Mycoplasma hominis Causes DNA Damage and Cell Death in Primary Human Keratinocytes.

Mycoplasma hominis can be isolated from the human urogenital tract. However, its interaction with the host remains poorly understood. In this study, we aimed to assess the effects of M. hominis infection on primary human keratinocytes (PHKs). Cells were quantified at different phases of the cell cycle. Proteins involved in cell cycle regulation and apoptosis progression were evaluated. The expression of genes encoding proteins that are associated with the DNA damage response and Toll-like receptor pathways was evaluated, and the cytokines involved in inflammatory responses were quantified. A greater number of keratinocytes were observed in the Sub-G0/G1 phase after infection with M. hominis. In the viable keratinocytes, infection resulted in G2/M-phase arrest; GADD45A expression was increased, as was the expression of proteins such as p53, p27, and p21 and others involved in apoptosis regulation and oxidative stress. In infected PHKs, the expression of genes associated with the Toll-like receptor pathways showed a change, and the production of IFN-γ, interleukin (IL) 1ß, IL-18, IL-6, and tumour necrosis factor alpha increased. The infection of PHKs by M. hominis causes cellular damage that can affect the cell cycle by activating the response pathways to cellular damage, oxidative stress, and Toll-like receptors. Overall, this response culminated in the reduction of cell proliferation/viability in vitro.

El diagnóstico de Infecciones de Transmisión Sexual por la técnica de biología molecular es la mejor estrategia para su diagnóstico oportuno y específico. Un caso clínico / The diagnosis of Sexually Transmitted Infections by the molecular biology technique is the best strategy for its timely and specific diagnosis. A clinical case

Resumen Se presenta el caso de un paciente a quien se le diagnosticó una Infección de Transmisión Sexual (ITS) por la técnica de PCR múltiple y en quién se logró por esta técnica, detectar cuatro agentes diferentes simultáneamente: Neisseria gonorreae, Mycoplasma hominis, Ureaplasma urealyticum/parvum y Trichomonas vaginalis, situación esta, que no hubiera sido posible utilizando el procedimiento estándar.

Summary Here we report the case of a patient with a Sexually Transmitted Disease (STI) in whom four different agents were detected by a multiple PCR technique: Neisseria gonorreae, Mycoplasma hominis, Ureaplasma urealyticum / parvum and Trichomonas vaginalis. This detection of multiple agents would not have been possible using conventional procedures.

Main etiological agents identified in 170 men with urethritis attended at the Fundação Alfredo da Matta, Manaus, Amazonas, Brazil,

Abstract Background: Sexually transmitted infections (STI) are a global public health problem. Urethritis are among the most common STIs, and can cause several complications and facilitate the transmission of the HIV virus. Objectives: To investigate the main etiologic agents of urethritis in 170 men treated at Fundação Alfredo da Matta. Methods: To identify the agents, urethral exudate and urine were collected. Gram and culture tests were performed in Thayer-Martin medium for Neisseria gonorrhoeae and polymerase chain reaction for Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, Ureaplasma urealyticum, Ureaplasma parvum, Mycoplasma hominis, Mycoplasma genitalium, and herpes simplex types 1 and 2. Results: N. gonorrhoeae were identified in 102 (60.0%) patients, C. trachomatis in 50 (29.4%), U. urealyticum in 29 (17.0%), M. genitalium in 11 (6.5 %), U. parvum in ten (5.9%), and M. hominis in seven (4.1%). Herpes simplex type 2 was diagnosed in 24 (21.6%) of the 111 patients who underwent PCR for this pathogen. In 69 cases there was co-infection; the most frequent were: N. gonorrhoeae and C. trachomatis in 21 (14.7%) patients; N. gonorrhoeae and C. trachomatis in 21 (12.4%) patients; N. gonorrhoeae and herpes simplex type 2 in 11 (6.5%), and N. gonorrhoeae and U. urealyticum in nine (5.3%). Study limitations: Not relevant. Conclusion: N. gonorrhoeae, C. trachomatis, U. urealyticum, and herpes simplex type 2 were the pathogens most frequently identified in the present study. The main coinfection found was N. gonorrhoeae and C. trachomatis. T. vaginalis and herpes simplex type 1 were not identified in any of the patients.

Main etiological agents identified in 170 men with urethritis attended at the Fundação Alfredo da Matta, Manaus, Amazonas, Brazil.

