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.

Micoplasma hominis y Ureaplasma spp en mujeres que consultan por infertilidad / Mycoplasma Hominis and Ureaplasma Spp. in women attending to infertility consultations

RESUMEN Introducción: Las infecciones por micoplasmas y ureaplasmas pueden producir fallos en la reproducción y vincularse con problemas de infertilidad femenina. Objetivo: Determinar la frecuencia de infecciones por Micoplasma hominis y Ureaplasma spp en mujeres que consultan por infertilidad e identificar si existe asociación entre las infecciones detectadas y los antecedentes de infecciones de transmisión sexual y enfermedad inflamatoria pélvica, procederes ginecológicos y síntomas de infecciones. Métodos: Se realizó un estudio descriptivo transversal, para evaluar muestras de exudados endocervicales de 175 mujeres, con edades entre 20 y 45 años, provenientes de la consulta de infertilidad del Instituto Nacional de Endocrinología, entre junio de 2016 y enero de 2017. Para la detección de micoplasmas urogenitales se utilizó el juego de reactivos Myco Well D-One. Se tuvieron en cuenta los aspectos éticos y se utilizó la prueba Chi Cuadrado para evaluar la significación estadística de las posibles asociaciones. Resultados: De las 175 muestras evaluadas, 102 (58,1 por ciento) mostraron la presencia de infecciones, de ellas 65 correspondieron a Ureaplasma spp (37,1 por ciento), 11 a Micoplasma hominis (6,2 por ciento), y 26 a asociaciones de Micoplasma hominis y Ureaplasma spp (14,8 por ciento). Se identificó asociación entre las infecciones detectadas y la presencia de antecedentes de infecciones de transmisión sexual y enfermedad inflamatoria pélvica, no así con relación a los procederes ginecológicos y síntomas de infecciones. Conclusiones: La frecuencia total de infecciones fue relativamente alta y la especie más frecuente el Ureaplasma spp. Las infecciones detectadas estuvieron asociadas a algunos de los factores estudiados(AU)

ABSTRACT Introduction: Infections caused by Mycoplasmas and Ureaplasmas may result in faults in the reproduction process and can be linked to female infertility. Objective: To determine the frequency of infection by Mycoplasma hominis and Ureaplasma spp. in women who attend to infertility consultations and if these are associated with a history of sexually transmitted infections and pelvic inflammatory disease, gynaecological procedures and symptoms of infections. Methods: A descriptive cross-sectional study was conducted to evaluate samples of endocervical swabs of 175 women between the ages of 20 to 45 years, from the Infertility consultation of the National Institute of Endocrinology, during June 2016 to January 2017. For the detection of urogenital mycoplasmas it was used the reagents kit Myco Well D-One. There were taken into account the ethical aspects and it was used the chi-square test to assess the statistical significance of the possible associations. Results: Of the 175 evaluated samples, 102 (58.1 percent) showed the presence of infections, 65 of them corresponded to Ureaplasma spp (37.1 percent), 11 to Mycoplasma hominis (6.2 percent), and 26 associations of Mycoplasma hominis and Ureaplasma spp (14.8 percent). It was identified association between the detected infections and the presence of a history of sexually transmitted infections and pelvic inflammatory disease, but not with the gynaecological procedures and the symptoms of infections. Conclusions: The total frequency of infection was relatively high and the most prevalent specie was the Ureaplasma spp. The detected infections were associated with some of the factors studied(AU)

Identificación de especies de Mycoplasma y de Ureaplasma diversum en rodeos lecheros de Argentina / Identification of species of Mycoplasma and Ureaplasma diversum from Argentinian dairy herds

