High rates of genital mycoplasma infection in the highlands of Papua New Guinea determined both by culture and by a commercial detection kit.

Duplicate vaginal swabs were collected from 100 women, and comparisons were made between an in-house broth-agar culture system and a commercially available kit, the Mycoplasma IST kit (bioMérieux), for the detection of Mycoplasma hominis and Ureaplasma urealyticum. There was good agreement between the two systems for detection of the genital mycoplasmas in terms of sensitivity, with values of > 92% being obtained. In terms of specificity, the mutual comparisons were less favorable, though specificity values of > 72% were obtained. Statistically there was no significant difference in the performance of the two tests (P < 0.1 for both M. hominis and U. urealyticum). While the broth-agar culture system was considerably less expensive than the kit, the Mycoplasma IST kit provided additional information on antibiotic susceptibilities and had the advantages of a shelf life of up to 12 months and not requiring the preparation of culture media. The prevalences of colonization obtained for M. hominis and U. urealyticum were extremely high in this randomly selected group of women from periurban and rural settlements in the Eastern Highlands of Papua New Guinea, being > or = 70% for M. hominis and > or = 78% for U. urealyticum. colonization with both genital mycoplasmas simultaneously was also very common, with > or = 60% of women being colonized by both M. hominis and U. urealyticum.     

Isolation of genital mycoplasmas and Chlamydia trachomatis in stillborn and neonatal autopsy material

Chlamydia trachomatis and the genital mycoplasmas are significantly prevalent in sexually active women. How these organisms may affect the outcome of pregnancy and the neonate was the principal thrust of this investigation. Placenta, liver, and lung tissue were cultured from Mycoplasma hominis, Ureaplasma urealyticum, Chlamydia trachomatis, and aerobic as well as anaerobic bacteria in 432 stillborn and neonatal autopsies. Genital mycoplasmas were isolated from 36 cases (8.3%). Acute chorioamnionitis and funisitis were present significantly more often in cases with genital mycoplasma than in those without these organisms. Isolation of genital mycoplasmas was not associated with an increased incidence of intrauterine fetal death, villitis, hyaline membrane disease, congenital anomalies, or polymorphonuclear leukocytes in alveolar spaces. Chlamydia trachomatis was not found in any of the sites sampled.     

Mycoplasma hominis and Ureaplasma urealyticum in different gynecologic diseases

The incidence of Mycoplasma hominis (M. hominis) y Ureaplasma urealyticum (U. urealyticum) was investigated in 113 endocervical samples obtained from women who were seen for different gynecological pathologies. Forty-seven (42%) patients were positive to these microorganisms; 26 cases (23%) were positive for M. hominis and 21 (19%; p = NS) for U. urealyticum. Average age was 32.1 +/- 7.7 years; the average number of sexual partners was 1.7 +/- 1.1. Eleven of 17 patients with 3 o more sexual partners were positive for Genital Mycoplasma (GM), and U. urealyticum was found more often in this group. A higher incidence of GM was found in women between 26 and 30 years (34%); 57.5% of the patients with positive cultures for GM had begun sexual activity before 20 years of age. M. Hominis was found in 61% of women with no parity and U. urealyticum in 71% of parous women. The cultures were positive in 10 of 14 patients with pelvic inflammatory diseases (PID). A cervical biopsy was taken from 52 cases and the diagnosis of cervical intraepithelial neoplasia (CIN) was made in 49 (94%) but only 24 of them were positive for GM (50%). Thirty-five patients suffered sterility, and 12 (34%) were positive for GM, however all positive cases consulted because of primary sterility. The conclusions obtained from this study are: 1) Near half of the patients was positive for GM and none of the species was predominant over the other. 2) The more sexual partners the higher was the incidence of GM, especially U. Urealyticum. 3) The lower the age of the first sexual intercourse the higher the probability of contamination with these microorganisms. 4) M. hominis was more common in nulliparous women and U. urealyticum was found more often in parous patients; the number of deliveries did not have influence in these findings. 5) A statistical significance between GM and PID was found (p = 0.03). 6) GM have no influence on spontaneous abortion. 7) No statistical significance was found between GM and the beginning and evolution of CIN. 8) No relation statistically significative was found between GM and sterility.     

