Frecuencia y detección molecular de Gardnerella vaginalis en una institución de tercer nivel / Frequency and molecular detection of Gardnerella vaginalis in a third level institute

Resumen Antecedentes: Los padecimientos vaginales son la razón más común para que las mujeres busquen atención médica, con una prevalencia global que oscila entre el 23 y el 29% en mujeres en edad reproductiva. La vaginosis bacteriana es una de las principales causas de estos padecimientos, y el agente etiológico más frecuentemente identificado es Gardnerella vaginalis, sin embargo su diagnóstico es difícil, ya que requiere de medios artificiales selectivos enriquecidos y diferenciales. Objetivo: Determinar la frecuencia de G. vaginalis mediante la amplificación de ácidos nucleicos empleando la reacción en cadena de la polimerasa (PCR) en muestras cervicovaginales de pacientes que asisten a un instituto de tercer nivel. Método: Se analizaron 121 muestras cervicovaginales para la detección molecular del ARN ribosomal 16S de G. vaginalis. Resultados: G. vaginalis. se detectó en 34 muestras, de estas, 23 fueron de mujeres embarazadas y 11 de no embarazadas. Conclusión: La PCR de punto final detectó tres veces más la presencia de G. vaginalis que el medio de cultivo artificial.

Abstract Background: Vaginal conditions are the most common reason for women to seek medical care, with an overall prevalence ranging from 23 to 29% in women of reproductive age. Bacterial vaginosis is one of the main causes of these conditions, and the most frequently identified etiological agent is Gardnerella vaginalis, however, its diagnosis is difficult since it requires enriched and artificial selective culture media. Objective: To determine the frequency of G. vaginalis by nucleic acid amplification using polymerase chain reaction (PCR) in cervicovaginal samples from patients attending a third level institute. Method: One hundred twenty-one cervicovaginal samples were analyzed for molecular detection of 16S ribosomal RNA from G. vaginalis. Results: G. vaginalis was detected in 34 samples, of these, 23 were from pregnant women and 11 from non-pregnant women. Conclusion: Endpoint PCR detected three times more the presence of G. vaginalis than artificial culture medium.

PCR múltiplex para el diagnóstico de microorganismos atípicos transmitidos sexualmente / Multiplex PCR testing for the atypical microorganisms diagnosis sexually transmitted

Resumen Antecedentes: Las infecciones de transmisión sexual son un problema de salud pública mundial. El análisis rutinario incluye solo pruebas microbiológicas y serológicas para el diagnóstico de patógenos. Los microorganismos atípicos como Chlamydia trachomatis y micoplasmas no son identificados debido a los requerimientos. Además, no es incluida Gardnerella vaginalis, aunque se asocia a la vaginosis bacteriana. Objetivo: Desarrollar una PCR múltiplex para el diagnóstico de C. trachomatis, micoplasmas y G. vaginalis. Método: Se estandarizó la PCR múltiplex utilizando oligonucleótidos para C. trachomatis (gen ompA, orf6 plasmídico), Mycoplasma/Ureaplasma y G. vaginalis (genes rRNA16s). Resultados: Se estandarizaron pruebas de PCR múltiplex para los microorganismos estudiados, optimizándose las concentraciones y condiciones de las reacciones múltiplex. Se obtuvieron PCR dúplex para C. trachomatis (ompA, orf6), Chlamydia/Gardnerella y Chlamydia/micoplasmas y tríplex para Chlamydia/Mycoplasma/Ureaplasma. También un cuádruplex para Chlamydia/Mycoplasma/Ureaplasma/Gardnerella. Los resultados fueron verificados por PCR e hibridación automática (HybriSpot 12) y análisis in silico. Conclusión: Se desarrollaron pruebas de PCR múltiplex con una alta sensibilidad y especificidad para la identificación de C. trachomatis, micoplasmas y G. vaginalis.

Abstract Background: Sexually transmitted infections are a global public health problem. Routine analysis includes microbiological and serological tests for the diagnosis of pathogens. Atypical microorganisms such as Chlamydia trachomatis and mycoplasmas are not determined due to the requirements for their identification. Furthermore, Gardnerella vaginalis is not included despite being associated with bacterial vaginosis. Objective: To develop a multiplex PCR to diagnose Chlamydia, mycoplasmas, and Gardnerella. Method: Standardization of multiplex PCR tests was carried out using oligonucleotides for the identification of Chlamydia (ompA gene, plasmid orf6), Mycoplasma/Ureaplasma and Gardnerella (rRNA16s genes). Results: Multiplex PCR tests were standardized for the microorganisms studied, optimizing the concentrations and conditions of the multiplex reactions. Duplex PCR was obtained for Chlamydia (ompA, orf6), Chlamydia/Gardnerella, and Chlamydia/mycoplasmas, and triplex PCR for Chlamydia/mycoplasmas. Also, a quadruplex for Chlamydia, Mycoplasma/Ureaplasma and Gardnerella. PCR and automatic hybridization verified the results obtained (HybriSpot 12) and in silico analysis. Conclusion: Multiplex PCR tests with high sensitivity and specificity were developed to identify C. trachomatis, mycoplasmas, and G. vaginalis.

Características clínicas y microbiológicas de la infección de vías urinarias bajas en población ambulatoria / Clinical and Microbiological Characteristics of Lower Urinary Tract Infections in an Outpatient Population

Introducción: El tratamiento de las infecciones del tracto urinario es casi siempre empírico, lo que genera una serie de problemas en la consulta diaria. Objetivo: Caracterizar clínica y microbiológicamente las infecciones de vías urinarias bajas no complicadas en pacientes de una clínica de primer nivel. Métodos: Se realizó un estudio transversal descriptivo. La identificación de las bacterias del cultivo de orina se efectuó por métodos establecidos. La prueba de susceptibilidad a los antimicrobianos se realizó por la técnica Kirby-Bauer. Se utilizó el programa estadístico SPSS versión 26, con la prueba de ji al cuadrado y un análisis multivariado discriminante. Se calculó también razón de momios con el programa Epi-Info. Resultados: Se incluyeron 270 pacientes, con frecuencia de 39,3 por ciento de cultivos positivos, y Escherichia coli como la especie predominante. Se identificaron, además, 31,3 por ciento de bacterias Gram positivas. Se presentó significancia estadística entre la infección urinaria y factores como el sexo, y la infección del tracto urinario previa en las mujeres. Se obtuvo 100 por ciento de cepas resistentes a ampicilina. En general, se obtuvieron porcentajes de resistencia altos en los antimicrobianos probados. Conclusiones: Escherichia coli fue la especie más frecuentemente aislada, sin embargo, existe una serie de microorganismos implicados en enfermedades del tracto genital como Gardnerella vaginalis, que parecen estar involucrados en la etiología de las infecciones del tracto urinario. Se identificaron factores de riesgo como el sexo biológico y las infecciones previas en mujeres. Se obtuvieron porcentajes de resistencia altos en los antimicrobianos probados(AU)