Varias especies de Mycoplasma y Ureaplasma diversum pueden causar enfermedades en el ganado bovino lechero, asociadas o no a manifestaciones clínicas. En nuestro país, ha sido detectada la presencia de solo tres especies de este grupo hasta el momento: Mycoplasma bovis, Mycoplasma californicum y Mycoplasma canadense. El objetivo del presente trabajo fue identificar otras especies de la familia Mycoplasmataceae. Se estudiaron treinta y cinco aislamientos compatibles con Mycoplasma spp. obtenidos a partir de diferentes muestras de bovinos, con o sin sintomatología clínica, provenientes de ocho rodeos ubicados en las provincias de Santa Fe, Córdoba, Buenos Aires y San Luis. Mediante el uso de reacciones en cadena de la polimerasa (PCR) específicas de especie se identificaron Mycoplasma bovigenitalum, Mycoplasma alkalescens, Mycoplasma bovirhinis y U. diversum, y mediante la amplificación y posterior secuenciación del espacio intergénico 16-23S ARNr se identificaron Mycoplasma arginini y M. californicum. La identificación de estas especies por primera vez en nuestro país es un hecho de Argentina relevancia, que representa un importante avance en el conocimiento para incluir estos patógenos en el diagnóstico diferencial de determinadas entidades clínico-patológicas de los bovinos de Argentina.

Several species of Mycoplasma and Ureaplasma diversum can cause diseases in dairy cattle, which can be associated or not with clinical manifestations. In our country, the presence of Mycoplasma bovis, Mycoplasma californicum and Mycoplasma canadense has been detected, being the only mycoplasma species identified so far. The objective of this study was to identify other species of the Mycoplasmataceae family. Thirty-five Mycoplasma spp.-like isolates obtained from different samples from cattle, with or without clinical symptoms, from eight herds located in the provinces of Santa Fe, Cordoba, Buenos Aires and San Luis were utilized in the present study. Through the use of species-specific polymerase chain reactions (PCR) Mycoplasma bovigenitalium, Mycoplasma alkalescens, Mycoplasma bovirhinis and U. diversum were identified and through amplification and further sequencing of the 16-23S rRNA intergenic spacer regions, Mycoplasma arginine and M. californicum were identified. The identification of these species represents an important advance in knowledge in order to include these pathogens in the differential diagnosis of certain clinical and pathological entities of cattle from Argentina.

Assessment of cow and farm level risk factors associated with Ureaplasma diversum in pasture-based dairy systems - A field study

ABSTRACT Potential risk factors for Ureaplasma diversum in the vaginal mucus of 1,238 dairy cows were included in a multivariate logistic regression model, based on the cow level (i.e., granular vulvovaginitis [+GVV], yearly milk production [4500 kg or more], pregnancy, predominance of Bos taurus [+Bos Taurus], score of corporal condition [at least 2.5], concomitant positivity for Escherichia coli [+E.coli]), and farm level i.e., milking room hygiene (-Milking room), dunghill location, and replacement female). Ureaplasma diversum was present in 41.1% of the samples. Independent risk factors for U. diversum were +GVV (odds ratio [OR], 1.31); +Mycoplasma spp (OR, 5.67); yearly milk production (4500 kg or more) (OR, 1.99); +Bos taurus (OR, 1.68); +E. coli (OR, 4.96); -milking room (OR, 2.31); and replacement females (OR, 1.89). Ureaplasma diversum vaginal colonization was strongly associated with Mycoplasma spp., E. coli, and number of pregnant cows.

Molecular characterization of ureaplasmas isolated from reproductive tract of goats and sheep from Brazil

Abstract Ovine/caprine ureaplasmas have not yet been assigned a species designation, but they have been classified into nine serotypes. Herein ureaplasmas were searched for in 120 samples of vulvo vaginal mucous from sheep and 98 samples from goats at 17 farms. In addition, semen samples were collected from 11 sheep and 23 goats. The recovered ureaplasma were from sheep and goats from animals without any reproductive disorder symptoms, but not all animals presented positive cultures. In sheep, 17 (68%) cultures of vulvovaginal mucous were positive for ureaplasma and 11 (27%) samples of semen presented positive cultures in animals with clinical signs of orchitis, balanoposthitis or low sperm motility. In goats four ureaplasma isolates were obtained from vulvovaginal mucus, but the semen samples were all negative. The isolates were submitted to Pulsed-field gel electrophoresis methodology and their 16S rRNA genes were sequenced. Fifty percent of ureaplasma recovered from sheep allowed for PFGE typing. Eleven isolates showed eight profiles genetically close to the bovine ureaplasmas. The 16S rRNA gene sequencing showed differences or similarities of isolates from sheep and goats, and the reference strains of bovine and human ureaplasma. Four clinical isolates from sheep were grouped separately. The studied ureaplasma isolates showed to be a diverse group of mollicutes.

