Molecular detection of Mycoplasma suis in extensive pig production systems in the State of Maranhão, northeast Brazil / Detecção molecular de Mycoplasma suis em suínos de criações extensivas no estado do Maranhão, nordeste do Brasil

Abstract Mycoplasma suis is a bacterium that causes hemoplasmosis in pigs. This agent is capable of adhering to the surface of porcine erythrocytes, inducing structural changes on these cells. In Brazil, there are few reports about the disease, its causal agent, and the economic impact of this pathogen on pig production systems and farm sanitation. The present study aimed to investigate the occurrence of M. suis in extensive swine farms located in the counties of Itapecuru Mirim, Santa Rita and Rosario, State of Maranhão, northeast Brazil. For such purpose, 64 blood samples of pigs from these facilities were tested for M. suis using a 16S rRNA gene-based quantitative real-time PCR (qPCR); 82.3%, 65.2% and 25% of blood samples of swine from farms in the cities of Itapecuru Mirim, Santa Rita and Rosario were positive for M. suis by qPCR, respectively. This study shows, for the first time, that M. suis circulates in pig populations from the state of Maranhão, Northeast Brazil.

Resumo Mycoplasma suis é uma bactéria que causa a hemoplasmose em suínos. Este agente é capaz de se aderir à superfície dos eritrócitos de suínos, ocasionando deformações estruturais nestas células. No Brasil, poucos são os relatos acerca do parasita, da infecção e de seus impactos econômicos nas esferas produtiva e sanitária. O objetivo deste estudo foi investigar, por meio da PCR em tempo real quantitativa (qPCR) baseada no gene 16S rRNA, a ocorrência de M. suis em 64 amostras de sangue de suínos de criações extensivas dos municípios de Itapecuru Mirim, Santa Rita e Rosário, localizados no estado do Maranhão. Foram obtidos um percentual de 82,3%, 65,2% e 25% de amostras positivas na qPCR para M. suis nos municípios de Itapecuru Mirim, Santa Rita e Rosário, respectivamente. Este estudo mostra que M. suis circula entre os suínos de criações extensivas no estado do Maranhão.

Mycoplasma pneumoniae: manifestaciones neurológicas y controversias diagnósticas. A propósito de un caso / Mycoplasma pneumoniae: neurologic manifestations and diagnostic controversies. A case report

Mycoplasma pneumoniae (Mp) es el agente causal de un 30% de las manifestaciones respiratorias de la población general. La neumonía ocupa el primer lugar dentro de este grupo. Las manifestaciones neurológicas representan las formas más frecuentes de presentación clínica extrapulmonar (40%). Las encefalitis y meningoencefalitis son las formas más habituales de sintomatología neurológica asociada a infección por Mp. La presentación de más de una variante clínica en un mismo paciente asociada a primoinfección por Mp es posible. El diagnóstico serológico plantea, habitualmente, controversias en su interpretación. A partir del caso de una niña de 7 años con inyección conjuntival, adenopatía cervical, rash descamativo y fotofobia con "pseudoedema de papila bilateral", que desarrolla durante su evolución parálisis facial periférica y meningitis aséptica, se analizarán las controversias que se plantean en relación con la interpretación diagnóstica asociada al compromiso neurológico por Mp.

Mycoplasma pneumoniae (Mp) is responsible for 30% of the respiratory manifestations of the general population. Pneumonia occupies the first place within this group. Among the extra-respiratory forms (40%), the neurological ones are the most frequent. Meningoencephalitis and aseptic meningitis are the most common. The presentation of more than one clinical variant in the same patient associated with primoinfection by Mp is possible. In relation to the serological diagnosis, controversies in interpretation sometimes occur. This is a 7-year-old girl with conjunctival injection, cervical adenopathy, photophobia with bilateral papilla pseudoedema, and scaly rash that develops peripheral facial paralysis and aseptic meningitis. We will discuss diagnostic controversies.

