Estudio multicéntrico de disfunción vaginal de la Red Nacional de Laboratorios BACOVA de la República Argentina: prevalencia, influencia de factores seleccionados, evaluación clínica y distribución de casos por región / Multicenter study on vaginal dysfunction of the National Network of BACOVA Laboratories in Argentina: prevalence, influence of selected factors, clinical evaluation and case distribution by region / Estudo multicêntrico de disfunção vaginal da Rede Nacional de Laboratórios BACOVA da República Argentina: prevalência, influência de fatores selecionados, avaliação clínica e distribuição dos casos por região

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Abstract The aims of this study were to update the prevalence of vaginal dysfunction (VD) in non-pregnant women of childbearing age, pregnant women and menopausal women, to analyse microbiological aspects, to evaluate the influence of parity and contraception in the vaginal microenvironment, to analyse case studies, and to compare the prevalence in the five regions of Argentina: Northeast, Northwest, Centre, Cuyo and South. Eight thousand three hundred twenty-four (8324) vaginal content samples of patients from 39 institutions, examined between May 2019 and June 2020, were prospectively and consecutively studied. The samples were analysed applying the standardized BAVACO-VIR methodology (study of the balance of vaginal content and genital inflammatory response). Of the 8324 samples, 5947 (71.5%) corresponded to women of childbearing age; 1627 (19.5%) to pregnant women and 750 (9.0%) to menopausal women. The most frequent basic vaginal state (BVS) in the three groups was BVS I with normal microbiota, accounting for 33.5% of the entire studied population. Moreover, 66.5% of vaginal dysfunction states were detected. In the pregnant women and menopausal women group there was a significant increase in the frequency of normal BVS I, in contrast with the group of women of childbearing age, where a significant increase of vaginosis and vaginitis was observed. In asymptomatic women, a predominance of BVS I was detected in the three groups. In symptomatic women, there was a predominance of BVS II and V in pregnant women, of BVS II, IV and V in women of childbearing age, and of BVS III and V in menopausal women. Asymptomatic VD was detected in 56.6% of pregnant women, 62.8% of women of childbearing age and 50.9% of menopausal women. A significantly positive association was detected between the presence of yeasts and BVS II and V and the presence of trichomonas and BVS V. The multiparity history variable showed a statistically significant positive association with BVS V and a negative association with BVS I. With regard to contraception, in women who used hormonal contraceptives it was observed that oral contraceptives increased the frequency of BVS I and II and decreased the frequency of BVS III, IV and V of VD; in those who used an intradermal device, there was a decrease in the frequency of BVS I, II and III and an increase in the frequency of BVS IV and V, whereas in women using injectable contraceptives, there was no association. The intrauterine device decreased the frequency of BVS I and II and increased the frequency of BVS V; the condom increased the frequency of BVS IV and the rhythm method decreased the frequency of BVS II. In BVS I, a decrease in the frequency of signs and symptoms was detected. The prevalence of VD in some regions accounted for values over 80%, a higher figure than that described at the national and international levels, which reflects the poor and insufficient action in Sexual and Reproductive Health. The high percentage of asymptomatic women with VD highlights the importance of studying the vaginal content, even in the absence of symptoms. A history of multiparity and contraception showed a link with vaginal function, reflecting the importance of considering this fact in a gynecological evaluation. Although the relationship of BVS with signs and symptoms does not constitute a diagnostic tool, it contributes to the understanding of pathogenic mechanisms.

