Detection of Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum (biovar 1) and Ureaplasma urealyticum (biovar 2) in patients with non-gonococcal urethritis using polymerase chain reaction-microtiter plate hybridization

AIM: To use polymerase chain reaction (PCR)-microtiter plate hybridization assays to detect Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum (biovar 1) and Ureaplasma urealyticum (biovar 2) in first-voided urine specimens from patients with non-gonococcal urethritis (NGU). METHODS: A total of 153 male patients with NGU, who visited one of 24 clinics in Japan, were recruited for this study. All were examined using PCR-microtiter plate hybridization assays for the presence of M. genitalium, M. hominis, U. parvum (biovar 1) and U. urealyticum (biovar 2) in first-voided urine specimens. They were also examined for the presence of Chlamydia trachomatis. RESULTS: Of these 153 patients, 73 (47.7%) were positive for C. trachomatis. Overall, the prevalence was 17.0% for M. genitalium, 16.3% for U. urealyticum (biovar 2), 7.8% for U. parvum (biovar 1) and 2.6% for M. hominis. In the 80 patients with non-chlamydial NGU, the prevalence of M. genitalium, U. urealyticum (biovar 2), U. parvum (biovar 1) and M. hominis was 23.8%, 18.8%, 8.8% and 2.6%, respectively. CONCLUSIONS: This study shows the prevalence of mycoplasmas and ureaplasmas in NGU in Japan. M. genitalium and U. urealyticum (biovar 2) might be pathogens of NGU and could be associated with persistent and recurrent urethritis. When patients with NGU are treated, such pathogens should be taken into account. This PCR-microtiter plate hybridization assay provides a useful method for diagnosing NGU caused by M. genitalium and U. urealyticum (biovar 2).     

Treatment of men with urethritis negative for Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum and Ureaplasma urealyticum

OBJECTIVE: Some patients with symptomatic non-gonococcal urethritis (NGU) are negative for Chlamydia trachomatis, mycoplasmas and ureaplasmas. The optimal antimicrobial chemotherapy for such NGU has not fully been elucidated, though many studies of antimicrobial chemotherapies for C. trachomatis-positive NGU have been performed. We assessed the efficacy of antimicrobial agents that are active against C. trachomatis on non-mycoplasmal, non-ureaplasmal and non-chlamydial NGU (NMNUNCNGU). METHODS: One hundred men whose first-pass urine samples were negative for C. trachomatis, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum, and Ureaplasma urealyticum were treated with levofloxacin, gatifloxacin, minocycline, or clarithromycin for 7 days. Urethritis symptoms and the presence of polymorphonuclear leukocytes (PMNL) in urethral smears were assessed before and after treatment. RESULTS: Eighty-eight (88.0%) of 100 men with NMNUNCNGU showed no signs of urethral inflammation after treatment, but two men complained of some symptoms of urethritis. Twelve (12.0%) of 100 men had significant numbers of PMNL in urethral smears, but five of these 12 men had no symptoms of urethritis. The efficacy for normalization of urethral smears was 90.7% for clarithromycin, 89.7% for levofloxacin, 87.5% for gatifloxacin, and 75.0% for minocycline. The 12 men who showed signs of urethral inflammation were retreated with levofloxacin, gatifloxacin, minocycline or clarithromycin for an additional 7 days. The 10 men who returned after the second treatment had negative urethral smears. CONCLUSION: Our present findings suggest that antimicrobial agents active against C. trachomatis are effective against NMNUNCNGU and that a 7-day treatment regimen with an appropriate antimicrobial agent may be sufficient to manage patients with NMNUNCNGU.     

