In Vitro Activities of the Benzoquinolizine Fluoroquinolone Levonadifloxacin (WCK 771) and Other Antimicrobial Agents against Mycoplasmas and Ureaplasmas in Humans, Including Isolates with Defined Resistance Mechanisms.

Levonadifloxacin (WCK 771) was evaluated against 68 type strains and clinical isolates of Mycoplasma genitalium, Mycoplasma hominis, Mycoplasma pneumoniae, and Ureaplasma spp. in comparison with moxifloxacin, levofloxacin, tetracycline, and azithromycin or clindamycin. Levonadifloxacin MICs were ≤0.5 µg/ml for M. genitalium MIC90s were 1 µg/ml for M. hominis, 0.125 µg/ml for M. pneumoniae, and 2 µg/ml for Ureaplasma spp. Levonadifloxacin merits further study for treating infections caused by these organisms.

Tetracycline and fluoroquinolone resistance in clinical Ureaplasma spp. and Mycoplasma hominis isolates in France between 2010 and 2015.

Objectives: As information on Ureaplasma spp. and Mycoplasma hominis resistance is currently limited, the aim of this study was to investigate the susceptibility of Ureaplasma spp. and M. hominis to tetracyclines and fluoroquinolones in France. Methods: The susceptibility of 1014 clinical isolates obtained in Bordeaux University Hospital (Bordeaux, France) between 2010 and 2015 was evaluated using two commercial kits, S.I.R. Mycoplasma (Bio-Rad) from 1 January 2010 to 5 October 2012 and MYCOFAST RevolutioN kit (ELITech Group) from 6 October 2012 to 31 December 2015. The MICs of isolates designated as resistant were determined using the broth microdilution assay. Additionally, the tet(M) gene and fluoroquinolone resistance-associated mutations were identified. Results: Among 831 Ureaplasma spp. isolates, the tetracycline, levofloxacin and moxifloxacin resistance rates were 7.5%, 1.2% and 0.1%, respectively. Among 183 M. hominis isolates, the resistance rates were 14.8%, 2.7% and 1.6% for tetracycline, levofloxacin and moxifloxacin, respectively. Over the 6 year period, no significant change in resistance to tetracycline or fluoroquinolones was observed. The tet(M) gene was found in all tetracycline-resistant isolates. All levofloxacin-resistant isolates harboured a mutation in the parC or parE genes. Isolates that were also resistant to moxifloxacin harboured an additional mutation in the gyrA gene. The MYCOFAST RevolutioN kit significantly overestimated levofloxacin and moxifloxacin resistance in Ureaplasma spp. isolates. Conclusions: Resistance to tetracycline and fluoroquinolones is limited in France in mycoplasmas but compared with a previous report in 1999-2002, a significant increase in tetracycline resistance among Ureaplasma spp. was observed. Ongoing monitoring of the antibiotic susceptibility of these urogenital mycoplasmas remains necessary.

Prevalence and Antimicrobial Susceptibility of Ureaplasma spp. and Mycoplasma hominis in Asymptomatic Individuals in Korea.

This study investigated the prevalence and antibiotic resistance of Ureaplasma spp. and Mycoplasma hominis isolated from asymptomatic individuals in Korea. Endocervical swabs from women and urine from men, from a total of 5,781 asymptomatic individuals, were analyzed using a Mycoplasma IST2 Kit. Of the 4,825 specimens tested from females, 486 (10.1%) were positive culture. In these positive specimens, 437 (9.1%) were positive only for Ureaplasma spp., 17 (0.4%) were positive only for M. hominis, and 32 (0.7%) were positive for both Ureaplasma spp. and M. hominis. In males, of the 956 tested specimens, only 4 (0.42%) were positive for Ureaplasma spp. and no M. hominis colonization was identified. In antimicrobial susceptibility tests, more than 93.2% of both M. hominis and Ureaplasma spp. was susceptible to tetracycline, doxycycline, josamycin, and pristinamycin. However, M. hominis isolates were found to be highly resistant to erythromycin, azithromycin, and clarithromycin (82.4%, 70.6%, and 76.5%, respectively). Ofloxacin and ciprofloxacin, which have recently exhibited increasing resistance rates, showed rates of 17.7% and 35.3%, respectively, in M. hominis, and 50.6% and 27.4%, respectively, in Ureaplasma spp. In conclusion, accurate antimicrobial susceptibility tests of the genital mycoplasmas should be conducted for each case to select the appropriate antibiotics. Fluoroquinolone-based drugs should be avoided in the initial treatment of urogenital mycoplasmas because of the increasing rate of resistance to quinolones, although the susceptibility to tetracycline remains high in Korea.