Evaluation of the Mycoplasma Plus and the SIR Mycoplasma kits for quantitative detection and antibiotic susceptibility testing of genital mycoplasma

We compared the results obtained with two commercially available systems (Diagnostics Pasteur) for the quantitative identification and the antibiotic susceptibility testing of the genital mycoplasmas. Ureaplasma urealyticum and Mycoplasma hominis with established methodologies, i.e. isolation on agar with enumeration by dilutions in broth medium and MIC determinations. The Mycoplasma Plus system, consisting of six cups, was designed for the identification and quantitation of genital mycoplasmas and the detection of yeasts. Used in parallel in 150 clinical specimens, it detected U. urealyticum in 42 out of 43 and M. hominis in 10 out of 11 specimens positive by the established methodology. The SIR Mycoplasma antibiogram, consisting of 16 cups, provided for the testing of 1 or 2 concentrations (micrograms/ml) of each of 8 antibiotics: doxycycline, minocycline and lymecycline (4-8); erythromycin (1-4); josamycin (2-8); clindamycin (2); pristinamycin (2); and ofloxacin (1-4). Using an inoculum of about 10(4)-10(5) organisms/ml, we found that major part of the results was in accord with those obtained with the MIC determined in broth for U. urealyticum and on agar for M. hominis. Strains intermediate or resistant to the tetracyclines were identified. Both systems seemed suitable for clinical laboratory use.     

Have Ureaplasma urealyticum and Mycoplasma hominis infections any significant effect on women fertility?

Ureaplasma urealyticum and Mycoplasma hominis are known as sexually transmitted agents. U. urealyticum and M. hominis jeopardize male fertility. However, it is unclear whether these infections significantly contribute to female infertility. In this controlled-study we aimed to establish whether M. hominis and U. urealyticum are risk factors for female fertility and prevalence of infection from these agents in patients attending our infertility clinic. Total 96 married women enrolled in this prospective study; the infertile (study) group consisted of 50 women and fertile (control) group comprised 46 women. The patients were searched about the presence of U. urealyticum and M. hominis by a micro-liquid culture method. The samples were collected from endocervical area with a dacron swab. 28 of 50 (56%) and 18 of 46 (39%) women were evaluated as positive for U. urealyticum culture in the study and control groups respectively. M. hominis was cultured from 4 of 50 (8%) women in the study group as no positive result in controls. There were no statistically significant differences between the groups for both agents (p>0.05), but the higher prevalence of U. urealyticum in infertile women gives emphasis to evaluate these agents in patients that have no any other etiological factor for infertility.     

Evaluation of the Mycoplasma Duo kit for the detection of Mycoplasma hominis and Ureaplasma urealyticum from urogenital and placental specimens

This study compares the Mycoplasma Duo kit for the detection of genital mycoplasmas with conventional culture using A7 differential agar for the detection of Mycoplasma hominis and Ureaplasma urealyticum in clinical samples. Detection of the mycoplasmas is based on the specific metabolic properties of each organism to hydrolyse either arginine or urea. The Mycoplasma Duo test showed a significantly higher detection rate than did culture, although many of the culture-negative results may have been due to the presence of bacterial overgrowth.     

Isolation of genital mycoplasmas from the blood of neonates and women with pelvic infection using conventional SPS-free blood culture media

Standard blood culture media used in our laboratory were tested for their ability to support the growth of Mycoplasma hominis and Ureaplasma urealyticum. Small inocula (approximately 10 colony forming units per ml) of both organisms grew in diphasic tryptone soya medium but not in any of several media containing sodium polyanetholesulphonate (SPS) including a modified Schaedler broth (RWH anaerobic medium) and two BACTEC media (6B and 7D). Both organisms were inhibited even by very low concentrations of SPS but grew well in the Royal Women's Hospital (RWH) anaerobic medium when SPS was omitted. During a 22-month period, routine "blind" plating of the aerobic blood cultures on to mycoplasma agar resulted in isolation of M. hominis or U. urealyticum from 12 women with postpartum or postoperative pelvic infection, and from 3 neonates. Genital mycoplasmas represented 35% of significant isolates from adult blood cultures.     