Detección molecular de agentes infecciosos de transmisión sexual en un grupo de hombres sintomáticos y su relación con la conducta sexual / Molecular detection of sexually transmitted agents in a symptomatic group of men and its relationship with sexual behavior

Background: Sexually transmitted infections (STIs) affect sexual and reproductive health of millions of men. Pathogens such as human papillomavirus (HPV), herpes simplex virus type 1 and 2 (HSV-1 y HSV-2), Chlamydia trachomatis,Mycoplasmagenitalium,Mycoplasma hominis and Ureaplasma urealyticum are associated with STIs. Aim: To detect pathogens associated with STIs in symptomatic men and its relationship with sexual behavior. Methodology: DNA was obtained from exfoliated cells of penis from 20 symptomatic men. Pathogens were detected using qPCR or PCR followed by reverse line blot. Sexual behavior was evaluated through a survey. Results: Two or more infectious agents were detected in 50% of samples. U. urealyticum was found in 25%, meanwhile C. trachomatis and M. hominis were detected in 15%. VHS-1, VHS-2 andM. genitalium were detected only in 5%. HPV was found in all samples. The most frequent HPV genotypes were VPH 16, 11, 70. There were no statistical link found between sexual behavior and the studied microorganisms Conclusion: Infectious agents associated with STIs were detected in symptomatic men. HPV was the most frequent pathogen and it was detected in multiple genotypes. It is necessary to increase the sample size to associate significantly the sexual behavior with the results.

Introducción: Las infecciones de transmisión sexual (ITS) afectan la salud sexual y reproductiva de millones de hombres. Patógenos como virus papiloma humano (VPH), virus herpes simplex (VHS-1 y VHS-2), Chlamydia trachomatis, Mycoplasma genitalium, Mycoplasma hominis y Ureaplasma urealyticum están asociados a ITS. Objetivo: Detectar patógenos asociados a ITS en hombres sintomáticos y relacionarlos con su conducta sexual. Metodología: Se obtuvo ADN de exfoliado celular del pene de 20 hombres sintomáticos de ITS. Los patógenos fueron detectados por RPC cuantitativa o RPC seguida de reverse line blot. La conducta sexual se evaluó mediante una encuesta. Resultados: En 50% de las muestras se detectaron dos o más agentes infecciosos; U. urealyticum fue detectado en 25% de los casos, mientras que C. trachomatis y M. hominis en 15%. VHS-1, VHS-2 y M. genitalium sólo en 5%. VPH se encontró en todas las muestras y los genotipos más frecuentes fueron VPH 16, 11, 70. No se encontró relación estadística entre los microorganismos estudiados y la conducta sexual de los encuestados. Conclusión: Se detectaron agentes infecciosos asociados a ITS en hombres sintomáticos, siendo VPH el más frecuente y encontrándose en múltiples genotipos. Es necesario aumentar el tamaño de muestra para asociar significativamente la conducta sexual a los resultados.

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.