BACKGROUND: Sexually transmitted infections (STI) are a global public health problem. Urethritis are among the most common STIs, and can cause several complications and facilitate the transmission of the HIV virus. OBJECTIVES: To investigate the main etiologic agents of urethritis in 170 men treated at Fundação Alfredo da Matta. METHODS: To identify the agents, urethral exudate and urine were collected. Gram and culture tests were performed in Thayer-Martin medium for Neisseria gonorrhoeae and polymerase chain reaction for Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, Ureaplasma urealyticum, Ureaplasma parvum, Mycoplasma hominis, Mycoplasma genitalium, and herpes simplex types 1 and 2. RESULTS: N. gonorrhoeae were identified in 102 (60.0%) patients, C. trachomatis in 50 (29.4%), U. urealyticum in 29 (17.0%), M. genitalium in 11 (6.5 %), U. parvum in ten (5.9%), and M. hominis in seven (4.1%). Herpes simplex type 2 was diagnosed in 24 (21.6%) of the 111 patients who underwent PCR for this pathogen. In 69 cases there was co-infection; the most frequent were: N. gonorrhoeae and C. trachomatis in 21 (14.7%) patients; N. gonorrhoeae and C. trachomatis in 21 (12.4%) patients; N. gonorrhoeae and herpes simplex type 2 in 11 (6.5%), and N. gonorrhoeae and U. urealyticum in nine (5.3%). STUDY LIMITATIONS: Not relevant. CONCLUSION: N. gonorrhoeae, C. trachomatis, U. urealyticum, and herpes simplex type 2 were the pathogens most frequently identified in the present study. The main coinfection found was N. gonorrhoeae and C. trachomatis. T. vaginalis and herpes simplex type 1 were not identified in any of the patients.

Examination of Ureaplasma urealyticum and Mycoplasma hominis in 4082 Chinese patients

The objective of this study was to analyze the infection rate and drug resistance of Ureaplasma urealyticum (UU) and Mycoplasma hominis (MH) in the genitourinary tract of Chinese patients. From December 2018 to June 2019, vaginal secretion or urinary secretion of outpatients in our hospital were selected for culture and drug sensitivity analysis of Ureaplasma urealyticum and Mycoplasma hominis. In 4082 Chinese samples, 1567 Mycoplasma were detected, a detection rate of 38.39%, among which 1366 cases were UU single positive, accounting for 33.47%, 15 cases were MH single positive, accounting for 0.36%, 186 cases were UU and MH mixed positive, accounting for 4.56%. The most affected age groups were 21-30 years and 31-40 years, accounting for 19.09 and 15.05%, respectively. The results of drug sensitivity showed that doxycycline, minocycline, josamycin, clarithromycin, and roxithromycin were more sensitive to mycoplasma infection. The distribution of Ureaplasma urealyticum and Mycoplasma hominis in the human genitourinary system and their sensitivity to antibiotics is different for sex and age groups.

Increased prevalence of endocervical Mycoplasma and Ureaplasma colonization in infertile women with tubal factor.

OBJECTIVE: Most women suffering from tubal factor infertility do not have a history of pelvic inflammatory disease, but rather have asymptomatic upper genital tract infection. Investigating the impacts of such infections, even in the absence of clinically confirmed pelvic inflammatory disease, is critical to understanding the tubal factor of infertility. The aim of this study was to investigate whether the presence of endocervical bacteria is associated with tubal factors in women screened for infertility. METHODS: This retrospective cross-sectional study involved 245 women undergoing hysterosalpingography (HSG), screened for endocervical colonization by Chlamydia trachomatis, Neisseria gonorrhea, Ureaplasma urealyticum and Mycoplasma hominis, as part of a routine female infertility investigation between 2016 and 2017. RESULTS: endocervical bacterial colonization by Chlamydia trachomatis, Ureaplasma urealiticum, Mycoplasma hominis and other bacteria corresponded to 3.7%, 9.0%; 5.7% and 9.8%, respectively. There was no colonization by Neisseria gonorrhea. The prevalence of tubal factor was significantly higher in patients with positive endocervical bacteria colonization, regardless of bacterial species. When evaluating bacteria species individually, the women who were positive for endocervical Mycoplasma hominis had significantly higher rates of tubal factor. Associations between endocervical bacterial colonization and tubal factor infertility were confirmed by multiple regression analysis adjusted for age and duration of infertility. CONCLUSION: Besides the higher prevalence of Mycoplasma and Ureaplasma infectious agents, the findings of this study suggest the possible association of endocervical bacterial colonization - not only Chlamydia trachomatis and Neisseria gonorrhea, but also Mycoplasma species with tubal performance.

Occurrence of Mycoplasmaspp. and Ureaplasmaspp. in genital specimens

Mycoplasmaspp. and Ureaplasmaspp. belong tohumans'genitourinary microbiota and sometimesare associated with infections of the genitourinarytract. The aim of this study was to evaluate the occurrence of Mycoplasmaspp. and Ureaplasmaspp. in genital specimens from patients of the 15thRegional de Saúde of ParanáState, Brazil, and to correlate the results with clinical and laboratory data.A retrospective cross-sectional study was conducted,based on the analysis of results of vaginal, endocervical, urine andurethral culture for mycoplasmas from patients attended in areference laboratory, from January 2009 to December 2016. We evaluated 2,475 results of culture for mycoplasmas. A total of 50.8% patients were positive for mycoplasmas. Of these, 76.8%had positive culture exclusively for Ureaplasmaspp. and 4.7% for Mycoplasmahominis. Both microorganisms were isolated in the microbiology culture of 18.5% of patients. Among the positive culture, 81.4% had significant concentrations.Bacterialvaginosis was the most common alteration observed in association with mycoplasmas.Thehigh positivity of cultures for mycoplasmas, especially Ureaplasmaspp. found in our study, highlightthe presence of these microorganisms in many of the genital tract disorders that can be sexually transmitted and, consequently, should not be neglected.