Introduction: The management of urinary tract infections is almost always empirical, which generates a series of problems in the daily consultation. Objective: To characterize, clinically and microbiologically, uncomplicated lower urinary tract infections in patients of a primary level clinic. Methods: A descriptive and cross-sectional study was carried out. Bacterial identification in urine culture was performed by established methods. Antimicrobial susceptibility testing was performed using the Kirby-Bauer technique. The statistical software SPSS (version 26) was used, with the chi squared test and multivariate discriminant analysis. Odds ratios were also calculated with the Epi-Info program. Results: A total of 270 patients were included, with a 39.3percent frequency of positive cultures and Escherichia coli as the predominant species. In addition, 31.3percent of Gram-positive bacteria were identified. There was statistical significance between urinary tract infection and factors such as sex or previous urinary tract infection in women. One result was 100percent of ampicillin-resistant strains. In general, high percentages of resistance were obtained for the tested antimicrobials. Conclusions: Escherichia coli was the most frequently isolated species; however, there is a number of microorganisms implicated in genital tract diseases, such as Gardnerella vaginalis, which appear to be involved in the etiology of urinary tract infections. Risk factors such as biological sex and previous infections in women were identified. High percentages of resistance were obtained for the tested antimicrobials(AU)

Actividad antimicrobiana in vitro del extracto de hojas de Bixa orellana L. contra bacterias anaerobias asociadas a la vaginosis bacteriana y Lactobacillus spp / In vitro antimicrobial activity of Bixa orellana L. leaves extract against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp.

Objetivos . Describir la actividad antimicrobiana in vitro del extracto metanólico de las hojas de Bixa orellana L. contra las bacterias anaerobias asociadas a la vaginosis bacteriana y Lactobacillus spp. Materiales y métodos . Se incluyeron en el estudio ocho cepas de referencia ATCC; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula y Lactobacillus crispatus, y 22 aislamientos clínicos; once aislados de Gardnerella vaginalis y once aislados de Lactobacillus. La susceptibilidad antimicrobiana se determinó mediante el método de difusión en agar. La concentración mínima inhibitoria (CMI) y la concentración bactericida mínima (CBM) fueron determinadas utilizando el método de dilución en agar y un método de dilución modificado, respectivamente. Resultados . Todas las cepas de referencia ATCC tuvieron un alto nivel de susceptibilidad al extracto, con excepción de P. vibia, V. parvula y L. crispatus. Interesantemente, los aislamientos clínicos de G. vaginalis y la cepa ATCC de G. vaginalis fueron los más susceptibles al extracto dados los bajos valores de CMI (1,0 - 2,0 mg/mL) y CBM (1,0 - 4,0 mg/mL), mientras que, los aislamientos clínicos de Lactobacillus spp. y la cepa ATCC de L. crispatus fueron los menos susceptibles debido a los altos valores de CMI (32,0 mg/mL) y CBM (≥ 32,0 mg/mL). Conclusiones . Los experimentos in vitro sugieren que el extracto posee propiedades antibacterianas selectivas dada su alta actividad contra bacterias anaerobias asociadas a vaginosis bacteriana y baja actividad contra especies de Lactobacillus.

Objective. To describe the in vitro antimicrobial activity of the methanolic extract of Bixa orellana L. leaves against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp. Materials and methods. Eight ATCC reference strains; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula, and Lactobacillus crispatus, and twenty-two clinical isolates; eleven Gardnerella vaginalis and eleven Lactobacillus strains, were included in the study. The antimicrobial susceptibility was determined by the agar diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by using agar dilution and a modified dilution plating method, respectively. Results. All ATCC reference strains showed high levels of susceptibility to the extract, except P. vibia, V. parvula and L. crispatus. Interestingly, all G. vaginalis clinical isolates and the G. vaginalis ATTC strain were the most susceptible to the extract, given their low MIC (1.0 - 2.0 mg/mL) and MBC (1.0 - 4.0 mg/mL) values, whereas, the Lactobacillus spp. clinical isolates and the L. crispatus ATCC strain were the least susceptible bacteria given their high MIC (32.0 mg/mL) and MBC (≥ 32.0 mg/mL) values. Conclusions. In vitro experiments suggest that the extract possesses selective antimicrobial properties given its high activity against bacterial vaginosis-associated anaerobic bacteria and low activity against Lactobacillus species.

Agents causing genital infections in routine cytological tests: frequency and characteristics of Papanicolaou smears / Agentes causadores de infecções genitais em exames citológicos de rotina: frequência e características dos esfregaços de papanicolaou

Urinary tract infections are responsible for most human infections, these are caused by bacteria, fungi, protozoa and associated microorganisms. The goal of this study was to determine the rate of vaginal infection-causing agents in routine cytological exams and also to evaluate the characteristics of positive tested Pap smears. A retrospective documental with descriptive aspect research was performed in a Clinical Pathology laboratory from Maceió-AL. The results of the Pap smears exams for Trichomonas vaginalis, Gardnerella vaginalis, Candida spp and HPV were arranged in a database as well as other data such as bacterial, protozoan, fungal and viral coinfections. The sample was composed by 18.645 women who have undergone Pap smear exams from 2013 to 2017. Of these analyzed exams, 27.4% in 2013, 10.9% in 2014, 10.6% in 2015, 15.2% in 2016 and 13.67% in 2017 were within normal range, however more than half of these exams presented some infections caused by unspecific or microbiological agents. By analyzing all the reports, 4.073 (21.84%) presented inflammations caused by some species of infectious agent with the following rate order: G. vaginalis and T. vaginalis. Furthermore, it was possible to confirm high rates of coinfection by and Candida spp. The rate of genital infections in this study highlights that there is a public health matter that must be controlled, which points a greater need for monitoring, guidance and actions towards greater awareness in order to prevent these problems.

