Lack of association between genital mycoplasmas and infertility

We studied the relation between colonization with Mycoplasma hominis and Ureaplasma urealyticum, and the results of infertility studies in 205 women with involuntary infertility of at least one year's duration. Isolation of M. hominis (but not of U. urealyticum) was significantly (P = 0.002) more common in patients with a history of pelvic inflammatory disease. However, no relation could be shown between these genital mycoplasmas and any of the following: evidence of prior pelvic inflammatory disease as determined by hysterosalpingography and laparoscopy; cervical inflammation; numbers and motility of spermatozoa on postcoital test; pyosemia; quality of cervical mucus; whether the cause of infertility was related to male or female factors, both, or neither; and occurrence and outcome of subsequent pregnancy. Mycoplasmas were cultured from only 10 of 203 endometrial biopsy specimens (4.9 per cent), and in no instance was inflammation associated with this finding. Out studies do not support a role for genital mycoplasmas in the cause of infertility.     

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.     

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.     

Occurrence of Ureaplasma parvum and Ureaplasma urealyticum in Women with Cervical Dysplasia in Katowice, Poland

The aim of this study was to evaluate the occurrence of genital mycoplasmas, especially Ureaplasma parvum and Ureaplasma urealyticum, in women with atypical squamous cells of undetermined significance (ASCUS), low grade squamous intraepithelial lesions (LSIL) and high grade squamous intraepithelial lesions (HSIL), compared to women with normal cytology living in Katowice, Poland. Two sterile swabs were used to obtain material from the posterior vaginal fornix of 143 women with squamous intraepithelial lesions and 39 healthy women: first for general bacteriology, second for detection of urogenital mycoplasmas using Mycoplasma IST2 kit. From each positive Mycoplasma IST2 culture DNA was isolated and PCR was performed for identification of U. parvum and U. urealyticum. Mycoplasma IST was positive in 34.1% cases. Urogenital mycoplasmas were demonstrated in women with HSIL significantly more often compared to women with LSIL, ASCUS, and with normal cytology. DNA of U. parvum was demonstrated in majority of Mycoplasma IST2-positive cases, U. urealyticum DNA-only in 9 (4.9%). Predominance of 3/14 serovars of U. parvum was demonstrated. U. urealyticum biovar 2 was present more often in women with squamous intraepithelial lesions.     

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.     

Vaginal colonization by genital mycoplasmas in pregnant and non-pregnant women

To compare vaginal colonization by genital micoplasmas in pregnant and non pregnant women and to determine the association between pregnancy and colonization by these microorganisms, samples of exocervix an endocervix from pregnant (n = 80) and non pregnant (n = 65) women, from two health centers of Maracaibo, Zulia State, Venezuela were processed. The Mycoplasma-Lyo kit (bioMérieux laboratories) was used for the culture and identification of genital micoplasmas. In pregnant women, prevalences of 10% for M. hominis and 26.25% for Ureaplasma spp. were found; 35.38% for M. hominis and 20% for Ureaplasma spp. in non-pregnant, were obtained. Among the pregnant, Ureaplasma spp. was the most frequently isolated micoplasma, in symptomatic and asymptomatic; while in the non pregnant group, M. hominis was more common among the symptomatic patients; only one case (1.54%) was an asymptomatic carrier of Ureaplasma spp. The highest positivity percentages were obtained in primigravidas (48.71%) and during the second gestational trimester (34.21%). No statistically significant differences were found between vaginal colonization by genital micoplasmas according to age, number of pregnancy and gestational trimester; but they were found between the presented symptomatology and vaginal colonization by genital micoplasmas. Genital micoplasmas were isolated from gravid women at approximately the same recovery rate as in non-pregnant women; being M. hominis the most frequently isolated in non-pregnant women and Ureaplasma spp. in the pregnant group.     

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.     

Association between preterm birth and vaginal colonization by mycoplasmas in early pregnancy

To examine the association between colonization by two newly classified species of genital ureaplasmas (Ureaplasma parvum and U. urealyticum) in early pregnancy and subsequent late abortion or preterm birth at <34 weeks of gestation, four species of genital mycoplasmas--Mycoplasma genitalium, M. hominis, U. parvum, and U. urealyticum--as well as Chlamydia trachomatis and Neisseria gonorrhoeae were examined by PCR-based methods in a prospective cohort study of 877 women with singleton pregnancies at <11 weeks of gestation. Antibiotics were used only in cases in which C. trachomatis and/or N. gonorrhoeae was detected. Multivariate logistic-regression analysis was used to assess independent risk factors after taking maternal low body weight and past history of preterm birth into account. M. genitalium, M. hominis, U. parvum, U. urealyticum, C. trachomatis, and N. gonorrhoeae were detected in 0.8%, 11.2%, 52.0%, 8.7%, 3.2%, and 0.1% of these 877 women, respectively. Twenty-one (2.4%) women experienced late abortion or preterm birth at <34 weeks of gestation. Three factors-detection of U. parvum in the vagina (odds ratio [OR], 3.0; 95% confidence interval [CI], 1.1 to 8.5); use of antibiotics, such as penicillin and cefatrizine, for incidental inflammatory complications before 22 weeks of gestation (OR, 4.2; 95% CI, 1.6 to 10.0); and past history of preterm birth (OR, 10.4; 95% CI, 2.7 to 40.5)-were independently associated with late abortion and preterm birth. In conclusion, vaginal colonization with U. parvum, but not U. urealyticum, is associated with late abortion or early preterm birth.     