Molecular investigation of hemotropic mycoplasms in human beings, dogs and horses in a rural settlement in Southern Brazil / Investigação molecular de espécies de micoplasmas hemotrópicos em cães, equinos e humanos de um assentamento rural do Sul do Brasil

SUMMARYThe aims of this study were to determine the prevalence of hemoplasmas in a rural Brazilian settlement's population of human beings, their dogs and horses, highly exposed to tick bites; to identify the tick species parasitizing dogs and horses, and analyze factors associated with their infection. Blood samples from 132 dogs, 16 horses and 100 humans were screened using a pan-hemoplasma SYBR green real-time PCR assay followed by a species-specific TaqMan real-time PCR. A total of 59/132 (44.7%) dog samples were positive for hemoplasmas (21 Mycoplasma haemocanisalone, 12 ' CandidatusMycoplasma haematoparvum' alone and 21 both). Only 1/100 (1.0%) human sample was positive by qPCR SYBR green, with no successful amplification of 16S rRNA or 23 rRNA genes despite multiple attempts. All horse samples were negative. Dogs >1 year of age were more likely to be positive for hemoplasmas ( p= 0.0014). In conclusion, although canine hemoplasma infection was highly prevalent, cross-species hemoplasma transmission was not observed, and therefore may not frequently occur despite overexposure of agents and vectors.

RESUMOOs objetivos deste estudo foram determinar a prevalência de hemoplasmas numa população restrita de cães, equinos e humanos altamente expostos a picadas de carrapatos em assentamento rural brasileiro; identificar as espécies de carrapatos parasitando cães e equinos, e analisar os fatores associados à infecção. Amostras de sangue de 132 cães, 16 cavalos e 100 humanos foram avaliadas utilizando um protocolo pan-hemoplasma em PCR quantitativas em tempo real (qPCR) com SYBR green, seguido de qPCR TaqMan espécie-específicos. Cinquenta e nove/132 (44,7%) cães foram positivos para hemoplasmas (21 Mycoplasma haemocanis, 12 ' Candidatus Mycoplasmahaematoparvum' e 21 para ambos). Uma amostra humana do total de 100 (1%) foi positiva pelo qPCR SYBR green, mas os genes 16S rRNA ou 23S rRNA não foram amplificados com sucesso, apesar de inúmeras tentativas. Todas as amostras de cavalos foram negativas. Cães > 1 ano apresentaram mais chance de serem positivos para hemoplasmas ( p= 0,0014). Concluindo, embora infecções por hemoplasmas caninos sejam altamente prevalentes, a transmissão de hemoplasmas entre espécies não foi observada, e desta forma podem não ocorrer de forma frequente apesar da alta exposição aos agentes e vetores.

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.

Eggshell apex abnormalities associated with Mycoplasma synoviae infection in layers

Eggs exhibiting eggshell apex abnormalities (EAA) were evaluated for changes in shell characteristics such as strength, thickness, and ultrastructure. Mycoplasma synoviae (MS) infection was confirmed by serological assay along with isolation of MS from the trachea and oviduct. Changes in eggshell quality were shown to be statistically significant (p < 0.01). We also identified ultrastructural changes in the mammillary knob layer by Scanning Electron Microscopy. While eggs may seem to be structurally sound, ultrastructural evaluation showed that affected eggs do not regain their former quality. In our knowledge, this is the first report describing the occurrence of EAA in Korea.

Cytauxzoon felis and 'Candidatus Mycoplasma haemominutum' coinfection in a brazilian domestic cat (Felis catus) / Co-infecção por Cytauxzoon felis e 'Candidatus Mycoplasma haemominutum' em um gato doméstico (Felis catus) no Brasil

This article describes the first detection of Cytauxzoon felis, using molecular techniques, in a naturally infected domestic cat from Brazil, South America. Coinfection with 'Candidatus Mycoplasma haemominutum' was also found. The molecular identification of the piroplasmid species was performed by Polymerase Chain Reaction (PCR) and sequencing analysis. A 284 pb fragment of the gene encoding the 18S ribosomal RNA region was amplified and showed 99% identity with other C. felis strains from North America. In addition, PCR-RFLP (restriction fragment length polymorphism) analysis, which amplifies a 595 bp fragment of the gene encoding 16S ribosomal RNA of some bacterial species, identified the co-infecting species as 'Candidatus M. haemominutum'.

