Antibiotic administration reduces the rate of intraamniotic inflammation in preterm prelabor rupture of the membranes.

BACKGROUND: Preterm prelabor rupture of the membranes (PPROM) is frequently complicated by intraamniotic inflammatory processes such as intraamniotic infection and sterile intraamniotic inflammation. Antibiotic therapy is recommended to patients with PPROM to prolong the interval between this complication and delivery (latency period), reduce the risk of clinical chorioamnionitis, and improve neonatal outcome. However, there is a lack of information regarding whether the administration of antibiotics can reduce the intensity of the intraamniotic inflammatory response or eradicate microorganisms in patients with PPROM. OBJECTIVE: The first aim of the study was to determine whether antimicrobial agents can reduce the magnitude of the intraamniotic inflammatory response in patients with PPROM by assessing the concentrations of interleukin-6 in amniotic fluid before and after antibiotic treatment. The second aim was to determine whether treatment with intravenous clarithromycin changes the microbial load of Ureaplasma spp DNA in amniotic fluid. STUDY DESIGN: A retrospective cohort study included patients who had (1) a singleton gestation, (2) PPROM between 24+0 and 33+6 weeks, (3) a transabdominal amniocentesis at the time of admission, and (4) intravenous antibiotic treatment (clarithromycin for patients with intraamniotic inflammation and benzylpenicillin/clindamycin in the cases of allergy in patients without intraamniotic inflammation) for 7 days. Follow-up amniocenteses (7th day after admission) were performed in the subset of patients with a latency period lasting longer than 7 days. Concentrations of interleukin-6 were measured in the samples of amniotic fluid with a bedside test, and the presence of microbial invasion of the amniotic cavity was assessed with culture and molecular microbiological methods. Intraamniotic inflammation was defined as a bedside interleukin-6 concentration ≥745 pg/mL in the samples of amniotic fluid. Intraamniotic infection was defined as the presence of both microbial invasion of the amniotic cavity and intraamniotic inflammation; sterile intraamniotic inflammation was defined as the presence of intraamniotic inflammation without microbial invasion of the amniotic cavity. RESULTS: A total of 270 patients with PPROM were included in this study: 207 patients delivered within 7 days and 63 patients delivered after 7 days of admission. Of the 63 patients who delivered after 7 days following the initial amniocentesis, 40 underwent a follow-up amniocentesis. Patients with intraamniotic infection (n = 7) and sterile intraamniotic inflammation (n = 7) were treated with intravenous clarithromycin. Patients without either microbial invasion of the amniotic cavity or intraamniotic inflammation (n = 26) were treated with benzylpenicillin or clindamycin. Treatment with clarithromycin decreased the interleukin-6 concentration in amniotic fluid at the follow-up amniocentesis compared to the initial amniocentesis in patients with intraamniotic infection (follow-up: median, 295 pg/mL, interquartile range [IQR], 72-673 vs initial: median, 2973 pg/mL, IQR, 1750-6296; P = .02) and in those with sterile intraamniotic inflammation (follow-up: median, 221 pg/mL, IQR 118-366 pg/mL vs initial: median, 1446 pg/mL, IQR, 1300-2941; P = .02). Samples of amniotic fluid with Ureaplasma spp DNA had a lower microbial load at the time of follow-up amniocentesis compared to the initial amniocentesis (follow-up: median, 1.8 × 104 copies DNA/mL, 2.9 × 104 to 6.7 × 108 vs initial: median, 4.7 × 107 copies DNA/mL, interquartile range, 2.9 × 103 to 3.6 × 107; P = .03). CONCLUSION: Intravenous therapy with clarithromycin was associated with a reduction in the intensity of the intraamniotic inflammatory response in patients with PPROM with either intraamniotic infection or sterile intraamniotic inflammation. Moreover, treatment with clarithromycin was related to a reduction in the load of Ureaplasma spp DNA in the amniotic fluid of patients with PPROM <34 weeks of gestation.

Association between genital mycoplasmas, acute chorioamnionitis and fetal pneumonia in spontaneous abortions.