Infecções do trato urinário são responsáveis pela maior parte das infecções humanas, sendo causadas por bactérias, fungos, protozoários, e por microrganismos associados. O objetivo deste trabalho foi determinar a frequência de agentes causadores de infecções vaginais nos exames citológicos de rotina, bem como avaliar as características dos esfregaços positivos. Foi realizada uma pesquisa do tipo documental retrospectiva de aspecto descritivo, com abordagem quantitativa, em um Laboratório de Patologia Clínica de Maceió-AL. Nesse sentido, os resultados dos exames colpocitológicos para Trichomonas vaginalis, Gardnerella vaginalis, Candida spp e HPV foram organizados em um banco de dados, contendo os resultados para estes agentes, bem como a coinfecção bacteriana, protozoária, fúngica e viral. A amostra foi composta por 18.645 mulheres que realizaram exame colpocitológico, no período de 2013 a 2017. Destes exames analisados 27,4% em 2013, 10,9% em 2014, 10,6% em 2015, 15,2% em 2016 e 13,67% em 2017, estavam dentro do limite da normalidade, porém mais que a metade desses exames analisados apresentou algumas infecções causadas por agentes inespecíficos ou microbiológicos. Do total de laudos analisados, 4.073 (21,84%), apresentaram inflamações causadas por algum tipo de agente infeccioso, com a seguinte ordem de frequência: G. vaginalis, e T. vaginalis. Além disto, foi possível confirmar a elevada frequência de coinfecção entre G. vaginalis e Candida spp. Os índices de infecções genitais nesse trabalho mostram que há um problema de saúde pública que deve ser controlado, apontando uma necessidade maior de acompanhamento, orientações e de ações para uma maior conscientização para prevenir esses problemas.

Agents causing genital infections in routine cytological tests: frequency and characteristics of Papanicolaou smears

Abstract Urinary tract infections are responsible for most human infections, these are caused by bacteria, fungi, protozoa and associated microorganisms. The goal of this study was to determine the rate of vaginal infection-causing agents in routine cytological exams and also to evaluate the characteristics of positive tested Pap smears. A retrospective documental with descriptive aspect research was performed in a Clinical Pathology laboratory from Maceió-AL. The results of the Pap smears exams for Trichomonas vaginalis, Gardnerella vaginalis, Candida spp and HPV were arranged in a database as well as other data such as bacterial, protozoan, fungal and viral coinfections. The sample was composed by 18.645 women who have undergone Pap smear exams from 2013 to 2017. Of these analyzed exams, 27.4% in 2013, 10.9% in 2014, 10.6% in 2015, 15.2% in 2016 and 13.67% in 2017 were within normal range, however more than half of these exams presented some infections caused by unspecific or microbiological agents. By analyzing all the reports, 4.073 (21.84%) presented inflammations caused by some species of infectious agent with the following rate order: G. vaginalis and T. vaginalis. Furthermore, it was possible to confirm high rates of coinfection by and Candida spp. The rate of genital infections in this study highlights that there is a public health matter that must be controlled, which points a greater need for monitoring, guidance and actions towards greater awareness in order to prevent these problems.

Resumo Infecções do trato urinário são responsáveis pela maior parte das infecções humanas, sendo causadas por bactérias, fungos, protozoários, e por microrganismos associados. O objetivo deste trabalho foi determinar a frequência de agentes causadores de infecções vaginais nos exames citológicos de rotina, bem como avaliar as características dos esfregaços positivos. Foi realizada uma pesquisa do tipo documental retrospectiva de aspecto descritivo, com abordagem quantitativa, em um Laboratório de Patologia Clínica de Maceió-AL. Nesse sentido, os resultados dos exames colpocitológicos para Trichomonas vaginalis, Gardnerella vaginalis, Candida spp e HPV foram organizados em um banco de dados, contendo os resultados para estes agentes, bem como a coinfecção bacteriana, protozoária, fúngica e viral. A amostra foi composta por 18.645 mulheres que realizaram exame colpocitológico, no período de 2013 a 2017. Destes exames analisados 27,4% em 2013, 10,9% em 2014, 10,6% em 2015, 15,2% em 2016 e 13,67% em 2017, estavam dentro do limite da normalidade, porém mais que a metade desses exames analisados apresentou algumas infecções causadas por agentes inespecíficos ou microbiológicos. Do total de laudos analisados, 4.073 (21,84%), apresentaram inflamações causadas por algum tipo de agente infeccioso, com a seguinte ordem de frequência: G. vaginalis, e T. vaginalis. Além disto, foi possível confirmar a elevada frequência de coinfecção entre G. vaginalis e Candida spp. Os índices de infecções genitais nesse trabalho mostram que há um problema de saúde pública que deve ser controlado, apontando uma necessidade maior de acompanhamento, orientações e de ações para uma maior conscientização para prevenir esses problemas.

Agents causing genital infections in routine cytological tests: frequency and characteristics of Papanicolaou smears / Agentes causadores de infecções genitais em exames citológicos de rotina: frequência e características dos esfregaços de papanicolaou

Urinary tract infections are responsible for most human infections, these are caused by bacteria, fungi, protozoa and associated microorganisms. The goal of this study was to determine the rate of vaginal infection-causing agents in routine cytological exams and also to evaluate the characteristics of positive tested Pap smears. A retrospective documental with descriptive aspect research was performed in a Clinical Pathology laboratory from Maceió-AL. The results of the Pap smears exams for Trichomonas vaginalis, Gardnerella vaginalis, Candida spp and HPV were arranged in a database as well as other data such as bacterial, protozoan, fungal and viral coinfections. The sample was composed by 18.645 women who have undergone Pap smear exams from 2013 to 2017. Of these analyzed exams, 27.4% in 2013, 10.9% in 2014, 10.6% in 2015, 15.2% in 2016 and 13.67% in 2017 were within normal range, however more than half of these exams presented some infections caused by unspecific or microbiological agents. By analyzing all the reports, 4.073 (21.84%) presented inflammations caused by some species of infectious agent with the following rate order: G. vaginalis and T. vaginalis. Furthermore, it was possible to confirm high rates of coinfection by and Candida spp. The rate of genital infections in this study highlights that there is a public health matter that must be controlled, which points a greater need for monitoring, guidance and actions towards greater awareness in order to prevent these problems.