Este artigo descreve a primeira detecção de Cytauxzoon felis em um gato doméstico naturalmente infectado no Brasil, América do Sul, através de técnicas moleculares. Também foi encontrada co-infecção com 'Candidatus Mycoplasma haemominutum'. A detecção molecular da espécie do piroplasmídeo foi realizada através da reação em cadeia pela polimerase (PCR) e sequenciamento. Um fragmento de 284 pb do gene codificador da região 18S do RNA ribossomal do parasito foi sequenciada e mostrou 99% de identidade com outros isolados de C. felis da América do Norte. Ademais, através da análise por meio de PCR-RFLP (Polimorfismo no comprimento de fragmentos de restrição), que amplifica um fragmento de 595 pb do gene codificador da porção 16 do RNA ribossomal de algumas espécies de bactérias, concluiu-se que a espécie com-infectante era 'Candidatus M. haemominutum'.

Micoplasma hominis, Ureaplasma urealyticum y bacterias aeróbicas en el semen de hombres que consultan por infertilidad / Micoplasma hominis, Ureaplasma urealyticum and aerobic bacteria present in the semen from men attending infertility service

Introducción: las infecciones en el semen humano pueden alterar la calidad espermática, y vincularse con problemas de infertilidad masculina. Objetivo: determinar la frecuencia de infecciones por Micoplasma hominis, Ureaplasma urealyticum y bacterias aeróbicas en el semen de hombres que consultan por infertilidad, e identificar si existe relación entre las infecciones encontradas y las alteraciones en las variables de calidad del semen. Métodos: se realizó un estudio descriptivo transversal, para evaluar muestras de semen de 140 hombres, con edades entre 20 y 45 años, provenientes de las consultas de infertilidad del Instituto Nacional de Endocrinología. Se realizó un espermograma completo, que incluyó leucocitospermia, siguiendo los lineamientos de la OMS, para determinar las variables cualitativas y cuantitativas del semen. Las muestras de semen fueron cultivadas en agar sangre y agar chocolate a 37° C en atmósfera de CO2 para investigar bacterias aeróbicas, y se utilizó un juego de reactivos (Mycoplasma System Plus) que permite realizar el cultivo, la identificación, el conteo semicuantitativo y el antibiograma de micoplasmas/ureaplasma urogenitales. Se tuvo en cuenta los aspectos éticos, y los resultados obtenidos se analizaron mediante cálculo de por cientos y la aplicación de la prueba de chi cuadrado. Resultados: de las 140 muestras de semen evaluadas, 58 (41,4 por ciento) mostraron la presencia de infecciones, de ellas 37 correspondieron a Ureaplasma urealyticum (25,7 por ciento), 2 a Micoplasma hominis (1,4 por ciento) y 19 a bacterias aeróbicas (13,8 por ciento ). Al comparar las variables cualitativas y cuantitativas del semen con los sujetos infectados y no infectados, no se observaron diferencias estadísticamente significativas en ninguna de las variables de calidad espermática evaluadas. Conclusiones: la frecuencia total de infecciones, en la muestra estudiada, fue relativamente alta, pero no asociada a alteraciones en las variables seminales(AU)

Introduction: human semen infections can alter the sperm quality and be associated to male infertility disorders. Objectives: to determine the frequency of infections from Micoplasma hominis, Ureaplasma urealyticum and other aerobic bacteria in the semen of men who attended the infertility service, and to identify whether there is some relation between the detected infections and the altered semen quality variables or not. Methods: a cross-sectional descriptive study was performed to evaluate semen samples from 140 men aged 20 to 45 years, who attended the infertility service at the National Institute of Endocrinology. According to the WHO guidelines, a complete spermiogram including leukocytospermia was performed in order to determine the qualitative and quantitative variables in the semen. The semen samples were cultured in blood agar and in chocolate agar at 37oC under CO2 environment to find out possible aerobic bacteria. To this end, a set of reagents known as Mycoplasma System Plus was used, allowing the culture, the identification, the semi-quantitative count and the antibiogram of urogenital mycoplasms/ureaplasms. The ethical aspects were allowed for; the results were analyzed through percentage estimations and the chi square test. Results: out of the 140 evaluated semen samples, 58 (41.4 percent) showed some infection, 37 of them were caused by Ureaplasma urealyticum (25.7 percent), 2 by Micoplasma hominis (1.4 percent) and 19 by the aerobic bacteria (13.8 percent). When making a comparison of the qualitative and quantitative variables of the semen from infected and non-infected subjects, there were not any statistically significant differences in the evaluated variables of the sperm quality. Conclusions: the total frequency of infections in the studied sample was relatively high, but was not associated to altered seminal variables(AU)