AIM: Ureaplasma parvum, Ureaplasma urealyticum and Mycoplasma hominis are also known as genital mycoplasmas. Acute chorioamnionitis is an inflammation of the placenta associated with miscarriage. We retrospectively evaluated a possible association between genital mycoplasmas detection, acute chorioamnionitis and fetal pneumonia from second and third trimester spontaneous abortions. METHODS: One hundred and thirty placenta and fetal lung samples were evaluated for histological examination. The placenta samples, along with corresponding fetal tracheo-bronchial aspirates, also underwent bacterial and fungal culture and real-time polymerase chain reaction (PCR) assay for the detection of genital mycoplasmas. RESULTS: Acute chorioamnionitis and pneumonia were diagnosed in 80/130 (61.5%) and 22/130 (16.9%) samples, respectively. Among samples positive for acute chorioamnionitis, the proportion of samples positive by real-time PCR and/or culture, was significantly higher than that of negative controls [54/80 (67.5%) vs. 26/80 (32.5%); P<0.001]. Ureaplasma parvum detection was significantly associated with acute chorioamnionitis compared to controls [9/11 (81.8%) vs. 2/11 (18.2%); P=0.019], as well as U. urealyticum [6/7 (85.7%) vs. 1/7 (14.3%); P=0.039]. Among tracheo-bronchial aspirates from abortions with pneumonia, the proportion of real-time PCR and/or culture positive samples was significantly higher than that of controls [13/22 (59.1%) vs. 9/22 (40.9%); P=0.029]. CONCLUSIONS: A strong association was found between acute histologic chorioamnionitis and microbial invasion with U. parvum and/or U. urealyticum.

Male non-gonococcal urethritis: From microbiological etiologies to demographic and clinical features.

OBJECTIVES: To detect microorganisms responsible for male acute urethritis and to define the microbiology of non-gonococcal urethritis. METHODS: The present study comprised 424 men with symptoms and signs compatible with acute urethritis. Their urethral swabs and first-voided urine underwent detection of the microorganisms. Demographic characteristics and clinical features of Mycoplasma genitalium-, Ureaplasma urealyticum-, Haemophilus influenza-, adenovirus- or Herpes simplex virus-positive monomicrobial non-gonococcal urethritis, or all-examined microorganism-negative urethritis in heterosexual men were compared with urethritis positive only for Chlamydia trachomatis. RESULTS: Neisseria gonorrhoeae was detected in 127 men (30.0%). In 297 men with non-gonococcal urethritis, C. trachomatis was detected in 143 (48.1%). In 154 men with non-chlamydial non-gonococcal urethritis, M. genitalium (22.7%), M. hominis (5.8%), Ureaplasma parvum (9.1%), U. urealyticum (19.5%), H. influenzae (14.3%), Neisseria meningitidis (3.9%), Trichomonas vaginalis (1.3%), human adenovirus (16.2%), and Herpes simplex virus types 1 (7.1%) and 2 (2.6%) were detected. Although some features of monomicrobial non-chlamydial non-gonococcal urethritis or all-examined microorganism-negative urethritis were significantly different from those of monomicrobial chlamydial non-gonococcal urethritis, most features were superimposed. CONCLUSIONS: Predicting causative microorganisms in men with non-gonococcal urethritis based on demographic and clinical features is difficult. However, the present study provides useful information to better understand the microbiological diversity in non-gonococcal urethritis, and to manage patients with non-gonococcal urethritis appropriately.

[Nonculture techniques of microorganisms determination in amniotic fluid in patients with preterm prelabor rupture of membranes]. / Nekultivacní stanovení mikroorganismu v plodové vode u pacientek s predcasným odtokem plodové vody.

OBJECTIVE: The preterm prelabor rupture of membranes is a serious obstetric complication that is frequently complicated by the presence of microorganisms in amniotic fluid. The aim of our work is to characterize current status of nonculture detection of microbial invasion into the amniotic cavity and the experience with the technique performed in University Hospital in Hradec Kralove. DESIGN: Original survey article. SETTING: Institute of Clinical Biochemistry and Diagnostics - molecular biology department, University Hospital Hradec Kralove. CONCLUSION: Application of nonculture techniques of microorganisms determination in amniotic fluid in patients with preterm prelabor rupture of membranes is currently available. According to the detection of genital mycoplasmas as the dominant pathogens in the amniotic fluid this technique should be regarded as the standard examination method in these patients.