Infecções do trato urinário são responsáveis pela maior parte das infecções humanas, sendo causadas por bactérias, fungos, protozoários, e por microrganismos associados. O objetivo deste trabalho foi determinar a frequência de agentes causadores de infecções vaginais nos exames citológicos de rotina, bem como avaliar as características dos esfregaços positivos. Foi realizada uma pesquisa do tipo documental retrospectiva de aspecto descritivo, com abordagem quantitativa, em um Laboratório de Patologia Clínica de Maceió-AL. Nesse sentido, os resultados dos exames colpocitológicos para Trichomonas vaginalis, Gardnerella vaginalis, Candida spp e HPV foram organizados em um banco de dados, contendo os resultados para estes agentes, bem como a coinfecção bacteriana, protozoária, fúngica e viral. A amostra foi composta por 18.645 mulheres que realizaram exame colpocitológico, no período de 2013 a 2017. Destes exames analisados 27,4% em 2013, 10,9% em 2014, 10,6% em 2015, 15,2% em 2016 e 13,67% em 2017, estavam dentro do limite da normalidade, porém mais que a metade desses exames analisados apresentou algumas infecções causadas por agentes inespecíficos ou microbiológicos. Do total de laudos analisados, 4.073 (21,84%), apresentaram inflamações causadas por algum tipo de agente infeccioso, com a seguinte ordem de frequência: G. vaginalis, e T. vaginalis. Além disto, foi possível confirmar a elevada frequência de coinfecção entre G. vaginalis e Candida spp. Os índices de infecções genitais nesse trabalho mostram que há um problema de saúde pública que deve ser controlado, apontando uma necessidade maior de acompanhamento, orientações e de ações para uma maior conscientização para prevenir esses problemas.

Associação entre Vaginose Bacteriana e Anormalidades Citológicas nos Exames Citopatológicos Analisados em um Laboratório Escola de Goiânia-GO / Association Between Bacterial Vaginosis and Cytological Abnormalities in Cytopathological Exams Analyzed in a Laboratory School of Goiânia-GO / Asociación entre Vaginosis Bacteriana y Anormalidades Citológicas en Exámenes Citopatológicos Analizados en una Escuela de Laboratorio de Goiânia-GO

Introdução: A Gardnerella vaginalis facilita a infecção pelo papilomavírus humano (HPV). Objetivo: Verificar a associação entre anormalidades citológicas e presença de Gardnerella vaginalis nos esfregaços cervicovaginais encaminhados ao Laboratório Clínico da Pontifícia Universidade Católica de Goiás (LAC/PUC Goiás) estratificadas por faixa etária. Método: Estudo transversal realizado no LAC/PUC Goiás entre janeiro de 2013 a dezembro de 2015. Para análises estatísticas, a variável idade foi categorizada em ≤39 anos e >40 anos, utilizando o programa IBM SPSS Statistics (Version 2.0, 2011®) para o teste de qui-quadrado (X²), com intervalo de confiança de 95% e valor p<0,05. Resultados: Foram analisados 4.558 exames citopatológicos, a maioria com presença de Lactobacillus spp. (46,97%). A prevalência dos agentes patogênicos foi a Gardnerella vaginalis (79,6%), seguida de Candida spp. (16,8%), Trichomonas vaginalis (2,2%), Herpes simplex (0,4 %) e Chlamydia trachomatis (0,1%). As anormalidades citológicas foram observadas em 9,1%, sendo atypical squamous cells of undetermined significance (ASC-US) 2,57%, low grade squamous intraepithelial lesion (LSIL) 1,78%, atypical squamous cells of undetermined significance cannot exclude high grade squamous intraepithelial lesion (ASC-H) 3,52%, high grade squamous intraepithelial lesion (HSIL) 1,08%, atypical endocervical cells, favor neoplastic (AGC-NEO) 0,22% e carcinoma 0,02%. Houve uma associação significante entre anormalidades citológicas graves e mulheres ≥40 anos, OR 3,01 (IC 95% 2,0-4,58) (p<0,0001). Mulheres ≤40 anos mostraram significância à presença de Gardnerella vaginalis (p<0,0004). Conclusão: Uma elevada prevalência de Gardnerella vaginalis foi encontrada associada com as anormalidades citológicas, principalmente em mulheres sexualmente ativas.

Introduction:Gardnerella vaginalis facilitates human papillomavirus (HPV) infection. Objective: To verify the association between cytological abnormalities and the presence of Gardnerella vaginalis in cervicovaginal smears sent to the Clinical Laboratory of the Pontifical Catholic University of Goiás (LAC/PUC Goiás) stratified by age range. Method: Cross-sectional study carried out at LAC/PUC Goiás from January 2013 to December 2015. For statistical analysis, the variable age was categorized as ≤39 years and >40 years, using the IBM SPSS Statistics program (Version 2.0, 2011®) for the chi-square test (X²), with a 95% confidence interval and p<0.05. Results:4,558 cytopathological exams were analyzed, most of them with the presence of Lactobacillus spp (46.97%). The prevalence of pathogens was Gardnerella vaginalis (79.6%), followed by Candida spp. (16.8%), Trichomonas vaginalis (2.2%), Herpes simplex (0.4%) and Chlamydia trachomatis (0.1%). Cytological abnormalities were observed in 9.1%, being atypical squamous cells of undetermined significance (ASC-US) 2.57%, low grade squamous intraepithelial lesion (LSIL) 1.78%, atypical squamous cells of undetermined significance cannot exclude high intraepithelial lesion (ASC-H) 3.52%, high grade squamous intraepithelial lesion (HSIL) 1.08%, atypical endocervical cells, neoplastic favor (AGC-NEO) 0.22% and carcinoma 0.02%. There was a significant association between severe cytological abnormalities and women >40 years old OR 3.01 (95% CI 2.0-4.58) (p<0.0001). Women ≤40 years old showed the presence of Gardnerella vaginalis (p<0.0004). Conclusion:A high prevalence of Gardnerella vaginalis was found and its association with cytological abnormalities, especially in sexually active women.