A Multiplex real-time PCR for detection of Mycoplasma gallisepticum and Mycoplasma synoviae in clinical samples from Brazilian commercial poultry flocks

Mycoplasma gallisepticum (MS) and Mycoplasma synoviae (MS) are important avian pathogens and cause economic losses to the poultry industry. Molecular biology techniques are currently used for a rapid detection of these pathogens and the adoption of control measures of the diseases. The aim of this study was to develop and validate a technique for simultaneous detection of MG and MS by multiplex real time polymerase chain reaction (PCR). The complete assay (Multiplex MGMS) was designed with primers and probes specific for each pathogen and developed to be carried out in a single tube reaction. Vaccines, MG and MS isolates and DNA from other Mycoplasma species were used for the development and validation of the method. Further, 78 pooled clinical samples from different poultry flocks in Brazil were obtained and used to determine the sensitivity and specificity of the technique in comparison to 2 real time PCR assays specific for MG (MG PCR) and MS (MS PCR). The results demonstrated an agreement of 100% (23 positive and 44 negative samples) between Multiplex MGMS and MG PCR in the analysis of 67 samples from MG positive and negative poultry flocks, and an agreement of 96.9% between Multiplex MGMS and MS PCR in the analysis of 64 samples from MS positive and negative poultry flocks. Considering the single amplification tests as the gold standard, the Multiplex MGMS showed 100% of specificity and sensitivity in the MG analysis and 94.7% sensitivity and 100% specificity in the MS analysis. This new assay could be used for rapid analysis of MG and MS in the poultry industry laboratories.

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.

Posible papel de Bacteroides fragilis enterotoxigénico en la etiología de la vaginitis infecciosa / Possible role of enterotoxigenic Bacteroides fragilis in the etiology of infectious vaginitis

La vaginitis es un trastorno ginecológico frecuente producido por distintas causas, algunas de las cuales permanecen desconocidas. Bacteroides fragilis es el anaerobio más importante en bacteriología clínica. Algunas cepas son enterotoxigénicas y se asocian con síndromes intestinales y extraintestinales. Recientemente han sido aisladas de pacientes con vaginitis. En este trabajo se planteó investigar la posible asociación de B. fragilis enterotoxigénico con la vaginitis infecciosa. Fueron procesadas 265 muestras de exudado vaginal. 202 de mujeres sintomáticas y 63 mujeres sanas. La identificación de los microorganismos se realizó por métodos convencionales. En 31,2% de las pacientes sintomáticas se identificaron: Gardnerella vaginalis, Candida albicans, Mobiluncus, Mycoplasma hominis, Ureaplasma urealyticum y Streptococcus agalactiae. En 27 pacientes sintomáticas y en 5 mujeres sanas se identificó B. fragilis. Estas cepas fueron cultivadas en medio líquido e incubadas durante 48 h a 36° C en anaerobiosis. La toxicidad en los sobrenadantes se ensayó en células HT-29. 18 cepas de B. fragilis aisladas de pacientes sintomáticas fueron enterotoxigénicas, ya que indujeron alteraciones en la monocapa celular y en las células. No se identificó en mujeres sanas (P<0,05). 77,7% de las cepas de B. fragilis enterotoxigénicas no se encontraron asociadas con otros patógenos específicos. Este hecho sugiere que pudiera ser un agente causante de vaginitis, ya que el efecto de la enterotoxina sobre la E-cadherina del epitelio vaginal podría facilitar la invasión y su posible papel patógeno en la vagina. Esta es la primera investigación que asocia a Bacteroides fragilis enterotoxigénico como posible causa de vaginitis infecciosa.