Prevalence of ureaplasma urealyticum, mycoplasma hominis and human papillomavirus coinfection in people attending a sexually transmitted infections (STI)/HIV reference centre in Salvador, Bahia, Brazil / Prevalência da coinfecção por ureaplasma urealyticum, mycoplasma hominis e papilomavírus humano em pessoas atendidas em centro de referência de doenças sexualmente transmissíveis (DST)/HIV em Salvador, Bahia

Introduction: Ureaplasma urealyticum and Mycoplasma hominis are frequently found at many women's and men's urogenital tract, and have been associated with non-gonococcal urethritis, cervicitis, infertility, chorioaminionitis and adverse pregnancy outcomes. Some studies show high prevalence of human papillomavirus (HPV) in patients with non-gonococcal urethritis, while also presenting high frequency of Ureaplasma urealyticum infection in women with cervicalcytology abnormalities and men with genital warts. Objectives: To evaluate the prevalence of Ureaplasma urealyticum, Mycoplasma hominis and HPV coinfection in people attending a sexually transmitted infections (STI)/HIV reference centre and to identify the risk factors associated. Methods: A cross-sectional study with patients aged >18 years, carried out for Ureaplasma urealyticum and Mycoplasma hominis from July 1st to December 31, 2015, in a STI/HIV reference centre from the State of Bahia, Brazil. Sociodemographic and clinical data were obtained from secondary data from patients' charts and laboratory findings, and analyzed using SPSS 20.0. Pearson's χ2 test or Fisher's exact test was used to evaluate categorical variables. HPV clinical diagnosis was considered positive as the presence of genital warts. Results: In this study, 849 patients were included ­ 196 men and 653 women. Of the sample, 51.4% was diagnosed with at least one of the two bacteria. The prevalence of Mycoplasma hominis infection was higher in coinfection (16.7%) than in isolated infection (2.2%). The prevalence of Ureaplasma urealyticum isolated infection was 32.4%. A strong association was found between the presence of genital warts and Ureaplasma urealyticum infection, with an estimated risk of 1.230 (p=0.014). Conclusion: Our findings suggest the need for further investigation for Ureaplasma urealyticum infection in patients presenting genital warts on physical examination. In addition, in this context, greater attention should be given to women and pregnant women.

Introdução: Ureaplasma urealyticum e Mycoplasma hominis são frequentemente encontrados no trato urogenital de homens e mulheres, e têm sido associados à ocorrência de uretrites não gonocócicas, cervicites, infertilidade, corioamnionite e outras patologias obstétricas. Alguns estudos mostraram alta prevalência de papilomavírus humano (HPV) em pacientes com uretrites não gonocócicas, bem como alta frequência de infecção por Ureaplasma urealyticum em mulheres com anormalidades na citologia cervical e homens apresentando verruga genital. Objetivos: Avaliar a prevalência da coinfecção por Ureaplasma urealyticum, Mycoplasma hominis e HPV em pessoas atendidas em um centro de referência de DST/HIV e identificar os fatores de risco associados. Métodos: Estudo transversal com pacientes maiores de 18 anos, testados para Ureaplasma urealyticum e Mycoplasma hominis entre 1º de julho e 31 de dezembro de 2015, em um centro de referência de DST/HIV da Bahia, Brasil. Os dados clínicos e sociodemográficos foram obtidos por coleta de dados secundários a partir dos prontuários e achados laboratoriais dos pacientes e analisados usando SPSS 20.0. O teste de χ2 Pearson ou teste exato de Fisher foram usados para avaliar as variáveis categóricas. O diagnóstico clínico do HPV foi considerado positivo quando houve presença de verruga genital. Resultados: Foram incluídos neste estudo, 849 pacientes, sendo 196 homens e 653 mulheres. Da amostra, 51,4% foi diagnosticada com infecção por pelo menos uma das duas bactérias. A prevalência de infecção por Mycoplasma hominis foi maior na coinfecção (16,7%) do que isoladamente (2,2%). A prevalência da infecção isolada por Ureaplasma urealyticum foi de 32,4%. Houve forte associação entre a presença de verruga genital e infecção por Ureaplasma urealyticum, com estimativa de risco de 1,230 (p=0,014). Conclusão: Nossos achados sugerem a necessidade de investigação adicional para a infecção por Ureaplasma urealyticum nos pacientes apresentando verruga genital ao exame físico. Além disso, nesse contexto, maior atenção deve ser dada a mulheres e gestantes.