Introducción:Gardnerella vaginalis facilita la infección por el virus del papiloma humano (VPH). Objetivo: Verificar la asociación entre anormalidades citológicas y la presencia de Gardnerella vaginalis en frotis cervicovaginales enviadas al Laboratorio Clínico de la Pontificia Universidad Católica de Goiás (LAC/PUC Goiás) estratificadas por grupo de edad. Método: Estudio transversal realizado en LAC/PUC Goiás desde enero de 2013 hasta diciembre de 2015. Para el análisis estadístico, la edad variable se clasificó como ≤39 años y >40 años, utilizando el programa IBM SPSS Statistics (Versión 2.0, 2011®) para la prueba de chi-cuadrado (X²), con un intervalo de confianza del 95% y p <0,05. Resultados: Se analizaron 4.558 exámenes citopatológicos. La prevalencia de Lactobacillusspp. con 46,97%. Los patógenos como Gardnerella vaginalis fueron 79,6%, Candidaspp. 16,8%, Trichomonas vaginalis 2,2%, Herpes simplex 0,4%, y Chlamydia trachomatis 0,1%. Se observaron anormalidades citológicas en 9,1%, con células escamosas atípicas de significado indeterminado (ASC-US) 2,57%, lesión intraepitelial escamosa de bajo grado (LSIL) 1,78%, células escamosas atípicas de significación indeterminada no pueden excluir lesión intraepitelial (ASC-H) 3,52%, lesión intraepitelial escamosa de alto grado (HSIL) 1,08%, células endocervicales atípicas, favor neoplásico (AGC-NEO) 0,22% y carcinoma 0,02%. Hubo una asociación significativa entre anormalidades citológicas severas y mujeres >40 años OR 3,01 (IC 95% 2,0-4,58) (p<0,0001). Las mujeres ≤40 años mostraron la presencia de Gardnerella vaginalis (p<0,0004). Conclusión: Se encontró una alta prevalencia de Gardnerella vaginalis y su asociación con anomalías citológicas, especialmente en mujeres sexualmente activas.

Infecciones en varones por gardnerella vaginalis- revisión bibliográfica

Introducción: Gardnerella vaginalis ha sido aislada de orina, semen, descarga uretral, hisopos endouretrales, rectales y del prepucio, en algunos casos asociada a manifestaciones clínicas; se han descrito uretritis y balanopostitis con diversos porcentajes. Los hallazgos en frotis rectal, semen y prepucio deben interpretarse cuidadosamente para otorgarles significado clínico. Ha quedado demostrado que el hombre la adquiere de sus parejas sexuales y en su forma cohesiva. Objetivo: Explorar el papel patógeno de G. vaginalis como causa de infección en los hombres. Método: La estrategia de búsqueda se realizó en: PubMed/ Medline, Scopus Cochrane Library, SciELO Lilacs, Redalyc; Google Scholar con proveedores como: EB-SCO y tesauros MeSH y DeCS. Resultados: Las diferentes publicaciones indicaron detección de G. vaginalis en muestras uretrales con reportes desde 1,5 %, 4,2 % hasta 14 % con manifestaciones clínicas de uretritis. Otros reportes indican 4,5 %, 5 %, 6,3 %, 7,2 % y 14,5 % sin uretritis. Dos estudios de infecciones del tracto urinario presentan porcentajes de 30,8 % y 67 % de G. vaginalis en hombres con síntomas urinarios. G vaginalis se reporta en frotis rectal, semen e hisopados del prepucio, pero sin clara atribución de etiología patógena. Conclusión: Es recomendable la búsqueda de G. vaginalis en hombres con uretritis no gonocócica, balanopostitis, e infecciones del tracto urinario por su probable significado patógeno, mientras que su papel en semen en pacientes con infertilidad y en el frotis rectal, requiere más estudios de investigación.

Introduction: Gardnerella vaginalis has been isolated from urine, semen, urethral discharge, and endourethral, rectal, and foreskin swabs, in some cases associated with clinical manifestations. Urethritis and balanoposthitis have been described in different percentages. Rectal swab, semen, and foreskin findings must be carefully interpreted for clinical significance. It has been demonstrated that man acquires it from her sexual partners in its cohesive form. Objective: To explore the role of G. vaginalis as a cause of infections in men. Method: Bibliography was searched through PubMed/ Medline, Scopus Cochrane Library, SciELO Lilacs, Redalyc; Google Scholar, and providers like EBSCO and thesauros MeSH y DeCS. Results: Several publications show the detection of G. vaginallis in urethral samples with reports of 1,5%, 4,2%, even 14% of urethritis. Some other reports show a 4,5%, 5%, 6,3%, 7,2% and 14,5% without urethritis. Two studies of urinary tract infections show a 30,8% and 67% of G. vaginalis in men with urinary symptoms. G. vaginalis is reported in rectal smears, semen, and foreskin swabs but without clear attribution of pathogenic etiology. Conclusion: The search for G. vaginalis in men with non-gonococcal urethritis, balanoposthitis, or urinary tract infections is recommended due to its probable pathogenic significance, while its role in patients with infertility and rectal smears requires further investigation, due to the presence in semen.

Clinical presentation, diagnosis and management of bacterial vaginosis: a hospital based cross-sectional study

Background: Bacterial vaginosis is most common cause of vaginal discharge. Clinical presentation varies from person to person. The management of bacterial vaginosis is largely syndromic and empirical, it is usually based on naked eye examination of vaginal discharge and that is unsatisfactory because the diagnostic accuracy is lost without microscopic examination. The modern management of bacterial vaginosis demands a specific diagnosis which is a combination of naked eye examination plus laboratory workup.Methods: The study was carried out on 183 females with complaint of vaginal discharge in the outpatient department of obstetrics and gynecology. Specimens were collected in outpatient department of Jhalawar Medical College in associated Zanana Hospital. The laboratory work was conducted in the department of microbiology in Jhalawar Medical College, Jhalawar, Rajasthan, India.Results: The prevalence of microbial positivity in our study was 79.9%. Bacterial vaginosis was the most common diagnosis seen in 86 (47.0%) cases. Maximum patients presented with complaint of copious amount of foul-smelling yellow discharge.Conclusions: The study concludes that it is important to know the various presentations, confirm the diagnosis by proper microbiological tests, and provide appropriate treatment to patients to prevent resistance and recurrence of bacterial vaginosis.