Identification of a gene in Mycoplasma hominis associated with preterm birth and microbial burden in intraamniotic infection.

OBJECTIVE: Microbial invasion of the amniotic cavity is associated with spontaneous preterm labor and adverse pregnancy outcome, and Mycoplasma hominis often is present. However, the pathogenic process by which M hominis invades the amniotic cavity and gestational tissues, often resulting in chorioamnionitis and preterm birth, remains unknown. We hypothesized that strains of M hominis vary genetically with regards to their potential to invade and colonize the amniotic cavity and placenta. STUDY DESIGN: We sequenced the entire genomes of 2 amniotic fluid isolates and a placental isolate of M hominis from pregnancies that resulted in preterm births and compared them with the previously sequenced genome of the type strain PG21. We identified genes that were specific to the amniotic fluid/placental isolates. We then determined the microbial burden and the presence of these genes in another set of subjects from whom samples of amniotic fluid had been collected and were positive for M hominis. RESULTS: We identified 2 genes that encode surface-located membrane proteins (Lmp1 and Lmp-like) in the sequenced amniotic fluid/placental isolates that were truncated severely in PG21. We also identified, for the first time, a microbial gene of unknown function that is referred to in this study as gene of interest C that was associated significantly with bacterial burden in amniotic fluid and the risk of preterm delivery in patients with preterm labor. CONCLUSION: A gene in M hominis was identified that is associated significantly with colonization and/or infection of the upper reproductive tract during pregnancy and with preterm birth.

Prevalence of genital mycoplasmas in asymptomatic male partners of women diagnosed as having chlamydial infections.

We examined 209 asymptomatic male partners of women diagnosed as having chlamydial infections for the prevalence of Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealyticum, and Ureaplasma parvum in their first-voided urine (FVU) by nucleic acid amplification tests. Quantification of leukocytes in FVU was performed by automated urine particle analyzers. Two (1.0%) men were positive for N. gonorrhoeae, and 92 (44.0%) were positive for C. trachomatis. In men negative for these pathogens, prevalences of M. genitalium, M. hominis, U. urealyticum, and U. parvum were 0.9%, 29.6%, 27.8%, and 20.1%, respectively, and 58.3% were positive for at least one species of the genital mycoplasmas. Leukocyte counts in FVU from 92 men positive for C. trachomatis were significantly greater than those from 115 men negative for C. trachomatis (p < 0.0001). However, there was no significant difference in leukocyte counts between 66 men positive for at least one species of M. hominis, U. urealyticum, and U. parvum and 48 men negative for all the species (p = 0.1657). The present population of asymptomatic male partners of women diagnosed as having chlamydial infections showed a low prevalence of M. genitalium infections but would be at high risk of being infected by the other genital mycoplasmas. However, it was still unclear whether these genital mycoplasmas would contribute to the development of inflammation of the male urethra. When these partners are negative for C. trachomatis and N. gonorrhoeae, the recommendation to presumptively treat them to disrupt transmission networks of the genital mycoplasmas would seem premature.

The Associations of Genital Mycoplasmas with Female Infertility and Adverse Pregnancy Outcomes: a Systematic Review and Meta-analysis.