Identificación de microorganismos vaginales en pacientes en trabajo de parto pretérmino / Identification of vaginal microorganisms of patients in preterm labor

Resumen OBJETIVO: Identificar los microorganismos vaginales más frecuentes en pacientes en trabajo de parto pretérmino, mediante el A.F. Genital System-Liofilchem®. MATERIALES Y MÉTODOS: Estudio descriptivo, prospectivo y transversal llevado a cabo en pacientes en trabajo de parto pretérmino atendidas en el servicio de Ginecología y Obstetricia de la Fundación Hospital Infantil Universitario de San José de Bogotá, entre julio de 2015 y febrero de 2016 de quienes se obtuvieron muestras de flujo del introito vaginal y se sembraron en el panel del A.F. Genital System-Liofilchem®, de acuerdo con las instrucciones del fabricante. Para el análisis de los datos se utilizó el programa estadístico Stata versión 13 (StataCorp®) y se implementó la prueba no paramétrica de Wilcoxon. RESULTADOS: Los microorganismos aislados con mayor frecuencia fueron: Staphylococcus aureus (89.1%), Ureaplasma urealyticum (43.4%) y Mycoplasma hominis (19.5%). De las muestras positivas para especies de micoplasma, 52.2% tuvo concentración mayor de 105 UFC/mL. De los agentes aislados, Ureaplasma urealyticum y Mycoplasma hominis mostraron resistencia de 100% para clindamicina y eritromicina, respectivamente. CONCLUSIONES: Los microorganismos vaginales representan un factor de riesgo de parto pretérmino. Ureaplasma urealyticum y Mycoplasma hominis muestran resistencia total a clindamicina y eritromicina.

Abstract OBJECTIVE: Determine the frequency of microorganisms present in the vagina of women in preterm labor. MATERIALS AND METHODS: Descriptive, prospective, cross-sectional study of a series of cases of 46 patients treated at the Fundación Hospital Infantil Universitario de San José de Bogotá for preterm labor, who were sampled from the vaginal introitus and planted on the A.F. Genital System-Liofilchem® panel. Genital System by Liofilchem®, according to the manufacturer's instructions. The statistical package Stata version 13 (StataCorp®) was used. The statistical analysis was descriptive, the nonparametric Wilcoxon test was run. RESULTS: The most isolated microorganism was Staphylococcus aureus with a frequency of 89.13%. Ureaplasma urealyticum was detected in 43.48% and Mycoplasma hominis in 19.57 Of the positive samples for genital Mycoplasmas, 52.2% showed a concentration >105 CFU/mL. Ureaplasma urealyticum isolates showed 100% resistance to clindamycin and 100% Mycoplasma hominis for erythromycin. CONCLUSIONS: Microorganisms that have been identified as risk factors for preterm delivery were identified in 93.5% of the vaginal discharge samples. For Ureaplasma urealyticum and Mycoplasma hominis, 100% resistance for clindamycin and erythromycin is identified.

Bacteriemia por Mycoplasma hominis: un agente etiologico subestimado / Mycoplasma hominis bacteremia: An underestimated etiological agent

Mycoplasma hominis es una bacteria de cultivo exigente y forma parte de la microbiota comensal de la zona urogenital en adultos. Puede ocasionar infecciones del tracto genitourinario, en particular en mujeres, e infecciones sistémicas en neonatos. Además, puede causar infecciones extragenitales graves, en especial en pacientes inmunocomprometidos. Describimos un caso de bacteriemia por M. hominis en una paciente inmunocompetente, luego de un legrado uterino por aborto incompleto. M. hominis está subestimado como agente etiológico de infecciones extragenitales debido a su difícil diagnóstico, ya que al carecer de pared celular no se visualiza por la coloración de Gram, requiere una incubación prolongada en atmósfera de anaerobiosis para su desarrollo y los métodos convencionales de detección pueden fallar. Este es el primer reporte de senal positiva en hemocultivo automatizado (BD BACTEC) con aislamiento de M. hominis.

Mycoplasma hominis is a fastidious bacterium, which usually colonizes the lower urogenital tract and may cause systemic infections in neonates and genital infections in adults. It can also be the cause of serious extra-genital infections, mainly in immunosuppressed or predisposed subjects. Case Presentation: We describe a case of bacteremia caused by M. hominis in a previously healthy woman after uterine curettage due to incomplete abortion. M. hominis could be an underestimated cause of bacteremia in immunocompetent patients. Mycoplasma organisms have fastidious growth requirements, are often difficult to culture on a cell-free medium and have no cell wall. The conventional method for detection may fail. This is the first report of M. hominis isolation from a positive automated blood culture (BD BACTEC, USA).

[Mycoplasma hominis bacteremia. An underestimated etiological agent]. / Bacteriemia por Mycoplasma hominis: un agente etiológico subestimado.

Mycoplasma hominis is a fastidious bacterium, which usually colonizes the lower urogenital tract and may cause systemic infections in neonates and genital infections in adults. It can also be the cause of serious extra-genital infections, mainly in immunosuppressed or predisposed subjects. CASE PRESENTATION: We describe a case of bacteremia caused by M. hominis in a previously healthy woman after uterine curettage due to incomplete abortion. M. hominis could be an underestimated cause of bacteremia in immunocompetent patients. Mycoplasma organisms have fastidious growth requirements, are often difficult to culture on a cell-free medium and have no cell wall. The conventional method for detection may fail. This is the first report of M. hominis isolation from a positive automated blood culture (BD BACTEC, USA).