Apoptosis in HEp-2 cells infected with Ureaplasma diversum

BACKGROUND: Bacterial pathogens have many strategies for infecting and persisting in host cells. Adhesion, invasion and intracellular life are important features in the biology of mollicutes. The intracellular location ofUreaplasma diversum may trigger disturbances in the host cell. This includes activation or inhibition of pro and anti-apoptotic factors, which facilitate the development of host damage. The aim of the present study was to associate U. diversum infection in HEp-2 cells and apoptosis induction. Cells were infected for 72hs with four U. diversum clinical isolates and an ATCC strain. The U. diversuminvasion was analyzed by Confocal Laser Scanning Microscopy and gentamicin invasion assay. The apoptosis was evaluated using pro-apoptotic and anti-apoptotic gene expression, and FITC Annexin V/Dead Cell Apoptosis Kit. RESULTS: The number of internalized ureaplasma in HEp-2 cells increased significantly throughout the infection. The flow cytometry analysis with fluorochromes to detect membrane depolarization and gene expression for caspase 2, 3 and 9 increased in infected cells after 24 hours. However, after 72 hours a considerable decrease of apoptotic cells was observed. CONCLUSIONS: The data suggests that apoptosis may be initially induced by some isolates in association with HEp-2 cells, but over time, there was no evidence of apoptosis in the presence of ureaplasma and HEp-2 cells. The initial increase and then decrease in apoptosis could be related to bacterial pathogen-associated molecular pattern (PAMPS). Moreover, the isolates of U. diversum presented differences in the studied parameters for apoptosis. It was also observed that the amount of microorganisms was not proportional to the induction of apoptosis in HEp-2 cells.

Detection of Ureaplasma spp. in semen samples from sheep in Brazil

A study was conducted to verify the presence of mycoplasmas and ureaplasmas DNA in sheep semen samples from the State of Pernambuco. The PCR assay was conducted of according with standard protocols with generic primers. Mollicutes DNA was detected in 26.0% and Ureaplasma spp. in 12.0% of semen samples.

Infecciones de transmisión sexual por Ureaplasma urealyticum y Mycoplasma hominis / Sexually-transmitted infections in Ureaplasma urealyticum and Mycoplasma hominis

Ureaplasma urealyticum y Mycoplasma hominis son bacterias anaerobias facultativas, que se asocian con la colonización e infección genital en los seres humanos adultos, aunque también pueden aislarse en individuos asintomáticos, lo que sugiere que pueden comportarse como patógenos oportunistas. El objetivo de este trabajo consistió en determinar la prevalencia de estos microorganismos en pacientes consultantes al Servicio de Enfermedades de Transmisión Sexual y Sida del Instituto Nacional de Epidemiología de la ciudad de Mar del Plata. Se realizó un estudio epidemiológico de tipo retrospectivo de 312 muestras de pacientes adultos de ambos sexos consultantes entre los años 2001 y 2006. La distribución por sexos fue 67.3% mujeres y 32.7% hombres, con edades comprendidas entre 15 y 71 años. La prevalencia de infección por Ureaplasma urealyticum fue de 51.9 % y la de Mycoplasma hominis de 25.6 %. La detección de estos microorganismos en forma simultánea para un mismo paciente fue de 30.2 %. La doxiciclina fue el antibiótico que mayor sensibilidad presentó frente a estos aislamientos. Candida albicans resultó el microorganismo más asociado a la presencia concomitante de U. urealyticum y M. hominis. De acuerdo con los datos obtenidos resulta necesario intensificar la detección de estos microorganismos potencialmente patógenos para monitorear su circulación en la población...

Ocorrência de Mollicutes e Ureaplasma spp. em surto de doença reprodutiva em rebanho bovino no Estado da Paraíba / Occurrence of Mollicutes and Ureaplasma spp. in outbreak of reproductive disease in cattle herds, State of Paraíba, Brazil