Infertility and vaginal microbiome: review study / Infertilidade e microbioma vaginal: estudo de revisão

Introduction: Infertility is an important public health problem and has many causal factors. Previous findings, based mainly on culture techniques, suggest an association between infertility and changes in the vaginal microbiome. The metagenomic approach allowed the discovery of new bacterial species, previously unidentified in the female genital tract, permitting a deeper knowledge of the role of vaginal microbiome in female reproductive health. Objective: To evaluate the association of changes in the vaginal microbiota with infertility, and its repercussions on the outcome of assisted reproduction techniques after a decade of the Human Microbiome Project. Methods: A systematic search was carried out in the MEDLINE database between September and November 2018, by selecting 14 studies, associating vaginal microbiome with infertility, or with results of assisted reproduction techniques. Results: The findings showed a higher prevalence of bacterial vaginosis, and increased microbial diversity in the vagina of infertile women. Regarding the success of assisted reproduction techniques, most studies did not show any significant association between bacterial vaginosis and reduction in pregnancy rates. Conclusion: Further studies are needed to better understand the influence of the balance of vaginal microorganism species on female reproductive health, addressing the microbiome composition in contexts beyond in vitro fertilization techniques.

Introdução: A infertilidade é um importante problema de saúde pública e possui muitos fatores causais. Achados prévios, baseados principalmente em técnicas de cultura, sugerem uma associação entre infertilidade e alterações do microbioma vaginal. A abordagem metagenômica permitiu a descoberta de novas espécies bacterianas, anteriormente não identificadas, no trato genital feminino, possibilitando um conhecimento mais aprofundado do papel do microbioma vaginal na saúde reprodutiva feminina. Objetivo: Avaliar a associação das alterações da microbiota vaginal com a infertilidade e suas repercussões no resultado das técnicas de reprodução assistida após uma década do Projeto Microbioma Humano. Métodos: Foi realizada uma busca sistemática na base de dados MEDLINE, entre setembro e novembro de 2018, e selecionados 14 estudos que associavam o microbioma vaginal com a infertilidade ou com resultados de técnicas de reprodução assistida. Resultados: Os achados evidenciaram uma maior prevalência de vaginose bacteriana e um aumento da diversidade microbiana na vagina de mulheres inférteis. Com relação ao sucesso das técnicas de reprodução assistida, a maioria dos estudos não evidenciou uma associação significante entre a vaginose bacteriana e a redução nas taxas de gravidez. Conclusão: São necessários novos estudos para melhor compreender a influência do equilíbrio das espécies de microrganismos vaginais na saúde reprodutiva feminina, abordando a composição do microbioma em contextos além das técnicas de fertilização in vitro.

Comparative analysis of virulence factors & biotypes of Gardnerella vaginalis isolated from the genital tract of women with & without bacterial vaginosis

Background & objectives: Bacterial vaginosis (BV) involves the presence of a thick vaginal multispecies biofilm, where Gardnerella vaginalis is the predominant species. The reason for an increase in the number of G. vaginalis which are usually present as normal flora of the female genital tract in cases of BV, is not known. Hence, the objective of the present study was to compare the biotypes and virulence factors of G. vaginalis isolated from the genital tract of women with and without BV. Methods: High vaginal swabs collected from 811 women of reproductive age were cultured. G. vaginalis isolates were biotyped and tested for adherence to vaginal epithelial cells, biofilm formation, agglutination of human red blood cells (RBCs), protease production, phospholipase production and surface hydrophobicity. Results: Of the isolates from women with BV, 83.3 per cent (60/72) showed good adherence, 78.4 per cent (58/74) produced biofilm, 82.9 per cent (63/76) produced phospholipase, 67.1 per cent (51/76) produced protease, 77.3 per cent (58/75) were positive for surface hydrophobicity and 61.6 per cent (45/73) were positive for haemagglutination of human RBC. In case of G. vaginalis from non-BV women, 25 per cent (15/60) isolates showed good adherence, 18.4 per cent (9/49) biofilm production, 35 per cent (21/60) phospholipase, 36.6 per cent (22/60) protease, 41.7 per cent (25/60) surface hydrophobicity and 10.1 per cent (6/59) agglutination of human RBCs. Maximum number of isolates belonged to biotypes 6, 2 and 3. Biotype 3 was more associated with non-BV rather than BV; biotype 6, 2 and 1 were more associated with cases of BV. Maximum virulence factors were expressed by biotypes 6, 2 and 1. Interpretation & conclusions: Virulence factors were more expressed by G. vaginalis isolates obtained from women with BV rather than from non-BV. Biotypes 6, 2 and 1 were more associated with cases of BV and expressed maximum virulence factors.

Identification of bacterial vaginosis-associated bacteria in male urethra: Co-occurrence of Atopobium vaginae and Gardnerella vaginalis

Aims@#Bacterial vaginosis (BV) is characterized by a transition in vaginal microflora from lactobacilli to anaerobic bacteria. Gardnerella vaginalis and Atopobium vaginae are considered the most responsible pathogens for the etiology of BV. Colonization of male urethra with BV-associated bacteria has been rarely investigated. The aim of this study was to investigate the differences in the presence of BV-associated bacteria in the healthy male urethra in regard to sexual exposure. @*Methodology and results@#The first-catch urine specimens, representative of urethral swabs, from 114 healthy male volunteers, were included in this study. Lactobacillus spp., L. crispatus, L. jensenii, L. gasseri, L. iners, G. vaginalis, A. vaginae, Peptoniphilus spp., P. lacrimalis, BVAB2, Mageeibacillus indolicus, Megasphaera type I, Mobiluncus mulieris, Leptotrichia/Sneathia, Corynebacterium spp., and Prevotella spp. were investigated using a PCR assay. The most frequently identified BV-associated bacteria were Lactobacillus spp., Peptoniphilus spp., and G. vaginalis. There was no association between any BV-associated bacteria and sexual exposure. There was statistically significant co-occurrence of A. vaginae and G. vaginalis in the MU of subjects independently of sexual exposure (p = 0.025). Also, there was a significant association between G. vaginalis and smoking (p = 0.023). @*Conclusion, significance and impact of study@#To the best of our knowledge, this is the first study reporting the co-occurrence of G. vaginalis and A. vaginae in the male urethra independently of sexual exposure.