Mycoplasmas in semen of chronic prostatitis patients

OBJECTIVE: To evaluate the occurrence of mycoplasmas in the semen of chronic prostatitis patients. MATERIAL AND METHODS: Genital mycoplasmas (Mycoplasma hominis, Mycoplasma genitalium, Ureaplasma urealyticum, Ureaplasma parvum) were sought in the semen of 121 chronic prostatitis patients [38 National Institutes of Health (NIH) category IIIa, 59 NIH category IIIb and 24 NIH category IV] and 40 controls. The commercially available kit Mycoplasma IST was applied to the semen samples of all 161 men, and polymerase chain reaction (PCR) to those of 60 randomly selected men. RESULTS: Ureaplasmas were found in all study groups (at frequencies ranging from 12% to 25%) using the Mycoplasma IST test, but M. hominis was found only in one NIH category IIIb patient. Using PCR, most of the ureaplasmas appeared to be U. parvum, which was found in all prostatitis groups (18% of NIH category IIIa, 15% of NIH category IIIb and 25% of NIH category IV patients) but not in the controls. M. genitalium was found in 18% of the NIH category IIIa patients. All of the mycoplasmas occurred significantly more frequently in prostatitis patients than in controls and in NIH category IIIa patients than in controls. CONCLUSION: Mycoplasmas occur more frequently in the semen of prostatitis patients than in that of healthy controls, with U. parvum being the most frequently occurring species.     

1186例慢性前列腺炎且不育患者前列腺按摩液中病原体检测及耐药性分析

目的:探讨慢性前列腺炎伴不育患者EPS中分离的细菌菌谱分布及衣原体和支原体感染情况,为临床提供用药依据。方法:对临床采集的慢性前列腺炎且不育患者EPS标本进行培养、鉴定、药物敏感试验,用荧光PCR法检测沙眼衣原体,并对所得结果进行统计学分析。结果:1 186例患者的EPS标本中,病原体总阳性率51.7%,其中革兰阳性球菌364例;革兰阴性杆菌20例;其他菌株5例;支原体157例,其中溶脲脲原体116例、人型支原体41例;沙眼衣原体DNA阳性67例。分离到的葡萄球菌对万古霉素全部敏感;对青霉素类耐药率最高,为76.9%~100%。无乳链球菌对红霉素及克林霉素的耐药率最高,为100%;对β-内酰胺类、氨基糖苷类、复方新诺明、利福平、万古霉素全部敏感。溶脲脲原体对环丙沙星耐药率最高,为59.5%;对交沙霉素、四环素、磷霉素耐药率最低,为1.7%。人型支原体对红霉素的耐药率最高,为100%;对强力霉素、磷霉素全部敏感。结论:慢性前列腺炎伴不育患者EPS中有很大部分可分离到细菌及支原体和衣原体,表明感染是引起男性不育的一个重要因素,分离菌株对不同抗菌药物耐药性有较大差异。     

The effect of bacterial contamination of semen on sperm chromatin integrity and standard semen parameters in men from infertile couples

A considerable proportion of male factor infertility cases are associated with inflammatory processes. The most common sexually transmissible agents causing sexually transmitted diseases in industrial countries are Chlamydia trachomatis, genital Ureaplasma and Mycoplasma spp. This study was undertaken to investigate whether these bacterial contaminants in semen affect sperm quality parameters and particularly DNA integrity (detected by sperm chromatin structure assay) in males from infertile couples (n = 293). The results showed that semen contaminations with the investigated bacterial species were not associated with sperm DNA fragmentation. However, contaminations with Mycoplasma spp. and C. trachomatis were associated with decreased sperm concentrations. Total sperm numbers in contaminated semen samples tended to be decreased, but not significantly. Mycoplasma had the highest adverse effect on sperm quality (concentration, motility, morphology and DNA condensation). Antibiotic therapy of the selected 47 men was successful in 55%, but semen quality parameters did not improve at least up to 3 months after the therapy. The presence of pathogenic bacteria in semen is primarily associated with low sperm production. Our data showed that Mycoplasma spp. contamination of semen had the highest adverse effect on sperm quality. Sperm chromatin integrity assessed by the presence of DNA breaks was not disturbed.     