Evaluación de la implementación de PCR Real Time en la detección de Chlamydia Trachomatis y Micoplasmas urogenitales en pacientes con diagnóstico de infertilidad / Evaluation of the implementation of Real Time PCR in the detection of Chlamydia Trachomatis and urogenital Mycoplasmas in patients diagnosed with infertility

La infección genital es una de las principales causas de infertilidad en la población mundial, afecta a cada una de las diversas partes de la anatomía reproductiva, tanto masculina como femenina. Los microrganismos de transmisión sexual como Neisseria gonorrohoeae (Ng), Trichomonas vaginalis (Tv), Chlamydia trachomatis (Ct), Mycoplasma hominis (Mh) y Ureoplasma urealiticum (Uu), se encuentran asociados a una disminución del potencial de fertilidad en parejas en edad reproductiva. Las infecciones por Neisseria gonorrrohoeae y Trichomonas vaginalis, generalmente tienen síntomas clínicos y con tratamientos adecuados son erradicadas completamente. Sin embargo, las infecciones por Chlamydia trachomatis, Mycoplasma hominis o Ureaplasma urealiticum, pueden transcurrir asintomáticas o subclínicas y por tal motivo la investigación de estos microorganismos está indicada antes de cualquier procedimiento y/o técnica de reproducción medicamente asistida. La Ct es una bacteria intracelular obligada que no crese en cultivos bacteriológicos convencionales. La infección en el hombre produce uretritis, prostatitis, epididimitis, que si no son tratadas a tiempo, pueden dañar al epitelio germinal del testículo ocasionando una infertilidad secundaria. En la mujer la infección inicialmente es en cuello uterino, con signos y síntomas de cervicitis, como ser, un cuello friable con secreción mucopurulenta y si la infección asciende del endocervix causando endometritis, salpingitis, abscesos tubaricos y peritonitis, se lo conoce como Enfermedad Inflamatoria Pélvica (EIP) pueden generar severas secuelas como infertilidad por obstrucción cicatrizal de las trompas y embarazos ectópicos. También puede afectar a la uretra femenina y causar piuria, disuria y aumento en la frecuencia urinaria. Las Ct son bacterias patógenas de transmisión sexual, vale decir que no forman parte de la flora habitual de la uretra distal del varón ni de la flora normal vaginal. En cambio los Mh y Uu, pueden estar como colonizantes de la uretra distal del hombre, y también pueden ser la causa de uretritis, prostatitis, epididimitis, cuyas consecuencias crónicas asintomáticas se conocen como obstrucciones tubaricas (conductos eferentes y deferentes, epidídimo), y sus efectos directos sobre el espermatozoide provocando enrollamiento de la cola espermática que afecta a su movilidad (aztenozoospermia). En la mujer, el Mh y el Uu, pueden colonizar la vagina o la uretra, y cuando la concentración de estos microorganismos se elevada y el microambiente lo permite suelen encontrarse asociadas a un "estado de vaginosis" o de una uretritis no gonocócica como es el caso de las infecciones por Uu. OBJETIVO Evaluar la utilidad de la reacción en cadena de la polimerasa en tiempo real (qPCR) en la detección de Ct, Mh y Uu en pacientes femeninos y masculinos que ingresen al laboratorio con un diagnóstico de infertilidad. Así como también, comprobar la posible asociación con otros parámetros evaluados en el laboratorio como el espermograma y el exudado vaginal. MATERIAL Y MÉTODO Se estudiaron 38 muestras de pacientes que ingresaron al laboratorio con diagnóstico de infertilidad (INFT); hisopados uretrales (hu), endocervicales (he), semen, orina de 1er porción, mediante qPCR. Las muestras endocervicales y vaginales se tomaron por duplicado con hisopos de dacrón, colocando uno de ellos en un tubo seco y el otro en medio conservante hasta el proceso de extracción de ADN (siguiendo algoritmo establecido para microrganismo de difícil crecimiento. Ver Anexo I, pág. 7). Todos los pacientes firmaron un consentimiento informado (CI) para el uso de sus resultados. Para la extracción del ADN bacteriano se utilizó un kit de extracción de ácidos nucleicos. RESULTADOS Del total de 38 pacientes de ambos sexos, dentro de un protocolo de fertilidad; 28 (74%) dieron no detectables y 10 (26%) detectables por qPCR. Siendo el Uu el de mayor frecuencia con un 80%, seguido de Mh con un 20%. Ct fue no detectable tanto en hombre como en mujeres. Los Uu fueron hallados en secreciones vaginales, las que al mismo tiempo fueron analizadas y evaluadas con el criterio de BACOVA en área de bacteriología clínica arrojando un Score +1 o +2, es decir, poseían un microbioma vaginal normal y equilibrado. DISCUSIÓN Dado el alto porcentaje de pacientes que dieron positivo para la detección de Uu, y que por métodos de bacteriología convencional no se obtuvo desarrollo; se recomienda el uso de qPCR en muestras de pacientes que se encuentran en el inicio de un protocolo de infertilidad y ante una inminente aplicación de técnicas de reproducción asistida