Vaginitis is a common gynecologic disorder. It is due to several causes, some even unknown. Bacteroides fragilis is the most important anaerobe in clinical bacteriology, some strains of this group are notable for being enterotoxigenic and they have been associated with intestinal and extraintestinal syndromes. They have recently been isolated from patients with vaginitis. The purpose of this study was to investigate a possible association of enterotoxigenic B. fragilis with infectious vaginitis. 265 samples of vaginal exudate were processed, 202 from symptomatic patients and 63 healthy women. The identification of the microorganisms was carried out by conventional methods. In 31.2% of symptomatic patients were identified: Gardnerella vaginalis, Mobiluncus, Candida albicans, Mycoplasma hominis, Ureaplasma urealyticum and Streptococcus agalactiae. B. fragilis was identified in 27 symptomatic patients and 5 healthy women. These strains were cultivated in liquid medium and incubated during 48 h at 36°C in anaerobe chambers. Supernatant activity was assayed in HT-29 cells. Eighteen B. fragilis strains isolated from symptomatic patients were enterotoxigenic, because induced alterations in target cell morphology. It was not identified in healthy women (P<0.05). 77.7% of enterotoxigenic B. fragilis strains were not associated with other specific pathogens. This fact suggests that enterotoxigenic B. fragilis could be a cause for vaginitis. The effect of enterotoxin on E-cadherin of vaginal epithelium could facilitate invasion and its possible pathogenic role in the vagina. This is the first report that associates enterotoxigenic Bacteroides fragilis as a possible cause of infectious vaginitis.

Meningoencephalitis associated with Mycoplasma pneumoniae / Meningoencefalite associada ao Mycoplasma pneumoniae

We report a case of a child with meningoencephalitis of atypical etiology. The patient developed the disease after an infection in the upper airways with unfavorable evolution. The clinical recovery was only possible after the administration of adequate antibiotic therapy for the etiological agent. This case report describes a child with meningoencephalitis of atypical etiology. The patient developed the disease after an infection in the superior airways with negative evolution. The clinical recovery was possible only after the introduction of adequate antibiotic therapy for the etiological agent.

Este relato de caso descreve uma criança com menignoencefalite de etiologia atípica. A paciente desenvolveu a doença após infecção de vias aéreas superiores, com evolução desfavorável. Houve recuperação clínica somente após introdução de antibioticoterapia adequada para o agente etiológico.

Simultaneous and rapid differential diagnosis of Mycoplasma genitalium and Ureaplasma urealyticum based on a polymerase chain reaction-restriction fragment length polymorphism.

Objectives: The aim of this investigation was to simultaneously detect and differentiate Mycoplasma genitalium and Ureaplasma urealyticum in female patients suffering from genital complications by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP). Materials and Methods : Genital swabs were taken from 210 patients. They were transported to the laboratory in phosphate-buffered saline. For PCR, samples were analysed with genus-specific MyUu-R and MyUu-F primers. This primer set, which was originally designed in our laboratory, amplified a 465 bp fragment (M. genitalium) and a 559 bp fragment (U. urealyticum). Samples containing a band of the expected sizes for the Mycoplasma strains were subjected to digestion with a restriction endonuclease enzyme of TaqI and Cac8I. Results: Of the 210 samples, a total of 100 (47.6%) samples were found to be positive for Mycoplasmas (seven M. genitalium isolates, 3.3%; and 89 U. urealyticum isolates, 42.4%), and coinfections with both species were detected in four samples (1.9%). The PCR-RFLP results showed that M. genitalium and U. urealyticum are different by enzyme patterns. Conclusion: PCR-RFLP offers a rapid and easily applicable protocol to simultaneous detection and differentiation of M. genitalium and U. urealyticum from clinical samples when specific primers and restriction enzymes are used.

Manifestaciones muco-cutáneas de la infección por Mycoplasma pneumoniae: Presentación de cuatro casos / Mucocutaneous findings of the Mycoplasma pneumoniae infection: Presentation of four cases

The mucocutaneous manifestations oí Mycoplasma pneumoniae infection appear in approximately 20 percent of all the infections produced by this microorganism. Maculopapular rash, vesicular or urticarial exanthemas, are frequent manifestations that can constitute Erythema multiforme or more rarely, Stevens - Johnson syndrome or epidermal toxic necrolisis. We describe the clinical evolution, diagnosis and treatment of four children with mucous and cu-taneous manifestations associated to infection by Mycoplasma pneumoniae and a review of the medical literature.