Ureaplasma urealyticum and Mycoplasma hominis in men attending for routine semen analysis. Prevalence, incidence by age and clinical settings, influence on sperm characteristics, relationship with the leukocyte count and clinical value

AIM: To investigate the clinical value of the screening of Ureaplasma urealyticum and Mycoplasma hominis in routine semen analysis. MATERIAL AND METHODS: Semen samples of 234 patients with several clinical settings (infertility, varicocele, spontaneous abortion, genital infections, undescended testicles, hemospermia, etc.) were distributed in three study groups: group 1--negative cultures; group 2--normal colonization (< or =10(3) colony-forming units (cfu)/ml), and group --pathogenic colonization (>10(3) cfu/ml). Frequency rates, incidence by age and clinical settings, association with abnormal sperm characteristics (density, vitality, motility and morphology) and with the leukocyte count were investigated. RESULTS: Prevalence of U. urealyticum was higher than M. hominis (< or = 10(3) cfu/ml: 28.2 vs. 24.8%; >10(3) cfu/ml: 20.5 vs. 13.3%). No difference was detected on the incidence of mycoplasmas by age and clinical settings, as well as in regard to the mean values of sperm density, vitality, motility, oval-headed sperm and leukocyte count (p > 0.05). CONCLUSION: In spite of the high incidence of mycoplasmas, not enough information was available regarding the influence of these microorganisms on the sperm quality and their relationship with the leukocyte count. Therefore, screening of U. urealyticum and M. hominis for routine semen analysis is not clinically relevant.     

Epididymal stereocilia in semen of infertile men: evidence of chronic epididymitis?

As free stereocilia in semen samples from infertile men could be related to a subclinical chronic infection of the seminal tract, we carried out a retrospective analysis of 14 files of infertile men with stereocilia in semen (group 1) and in two control groups: 30 infertile men from the same study population (group 2) and 40 fertile sperm donors (group 3). Clinical, bacteriological, sonographic and microscopic findings associated with these semen samples were compared. Epididymal cysts were found in two of 14 men in group 1 and in four of 28 in group 2. Positive semen cultures for Chlamydia trachomatis, Mycoplasma hominis or Ureaplasma urealyticum (chi(2) = 8.20; P = 0.002; OR: 7.22) and varicocele (chi(2) = 8.96; P = 0.002; OR: 2.25) were associated to the presence of free stereocilia in semen. These data suggest that a silent chronic infection of the epididymis by C. trachomatis, M. hominis, U. urealyticum and/or inflammation produced by varicocele might be the cause of stereocilia loss in the semen.     

Azithromycin treatment for nongonococcal urethritis negative for Chlamydia trachomatis,Mycoplasma genitalium,Mycoplasma hominis,Ureaplasma parvum,and Ureaplasma urealyticum

Some patients with nongonococcal urethritis (NGU) are negative for Chlamydia trachomatis, mycoplasmas, and ureaplasmas. The optimal antimicrobial chemotherapy for such NGU has not fully been clarified. We assessed the efficacy of azithromycin for treatment of nonmycoplasmal, nonureaplasmal, nonchlamydial NGU (NMNUNCNGU). Thirty‐eight men whose first‐pass urine was negative for Chlamydia trachomatis, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum, and Ureaplasma urealyticum were treated with a single dose of 1 g azithromycin. Urethritis symptoms and polymorphonuclear leukocytes in urethral smears or in first‐pass urine were assessed before and after treatment with azithromycin. Thirty‐two (84.2%) of the 38 men with NMNUNCNGU showed no signs of urethral inflammation after treatment. The efficacy of this azithromycin regimen was comparable to that of the 7‐day regimen of levofloxacin, gatifloxacin, minocycline, or clarithromycin reported previously. A single dose of 1 g azithromycin, which is effective not only for NGU due to specific pathogens but also for NMNUNCNGU, is an appropriate treatment for NGU.     

Genital Mycoplasma infections--clinical aspects, diagnosis and therapy

Ureaplasma urealyticum und Mycoplasma hominis are frequently isolated from the urogenital tract of sexually active persons. Their pathogenetic role in urogenital infections and infertility are still controversial. U. urealyticum can cause urethritis and most likely also chronic prostatitis. M. hominis can cause cystitis, salpingitis, and postpartal fever in women and meningitis in newborns. M. genitalium was recently isolated from patients with urethritis and salpingitis. The diagnosis is made by the cultivation of mycoplasmas in high concentrations and rising antibody titers in the serum. As in chlamydial infections, the therapy of first choice is tetracyclin. M. hominis can also be treated with clindamycin, U. urealyticum with erythromycin.     

