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.

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.

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.

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

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