The roles of genital mycoplasmas including Mycoplasma genitalium (M. genitalium), Mycoplasma hominis (M. hominis), Ureaplasma urealyticum (U. urealyticum), and Ureaplasma parvum (U. parvum) in reproductive diseases are equivocal. To investigate whether genital mycoplasmas are risk factors of female infertility and adverse pregnancy outcomes, we performed a systematic review and meta-analysis. Electronic databases were searched for related studies. A random-effects model or fixed-effects model was employed to generate forest plots. Pooled odd ratios (ORs) with 95% confidence intervals (CIs) were applied to measure the strength of associations. Meanwhile, heterogeneity was evaluated by H statistic and I2 statistic, and publication bias was explored by funnel plots based on Egger's test and Begg's test. The search yielded 2054 relevant records, and 35 articles were ultimately included for meta-analysis. M. genitalium was a significant risk factor for female infertility (OR, 13.03 [95% CI, 3.46-48.98]) and preterm birth (PTB) (OR, 1.81 [95% CI, 1.17-2.80]), but not for spontaneous abortion (SA) (OR, 0.58 [95% CI, 0.25-1.35]). M. hominis can significantly increase the potential risk of female infertility (OR, 1.56 [95% CI, 1.02-2.38]), SA (OR, 9.14 [95% CI, 4.14-20.18]), stillbirth (OR, 3.98 [95% CI, 1.39-11.36]), and premature rupture of membranes (PROM) (OR, 1.79 [95% CI, 1.26-2.55]), but was not associated with PTB (OR, 1.29 [95% CI, 0.78-2.15]). U. urealyticum had no significant risk effect on female infertility (OR, 0.68 [95% CI, 0.42-1.11]). Coinfections of M. hominis and Ureaplasma were significantly associated with female infertility, SA, and stillbirth, but not with PROM. On the basis of current evidences, this meta-analysis supports that M. genitalium is a risk factor for female infertility and PTB; M. hominis is a potential risk factor for female infertility, SA, stillbirth, and PROM; U. urealyticum has no significant association with female infertility; and the relationship of U. parvum with female infertility and adverse pregnancy outcomes needs to be paid more attention to and remains to be further revealed.

Human miscarriage and infection in Tunisia: Role of Mycoplasma hominis and high Waddlia seroprevalence.

INTRODUCTION: Miscarriage is one of the most common adverse pregnancy outcomes. The aim of this study was to investigate the relationship between miscarriage in humans and infections caused by zoonotic bacteria and genital pathogens. METHODOLOGY: Cervicovaginal swabs and placenta samples from 132 women with miscarriage (patient group: PG), and cervicovaginal swabs from 54 women with normal pregnancy (control group:CG), were subjected to bacteriological culture and real time PCRs detecting Coxiella burnetii, Brucella spp, Mycoplasma hominis, Mycoplasma genitalium, Ureaplasma urealyticum, Chlamydia trachomatis, Waddlia chondrophila and Parachlamydia acanthamoebeae DNA. Serology of C. burnetii, C. trachomatis and W. chondrophila was also performed. RESULTS: Placenta samples were positive for E. coli, S. agalactiae, U. urealyticum, M. hominis and C. trachomatis in 4.7%, 3.1%, 3.1%, 0.7% and 0.7% of cases, respectively. For cervicovaginal swabs, M. hominis was more frequently detected among PG than CG with a significant statistical difference (p = 0.02). C. trachomatis was detected in 3.3% and 5.5% among PG and CG, respectively. U. urealyticum DNA was detected with high percentages in the two groups. Samples from both groups showed negatives results for C. burnetii, Waddlia, and Brucella qPCRs. A high rate of W. chondrophila seroprevalence (42%) was noted with significant difference among women with early miscarriage. CONCLUSIONS: C. trachomatis, S. agalactiae and M. hominis may play a role in miscarriage. However, the full characterization of the vaginal flora using other technologies such as NGS-based metagenomics is needed to clarify their role in miscarriage. Finally, further investigations should be performed to explain high W. chondrophila seroprevalence.

Prevalence and antimicrobial susceptibility of Ureaplasma species and Mycoplasma hominis in Greek female outpatients, 2012-2016.

Mycoplasma hominis and Ureaplasma species are opportunistic pathogens associated with urogenital infections, complications during pregnancy and postpartum infections. Appropriate empirical antimicrobial treatment is necessary to achieve an optimal therapeutic outcome. This study evaluated the prevalence and the antimicrobial susceptibility of Mycoplasma hominis and Ureaplasma spp. isolated from 1,008 endocervical samples of outpatients in Crete, Greece, during a five-year period (2012-2016), using the commercially available Mycoview kit (Zeakon diagnostics, France). Ureaplasma spp. was isolated from 116 patients (11.5%), M. hominis from 6 (0.6%), while coinfection with both mycoplasmas was demonstrated in 17 (1.7%). All Ureaplasma strains were susceptible to josamycin and doxycycline. Doxycycline, minocycline and ofloxacin were the most potent antibiotics against M. hominis. Docycycline was proved the most active and is still the drug of choice for the treatment of genital mycoplasma infections. Local surveillance to monitor changes in antimicrobial susceptibilities is necessary to guide treatment strategies.