Screening for bacterial pathogens in semen samples from infertile men with and without leukocytospermia

We aimed to study the correlation between leukocyte counts in semen and bacterial pathogens in seminal samples of infertile men, and to establish the minimum leukocyte count associated with significant bacteriospermia. A total of 116 patients who underwent evaluation of fertility were investigated using routine semen analysis according to the guidelines of the WHO and bacterial pathogens analysis by culture and in-house PCR assay. The overall prevalence of bacteriospermia in semen samples was 56.9% independent of the presence of leukocytes. The most common bacterial species detected were Chlamydia trachomatis (41.4%), Ureaplasma urealyticum (15.5%) and Mycoplasma hominis (10.3%). The receiver operating characteristic curve analysis demonstrated that the sensitivity/specificity for detecting bacteria at a cut off level of ≥1 × 10⁶ leukocytes per ml (which is the WHO defined level for leukocytospermia) was 20.3%/81.5%. The highest sensitivity/specificity ratio was found in semen samples with a cut-off level of ≥0.275 × 10⁶ leukocytes per ml, which is best shown with the odds ratio of 2.47. A significant correlation was found between bacteriospermia and leukocytospermia at the cut-off level of ≥0.275 × 10⁶ leukocytes per ml of semen samples ( P  =   0.032). We proposed that this is a possible new cut-off level to predict the presence of bacteria in semen of infertile men.     

泌尿生殖道炎症病人支原体感染现况及耐药性分析

目的 :了解泌尿生殖道炎症病人支原体感染现况及耐药性情况。　方法 :统计 1999～ 2 0 0 3年门诊诊断为非淋菌性尿道炎 (NGU)、慢性前列腺炎及盆腔炎等泌尿生殖道疾病病人 30 5 5例 ,分析其支原体培养及药物敏感试验结果。　结果 :30 5 5例病人共检出支原体感染 992例 ( 32 .5 % ) ,5年间差异无显著性 (P <0 .0 5 )。其中单纯溶脲脲原体 (Uu)感染 70 1例 ( 70 .7% ) ,单纯人型支原体 (Mh)感染 4 4例 ( 4 .4 % ) ,Uu和Mh混合感染 2 4 7例 ( 2 4 .9% ) ,单纯Uu感染远高于混合感染和单纯Mh感染 (P <0 .0 1)。单纯Uu感染计数≥ 10 4cfu/ml占 76 .7% ,而单纯Mh感染者≥10 4cfu仅占 18.2 %。对强力霉素、原始霉素、交沙霉素和四环素敏感性较高 ,分别为 94 .3%、96 .6 %、86 .5 %和97.4 % ,对红霉素和氧氟沙星敏感性仅 5 4 .8%和 2 9.4 %。　结论 :在对病人进行支原体检测时 ,应同时进行Uu和Mh的检测 ,并做支原体药物敏感试验 ,以合理选择抗生素。     

Physiological integrity of human sperm in the presence of Ureaplasma urealyticum

Several species of Mycoplasma have been isolated from the human genital tract, the most common being M. hominis and Ureaplasma urealyticum. A causal relationship between such infections and sperm dysfunction and infertility has yet to be established. It was the purpose of this study to examine the effects of U. urealyticum infection on the function of sperm as assessed by seminal fluid analysis (SFA), in vitro penetration of bovine cervical mucus (BCMP), and the hamster sperm penetration assay (SPA). No significant differences were noted in the SFA of infected and uninfected samples, either fresh or frozen, fertile or infertile. In addition, no differences were noted in the BCMP or SPA. In sperm from U. urealyticum-infected individuals the basic physiological mechanisms underlying mucus penetration and ovum fertilization seem intact.     

