Ureaplasma parvum undergoes selection in utero resulting in genetically diverse isolates colonising the chorioamnion of fetal sheep

Ureaplasmas are the microorganisms most frequently isolated from the amniotic fluid of pregnant women and can cause chronic intrauterine infections. These tiny bacteria are thought to undergo rapid evolution and exhibit a hypermutatable phenotype; however, little is known about how ureaplasmas respond to selective pressures in utero. Using an ovine model of chronic intra-amniotic infection, we investigated if exposure of ureaplasmas to sub-inhibitory concentrations of erythromycin could induce phenotypic or genetic indicators of macrolide resistance. At 55 days gestation, 12 pregnant ewes received an intra-amniotic injection of a non-clonal, clinical U. parvum strain, followed by: (i) erythromycin treatment (IM, 30 mg/kg/day, n=6); or (ii) saline (IM, n=6) at 100 days gestation. Fetuses were then delivered surgically at 125 days gestation. Despite injecting the same inoculum into all ewes, significant differences between amniotic fluid and chorioamnion ureaplasmas were detected following chronic intra-amniotic infection. Numerous polymorphisms were observed in domain V of the 23S rRNA gene of ureaplasmas isolated from the chorioamnion (but not the amniotic fluid), resulting in a mosaic-like sequence. Chorioamnion isolates also harboured the macrolide resistance genes erm(B) and msr(D) and were associated with variable roxithromycin minimum inhibitory concentrations. Remarkably, this variability occurred independently of exposure of ureaplasmas to erythromycin, suggesting that low-level erythromycin exposure does not induce ureaplasmal macrolide resistance in utero. Rather, the significant differences observed between amniotic fluid and chorioamnion ureaplasmas suggest that different anatomical sites may select for ureaplasma sub-types within non-clonal, clinical strains. This may have implications for the treatment of intrauterine ureaplasma infections.

The characterisation of Ureaplasma species isolated from the Chorioamnion of women who deliver late Preterm

This thesis investigated, for the first time, the prevalence of Ureaplasma species infection within the placentae of women who delivered in the late preterm stages of pregnancy. The presence of these microorganisms was associated with either severe inflammation within the placenta or, for some women, there were no pregnancy complications and these women delivered at term. Ureaplasmas are able to vary their surface exposed lipoproteins and we demonstrated that different host immune responses were generated in vivo to different sized surface lipoproteins. This may explain why ureaplasma infections do not always result in adverse pregnancy and neonatal outcomes.

Placental infection with Ureaplasma species is associated with histologic chorioamnionitis and adverse outcomes in moderately preterm and late-preterm infants

Objective The human Ureaplasma species are the microbes most frequently isolated from placentae of women who deliver preterm. The role of Ureaplasma species has been investigated in pregnancies at <32 weeks of gestation, but currently no studies have determined the prevalence of ureaplasmas in moderately preterm and late-preterm (hereafter, “moderate/late preterm”) infants, the largest cohort of preterm infants. Methods Women delivering moderate/late preterm infants (n = 477) and their infants/placentae (n = 535) were recruited, and swab specimens of chorioamnion tissue, chorioamnion tissue specimens, and cord blood specimens were obtained at delivery. Swab and tissue specimens were cultured and analyzed by 16S ribosomal RNA polymerase chain reaction (PCR) for the presence of microorganisms, while cord blood specimens were analyzed for the presence of cytokines, chemokines, and growth factors. Results We detected microorganisms in 10.6% of 535 placentae (443 were delivered late preterm and 92 were delivered at term). Significantly, Ureaplasma species were the most prevalent microorganisms, and their presence alone was associated with histologically confirmed chorioamnionitis in moderate/late preterm and term placentae (P < .001). The presence of ureaplasmas in the chorioamnion was also associated with elevated levels of granulocyte colony-stimulating factor (P = .02). Conclusions These findings have important implications for infection and adverse pregnancy outcomes throughout gestation and should be of major consideration for obstetricians and neonatologists.