Clinical aspects of pelvic inflammatory disease

Pelvic inflammatory disease (PID) is a common and poorly managed condition. Untreated or inadequately treated, it leads to tubal infertility, ectopic pregnancy and chronic pelvic pain. Diagnostic difficulties are compounded by the wide variety of clinical presentations and the insensitivity and poor specificity of laboratory tests. Better recognition of mild and atypical disease needs a high index of suspicion whenever young, sexually active women present with gynaecological symptoms. Laparoscopy supplemented by microbiological tests and fimbrial minibiopsy should be regarded as the diagnostic 'gold standard' for research studies; new studies are required to identify techniques which might reduce under- and over-diagnosis. Early treatment reduces the risk of an adverse effect on fertility. Any therapeutic regimen selected should be effective against the common aetiological agents Chlamydia trachomatis, Neisseria gonorrhoeae, genital mycoplasmas and aerobic and anaerobic bacteria. Since at least 60% of cases of PID can be attributed to infection with a sexually transmitted organism, partner notification forms an essential part of management.      

Morphology of human Fallopian tubes after infection with Mycoplasma genitalium and Mycoplasma hominis--in vitro organ culture study

BACKGROUND: Female infertility can be caused by scarring and occlusion of the Fallopian tubes. Sexually transmitted bacteria can damage the delicate epithelial layer of human Fallopian tubes (HFT). Genital mycoplasmas are associated with human reproductive failure. Yet, there is not enough evidence that mycoplasmas can cause tubal factor infertility. We analysed the effects of infections with Mycoplasma hominis and Mycoplasma genitalium on the HFT epithelium and compared them with the effects of infections with genital pathogens: Chlamydia trachomatis and Neisseria gonorrhoeae. METHODS: We used an in vitro model in which pieces of normal HFT were infected with different bacteria, and the outcome of the infections was analysed by scanning electron microscopy (SEM) and confocal microscopy. RESULTS: The presence of M. hominis did not cause any morphological changes of the epithelium of HFT. Noticeable changes in the morphology of the ciliated cells were observed in M. genitalium-infected tissue. Five days post-infection, the cilia were abnormally swollen and some of the ciliated cells fell off the epithelium. These effects could be inhibited by pre-incubation of M. genitalium with antibody directed against the C-terminal part of the adhesion protein MgPa before infection of HFT organ culture. CONCLUSION: We have shown that the presence of M. genitalium, but not M. hominis, in the HFT organ culture affected the epithelium and resulted in cilia damage. The effect of infection with M. genitalium on the HFT was, however, very moderate when compared with the extensive damage of the epithelium caused by N. gonorrhoeae or C. trachomatis.     

Isolation of mycoplasmas from female genitalia

The author tested ways of collection, transport and storage of material for mycoplasmatological examination. The use of tampons on a stick during transport in urea substrate medium proved useful. The site of maximum occurrence of mycoplasmas was the posterior vaginal vault. On examination of the vaginal secretion of 804 women Mycoplasma hominis was isolated in 29.6% and Ureaplasma urealyticum in 65.2% of the cases. Concurrent isolation of Mycoplasma hominis and Ureaplasma urealyticum was recorded in 22.1% of the women. The results of the examination do not suggest the participation of mycoplasmas in the development of aminocolpitis. In pregnant women there is a greater probability of colonization of the vagina by U. urealyticum.     

Methods of studying mycoplasma infections

Mycoplasmas are most often responsible for respiratory and genital infections. At present, diagnosis is carried out by serology for infections caused by M. pneumoniae and by culture for infections due to genital mycoplasmas. For M. pneumoniae, new prospects may lead to a rapid diagnosis, detection by molecular hybridization and immunological detection. Also, the research of specific antibodies should benefit from a better knowledge of the major antigens. Culture of the genital mycoplasmas, U. urealyticum and M. hominis is simple, but the interpretation of their presence is difficult because they may be recovered in a commensal condition. The envisaged advances should lead to a better assessment of their pathogenicity. The role of M. genitalium, a species related to M. pneumoniae recently discovered in respiratory specimens, should be better determined by sensitive techniques developed to distinguish it from M. pneumoniae.     

解脲脲原体感染研究进展

解脲脲原体属于支原体科中的脲原体属,与男女性生殖道炎症、HIV感染、不孕不育、不良妊娠结局以及新生儿疾病等有关。该文从以上几个方面就解脲脲原体与人类健康的关系进行阐述。     

Genital mycoplasma infection. Intrauterine infection: pathologic study of the fetus and placenta.