Em março de 2012 foi diagnosticado um surto de doença reprodutiva em rebanho bovino no Estado da Paraíba, Brasil. Foram examinadas 32 vacas e dois touros da raça Girolando. As vacas apresentaram sinais de doença reprodutiva como repetição de cio, vulvovaginite granular, infertilidade e abortos. As amostras de suabes vaginais e prepuciais foram colhidas e submetidas a isolamento bacteriano e PCR. As reações da PCR para Mollicutes e Ureaplasma spp. foram realizadas com os iniciadores MGSO-GPO3 e UGP'F-UGP'R, respectivamente. Na Nested PCR para Ureaplasma diversum, os iniciadores usados foram UD1, UD2, UD3 e UD4. Para isolamento bacteriano, as amostras foram diluídas de 10-1 até 10-5, semeadas em meio "UB", líquido e placa, sendo incubadas por até 21 dias a 37ºC em jarra de microaerofilia. A frequência de Mollicutes detectada na PCR foi de 65,6% e para Ureaplasma spp. foi de 50,0%, enquanto que para U. diversum foi de 15,6%. No isolamento a frequência de Mollicutes foi de 57,1% e para Ureaplasma spp. foi de 28,6%. No ágar "UB" foi visualizado o crescimento misto de Mycoplasma spp. e Ureaplasma spp. em seis amostras. Foi confirmado o envolvimento de micro-organismos da Classe Mollicutes em surto de doença reprodutiva em vacas no sertão paraibano.

In March of 2012 was investigated a reproductive disease outbreak in cattle herds from Paraíba State, Brazil. Were examined 32 cows and two bulls Giroland breed. The cows showed signs and symptoms of reproductive failure such as repeat breeding, granular vulvovaginitis, infertility and abortions. Vaginal and preputial mucous samples were collected for analysis by PCR and isolation. The PCR reactions for Mollicutes and Ureaplasma spp. were realized with primers MGSO and GPO3, and UGP'F and UGP'R respectively. The nested PCR assay for Ureaplasma diversum was realized with primers UD1, UD2, UD3 and UD4. For bacteriologic isolation, obtained samples were diluted up to 10-1 at 10-5, inoculated into liquid and solid "UB" medium, and incubated for up to 21 days, at 37ºC in microaerophilie jar. In the PCR reactions the frequency of Mollicutes detected in the analyzed vaginal mucous samples was 65.6, for Ureaplasma spp. was 50.0, while for U. diversum was 15.6. The frequency for isolation of Mollicutes was of 57.1 and for Ureaplasma spp. was of 28.6. In the UB agar was visualized growth of Mycoplasma spp. and Ureaplasma spp., associated in six of the samples. In the cows the presence of Mollicutes and Ureaplasma spp. was confirmed for the reproductive disease outbreak in the semiarid region of Paraiba.

Mycoplasmateceae species are not found in fallopian tubes of women with tubo-peritoneal infertility

BACKGROUND: The role of mycoplasmas on the development and sequelae of pelvic inflammatory disease remains controversial. The objective of the present study is to correlate directly the presence of Mycoplasmateceae through polimerase chain reaction (PCR) determinations in cervix and Fallopian tubes of infertile patients with tubo-peritoneal factor diagnosed through laparoscopy. METHODS: Thirty patients with tubo-peritoneal infertility and 30 normal fertile patients were included in the study; cervical samples and tubal flushings were obtained during laparoscopy. PCR determinations for the detection of genetic material of Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealiticum, Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis in cervix and tubal flushings were performed. RESULTS: No Mycoplasmataceae species as "only" microorganisms were found in tubal flushings of tubo-peritoneal infertility patients, whereas three (10%) fertile patients with normal tubes were positive for mycoplasma presence. This difference was not significant (p = 0.237). Among the 30 patients suffering from tubal infertility diagnosed through laparoscopy, Mycoplasmatecae species were not detected in the Fallopian tubes by PCR determinations, while in normal tubes from fertile patients these and other microorganisms could be found without distorting tubal anatomy. CONCLUSION: Mycoplasmateceae species were not detected in Fallopian tubes of women with tubo-peritoneal infertility.