The importance and frequency of mixed infections with Chlamydia trachomatis and mycoplasmas in acute gonococcal urethritis

Chlamydia trachomatis, Ureaplasma urealyticum and Mycoplasma hominis are frequently involved in gonococcal urethritis. We investigated 143 male White and Black patients with gonococcal urethritis (average age 22,5 years). Of these 29% had coexisting Chlam. trachomatis infection, 34% U. urealyticum infection and 13% Mycoplasma hominis infection. Conventional penicillin therapy did not affect Chlam. trachomatis, U. urealyticum or Mycoplasma hominis, which persisted in the lower urogenital tract, causing a so-called 'post-gonococcal urethritis.' Additional therapy with tetracycline or erythromycin was successful in most cases.     

IgA antibodies to Chlamydia trachomatis and seminal parameters in asymptomatic infertile males

Prevalence of IgA antibodies to Chlamydia trachomatis in semen samples from infertile men was estimated and its clinical meaning is discussed. The ejaculate of 102 infertile men without any symptom of genital infection was studied, and seminal alterations were classified according to WHO criteria. Antichlamydial IgA antibodies were detected using a solid-phase, enzyme-linked immunoassay (ImmunoComb II Chlamydia trachomatis monovalent IgA) and related to sperm count, motility and membrane integrity, seminal leucocyte count, and past history of sexually transmitted disease (STD). Prevalence of IgA antibodies to C. trachomatis was 23%. There was no relationship between IgA antibodies and the sperm variables or leucocyte count in semen. However, a strong association between antichlamydial IgA antibodies and the antecedent of STD was found (p < .005; OR = 6). IgA antibodies to C. trachomatis did not cause alterations in sperm function and they were not associated with inflammatory response. However, these antibodies in semen of asymptomatic infertile men would indicate a risk of C. trachomatis infection for the couples of those patients.     

Association of ureaplasma urealyticum with abnormal reactive oxygen species levels and absence of leukocytospermia

PURPOSE: Ureaplasma urealyticum is a commensal of the lower genitourinary tract of many sexually active adults. The organism is more common in partners of infertile than fertile marriages. We conducted a prospective study at our tertiary care center to confirm a possible association between U. urealyticum and abnormal sperm function parameters. MATERIALS AND METHODS: A total of 50 consecutive male patients seeking general urology consultation for lower urinary tract symptoms characteristic of chronic prostatitis were evaluated. Urine and semen localization cultures were performed with additional semen cultures for U. urealyticum, Chlamydia trachomatis and Mycoplasma hominis. Specimens from 21 healthy men were used as controls. Specimens were analyzed by a computer assisted semen analyzer, and verified manually for concentration, percent motility and morphology. Leukocytospermia was measured by the Endtz test. Semen specimens were also analyzed for reactive oxygen species (ROS), acrosome reaction and mannose binding assay. RESULTS: Of the patients 17 had positive U. urealyticum cultures and the other cultures were negative. Patients with U. urealyticum had significantly higher ROS levels (log [ROS + 1] = 2.52 +/- 0.25) than those without U. urealyticum (1.49 +/- 0.20, p = 0.002) or controls (1.31 +/- 0.19, p = 0.002). Leukocytospermia was detected in only 1 of the 17 (6%) positive specimens and 4 (12%) negative specimens. CONCLUSIONS: Seminal ROS levels are elevated among patients with U. urealyticum. ROS induces lipid peroxidation, which reduces membrane fluidity and sperm fertilization capability, and may be the mechanism by which U. urealyticum impairs sperm function. Absence of leukocytospermia does not exclude U. urealyticum.     

Ureaplasma urealyticum and mycoplasma hominis in male urethritis]

We conducted a multi-center clinical study to evaluate the role of Ureaplasma urealyticum and Mycoplasma hominis in male urethritis. The incidence of each organism in first-voided urine samples of 160 male urethritis patients, including 28 with gonococcal and 126 with non-gonococcal urethritis, was investigated. U. urealyticum and M. hominis were isolated from 13.6% and 6.5%, respectively, of the urine samples, and the concentration of each mycoplasma exceeded 10(3) ccu/ml (color changing units/ml) in 5.2% and 3.9%, respectively. Among 64 patients with non-gonococcal non-chlamydial urethritis, U. urealyticum and M. hominis concentration exceeding more than 10(3) ccu/ml were detected in only two and one samples, respectively. The incidence among urethritis patients with a concentration of either mycoplasma exceeding 10(3) ccu/ml was not significantly greater than that among subfertile males without urethritis. The findings of the present study suggest that, although mycoplasmas may cause urethritis in some patients, the incidence of urethritis due to U. urealyticum or M. hominis is low among patients with non-gonococcal, non-chlamydial urethritis.     