Desempenho da citologia em meio líquido na identificação de agentes microbiológicos cérvico-vaginais / Liquid medium cytology performance in identification of cervico-vaginal microbiological agentes

Verificar o desempenho da citologia em meio líquido na identificação de agentes microbiológicos cérvico-vaginais em relação à citologia convencional. Métodos: Estudo analítico e transversal realizado no Laboratório de Citopatologia de um centro universitário. Foram realizadas coletas citopatológicas pelas duas metodologias: convencional e em meio líquido (Liqui-PREP®), além de coleta microbiológica. Resultados: Foram avaliadas 67 amostras cérvico-vaginais pelas duas técnicas citopatológicas, verificando-se que a citologia convencional identificou 26 esfregaços com microrganismos de interesse clínico, enquanto que a citologia em meio líquido identificou 20 casos, sendo observada boa concordância entre as técnicas na identificação de Gardnerella vaginalis e Candida spp. A citologia convencional também evidenciou mais casos de esfregaços com Trichomonas vaginalis e ocorreu boa concordância entre as técnicas citopatológicas e o exame microbiológico na identificação de Gardnerella vaginalis. Conclusão: Observou-se boa concordância entre as técnicas citopatológicas na identificação de Candida spp. e Gardnerella vaginalis, assim como entre as duas técnicas frente ao exame microbiológico na evidência de Gardnerella vaginalis.

Retrospective Study on the Association between Maternal Macronutrient Intake and Urogenital Infections during Pregnancy in a Swiss Cohort

Aims: Urogenital infections during pregnancy have been linked to adverse pregnancy outcomes. The objective of this study was to test the hypothesis that macronutrient intake and relative macronutrient contribution to diet is related to the risk of developing urogenital infections during pregnancy. Study Design: This is a retrospective single center cohort study. Place and Duration of Study: Outpatient Clinic of Obstetrics at the University Hospital Zurich, Switzerland; between January 2009 and December 2010. Methodology: We included 774 pregnant women of ages ranging from 16 to 47 years with data on urogenital infections and diet history. A diet history of these pregnant women based on food intake during the last seven days was collected in a nutritional counselling program. Diet information of these same women was matched with vaginal/urinary/cervical specimens collected within 90 days (range) prior to the nutrition assessment. The pathogens analyzed included Gram-negative rods, Gram-positive rods, Gram-positive cocci (including group B Streptococcus), Gardnerella vaginalis, Chlamydia trachomatis, and Candida spp. The covariates were maternal age, body mass index (BMI), origin, and parity. Crude and adjusted odds ratios (ORs) were determined by logistic regression. Results: Among the 774 pregnant women, 47.7% had some kind of infection. High fat intake was positively associated with Gardnerella vaginalis (adjusted OR=3.6; 95% confidence interval (CI)=1.3–10; p=0.01). No association was seen between macronutrients or their distribution and other pathogens. However, significant associations were found between infections and covariates. Conclusions: Findings suggested that increased dietary fat intake is associated with vaginal infections, thereby predisposing women to adverse pregnancy outcomes. This signified the importance of appropriate diet during pregnancy.

Fatores de risco associados ao desenvolvimento de alterações cervicouterinas em mulheres que realizam exame citopatológico / Risk factors associated to the development of changes in the cervix of uterus in women undergoing a cytopathological examination

Introdução: O câncer de colo do útero ainda é um sério problema de saúde pública em nosso país. Aparece em terceiro lugar entre os tipos de câncer mais comum entre as mulheres. Vários são os fatores envolvidos na etiologia desse câncer e suas lesões precursoras. Objetivo: Verificar os principais cofatores associados às alterações cervicouterinas. Casuística e Métodos: Trata-se de um estudo analítico e documental, realizado em um laboratório do município de Surubim-PE, incluindo resultados de exames citopatológicos do período de janeiro de 2014 a dezembro de 2015. Os dados foram avaliados pelo programa PRISM versão 7.0, por meio do teste Qui-quadrado, com intervalo de confiança 95% e razão de prevalência (RP). Resultados: Foram incluídos 1.200 resultados de exames de mulheres na faixa etária entre 14 e 78 anos. Quarenta e três por cento das mulheres pertenciam à faixa etária entre 31 e 47 anos e 86,6% haviam realizado seu último exame citopatológico a menos de três anos. A faixa etária entre 14 e 30 anos esteve associada a um maior risco para o desenvolvimento das anormalidades cervicais (p< 0,001 e OR 2,9). Mulheres que realizaram o exame citopatológico a menos de três anos apresentaram uma maior associação com a presença de lesões intraepiteliais escamosas de baixo grau (LSIL) e atipias escamosas (p < 0,0001 e OR 11,3). A lesão intraepitelial escamosa de baixo grau foi a anormalidade cervical de maior frequência, correspondendo a 46,3%. Mulheres com presença de Gardnerella vaginalis e/ou Mobiluncus spp. apresentaram associação com a presença de anormalidades cervicais (p< 0,0001). Conclusão: Mulheres abaixo de 30 anos, que realizaram exame citopatológico a menos de três anos ou apresentam Gardnerella vaginalis e/ou Mobiluncus spp.devem ser acompanhadas quanto aos possíveis riscos associados ao desenvolvimentode alterações cervicouterinas.

Introduction: Cervical cancer is still a serious public health problem in our country. It appears third among the most common types of cancer among women. Several are the factors involved in the etiology of this cancer and its precursor lesions. Objective: Verify the main cofactors associated with the changes in the cervix of the uterus. Patients and Methods:This is an analytical and documentary study, carried out in a laboratory in the municipality of Surubim-PE, including results of cytopathological exams from January 2014 to December 2015. Data were evaluated using the software PRISM version 7.0, by the Chi-square test, with 95% confidence interval and prevalence ratio (PR). Results: We included 1,200 tests results of women in the age between 14 and 78 years. Forty-three percent of the women were in the age group from 31 to 47 years and 86.6% had performed their last cytopathological examination less than three years. The age group ranging from 14 to 30 years was associated with a higher risk for the development of cervical abnormalities (p< 0.001 and OR 2.9). Women who underwent cytopathological examination less than three years presented a greater association with the presence of low squamous intraepithelial lesions (LSIL) and squamous atypia (p< 0.0001 and OR 11.3). The low-grade squamous intraepithelial lesion was the most frequent cervical abnormality, corresponding to 46.3%. Conclusion: Women under 30 years, who performed cytopathological examination at less than three years or present Gardnerella vaginalis and / or Mobiluncus spp. should be accompanied for the possible risks associated with the development of the changes in the cervix of uterus.