Avaliação da taxa de isolamento de micoplasmas do trato urogenital por um método comercial em comparação com meios de cultura produzidos no próprio laboratório / Assessing the rate of isolation of mycoplasma treatment of urogenital by a business method in compared with media culture produced in itself laboratory

Este trabalho procurou avaliar a taxa de isolamento de Mycoplasma hominis e Ureaplasma urealiticum/U. parvum emamostras de conteúdo vaginal de 84 mulheres atendidas em um laboratório de analises clínica de Blumenau – SC, por dois meios de cultura de procedência diferentes. Os resultados demonstraram 50,0 % de positividade para Ureaplasma urealiticum/U. parvum e 20,24% de positividade para Micoplasma hominis por cultura em meio feito em nosso laboratório (P< 0,0001) e 16,67% de positividade para U. urealiticum e 8,33% de positividade para M. hominis por cultura em meio comercial (P< 0,0001). As amostras que foram cultivadas por meio próprio e obtiveram resultado positivo foram também analisadas por diluições seriadas para identificar a quantidadedestes microrganismos expresso em C.C.U. (color changing units). Os resultados demonstraram que 52,94% destas amostras apresentaram C.C.U. maior que 104 para M. hominis e 42,86% apresentaram C.C.U. maior que 103 para U. urealyticum, sendo estesos níveis considerados clinicamente significativos para que a presença destes microrganismos seja implicada na etiologia dos sintomas. No presente trabalho discutiu-se as possíveis causas desta diferença de positividade entre os dois métodos utilizados.

Detecção de Mycoplasma spp. e Ureaplasma diversum em vacas com distúrbios reprodutivos / Detection of Mycoplasma spp. and Ureaplasma diversum in cown with reproductive disorders

Foram utilizadas 112 amostras de muco vulvovaginal, coletados de vacas com distúrbios reprodutivos, para pesquisa de Mycoplasma e Ureaplasma. Para isolamentos, foram usados meios específicos para micoplasmas (SP-4) e para ureaplasmas. PCR genérica, PCR específica para Mycoplasma bovis e nested-PCR em tubo único para Ureaplasma diversum foram realizados com os DNAs extraídos das amostras. Mycoplasma spp. e U. diversum foram detectados em 12,5 e 25,0 por cento, respectivamente. A PCR genérica resultou em reações positivas em 63,4 por cento das amostras transportadas em SP-4 e em 69,6 por cento das transportadas em meio de ureaplasma. M. bovis foi detectado, na PCR específica, em 9,8 por cento das amostras e U. diversum, na nested-PCR, em 37,5 por cento. Houve maior sensibilidade na metodologia da PCR quando comparada à técnica de cultivo para Mycoplasma e Ureaplasma

In the study, 112 samples of vulvovaginal mucus of cows bearing reproductive disturbance were investigated for Mycoplasma and Ureaplasma. Specific media for the culture of mycoplasmas (SP-4) and ureaplasmas were used. PCR with general primers, PCR specific for Mycoplasma bovis, and nested-PCR in a single tube for Ureaplasma diversum were performed to detect DNA of the sample. Mycoplasma spp. and U. diversum were isolated in 12.5 and 25.0 percent, respectively. With generic PCR, positive reaction was obtained in 63.4 percent of the samples transported in SP-4 and 69.6 percent in ureaplasma medium. M. bovis was detected in 9.8 percent of samples and nested-PCR in a single tube for U. diversum resulted in 35.0 percent of positive reaction. Results demonstrated increased sensitivity of PCR methodology compared with culture technique applied to the search of microorganisms of Mycoplasma and Ureasplasma genera

Diagnóstico de Mycoplasma hominis, Ureaplasma parvum y Ureaplasma urealyticum en pacientes con vaginosis bacteriana / Diagnosis of Mycoplasma hominis, Ureaplasma parvum y Ureaplasma urealyticum in patients with bacterial vaginosis

Se realizó un estudio observacional descriptivo, para determinar la frecuencia de aislamientos de Mycoplasma hominis, Ureaplasma parvum y Ureaplasma urealyticum en pacientes con vaginosis bacteriana, donde se incluyeron 296 pacientes con secreción vaginal, atendidas en consultas médicas en 2 hospitales. El diagnóstico se llevó a cabo según los criterios Amsel, y a las mujeres positivas a esta enfermedad se les tomó muestra de exudado endocervical para el diagnóstico por métodos convencionales para M. hominis y Ureaplasma spp. Por la reacción en cadena de la polimerasa se identificaron U. parvum y U. urealyticum. Se diagnosticó vaginosis bacteriana en 30,1 por ciento de las pacientes, y en 77,5 por ciento se detectó la presencia de los micoplasmas urogenitales estudiados. M. hominis fue la especie más frecuente encontrada (71 por ciento), mientras que U. parvum y U. urealyticum se identificaron en 23,2 y 5,8 por ciento, respectivamente. En las mujeres con vaginosis bacteriana se debe realizar el diagnóstico de micoplasmas y ureaplasmas, lo que permitiría instaurar un adecuado control terapéutico, y ayudaría a evitar futuras enfermedades en el tracto genital.