A twelve-year retrospective analysis of prevalence and antimicrobial susceptibility patterns of Ureaplasma spp. and Mycoplasma hominis in the province of Lower Silesia in Poland.

OBJECTIVE: Genital mycoplasmas are opportunistic pathogens that have been associated with urogenital infections in humans. Only a few groups of antimicrobials are available for treatment of urogenital tract infections caused by genital mycoplasmas. However, emerging resistance of mycoplasmas to antimicrobial agents has been reported worldwide. The aim of the study was a retrospective analysis of the prevalence and antimicrobial susceptibility patterns of M. hominis and Ureaplasma spp. in patients with urogenital tract infections during a twelve-year period between 2003 and 2015. STUDY DESIGN: Mycoplasma IST2 test was used for the detection, enumeration, identification and antimicrobial susceptibility testing of genital mycoplasmas in 1182 samples from 778 women and 404 men with genitourinary tract infection. Indicative enumeration in the test determines whether the mycoplasma count in the sample is equal or higher than the threshold set at 104 colony forming units. RESULTS: A total of 152 (12.8%) samples were found to be positive for genital mycoplasmas. M. hominis was detected only in three samples and Ureaplasma spp. in 141 samples. Both, M. hominis and Ureaplasma spp. were detected in the remaining eight samples. In the analyzed period between 2003 and 2015, a gradually increasing resistance of ureaplasmas to ciprofloxacin and clarithromycin and decreasing resistance to ofloxacin, erythromycin and tetracycline were observed. Pristinamycin, josamycin and doxycycline were most active against Ureaplasma spp. In contrast, fluoroquinolones had the lowest efficacy against Ureaplasma spp. and as many as 116 (82.3%) and 77 (54.6%) of Ureaplasma spp. isolates were resistant to ciprofloxacin and ofloxacin, respectively. M. hominis isolates were uniformly resistant to azithromycin, clarithromycin and erythromycin but susceptible to josamycin, ofloxacin, doxycycline and pristinamycin. One-third of these isolates were resistant to ciprofloxacin and tetracycline. CONCLUSION: In the study Ureaplasma spp. and M. hominis were detected with relatively low frequency in comparison with other studies however, most of these isolates were resistant to ciprofloxacin indicating the need for better management of ciprofloxacin prescription. Important limitations of Mycoplasma IST2 assay concerning antimicrobial susceptibility testing and divergences between breakpoints in the test and EUCAST guidelines point the need to introduce new methodologies to improve evaluation of resistant strains at our region.

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. METHODS: This 21-month multicentre observational study was conducted at four Italian clinical microbiology laboratories. Women reporting symptoms of vaginitis/cervicitis, or with history of infertility, pregnancy, miscarriage or preterm birth were included. Detection of Ureaplasma parvum, Ureaplasma urealyticum, Mycoplasma hominis, Mycoplasma genitalium was performed from cervical swabs by means of a commercially available multiplex real-time PCR. RESULTS: a total of 1761 women fulfilled the inclusion criteria and were included in the study. The overall prevalence was: U. parvum 38.3%, U. urealyticum 9%, M. hominis 8.6% and M. genitalium 0.6%. The proportion of foreign patients positive for U. parvum was significantly higher compared to Italian patients (37% vs 30.1%, p = 0.007) and also for overall mycoplasma colonization (53.4% vs 45.8%, p = 0.011). The number of symptomatic patients positive for M. hominis was significantly higher than that of negative controls (2.9% vs 1%, p = 0.036). A significant positive trend in mycoplasma colonization was found in relation to the pregnancy week for U. urealyticum (p = 0.015), M. hominis (p = 0.044) and for overall mycoplasma colonization (p = 0.002). CONCLUSION: multiplex RT-PCR can be a valuable tool to evaluate the real epidemiology of cervical mycoplasma colonization.

Polybacterial stimulation suggests discrete IL-6/IL-6R signaling in human fetal membranes: Potential implications on IL-6 bioactivity.