Micoplasmas urogenitales como causa de infertilidad femenina. Hospital Ginecobstétrico Provincial de Matanzas. 2014-2015 / Urogenital micoplasmas as a cause of feminine infertility. Provincial Gynecoobstetric Hospital of Matanzas. 2014-2015

Introducción: los micoplasmas urogenitales (Mycoplasma hominis, Ureaplasma urealyticum), a pesar de formar parte de la flora habitual de vagina, se encuentran entre las especies bacterianas más frecuentemente involucradas en la infertilidad de la pareja. Objetivos: determinar la incidencia de micoplasmas urogenitales en muestras de exudados endocervicales de pacientes, de la Consulta Provincial de Atención a la Pareja Infértil, clasificar la severidad de la infección detectada e identificar la sensibilidad-resistencia a diferentes antimicrobianos de los micro-organismos aislados Materiales y métodos: Se efectuó estudio observacional descriptivo transversal en el Hospital Gineco-Obstétrico Docente Provincial “Dr. Julio Rafael Alfonso Medina”, de Matanzas, entre los meses de noviembre de 2014-enero de 2015. El universo estuvo constituido por las 117 pacientes que cumplieron los criterios de inclusión y exclusión. Resultados: el 56,4 % de las muestras analizadas fueron positivas, siendo la especie más frecuente el Ureaplasma urealyticum. Predominaron las infecciones leves, en un 59,09 %. El síntoma más referido fue leucorrea con 42,73 %. Ureaplasma urealyticum mostró mayor resistencia frente a ofloxacino, con un 82 %. No se encontró resistencia de Mycoplasma hominis frente a los antimicrobianos usados en la investigación. Las coinfecciones fueron más resistentes a azitromicina (100 %), ofloxacino (90 %), y eritromicina (80 %). Conclusiones: el microorganismo más aislado fue Ureaplasma urealyticum. El síntoma más frecuente fue la leucorrea. Predominaron las infecciones leves. Ureaplasma urealyticum muestra mayor resistencia a los antimicrobianos que Mycoplasma hominis. Ambos son altamente sensibles a pefloxacino y minociclina.

Background: urogenital mycoplasmas (Mycoplasma hominis, Ureaplasma urealyticum) are among the most frequent bacterial species involved in the couple infertility, although they are part of the vagina regular flora. Aims: determining the incidence of urogenital mycoplasmas in the samples of endocervical exudates of patients of the Provincial Consultation of Attention to Infertile Couple; classifying the severity of the detected infection and; identifying the isolated microorganisms’ sensibility-resistance to different antimicrobials. Materials and methods: a cross-sectional, descriptive, observational study was carried out in the Provincial Teaching Gyneco-obstetric Hospital “Dr. Julio Rafael Alfonso Medina” of Matanzas in the period from November 2014 to January 2015. The universe was formed by the 117 female patients who fulfilled the criteria of inclusion and exclusion. Outcomes: 56.4 % of the analyzed samples were positive, being Ureaplasma urealyticum the most frequent specie. Light infections predominated, in 59.09 %. The most referred symptom was leucorrhea with 42.73 %. Ureaplasma urealyticum showed higher resistance toward ofloxacin, with 82 %. There was not resistance of Mycoplasma hominis toward the antimicrobials used in the research. The co-infections were more resistant to azythromycin (100 %), ofloxacin (90 %), and erythromycin (80 %). Conclusions: Ureaplasma urealyticum was the most isolated microorganism. The most frequent symptom was leucorrhea. Light infections predominated. Ureaplasma urealyticum shows higher resistance to antimicrobials than Micoplasma hominis. Both are highly sensible to pefloxacin and minocycline.

Utilidad de la reacción de polimerasa en cadena convencional para la detección de Mycoplasma hominis, Ureaplasma spp. y Trichomonas vaginalis en muestras genitales de mujeres en consulta ambulatoria / Usefulness of conventional polymerase chain reaction for the detection of Mycoplasma hominis, Ureaplasma spp. and Trichomonas vaginalis in female outpatient's genital samples

Introduction: Trichomonas vaginalis, Mycoplasma hominis and Ureaplasma spp. are microorganisms responsible for genitourinary and pregnancy pathologies. Nucleic acid amplification methods have shown several advantages, but have not been widely studied for the detection of these microorganisms. Aim: To implement a conventional polymerase chain reaction (PCR) for the detection of the microorganisms and to compare its results versus the methods currently used at our laboratory. Material and Methods: 91 available samples were processed by PCR, culture (M. hominis y Ureaplasma spp.) and wet mount (T vaginalis). Results were compared and statistically analyzed by kappa agreement test. Results: 85, 80 and 87 samples resulted in agreement for the detection of M. hominis, Ureaplasma spp. y T. vaginalis, respectively. For M. hominis and Ureaplasma spp., agreement was substantial, whereas for T. vaginalis it was moderate, however, for the latter, PCR detected more cases than wet mount. Conclusion: We recommend the implementation of PCR for detection of T. vaginalis whereas culture kit is still a useful method for the other microorganisms.