An observational descriptive study to determine the frequency of Mycoplasma hominis, Ureaplasma parvum and Ureaplasma urealyticum isolates in patients with bacterial vaginosis was carried out in 296 patients who had vaginal secretion and were seen at two hospitals. The diagnosis was based on Amsel´s criteria. Endocervical swabs were taken from women positive to this disease for M. hominis and Ureaplasma spp. diagnosis by traditional methods. Polymerase chain reaction identified U. parvum and U. urealyticum. Bacterial vaginosis was diagnosed in 30.1 percent of females, and in 77.5 percent of them the studied urogenital mycoplasmas were present. M. hominis was the most common species (71 percent) whereas U. parvum and Urealyticum were detected in 23.2 percent and 5.8 percent of cases respectively. The diagnosis of Mycoplasmas and ureaplasmas should be performed in females with bacterial vaginosis, which will allow applying adequate therapeutic control and avoiding future pathologies in the genital tract.

Isolates of Ureplasma diversum genotyped by single-enzyme amplified length polymorphism

Isolates of Ureaplasma diversum recovered from bovines with reproductive disorders and healthy ones of four premises were compared by SE-AFLP. Twenty-eight SE-AFLP profiles without monomorphic fragments were obtained. The ureaplasma studied were divided in clusters A and B. Cluster A was divided in subclusters A1 and A2, while A1 was divided in subclusters A1a and A1b. Cluster B grouped only the reference strains. The clusters obtained were not associated with the reproductive disorders. The dendrogram obtained showed high heterogeneity among the studied ureaplasmas and indicated a low genomic stability as detected in other species of microorganisms of class Mollicutes.

Cepas de referência e 30 estirpes de Ureaplasma diversum isoladas do muco vaginal de bovinos apresentando ou não distúrbios reprodutivos, de quatro diferentes propriedades, foram comparadas por meio da metodologia da SE-AFLP (single-enzyme amplified fragment length polymorphism). Foram obtidos 28 perfis, com ausência de fragmentos monomórficos. No dendrograma, as amostras foram divididas em grupos A e B. O grupo A foi subdividido em A1 e A2 e o A1 dividiu-se em A1a e A1b. As amostras de referência formaram o grupo B. Não houve diferenciação entre as estirpes isoladas de animais doentes ou sadios. Evidenciou-se grande heterogeneidade entre os ureaplasmas estudados indicando baixa estabilidade genômica, como detectado em outras espécies dos microrganismos da classe Mollicutes.

Aplicación de método molecular en la detección de Mycoplasma genitalium en hombres y en mujeres embarazadas / Molecular detection of Mycoplasma genitalium in men and pregnant women