解脲支原体感染与男性不育

用双抗体夹心ＥＬＩＳＡ法检测解脲支原体，并与经典的培养法作了比较。３９例生育男性精浆解脲支原体ＥＬＩＳＡ法和培养法的阳性率分别为１０．３％和７．７％，１６４例不育者精浆解脲支原体两法阳性率分别为３５．４％和３１．７％，生育者与不育者解脲支原体阳性率差异非常显著（Ｐ＜０．００５）；ＥＬＩＳＡ法与培养法符合率为８６．５％。精子畸形率超过３０％及精子密度偏低（＜０．２亿／ｍｌ）的不育者，解脲支原体阳性率分别高达６９．２％和５６．５％。解脲支原体感染可导致精子形态与功能异常，从而引起男性不育。     

解脲支原体感染与男性不育

用高度纯化的人解脲支原体免疫家兔,获得兔抗人解脲支原体抗血清,以此抗血清(一抗)及羊抗兔 IgG 荧光抗体(二抗),分别处理20例正常生育男性和20例精液解脲支原体培养阳性的不明原因不育男性的精子,并在荧光显微镜下观察。结果显示:在不育男性部分精子的头部和(或)尾部有特异性的较强荧光结合,表明有解脲支原体吸附,并有大量畸形精子,多数为卷尾畸形。此外,用比浊法检测了正常生育男性与上述不育男性精液中快速运动相精子的百分含量(FRM)及平均速度(VRM)。不育男性组比生育组的 FRM 及 VRM 都显著降低(分别为 P<0.01和 P<0.001)。本研究首次用特异性方法证实解脲支原体吸附于不育男性精子表面,提示解脲支原体感染可能通过影响精子的形态、精子的运动和精卵识别过程而导致男性不育。     

Screening for Chlamydia trachomatis and Ureaplasma urealyticum infection in semen from asymptomatic male partners of infertile couples prior to in vitro fertilization

In a prospective study, the prevalence of infection with Chlamydia trachomatis and Ureaplasma urealyticum was evaluated in the semen of 92 asymptomatic male partners of infertile couples using polymerase chain reaction and culture, respectively. The results were compared with the detection of serologically specific antibodies. U. urealyticum and C. trachomatis were detected in 12 (13%) and 10 (10.8%) of the tested ejaculates, respectively. One mixed infection was detected. No correlation was found between detection of the pathogens in ejaculates and the presence of specific antibodies in serum. This study therefore confirms the limited diagnostic value of serological analysis to ascertain infection with C. trachomatis or U. urealyticum. The high frequency of detection of these pathogens among asymptomatic male partners of infertile couples emphasizes their potential role in the impairment of male fertility, and the need for sensitive and specific detection methods to prevent infection of the early embryo when using new reproductive techniques such as zona pellucida hatching or intracytoplasmic microinjection.     

Microbial flora in semen of infertile African men at Garankuwa hospital: Die mikrobielle Flora im Sperma von kinderlosen afrikanischen Männern aus dem Garankuwa-Hospital

This study was undertaken to determine the frequency of bacteria and Mycoplasmas in semen of infertile African men and to compare abnormal semen parameters with the presence of these organisms. Routine semen analysis were performed on 100 patients, and semen and first-voided urine samples were cultured. Mycoplasma hominis was cultured in 28% of seminal fluid and Ureaplasma urealyticum in 42%. These organisms were both present in 22% of cultures. Bacteriological cultures were positive in 6 urine and 21 seminal specimens. The frequency of tail abnormalities in the Mycoplasma positive group was greater as compared to normal values. From these it would appear that culture for Ureaplasma should probably be performed routinely at the Andrology Clinic.