The polybacterial invasion of the amniotic cavity and risk of preterm birth is often due to cervicovaginal bacteria such as genital mycoplasmas (Mycoplasma hominis and Ureaplasma urealyticum) and Gardnerella vaginalis. The most studied biomarker associated with preterm birth is interleukin-6 (IL-6), a pleiotropic cytokine that performs different functions based on classical or trans-signaling mechanisms. This study evaluated the changes in IL-6 and IL-6 function associated accessory molecules by human fetal membranes to determine the functional availability of IL-6 assessment in an in vitro model of polybacterial infection. Fetal membranes were treated with LPS or heat-inactivated genital mycoplasmas and G. vaginalis alone or in combination. IL-6 and its soluble receptors (sgp130, sIL-6R) were assessed in conditioned medium by immunoassays and membrane-bound receptors were evaluated in the tissue using immunohistochemistry and RT-PCR. Data from protein and gene expression were evaluated using linear mixed effects models. Data from immunohistochemistry were evaluated using one-way analysis of variance followed by the Tukey test. Genital mycoplasmas alone, or in combination, inhibited IL-6 trans-signaling with increased sgp130 production. G. vaginalis activated the classical IL-6 signaling pathway, as did LPS. Polybacterial treatment resulted in a balanced response with neither pathway being favored. The increase in IL-6 production by fetal membranes in response to infection is likely a non-specific innate response and not an indicator of a functional mediator of any labor-inducing pathways. This suggests that correlating the risk of adverse pregnancy outcomes and designing interventions based on IL-6 levels without considering soluble receptors may be an ineffective strategy.

Adverse pregnancy and neonatal outcomes associated with Neisseria gonorrhoeae, Mycoplasma genitalium, M. hominis, Ureaplasma urealyticum and U. parvum: a systematic review and meta-analysis protocol.

INTRODUCTION: Several bacterial sexually transmitted and genital mycoplasma infections during pregnancy have been associated with poor pregnancy and perinatal outcomes. Comprehensive and systematic information about associations between sexually transmitted infections (STI) and genital infections in pregnancy and adverse perinatal outcomes is needed to improve understanding about the evidence for causal associations between these infections and adverse pregnancy and neonatal outcomes. Our primary objective is to systematically review the literature about associations between: (1) Neisseria gonorrhoeae in pregnancy and preterm birth; (2) Mycoplasma genitalium in pregnancy and preterm birth; (3) M. hominis, Ureaplasma urealyticum and/or U. parvum in pregnancy and preterm birth. METHODS AND ANALYSIS: We will undertake a systematic search of Medline, Excerpta Medica database and the Cochrane Library and Cumulative Index to Nursing and Allied Health Literature. Following an initial screening of titles by one reviewer, abstracts will be independently assessed by two reviewers before screening of full-text articles. To exclude a manuscript, both reviewers need to agree on the decision. Any discrepancies will be resolved by discussion, or the adjudication of a third reviewer. Studies will be included if they report testing for one or more of N. gonorrhoeae, M. genitalium, M. hominis, U. urealyticum and/or U. parvum during pregnancy and report pregnancy and/or birth outcomes. In this review, the primary outcome is preterm birth. Secondary outcomes are premature rupture of membranes, low birth weight, spontaneous abortion, stillbirth, neonatal mortality and ophthalmia neonatorum. We will use standard definitions, or definitions reported by study authors. We will examine associations between exposure and outcome in forest plots, using the I2 statistic to examine between study heterogeneity. Where appropriate, we will use meta-analysis to combine results of individual studies. ETHICS AND DISSEMINATION: This systematic review of published literature does not require ethical committee approval. Results of this review will be published in a peer reviewed, open access journal. PROSPERO REGISTRATION NUMBER: CRD42016050962.

Combinations and loads of bacteria affect the cytokine production by fetal membranes: An in vitro study.

PROBLEM: The polybacterial invasion and inflammation of the amniotic cavity is a common scenario in PTB, and then, we analyzed the cytokine production by human fetal membranes to better understand the host response to polybacterial infections. METHOD OF STUDY: Fetal membranes were treated by heat-inactivated genital mycoplasmas and Gardnerella vaginalis at 103 or 106 colony/color-forming units/mL alone or in combination. Cytokines/receptors were measured in the medium by immunoassays. RESULTS: Stimulation of genital mycoplasmas did not increase the proinflammatory cytokines, except Ureaplasma urealyticum that increased IL-8 levels. However, U. urealyticum and Mycoplasma hominis significantly increased IL-10 and IL-13 levels. G. vaginalis alone or in combination with genital mycoplasmas showed an increased proinflammatory and anti-inflammatory cytokines. CONCLUSIONS: G. vaginalis sustain a proinflammatory response in the fetal membranes in vitro, while genital mycoplasmas induce a strong control of the inflammatory response. The ability of genital mycoplasmas to control the proinflammatory response may favor their survival in the upper genital tract.