Prevalence and antimicrobial resistance of Ureaplasma spp. and Mycoplasma hominis isolated from semen samples of infertile men in Shanghai, China from 2011 to 2016.

This study was conducted to estimate the prevalence and antimicrobial resistance rate of Ureaplasma spp. and Mycoplasma hominis that were isolated from the semen samples of infertile males in Shanghai, China from 2011 to 2016. A total of 5016 infertile males and 412 healthy male controls were examined. The cultivation, identification, and antimicrobial susceptibilities of Ureaplasma spp. and M. hominis were assessed by using a Mycoplasma IST kit that was performed in parallel to selective solid agar cultivation. The positive rate of genital Mycoplasma infections in infertile men from 2011 to 2016 was 30-55%, which initially decreased during the first four years and then increased in the last two. Two distinct high-risk age ranges of Mycoplasma infections were observed: 26-30 years (37.8%) and 31-35 years (30.7%). Semisynthetic tetracyclines and macrolide antibiotics were the most effective agents against Ureaplasma spp. Among the fluoroquinolones, sparfloxacin and levofloxacin were also effective. Antibiotic resistance of Ureaplasma spp. against tetracyclines and macrolide antibiotics in the last six years did not vary significantly. However, the rate of resistance to fluoroquinolones (except norfloxacin) and spectinomycin decreased in the last two years. The rate of genital Mycoplasma presence in infertile patients between the ages of 26 and 35 years in Shanghai was high. The prevalence of genital Mycoplasma decreased during the first four years and then increased, with a peak in 2016. Doxycycline, minocycline, josamycin, and sparfloxacin can be recommended for first-line empirical treatment of Mycoplasma infections in infertile men in Shanghai, China.

Biochanin A partially restores the activity of ofloxacin and ciprofloxacin against topoisomerase IV mutation-associated fluoroquinolone-resistant Ureaplasma species.

PURPOSE: This study aims to investigate the synergistic antimicrobial activity of four phytoalexins in combination with fluoroquinolones against Ureaplasma spp., a genus of cell wall-free bacteria that are intrinsically resistant to many available antibiotics, making treatment inherently difficult. METHODOLOGY: A total of 22 958 urogenital tract specimens were assessed for Ureaplasma spp. identification and antimicrobial susceptibility. From these, 31 epidemiologically unrelated strains were randomly selected for antimicrobial susceptibility testing to determine the minimum inhibitory concentration (MIC) of four fluoroquinolones and the corresponding quinolone resistance-determining regions (QRDRs). Synergistic effects between fluoroquinolones and four phytoalexins (reserpine, piperine, carvacrol and biochanin A) were evaluated by fractional inhibitory concentration indices (FICIs). RESULTS: Analysis of the QRDRs suggested a vital role for the mutation of Ser-83→Leu in ParC in fluoroquinolone-resistant strains, and the occurrence of mutations in QRDRs showed significant associations with the breakpoint of levofloxacin. Moreover, diverse synergistic effects of the four phytoalexins with ofloxacin or ciprofloxacin were observed and biochanin A was able to enhance the antimicrobial activity of fluoroquinolones significantly. CONCLUSION: This is the first report of the antimicrobial activity of biochanin A in combination with fluoroquinolones against a pathogenic mycoplasma, and opens up the possibility of using components of biochanin A as a promising therapeutic option for treating antibiotic-resistant Ureaplasma spp. infections.

In Vitro Activities of Gepotidacin (GSK2140944) and Other Antimicrobial Agents against Human Mycoplasmas and Ureaplasmas.

Gepotidacin, a novel first-in-class triazaacenaphthylene topoisomerase II inhibitor, was tested against 85 type strains and clinical isolates of Mycoplasma pneumoniae, Mycoplasma hominis, Mycoplasma genitalium, Ureaplasma parvum, and Ureaplasma urealyticum in comparison to levofloxacin, moxifloxacin, azithromycin or clindamycin, and tetracycline. Gepotidacin MIC90s (µg/ml) were 0.125 (M. pneumoniae), 0.032 (M. genitalium), 2 (M. hominis), and 8 (Ureaplasma species). Gepotidacin activity was not affected by resistance to fluoroquinolones, tetracyclines, or macrolides in the strains tested. Gepotidacin merits further study for treating infections caused by these organisms.