Detection of antibodies to Ureaplasma urealyticum in pregnant women by enzyme-linked immunosorbent assay using membrane antigen and investigation of the significance of the antibodies.

Optimal conditions of a microenzyme-linked immunosorbent assay using a group-specific membrane antigen of Ureaplasma urealyticum serotype 7 were established with rabbit antisera and applied for the evaluation of immunoglobulin M (IgM) and IgG antibodies in 139 serum specimens from pregnant women between 26 and 38 weeks of gestation, and the assay was compared with microorganism culture and investigated to determine the role of U. urealyticum in perinatal morbidity and mortality. U. urealyticum was isolated from 75 (54%) of 139 patients; 40 had a colonization greater than or equal to 10(6) cells per ml of swab (29%); 64 (85%) of 75 culture-positive patients had IgG antibodies (absorbance mean, 0.650), versus 4 (6%) of 64 culture-negative patients (absorbance mean, 0.103) (P less than 0.001). There was no cross-reactivity with Chlamydia trachomatis infection from patients from whom no mycoplasmas were isolated, but this cross-reactivity occurred in 24% of patients with other mycoplasma infections. There was a good correlation between quantitative evaluation of U. urealyticum colonization and antibody level (P less than 0.05). However, IgM antibody was found in 30% of culture-positive patients but also in 25% of the culture-negative group. Frequency of U. urealyticum colonization was greater in unmarried young women (less than 25 years old) with a history of genital infection, and a significantly greater frequency was detected in patients who smoked (P less than 0.01) and had a lower socioeconomic status (P less than 0.001). A lower infant birth weight was more associated with U. urealyticum colonization greater than or equal to 10(5) cells per ml. The enzyme-linked immunosorbent assay provided an additional means to diagnose and evaluate U. urealyticum infection in pregnant women.     

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.     

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.     

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.     

Antibiotic sensitivity of Mycoplasma pathogenic for man

Human pathogen mycoplasmas (Mycoplasma pneumoniae, Mycoplasma hominis, Ureaplasma urealyticum) are intrinsically resistant to antibiotics which inhibit the cell wall biosynthesis (beta-lactams, vancomycin, bacitracin), to polymyxins, rifamycins, sulfonamides, trimethoprim, 5-nitroimidazoles, nitrofurans and to presently available quinolones. These three species are moderately susceptible to aminoglycosides, susceptible to chloramphenicol and highly susceptible to tetracyclines. M. pneumoniae is susceptible to macrolides, lincosamins and streptogramins. M. hominis is resistant to early macrolides (erythromycin, oleandomycin, spiramycin) and susceptible to new macrolides (josamycin, midecamycin, rosaramicin), lincosamins and streptogramins. U. urealyticum is resistant to lincosamins and susceptible to macrolides and streptogramins. Discordant results from various reports can be explained by differences in methods and breakpoint concentration values. In M. pneumoniae species, two strains resistant to macrolides and lincosamins have been described. In M. hominis species, one strain resistant to tetracyclines and another one resistant to tetracyclines and chloramphenicol have been reported. Two to ten percent of U. urealyticum strains are resistant to tetracyclines. These resistances are likely to be plasmid-mediated.     

Prevalence of cervical colonization by Ureaplasma parvum,Ureaplasma urealyticum,Mycoplasma hominis and Mycoplasma genitalium in childbearing age women by a commercially available multiplex real-time PCR: An Italian observational multicentre study

Background

Mycoplasmas are frequently isolated from the genital tract. New molecular PCR-based methods for the detection of mycoplasmas can better define the real epidemiology of these microorganisms. The aim of this study was to evaluate the prevalence of mycoplasmas in a population of childbearing age women by means of PCR.

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.     

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.     

In vitro sensitivity to antibiotics of genital mycoplasmas isolated in Toulouse. Study of new molecules (macrolides and quinolones)]

The minimal metabolism-inhibiting concentrations (MMC) of 11 antibiotics were determined for 40 strains each of M. hominis and U. urealyticum using a terminal color change broth method. All strains were recovered in 1990. Resistance to tetracycline (MMC greater than 8 mg/l) was found for 12.5% of strains of M. hominis and U. urealyticum, as compared with 5% in 1985. Rokitamycin was the most active macrolide against M. hominis (MMC 90: 0.06 mg/l). U. urealyticum strains were susceptible to all the macrolides tested, with the greatest activities being seen for rokitamycin and clarithromycin (MMC 90: 0.06 mg/l and 0.12 mg/l respectively). Sparfloxacin was the most active quinolone against both species. Human clinical trials designed to evaluate these new molecules for the treatment of mycoplasmal and ureaplasmal genital infections are warranted.     

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.     

Occurrence of mycoplasmas in urinary tracts of patients with acute pyelonephritis

Mycoplasma hominis was isolated from the upper urinary tract in 7 of 80 patients with acute pyelonephritis and from 0 of 60 patients with noninfectious diseases of the urinary tract, a significant difference. In four cases M. hominis was isolated in pure culture, in one it was isolated together with Ureaplasma urealyticum, and in two it was isolated with bacteria. U. urealyticum was isolated from the upper urinary tract of five patients, all with acute pyelonephritis; this was not significantly different from the control group.