Introducción: Trichomonas vaginalis, Mycoplasma hominis y Ureaplasma spp. son microorganismos causantes de patología genito-urinaria y durante el embarazo. Los métodos de amplificación de ácidos nucleicos han demostrado numerosas ventajas, pero no han sido ampliamente estudiados para la detección de estos microorganismos. Objetivo: Implementar una reacción de polimerasa en cadena convencional (RPC) para su detección y comparar sus resultados con los métodos actuales de nuestro laboratorio. Material y Métodos: Se procesaron 91 muestras mediante RPC, cultivo (M. hominis y Ureaplasma spp.) y observación microscópica al fresco (T. vaginalis). Los resultados fueron comparados y analizados estadísticamente mediante el test de concordancia kappa. Resultados: 85, 80 y 87 muestras tuvieron resultados concordantes para la detección de M. hominis, Ureaplasma spp. y T. vaginalis, respectivamente. Para M. hominis y Ureaplasma spp. el nivel de concordancia fue considerable mientras que para T. vaginalis fue moderado; sin embargo, para esta última, la RPC detectó más casos que la microscopia al fresco. Conclusión: Se recomienda la implementación de la RPC para la detección de T. vaginalis. Para M. hominis y Ureaplasma spp. el kit de cultivo continúa siendo un buen método.

High rates of double-stranded RNA viruses and Mycoplasma hominis in Trichomonas vaginalis clinical isolates in South Brazil.

Trichomonas vaginalis is the etiological agent of trichomoniasis, the most common non-viral sexually transmitted disease (STD) in world, with 276.4 million new cases each year. T. vaginalis can be naturally infected with Mycoplasma hominis and Trichomonasvirus species. This study aimed to evaluate the prevalence of T. vaginalis infected with four distinct T. vaginalis viruses (TVVs) and M. hominis among isolates from patients in Porto Alegre city, South Brazil. An additional goal of this study was to investigate whether there is association between metronidazole resistance and the presence of M. hominis during TVV infection. The RNA expression level of the pyruvate ferredoxin oxidoreductase (PFOR) gene was also evaluated among metronidazole-resistant and metronidazole-sensitive T. vaginalis isolates. A total of 530 urine samples were evaluated, and 5.7% samples were positive for T. vaginalis infection. Among them, 4.51% were isolated from female patients and 1.12% were from male patients. Remarkably, the prevalence rates of M. hominis and TVV-positive T. vaginalis isolates were 56.7% and 90%, respectively. Most of the T. vaginalis isolates were metronidazole-sensitive (86.7%), and only four isolates (13.3%) were resistant. There is no statistically significant association between infection by M. hominis and infection by TVVs. Our results refute the hypothesis that the presence of the M. hominis and TVVs is enough to confer metronidazole resistance to T. vaginalis isolates. Additionally, the role of PFOR RNA expression levels in metronidazole resistance as the main mechanism of resistance to metronidazole could not be established. This study is the first report of the T. vaginalis infection by M. hominis and TVVs in a large collection of isolates from South Brazil.

Molecular mechanism of fluoroquinolones resistance in Mycoplasma hominis clinical isolates.

To evaluate the molecular mechanism of fluoroquinolones resistance in Mycoplasma hominis (MH) clinical strains isolated from urogenital specimens. 15 MH clinical isolates with different phenotypes of resistance to fluoroquinolones antibiotics were screened for mutations in the quinolone resistance-determining regions (QRDRs) of DNA gyrase (gyrA and gyrB) and topoisomerase IV (parC and parE) in comparison with the reference strain PG21, which is susceptible to fluoroquinolones antibiotics. 15 MH isolates with three kinds of quinolone resistance phenotypes were obtained. Thirteen out of these quinolone-resistant isolates were found to carry nucleotide substitutions in either gyrA or parC. There were no alterations in gyrB and no mutations were found in the isolates with a phenotype of resistance to Ofloxacin (OFX), intermediate resistant to Levofloxacin (LVX) and Sparfloxacin (SFX), and those susceptible to all three tested antibiotics. The molecular mechanism of fluoroquinolone resistance in clinical isolates of MH was reported in this study. The single amino acid mutation in ParC of MH may relate to the resistance to OFX and LVX and the high-level resistance to fluoroquinolones for MH is likely associated with mutations in both DNA gyrase and the ParC subunit of topoisomerase IV.