Mycoplasma infection was present in the fetuses from three spontaneous abortions and in one second-trimester newborn. Gross examination revealed in most cases a severely infected placenta and membranes, with a fetus of normal appearance. The fetal infection presumably followed placental involvement and appeared to have been acquired shortly prior to delivery. Genital mycoplasmas, Ureaplasma urealyticum or Mycoplasma hominis, were isolated from the placentas and the fetal tissues, and from the genital tracts of the mothers. Isolation of mycoplasmas from the liver indicated that bloodstream dissemination of these organisms occurred in the fetus. In the fetus, the pathologic changes were variable. Lesions were identified in the lung by scanning electron microscopy of the bronchial tree in two cases and were accompanied by interstitial pneumonia. An abnormally dilated left ventricle suggestive of cardiomyopathy was observed in one case.     

Effect of experimental genital mycoplasmosis on production of matrix metalloproteinases in membranes and amniotic fluid of Sprague-Dawley rats

PROBLEM: Preterm, premature rupture of membranes (PPROM) is a dire pregnancy outcome that is frequently associated with infection by the genital mycoplasmas, Mycoplasma hominis, Ureaplasma parvum, and U. urealyticum. One potential mechanism by which these microorganisms may cause PPROM is by increasing the concentration of matrix metalloproteinases (MMPs) in the membranes and amniotic fluid. We tested this hypothesis in a well-defined model system of genital infection with M. pulmonis, a natural reproductive pathogen of rats. METHOD OF STUDY: Timed-pregnant, specific pathogen-free, Sprague-Dawley rats were infected with 10(7) CFU M. pulmonis at gestation day (gd) 14. Controls received an equivalent volume (100 microL) of sterile medium. At gd 18, rats were euthanized, and membranes and amniotic fluids were harvested and stored at -70 degrees C until analysis. Proteinase activity of amniotic fluid and membranes was resolved on discontinuous 7.5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis gelatin zymography gels. Band intensity was determined using a digital gel documentation system and the manufacturer's software (Kodak). RESULTS: Gelatinolytic activity associated with a band similar in molecular weight to ProMMP-9 (92 kDa, the inactive precursor of MMP-9) was significantly increased in amniotic fluids and membranes harvested from M. pulmonis-treated pups at gd 18 when compared with tissues harvested from control pups. Both ProMMP-9 and ProMMP-2 (72 kDa, the inactive precursor of MMP-2) were increased in infected animals at gd 21. CONCLUSION: Our study suggests that the genital mycoplasmas can increase MMP-9 production in vivo.     

Experimental Infection of the Genital Tract of Female Grivet Monkeys by Mycoplasma hominis: Effects of Different Routes of Infection

In a previous study we produced acute salpingitis and parametritis in grivet monkeys by inoculation of Mycoplasma hominis directly into the uterine tubes. With the purpose of examining in the same animal model the effect of more natural routes of infection, six female grivet monkeys were inoculated, two by two, with M. hominis by the following methods: (experiment A) into the uterine cavity through the cervical canal; (experiment B) into the uterine cavity by the same route, but after ligature of the isthmus of the uterine tubes had been performed and followed by curettage of the endometrium; and (experiment C) into the cervical epithelium. Whereas the animals in experiment A developed no or only very slight signs of infection, all monkeys in experiments B and C developed pronounced gross and microscopic inflammatory lesions of the uterine tubes and parametria, together with a significant antibody response. It is concluded that ascending M. hominis infection of the genital tract must be preceded by mechanical injury of the epithelial barrier, and that subsequent spread of the infection occurs via blood and lymph vessels rather than by the canalicular route. The inflammatory genital tract disease produced experimentally with M. hominis in grivet monkeys is very similar in its pathogenesis and pathology to the type of salpingitis caused in humans by microorganisms other than Neisseria gonorrhoeae and Chlamydia trachomatis.     

Chorioamnionitis and colonization of the newborn infant with genital mycoplasmas.

To study the role of Mycoplasma hominis and T-mycoplasmas (Ureaplasma urealyticum) in chorioamnionitis, we obtained culture from 249 puerperal women and their babies. The placentas were examined histologically. Infants whose placentas showed inflammation (chorioamnionitis) had cultures positive for T-mycoplasmas more frequently (37.5 per cent) than those with normal placentas (19.0 per cent) (P = 0.021). Colonization with M. hominis was found in 16.0 per cent of the babies and was not significantly associated with chorioamnionitis. Material colonization with mycoplasmas was more frequent (73.4 per cent) and was not correlated with placental inflammation. We conclude that a substantial proportion of cases of chorioamnionitis may be caused by prenatal infection with T-mycoplasmas. The fact that these organisms are not highly virulent could explain the frequent finding of inflammed placentas from otherwise normal pregnacies. No adverse clinical effects of the placental lesions or of mycoplasmal colonization could be detected in this small study.     