Ureaplasma species and Mycoplasma hominis in cervical fluid of pregnancies complicated by preterm prelabor rupture of membranes.

OBJECTIVE: To evaluate Ureaplasma species and Mycoplasma hominis DNA in the cervical fluid and their association with microbial invasion of the amniotic cavity (MIAC) and/or histological chorioamnionitis (HCA) in pregnancies complicated by preterm prelabor rupture of membranes (PPROM). STUDY DESIGN: A prospective study of 68 women with singleton pregnancies complicated by PPROM between 24(0/7) and 36(6/7) weeks was conducted. Cervical fluid and amniotic fluid were collected from all women at the time of admission. The Ureaplasma species and Mycoplasma hominis DNA in the cervical fluid were identified using specific real-time PCR. RESULTS: Ureaplasma species and Mycoplasma hominis DNA were identified in 59% (40/69) of the cervical fluid samples. Women with the presence of Ureaplasma species DNA with and without Mycoplasma hominis DNA in the cervical fluid had a higher rate of MIAC alone [35% (14/40) versus 11% (3/28); p = 0.02] and a higher rate of the presence of both MIAC and HCA [30% (12/40) versus 4% (1/28); p = 0.01] than women without Ureaplasma species and Mycoplasma hominis DNA in the cervical fluid. CONCLUSIONS: The presence of Ureaplasma species DNA with and without Mycoplasma hominis DNA in the cervical fluid is associated with a higher risk of MIAC or MIAC and HCA together in pregnancies complicated by PPROM.

Gastric fluid versus amniotic fluid analysis for the identification of intra-amniotic infection due to Ureaplasma species.

OBJECTIVE: Early neonatal sepsis is often due to intra-amniotic infection. The stomach of the neonate contains fluid swallowed before and during delivery. The presence of bacteria as well as neutrophils detected by culture or Gram stain of the gastric fluid during the first day of life is suggestive of exposure to bacteria or inflammation. We undertook this study to determine the relationship between gastric fluid analysis and amniotic fluid obtained by transabdominal amniocentesis in the detection of Ureaplasma species, the most frequent microorganisms responsible for intra-amniotic infection. MATERIALS AND METHODS: The study population consisted of 100 singleton pregnant women who delivered preterm neonates (<35 weeks) within 7 days of amniocentesis. Gastric fluid of newborns was obtained by nasogastric intubation on the day of birth. Amniotic fluid and gastric fluid were cultured for genital Mycoplasmas, and polymerase chain reaction (PCR) for Ureaplasma species was performed. Intra-amniotic inflammation was defined as an elevated amniotic fluid matrix metalloproteinase-8 concentration (>23 ng/mL). RESULTS: (1) Ureaplasma species were detected by culture or PCR in 18% (18/100) of amniotic fluid samples and in 5% (5/100) of gastric fluid samples; (2) among the amniotic fluid cases positive for Ureaplasma species, these microorganisms were identified in 27.8% (5/18) of gastric fluid samples; (3) none of the cases negative for Ureaplasma species in the amniotic fluid were found to be positive for these microorganisms in the gastric fluid; (4) patients with amniotic fluid positive for Ureaplasma species but with gastric fluid negative for these microorganisms had a significantly higher rate of intra-amniotic inflammation, acute histologic chorioamnionitis, and neonatal death than those with both amniotic fluid and gastric fluid negative for Ureaplasma species; and (5) no significant differences were observed in the rate of intra-amniotic inflammation, acute histologic chorioamnionitis, and neonatal death between patients with amniotic fluid positive for Ureaplasma species but with gastric fluid negative for these microorganisms and those with both amniotic fluid and gastric fluid positive for Ureaplasma species. CONCLUSIONS: Gastric fluid analysis has 100% specificity in the identification of intra-amniotic infection with Ureaplasma species. However, the detection of Ureaplasma species by culture or PCR in the gastric fluid of neonates at birth did not identify these microorganisms in two-thirds of cases with microbial invasion of the amniotic cavity. Thus, amniotic fluid analysis is superior to that of gastric fluid in the identification of intra-amniotic infection.