El compromiso muco-cutáneo de la infección por Mycoplasma pneumoniae se presenta en aproximadamente 20 por ciento de todas las enfermedades producidas por este microorganismo. Frecuentemente se manifiesta con lesiones máculo-papulares, vesiculosas o urticariales, que pueden constituir el eritema multiforme, más raramente síndrome de Stevens-Johnson o necrosis epidérmica tóxica. Describimos la evolución clínica, diagnóstico y el tratamiento administrado a cuatro niños que presentaron manifestaciones de piel y mucosas en relación a la infección por Mycoplasma pneumoniae. Se efectuó además una revisión de la literatura médica.

Diagnóstico de Mycoplasma genitalium por amplificación de los genes MgPa y ARN ribosomal 16S / Diagnosis of Mycoplasma genitalium by MgPa and rRNA 16S gene amplification

OBJETIVO: El microorganismo Mycoplasma genitalium se ha relacionado con la uretritis no gonocócica (UNG). La técnica de PCR se ha convertido en el principal método de detección de este patógeno. En consecuencia, debe aplicarse un método de diagnóstico mediante la amplificación de fragmentos de ADN por la técnica PCR. MATERIAL Y MÉTODOS: Se seleccionaron los cebadores MGF-MGR y MgPaF-MgPaR, complementarios de los genes de ARNr 16S y MgPa de M. genitalium, respectivamente. Se efectuaron ensayos de especificidad y sensibilidad y se estudiaron muestras clínicas. RESULTADOS: La PCR con cada grupo de cebadores utilizado fue específica sólo para M. genitalium y la sensibilidad fue mayor con el grupo de cebadores MGF-MGR. En el estudio de 34 muestras clínicas, 18.5 por ciento fue positivo a M. genitalium y se encontró un mayor número de muestras positivas al utilizar los cebadores MgPaF-MgPaR. CONCLUSIONES: Debe aplicarse en la práctica clínica el diagnóstico de M. genitalium mediante la amplificación del ADN por PCR en los pacientes con UNG.

OBJECTIVE: Mycoplasma genitalium has been associated with nongonococcal urethritis (NGU). Diagnosis by PCR has become the primary detection method for this organism. Thus, diagnosis by DNA amplification using the PCR technique should be utilized. MATERIAL AND METHODS: GMF/GMR and MgpF/MgpR primer pairs, complementary to the M. genitalium 16S rRNA and MgPa genes, respectively, were selected. Specificity and sensibility assays were conducted and clinical samples were studied. RESULTS: The PCR with each primer pair was specific only for M. genitalium, and the sensibility was higher with the GMF/GMR primers. In the study of 34 clinical samples, 18,5 percent were positive for M. genitalium, with more positive samples when the MgpF/MgpR primers were used. CONCLUSIONS: DNA amplification by PCR should be applied in clinical practice to the diagnosis of M. genitalium in patients with NGU should using.

Detección de Mycoplasma genitalium y correlación con manifestaciones clínicas en una población del estado Zulia, Venezuela / Mycoplasma genitalium detection and correlation with clinical manifestations in population of the Zulia State, Venezuela

Diverse studies demonstrate an association between Mycoplasma genitalium and urogenital pathologies. The aim of this study was to investigate the prevalence of M. genitalium in patients attending gynecological evaluation in private clinics (n = 172). DNA amplification assays of the genes 16S rRNA and MgPa were utilized. The prevalence of M. genitalium in the study population was 7.5 percent. M. genitalium was detected in 12.1 percent and 4.1 percent of the symptomatic and asymptomatic patients, respectively (p = 0.047). The infection was diagnosed in patients with cervicitis (17.2 percent) and mucopurulent secretion (16.6 percent) and the highest prevalence of infections was registered in the 31-40 years age group. No significant association between the presence of M.genitalium and individual clinical manifestations or the patients age was showed (p > 0.05). The high prevalence of M. genitalium infections, mostly in patients with clinical manifestations showed in this study, warrants the application of diagnostic strategies in the population to investigate the clinical meaning of these microorganisms and to reevaluate therapeutic schemes against non-gonococcal and non-chlamydial infections.