Gardnerella vaginalis in perinatology: An overview of the clinicopathological correlation

@#Gardnerella vaginalis (GV) is a facultatively anaerobic gram-variable bacillus and is the major organism involved in bacterial vaginosis. GV-associated bacterial vaginosis has been associated with adverse pregnancy outcomes include preterm parturition and subclinical chorioamnionitis. Inflammatory response induced by GV presents paediatric problems as well. Studies had shown that increased levels of proinflammatory cytokines include TNF-α, IL-1β and IL-6 following fetal inflammatory response syndrome secondary to GV-induced intrauterine infection may result in the development of periventricular leukomalacia and bronchopulmonary dysplasia in the infected fetus. There is increasing evidence that GV-associated BV infection serves as a risk factor for long-term neurological complications, such as cerebral palsy and learning disability. GV is fastidious and could elude conventional detection methods such as bacterial cultures. With current more sophisticated molecular biology detection methods, its role and pathogenic effects have been shown to have a greater impact on intrauterine inflammation and fetal/neonatal infection. This review gives an overview on the characteristics of GV and its virulence properties. Its detrimental role in causing unfavourable GV-related perinatal outcomes, with emphasis on the possible mechanistic pathways is discussed. The discovery of disease mechanisms allows the building of a strong platform where further research on innovative therapies can be based on, for instance, an anti-TLR monoclonal antibody as therapeutic agent to halt inflammation-precipitate adverse perinatal outcomes.

Gardnerella vaginalis in perinatology: An overview of the clinicopathological correlation

@#Gardnerella vaginalis (GV) is a facultatively anaerobic gram-variable bacillus and is the major organism involved in bacterial vaginosis. GV-associated bacterial vaginosis has been associated with adverse pregnancy outcomes include preterm parturition and subclinical chorioamnionitis. Inflammatory response induced by GV presents paediatric problems as well. Studies had shown that increased levels of proinflammatory cytokines include TNF-α, IL-1β and IL-6 following fetal inflammatory response syndrome secondary to GV-induced intrauterine infection may result in the development of periventricular leukomalacia and bronchopulmonary dysplasia in the infected fetus. There is increasing evidence that GV-associated BV infection serves as a risk factor for long-term neurological complications, such as cerebral palsy and learning disability. GV is fastidious and could elude conventional detection methods such as bacterial cultures. With current more sophisticated molecular biology detection methods, its role and pathogenic effects have been shown to have a greater impact on intrauterine inflammation and fetal/neonatal infection. This review gives an overview on the characteristics of GV and its virulence properties. Its detrimental role in causing unfavourable GV-related perinatal outcomes, with emphasis on the possible mechanistic pathways is discussed. The discovery of disease mechanisms allows the building of a strong platform where further research on innovative therapies can be based on, for instance, an anti-TLR monoclonal antibody as therapeutic agent to halt inflammation-precipitate adverse perinatal outcomes.

Generation of dimeric single-chain antibodies neutralizing the cytolytic activity of vaginolysin

Background: Gardnerella vaginalis is a bacterial vaginosis (BV)-associated vaginal bacterium that produces the toxin vaginolysin (VLY). VLY is a pore-forming toxin that is suggested to be the main virulence factor of G. vaginalis. The high recurrence rate of BV and the emergence of antibiotic-resistant bacterial species demonstrate the need for the development of recombinant antibodies as novel therapeutic agents for disease treatment. Single-chain variable fragments (scFvs) generated against VLY exhibited reduced efficacy to neutralize VLY activity compared to the respective full-length antibodies. To improve the properties of scFvs, monospecific dimeric scFvs were generated by the genetic fusion of two anti-VLY scFv molecules connected by an alpha-helix-forming peptide linker. Results: N-terminal hexahistidine-tagged dimeric scFvs were constructed and produced in Escherichia coli and purified using metal chelate affinity chromatography. Inhibition of VLY-mediated human erythrocyte lysis by dimeric and monomeric scFvs was detected by in vitro hemolytic assay. The circulating half-life of purified scFvs in the blood plasma of mice was determined by ELISA. Dimeric anti-VLY scFvs showed higher neutralizing potency and extended circulating half-life than parental monomeric scFv. Conclusions: The protein obtained by the genetic fusion of two anti-VLY scFvs into a dimeric molecule exhibited improved properties in comparison with monomeric scFv. This new recombinant antibody might implement new possibilities for the prophylaxis and treatment of the diseases caused by the bacteria G. vaginalis.

An update on Gardneralla vaginalis associated bacterial vaginosis in Malaysia

Objeetive:To update the status of Gardnerella vaginalis (G.vaginalis) as a causative agent of bacterial vaginosis (BV) in Malaysia and to define its epidemiology,metronidazole resistance and virulence properties.Methods:It is a single-centre (Gynaecology clinic at the Hospital Kuala Lumpur,Malaysia) prospective study with laboratory-based microbiological follow up and analyses.Vaginal swabs collected from the patients suspected for BV were subjected to clinical BV diagnosis,isolation and identification of G.vaginalis,metronidazole susceptibility testing,vaginolysin and sialidase gene PCR,Piot's biotyping and amplified ribosomal DNA restriction analysis genotyping.Results:Among the 207 patients suspected for BV,G.vaginalis was isolated from 47 subjects.G.vaginalis coexisted with Trichomonas vaginalis and Candida albicans in 26 samples.Three G.vaginalis isolates were resistant to metronidazole.Biotyping revealed 1 and 7 as the common types.Amplified ribosomal DNA restriction analysis genotype Ⅱ was found to be more common (n =22;46％) than Ⅰ (n =12;25.53％) and Ⅲ (n =13;27.6％).All genotype Ⅰ and Ⅲ isolates carried the sialidase gene,while 91.6％ and 84.6％ contained the vaginolysin gene.Genotype Ⅰ was significantly associated with postgynaecological surgical complications and abortions (P =0.002).Conclusions:The existence of pathogenic G.vaginalis clones in Malaysia including drug resistant strains should not be taken lightly and needs to be monitored as these may bring more complications especially among women of child bearing age and pregnant women.