Drift of tetM determinant in urogenital microbiocenosis containing mycoplasmas during treatment with a tetracycline antibiotic

We studied the correlation between genetic transfer of tetM determinant in Tn916 conjugative transposon by urogenital mycoplasmas (Mycoplasma hominis and Ureaplasma urealyticum) and changes in the bacterial repertoire during treatment with a tetracycline antibiotic. Basic conditions favoring the nonspecific transfer of tetM determinant into mollicute cells are determined and the allele polymorphism of tetM determinant in clinical strains of M. hominis and U. urealyticum is evaluated. The structure of tetM gene in clinical mycoplasma and ureaplasma strains is characterized by a peculiar mosaic pattern and differs from all previously described alleles of this gene. The results suggest that tetracycline resistance in mollicutes is determined by mechanisms alternative to genetic transfer of tetM determinant.     

PCR法检测不孕症妇女二种病原体(CT,UU)结果分析

目的 :探讨不孕症患者沙眼衣原体 (CT)和解脲支原体 (UU)感染的相关性。方法 :采用聚合酶链反应 (PCR)技术对 1 31 6例不孕症患者及 1 84例正常对照者进行宫颈沙眼衣原体 (CT)和解脲支原体 (UU)检测。结果 :不孕组宫颈分泌物的CTDNA和UUDNA阳性检出率显著高于对照组 (p <0 0 1 )。 结论 :对不孕症患者 ,尤其对原因不明的不孕症、继发性不孕症患者 ,应将生殖道CT及UU检测列为常规     

Systematic screening tests for Chlamydia trachomatis, Mycoplasma hominis and Ureaplasma urealyticum in urogenital specimens of infertile couples

We conducted a prospective study on 100 couples consulting for infertility at the teaching Hospital of Tours, with the scope to determine if there is a benefit for systematic screening of Chlamydia trachomatis, Mycoplasma hominis and Ureaplasma urealyticum among genito-urinary specimen when exploring couples infertility. C. trachomatis was detected by PCR on sperm, endocervix and urine specimen. M. hominis and U. urealyticum were detected by culture on A7 agar medium and with minigaleries on sperm and endocervix specimen. Standard cultures were also performed on sperm, endocervix, vaginal and urine specimen. Only one specimen (sperm) was positive for C. trachomatis. Three percent of the specimen were positive for U. urealyticum (from which 2,5% of the sperm specimen). No specimen was positive for M. hominis. Our results show that screening of C. trachomatis, M. hominis and U. urealyticum is not systematically required for among check up of infertile couples, given the prevalence of chlamydiosis among the population studied. However, it would be interesting to perform it on a targeted population, according to anamnestic or clinical criteria. In addition, an important modification of vaginal flora was observed in 12% of cases, and 2 vaginosis were diagnosed; the putative consequences of this disequilibrium has to be further investigated.     

Cytokine concentrations in seminal plasma from subfertile men are not indicative of the presence of Ureaplasma urealyticum or Mycoplasma hominis in the lower genital tract

The inflammatory response to the presence of Ureaplasma urealyticum or Mycoplasma hominis in the lower genital tract of subfertile men without any signs or symptoms of infection was investigated by measuring the concentrations of interleukin (IL)-6, IL-8, tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) in seminal plasma. Semen samples were collected from 30 culture-positive subfertile males and 23 culture-negative subfertile males. Enzyme-linked immunosorbent assays showed that IL-8 was present in relatively high concentrations (0.12-4.8 ng/ml) in all semen samples investigated. In contrast, the other cytokines were only detectable in 72% (IFN-gamma), 44% (IL-6) and 19% (TNF-gamma) of the samples and were present in relatively low concentrations (1-410 pg/ml). Seminal plasma cytokine concentrations were similar in samples from culture-positive and culture-negative males. These data strongly indicate that the presence of U. urealyticum or M. hominis in the lower genital tract of subfertile males reflects a silent colonisation rather than infection.