The common vaginal commensal bacterium Ureaplasma parvum is associated with chorioamnionitis in extreme preterm labour

OBJECTIVE: To assess the association of vaginal commensal and low grade pathogenic bacteria including Ureaplasma parvum, Ureaplasma urealyticum, Mycoplasma hominis, Mycoplasma genitalium, Group B Streptococcus (GBS), and Gardnerella vaginalis, in women who delivered preterm at less than 37 weeks gestation in the presence or absence of inflammation of the chorioamnionitic membranes.

METHODS: A case control study involving women who delivered before 37 weeks gestation with and without inflammation of chorioamnionitic membranes. A total of 57 placental samples were histologically examined for polymorphonuclear leukocyte infiltration of placental tissue for evidence of chorioamnionitis, and by type-specific nucleic acid amplification for evidence of infection with one or more of the target bacteria. Demographic data was collected for each mother.

RESULTS: Amongst the 57 placental samples, 42.1% had chorioamnionitis and 24.6% delivered in the second trimester of pregnancy; U. parvum, U. urealyticum, G. vaginalis and GBS were all detected in the study with respective prevalence of 19.3%, 3.5%, 17.5% and 15.8%; M.genitalium and M. hominis were not detected. U. parvum was significantly associated with chorioamnionitis (p value = 0.02; OR 5.0; (95% CI 1.2-21.5) and was more common in women who delivered in the second (35.7%) compared to the third trimester of pregnancy (13.9%). None of the other bacteria were associated with chorioamnionitis or earlier delivery and all G.vaginalis positive women delivered in the third trimester of pregnancy (p value 0.04).

CONCLUSIONS: The detection of U. parvum in placental tissue was significantly associated with acute chorioamnionitis in women presenting in extreme preterm labour.

1.° Martyrologium monasterii sancti Martialis, ab illo diversum quod Usuardi nomine circumfertur. — 2.° Necrologium ejusdem monasterii : item societates cum aliis coenobiis initae, Burguliensi nempè, Malliacensi, Usercensi et sancti Florentii. — Hujusce codicis pars duodecimo, pars decimo tertio saeculo videtur exarata.

Sancti Martialis Lemovicensis

1.° Martyrologium ab Augusti die decima quinta initium ducens, nec ab illo diversum quod Adonis nomine circumfertur. — 2.° Proverbia, Ecclesiastes et Canticum canticorum. — Hujus codicis pars decimo, pars undecimo saeculo videtur exarata.

Sancti Martialis Lemovicensis

Estudio de la ultraestructura seminal y de los parámetros seminológicos, en pacientes infértiles con PCR positivo para Ureaplasma urealyticum

Tesis (Maestría en Ciencias con Especialidad en Morfología) UANL

AGAR MEDIUM FOR HUMAN UREAPLASMA UREALYTICUM

The authors wish to report a simple medium for the isolation and cultivation of Ureaplasma urealyticum starting from clinical materials. This medium induced growth in eleven of the twelve different serotypes of Ureaplasma urealyticum studied.

First-void versus midstream urine culture for detection of Ureaplasma urealyticum in the urinary tract

Quantitative Ureaplasma urealyticum culture was performed on clean first-void and midstream urine to evaluate the presence of these mollicutes in the urinary tract. The results, expressed as color changing units (CCU), showed that 14 (63%) of the 22 Ureaplasma urealyticum positive patients yielded counts equal to or higher that 10(7) CCU/mL for both the initial and the middle urine specimens. No abnormal chemical or microscopic findings (protein content, leukocyte numbers) were observed. The occurrence of U. urealyticum in midstream urine samples, even when numbers are considered, may be no more than a guide to the presence of ureaplasmas in the urinary tract.