Diversos estudios demuestran una asociación entre Mycoplasma genitalium y patologías urogenitales. El objetivo de este trabajo fue investigar la prevalencia de infecciones por M. genitalium en pacientes atendidas en clínicas privadas (n = 172). Se utilizaron ensayos de amplificación de genes 16S rARN y MgPa. La prevalencia de M. genitalium en esta población fue 7,5 por ciento. Mycoplasma genitalium fue detectado en 12,1 y 4,1 por ciento) de las pacientes sintomáticas y asintomáticas, respectivamente (p = 0,047). La infección se diagnosticó en pacientes con cervicitis (17,2 por ciento) y con secreción mucopurulenta (16,6 por ciento) y la mayor prevalencia de infecciones se registró en el grupo etario de 31 a 40 años. No se encontró asociación significativa entre la presencia de M. genitalium y manifestaciones clínicas individuales o edad de las pacientes (p > 0,05). La alta prevalencia de infecciones por M. genitalium, principalmente en pacientes con manifestaciones clínicas demostrada en este estudio, demanda la aplicación de estrategias diagnósticas en la población para investigar el significado clínico de estos microorganismos y reevaluar esquemas terapéuticos contra infecciones no gonocóccicas y no clamidiales.

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.

Experimental pathogenicity evaluation of Mycoplasma canadense from bovine mastitis in vitro and in vivo laboratory models.

Mycoplasma canadense, a clinical isolate from milk of a mastitic buffalo, was experimentally tested for its pathogenic potential in hamster tracheal ring and rabbit fallopian tube explant organ cultures (in vitro) and rat and rabbit mammary gland (in vivo) models. The activity percentage reduction in M. canadense infected hamster tracheal rings was 99.1% in comparison to 16.4% in control rings. Mycoplasma canadense, also induced complete ciliostasis at 11-day post-infection in rabbit fallopian tube explants. Histopathological lesions in these infected organ cultures were loss of cilia, desquamation or denudation of epithelium, infiltration of inflammatory cells and proliferation of macrophages as well as oedema in lamina propria. At the end of the experiments, M. canadense organisms were reisolated in pure colonies from the infected but not the control organ cultures. In the rat and rabbit mammary glands, M. canadense organisms persisted upto 6-day and 7-day postinfection, respectively and caused histopathological changes suggestive of subacute to chronic mastitis during the experimental period. The results indicate that the tested M. canadense clinical isolate was virulent.

Prevalence of Ureaplasma urealyticum and Mycoplasma genitalium in men with non-gonococcal urethritis

This study in Teheran, Islamic Republic of Iran, investigated the prevalence of Ureaplasma urealyticum and Mycoplasma species in men with non-gonococcal urethritis. Urethral swab samples were collected from 125 cases and 125 healthy men as a control group. The samples were then investigated by culture methods. The rates of detected bacteria in case and control groups were 19.2% and 7.2% for U. urealyticum, 7.2% and 0.8% for M. genitalium, and 2.4% and 1.6% for M. hominis respectively. Statistical analysis showed a significant difference between case and control groups in the prevalence of U. urealyticum and M. genitalium but not M. hominis. It is concluded that in men, U. urealyticum and M. genitalium may have an etiologic role in non-gonococcal urethritis

Experimental mastitogenicity of Mycoplasma capricolum subsp. capripneumoniae for rabbit mammary glands.

Mycoplasma capricolum subsp. capripneumoniae of cow-udder origin was tested in rabbit mammary-glands for its mastitogenic capability. Establishment of mycoplasma organisms and presence of histopathological lesions in mammary glands were the parameters for describing mastitogenic potential. The reisolation of injected Mycoplasma capricolum subsp. capripneumoniae organisms in the pure form from the infected glands along with the occurrence of histopathological changes were suggestive of mastitis during the entire 8-days period of observation. Rabbit mammary-gland is recommended as a potential in vivo experimental laboratory model to screen the mastitogenic potential of mycoplasmas of animal-udder origin.