Mycoplasma genitalium es un patógeno oportunista del tracto genital. En el hombre es causa de uretritis, en tanto que en mujeres ha sido implicado en la etiología de cervicitis y de enfermedad inflamatoria pelviana (EIP). El objetivo de este estudio fue determinar la prevalencia de M. genitalium en pacientes masculinos con uretritis y en muestras vaginales de mujeres embarazadas. Se obtuvo muestras de secreción uretral en 37 pacientes con uretritis y de muestras vaginales de 50 consecutivas mujeres embarazadas, determinándose la presencia de M. genitalium mediante reacción de polimerasa en cadena (RPC). Las muestras de secreción uretral fueron también evaluadas en busca de Chlamydia trachomatis, Neisseria gonorrhoeae y Ureaplasma sp en tanto que en las de origen vaginal se investigó la microbiota y presencia de micoplasmas de tipo genital. Veintitrés casos fueron clasificados como uretritis no gonocóccica (UNG) y 14 como enfermedad gonocóccica. M. genitalium fue detectado en 3 de 23 (13,04 por ciento) varones con UNG; en dos casos asociado a Ureaplasma sp, y en un paciente como agente único. C. trachomatis fue detectado en 7 pacientes con UNG y en uno con gonorrea. Ureaplasma sp fue aislado en 13 (35,1 por ciento) pacientes, 8 casos de UNG y en 5 con gonorrea. El microorganismo fue detectado también en 6 (15 por ciento) de 40 mujeres; en 5 casos en presencia de microbiota normal (score de Nugent 0-3), y en un caso en presencia de vaginosis bacteriana. Ureaplasma spp fue aislado en las seis muestras positivas. En conclusión, este estudio demuestra que M. genitalium debe ser también considerado en la etiología de la UNG así como en el tracto genital inferior en la mujer embarazada, en presencia de una microbiota vaginal normal.

Ureaplasma spp. sindromas clínicos e diagnóstico laboratorial / Ureaplasma spp. clinical syndromes and laboratory diagnosis

Os ureaplasmas continuam a ser um grupo de microrganismos subvalorizado pela maioria dos profissionais de saúde, incluindo os responsáveis pelo seu diagnóstico. As principais causas desta situação são o seu crescimento fastidioso, a falta de meios comerciais e a ausência de procedimentos para um diagnóstico rápido. Este artigo pretende ser uma revisão sobre o papel dos ureaplasmas nas doenças humana e dos métodos mais utilizados no seu diagnóstico, incluindo as tecnologias mais recentes, indicando também quais os produtos mais adequados.

Mycoplasmas hominis y ureaplasma urealyticum en diferentes patologías ginecológicas / Mycoplasma hominis and ureaplasma urealyticum in different gynecological pathologies

Se investigo la frecuencia de Mycoplasma hominis (M.hominis) y Ureaplasma urealyticum (U.urealyticum) en muestras obtenidas del canal endocervical de 113 mujeres que resultaron por diferentes patologías ginecológicas. Cuarenta y siete pacientes (42 por ciento) resultaron positivas a la presencia de estos dos microorganismos, siendo 26 casos (23 por ciento) positivos a M. hominis y 21 (19 por ciento; p=NS) a U. urealyticum. La edad promedio fue de 32,1 ñ

Factores de riesgo para la colonización por micoplasmas genitales / Risk factors for colonization with genital mycoplasmas

Con el propósito de determinar los factores de riesgo para colonización por micoplasmas genitales, se procesaron 93 muestras de origen urogenital, provenientes de pacientes que acudieron a la Consulta de Microbiología de la Policlínica San Luis de Maracaibo entre marzo y diciembre de 1997. El cultivo, aislamiento y recuento de estos microorganismos, se realizó de acuerdo con la metodología recomendada por Biomerieux, a través de su equipo comercial, Mycoplasma-Lyo. Las frecuencias relativas de micoplasmas genitales fueron comparadas a través de la prueba de "z" para determinar diferencias entre proporciones de la población, con un porcentaje de significacia del 5 por ciento (alfa=0,05). Se obtuvo una proporción de 39,92 por ciento para M.hominis en mujeres y del 4.30 por ciento y 21.54 por ciento para U.urealyticum en el sexo masculino y femenino, respectivamente. No hubo aislamiento de M.fermentans. La edad, el sexo, la actividad sexual (sobre todo si es de inicio precoz), la promiscuidad sexual, la raza, el nivel socioeconómico y abortos previos, constituyen factores de riesgo para la colonización genital por micoplasmas; a diferencia de la anticoncepción, embarazos previos, instrumentación, alteraciones bioquímicas, fisiológicas y/o funcionales, así como los tratamientos previos, que al parecer, no son factores predisponentes para la colonización por estos microorganismos