Associação entre presença de Mycoplasma hominis e Ureaplasma urealyticum e níveis de citocinas pró e antiinflamatórias no líquido amniótico de gestação de termo

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Correlação entre presença de Mycoplasma hominis e Ureaplasma urealyticum e níveis IL-6 e IL-10 no líquido amniótico de gestantes com trabalho de parto prematuro

O trabalho de parto prematuro (TPP) é uma intercorrência obstétrica com etiologia multifatorial, que tem como principal complicação a prematuridade. A infecção da cavidade amniótica (CA) é um fator associado ao TPP e, nos últimos anos, inúmeros trabalhos têm demonstrado a presença de diferentes espécies bacterianas no líquido amniótico (LA) de pacientes em TPP. Dentre essas, destacam-se duas espécies de micoplasmas genitais, Mycoplasma hominis e Ureaplasma urealyticum, que embora sejam freqüentemente detectadas no LA ainda são pouco estudadas quanto à sua relação com essa complicação obstétrica. Avaliar a infecção na cavidade amniótica por M. hominis e U. urealyticum bem como determinar os níveis de IL-6 e IL-10 no líquido amniótico de gestantes com trabalho de parto prematuro. Foi realizado estudo prospectivo com 20 gestantes em TPP, atendidas no Serviço de Obstetrícia do Hospital das Clínicas da Faculdade de Medicina de Botucatu, UNESP. O grupo controle foi constituído de 20 gestantes com indicação para amniocentese transabdominal para avaliação da maturidade fetal. Foram obtidas amostras do líquido amniótico e das membranas corioamnióticas de todas as pacientes incluídas no estudo. A pesquisa de M.hominis e U.urealyticum no LA foi realizada empregando-se a técnica de reação em cadeia da polimerase (PCR) e os níveis de IL-6 e IL-10 quantificados por ensaio imunoenzimático (ELISA). Os dados obtidos referentes às características maternas, infecção da cavidade amniótica e concentração de citocinas no LA foram submetidos ao teste z de proporção e ao teste de Mann-Whitney e o nível de significância adotado foi de 5%. A incidência de TPP no período do estudo foi de 5,8%. No grupo TPP, a pesquisa de invasão microbiana da CA foi positiva para... (Resumo completo, clicar acesso eletrônico abaixo)

Antibiotic Susceptibility and Sequence Type Distribution of Ureaplasma Species Isolated from Genital Samples in Switzerland.

Antibiotic resistance in Ureaplasma urealyticum/Ureaplasma parvum and Mycoplasma hominis is an issue of increasing importance. However, data regarding the susceptibility and, more importantly, the clonality of these organisms are limited. We analyzed 140 genital samples obtained in Bern, Switzerland, in 2014. Identification and antimicrobial susceptibility tests were performed by using the Mycoplasma IST 2 kit and sequencing of 16S rRNA genes. MICs for ciprofloxacin and azithromycin were obtained in broth microdilution assays. Clonality was analyzed with PCR-based subtyping and multilocus sequence typing (MLST), whereas quinolone resistance and macrolide resistance were studied by sequencing gyrA, gyrB, parC, and parE genes, as well as 23S rRNA genes and genes encoding L4/L22 ribosomal proteins. A total of 103 samples were confirmed as positive for U. urealyticum/U. parvum, whereas 21 were positive for both U. urealyticum/U. parvum and M. hominis. According to the IST 2 kit, the rates of nonsusceptibility were highest for ciprofloxacin (19.4%) and ofloxacin (9.7%), whereas low rates were observed for clarithromycin (4.9%), erythromycin (1.9%), and azithromycin (1%). However, inconsistent results between microdilution and IST 2 kit assays were recorded. Various sequence types (STs) observed previously in China (ST1, ST2, ST4, ST9, ST22, and ST47), as well as eight novel lineages, were detected. Only some quinolone-resistant isolates had amino acid substitutions in ParC (Ser83Leu in U. parvum of serovar 6) and ParE (Val417Thr in U. parvum of serovar 1 and the novel Thr417Val substitution in U. urealyticum). Isolates with mutations in 23S rRNA or substitutions in L4/L22 were not detected. This is the first study analyzing the susceptibility of U. urealyticum/U. parvum isolates in Switzerland and the clonality outside China. Resistance rates were low compared to those in other countries. We hypothesize that some hyperepidemic STs spread worldwide via sexual intercourse. Large combined microbiological and clinical studies should address this important issue.