Microbial load of umbilical cord blood Ureaplasma species and Mycoplasma hominis in preterm prelabor rupture of membranes.

OBJECTIVE: To evaluate Ureaplasma species and M. hominis DNA in the umbilical cord blood and its correlation with its microbial load in the amniotic fluid, as a measure of microbial burden in fetal inflammatory response and neonatal outcome in pregnancies complicated by preterm prelabor rupture of membranes (pPROM). STUDY DESIGN: A retrospective study of 158 women with singleton pregnancies complicated by pPROM between 24(0/7) and 36(6/7) weeks was conducted. Amniotic fluid was obtained from all women by transabdominal amniocentesis, and umbilical cord blood was obtained by venipuncture from umbilical cords immediately after the delivery of the neonates. The Ureaplasma species and M. hominis DNA was quantitated using absolute quantification techniques. RESULT: Ureaplasma species and M. hominis DNA was identified in 9% of the umbilical cord blood samples. No correlation between the amniotic fluid and umbilical cord blood microbial load was observed. The presence of Ureaplasma species and M. hominis DNA in the umbilical cord blood had no impact on short-term neonatal morbidity. CONCLUSIONS: A high microbial load of genital mycoplasma Ureaplasma species DNA in the umbilical cord in pregnancies complicated by pPROM is not associated with a high fetal inflammatory response and is therefore not associated with serious neonatal morbidity.

Genital Mycoplasmas in the Newborn Infants: Colonization, Prevalence and Clinical Significance

PURPOSE: The genital mycoplasmas: Ureaplasma urealyticum and Mycoplasma hominis have recently assumed an increasing importance as neonatal pathogens especially in preterm infants. The aim of the present study was to determine the prevalence of infections with these organisms in newborn infants who were admitted in Neonatal Intensive Care Unit(NICU) and who were suspected having infection in newborn nursery METHODS: Sixty four inborns who were hospitalized in the NICU of Seoul National University Children's Hospital and Fourty seven term newborns who were born in Seoul National University Hospital and were evaluated for sepsis in the nursery due to high risk mother and baby's clinical symptoms from May 1994 through August 1994 were included in this study. Blood during the first hour of life and throat swabs during the second hours of life of the baby were collected. Tracheal aspirates were collected in the mechanically ventilated infants during the first day of life. Transport media for genital mycoplasma, urea and mycoplasma broth, and urea agar were used for isolation of genital mycoplasmas. RESULTS: In 64 inborns of NICU included in this study U. urealyticum was isolated in five(11.1%) out of 45 throat swab cultures, one(1.7%) out of 60 blood cultures and one(7.1%) out of 14 tracheal aspirates. M. hominis was isolated in 2 throat cultures. So total 14.3% of these infants harbored genital mycoplasmas. Among 47 term newborns included in this study from nursery, U. urealyticum was isolated 3(8.6%) out of 35 throat swab cultures and one (2.2%) out of 45 blood cultures shortly after birth. M. hominis was not isolated among them. Genital mycoplasma-positive infants in NICU had lesser gestatonal age and lower birth weight than genital mycoplasma- negative infants. Isolation of genital mycoplasmas was also associated with maternal clinical chorioamnionitis. No evidences that neonatal disease such as suspected sepsis, chronic lung disease of prematurity and neonatal outcome such as duration of hospital stay and mortality were related to genital mycoplasmas were noted. Cultures for genital mycoplasmas in amniotic fluid were performed in 26 preterm infants' mothers whose babies hospitalized in NICU and 11 had positve results. Acquisition rate of genital mycoplasmas in their neonates was 54.5%(6/11). One preterm infant who harbored U. urealyticum in the blood exhibited suspected congenital pneumonia. CONCLUSIONS: Our results demonstrate that colonization with genital mycoplasmas is not uncommon in the newborn infants in Korea and genital mycoplasma-positive preterms had lesser gestational age and lower birth weight than genital mycoplasma- negative preterms. Further study about the relation of genital mycoplasmas to neonatal morbidity will be needed.