Molecular mechanism of fluoroquinolones resistance in Mycoplasma hominis clinical isolates

To evaluate the molecular mechanism of fluoroquinolones resistance in Mycoplasma hominis (MH) clinical strains isolated from urogenital specimens. 15 MH clinical isolates with different phenotypes of resistance to fluoroquinolones antibiotics were screened for mutations in the quinolone resistance-determining regions (QRDRs) of DNA gyrase (gyrA and gyrB) and topoisomerase IV (parC and parE) in comparison with the reference strain PG21, which is susceptible to fluoroquinolones antibiotics. 15 MH isolates with three kinds of quinolone resistance phenotypes were obtained. Thirteen out of these quinolone-resistant isolates were found to carry nucleotide substitutions in either gyrA or parC. There were no alterations in gyrB and no mutations were found in the isolates with a phenotype of resistance to Ofloxacin (OFX), intermediate resistant to Levofloxacin (LVX) and Sparfloxacin (SFX), and those susceptible to all three tested antibiotics. The molecular mechanism of fluoroquinolone resistance in clinical isolates of MH was reported in this study. The single amino acid mutation in ParC of MH may relate to the resistance to OFX and LVX and the high-level resistance to fluoroquinolones for MH is likely associated with mutations in both DNA gyrase and the ParC subunit of topoisomerase IV.

Molecular mechanism of fluoroquinolones resistance in Mycoplasma hominis clinical isolates

To evaluate the molecular mechanism of fluoroquinolones resistance in Mycoplasma hominis (MH) clinical strains isolated from urogenital specimens. 15 MH clinical isolates with different phenotypes of resistance to fluoroquinolones antibiotics were screened for mutations in the quinolone resistance-determining regions (QRDRs) of DNA gyrase (gyrA and gyrB) and topoisomerase IV (parC and parE) in comparison with the reference strain PG21, which is susceptible to fluoroquinolones antibiotics. 15 MH isolates with three kinds of quinolone resistance phenotypes were obtained. Thirteen out of these quinolone-resistant isolates were found to carry nucleotide substitutions in either gyrA or parC. There were no alterations in gyrB and no mutations were found in the isolates with a phenotype of resistance to Ofloxacin (OFX), intermediate resistant to Levofloxacin (LVX) and Sparfloxacin (SFX), and those susceptible to all three tested antibiotics. The molecular mechanism of fluoroquinolone resistance in clinical isolates of MH was reported in this study. The single amino acid mutation in ParC of MH may relate to the resistance to OFX and LVX and the high-level resistance to fluoroquinolones for MH is likely associated with mutations in both DNA gyrase and the ParC subunit of topoisomerase IV.(AU)

Hallazgos de Micoplasma hominis y Ureaplasma urealyticum en mujeres con infecciones urogenitales / Mycoplasma hominis and Ureaplasma urealyticum found in women with urogenital infections

Fundamento: los micoplasmas constituyen uno de los agentes bacterianos implicados en infecciones urogenitales. Entre las especies de importancia médica se citan: Ureaplasma urealyticum, Micoplasma genitalium, y Micoplasma hominis. Objetivo: determinar la frecuencia de micoplasmas en mujeres con infecciones urogenitales, las asociaciones con otros agentes biológicos y la resistencia antimicrobiana a drogas potentes. Método: el estudio se realizó en el laboratorio de Microbiología del Centro Provincial de Higiene, Epidemiología y Microbiología de Camagüey. El universo fue de 655 pacientes y la muestra de 135. Se utilizó el Kitt diagnóstico de micoplasma de los laboratorios italianos Lio FILCHEM. Resultados: se identificaron 135 aislamientos de micoplasma. Las especies más frecuentes fueron: Ureaplasma urealyticum (45,9 %), Micoplasma hominis (8,9 %), y el complejo Micoplasma hominis + Ureaplasma urealyticum (45,2 %). El 55,5 % de las especies tuvieron asociaciones con Chlamydias, Gardnerella vaginalis, Trichomonas vaginalis y levaduras. Hubo emergencia de cepas resistentes en mayor o menor grado frente a ofloxacina (33,3 %), eritromicina (17,8 %), claritromicina (11,2 %), y otros. Conclusiones: se comprobó la afectación de especies de micoplasma en las mujeres estudiadas, el poder de asociación de esta bacteria con otros patógenos genitales y la emergencia de cepas resistentes hacia antimicrobianos de uso terapéutico.

Background: mycoplasmas constitute one of the bacterial agents involved in the urogenital infections. Among the species of medical relevance can be mentioned Ureaplasma urealyticum, Mycoplasma genitalium, and Mycoplasma hominis. Objective: to determine the frequency of appearance of mycoplasmas in women with urogenital infections, the association with other biological agents, and the antimicrobial resistance to powerful drugs. Method: the study was conducted at the Microbiology Laboratory of the Provincial Center of Hygiene, Epidemiology, and Microbiology of Camagüey. The universe was composed of 655 patients and the sample included 135. The diagnostic kit of micoplasma from the Italian laboratories Lio FILCHEM was used. Results: 135 isolations of micoplasma were identified. The most frequent species were Ureaplasma urealyticum, Micoplasma hominis, and the complex of Micoplasma hominis + Ureaplasma urealyticum. The 55.5 % percent of the species were associated to Chlamydia, Gardnerella vaginalis, Trichomonas vaginalis and yeasts. There was an emergence of strains resistant to ofloxacin, erythromycin, clarithromycin and others in a greater or lesser degree. Conclusions: the affectation of mycoplasma species in the studied women, the power of association of these bacteria with other genital pathogen, and the emergence of resistant strains to therapeutic antimicrobial, were confirmed.