Resumo Os objetivos deste estudo foram: atualizar a prevalência de disfunção vaginal (DV) em mulheres não grávidas em idade fértil (MEF), mulheres grávidas (EMB) e mulheres menopausadas (MPN), analisar aspectos microbiológicos, avaliar a influência da paridade e contracepção no microambiente vaginal, analisar quadros clínicos e comparar as prevalências das 5 regiões do nosso país: Nordeste, Noroeste, Centro, Cuyo e Sur. 8324 conteúdos vaginais de pacientes de 39 instituições, que compareceram entre maio de 2019 e junho de 2020, foram estudados prospectiva e facilitadoconsecutivamente. As amostras foram analisadas aplicando-se a metodologia padronizada BACOVA-ERIGE (estudo do equilíbrio do conteúdo vaginal e resposta inflamatória genital). Das 8324 amostras de conteúdo vaginal, 5947 (71,5%) corresponderam às MEF; 1627 (19,5%) às EMB e 750 (9,0%) às MNP. O estado vaginal básico (EVB) mais frequente nos três grupos foi EVB I de microbiota normal e representou 33,5% de toda a população global. Foram detectados 66,5% dos estados de disfunção vaginal. No grupo EMB e MNP, um aumento significativo na frequência de EVB I normal foi registrado, em contraste com o grupo MEF, onde foi reconhecido um aumento significativo de EVB de vaginose e vaginite. Em mulheres assintomáticas, foi detectado predomínio de EVB I nos três grupos. Em mulheres sintomáticas foi detetado: nas EMB, predomínio de EVB II e V; nas MEF, predominância de EVB II, IV e V, e nas MNP, predominância de EVB III e V. 56,6% das EMB, 62,8% das MEF e 50,9% das MNP apresentaram DV na ausência de sintomas. Foi detectada associação significativamente positiva entre a presença de leveduras e EVB II e V e a presença de tricomonas e EVB V. A variável antecedente de multiparidade apresentou associação positiva estatisticamente significativa com EVB V e associação negativa com EVB I. Em relação à contracepção, observou-se em mulheres que usavam anticoncepcionais hormonais, que os anticoncepcionais orais aumentaram a frequência de EVB I e II e diminuíram a frequência de EVB III, IV e V de DV; Em mulheres com dispositivo intradérmico, foi observada uma diminuição na frequência de EVB I, II e III e um aumento na frequência de EVB IV e V, e finalmente em mulheres com anticoncepcionais injetáveis, nenhuma associação foi demonstrada. O dispositivo intrauterino diminuiu a frequência de EVB I e II e aumentou a frequência de EVB V; o preservativo aumentou a frequência de EVB IV e o método do ritmo diminuiu a frequência de EVB II. Na EVB I, foi detectada diminuição da frequência de sinais e sintomas. A prevalência de DV em algumas regiões ultrapassou valores de 80%, valor superior ao descrito a nível nacional e internacional, o que reflete a atuação precária e insuficiente em Saúde Sexual e Reprodutiva. O alto percentual de mulheres assintomáticas com DV demonstra a importância da realização do estudo do conteúdo vaginal, mesmo na ausência de sintomas. A história de multiparidade e contracepção mostrou ligação com a função vaginal, refletindo a importância da sua consideração na avaliação ginecológica. Embora a relação da EVB com os sinais e sintomas não constitua uma ferramenta diagnóstica, ela contribui para o entendimento dos mecanismos patogênicos.

Respuesta inflamatoria genital en la detección de alteraciones por virus del papiloma humano / The inflammatory genital reaction in the detection of alterations by human papilloma virus / Reagào inflamatoria genital na detecgào de alteragòes por Papilomavírus Humano

Disfunción vaginal (DV) (vaginosis/vaginitis) es el síndrome genérico de mayor prevalencia, alcanzando el 50% de todas las mujeres en edad fértil (sintomáticas y asintomáticas). El virus del Papiloma Humano (HPV) se detecta en 30 a 40% de mujeres en edad fértil (sintomáticas y asintomáticas) y se asocia a alteraciones pre-neoplásicas y a carcinoma invasor del cuello uterino. El diagnóstico sindrómico de DV y alteraciones inducidas por HPV es ineficiente y en la actualidad la morfología (macro y microscópica) es el gold standard, pero requiere ordenamiento. El Estudio del Contenido Vaginal es la prueba de laboratorio bacteriológico de mayor solicitud luego del urocultivo. BACOVA normatiza el diagnóstico de vaginosis/vaginitis y ERIGE aumenta el valor predictivo de células que alertan sobre alteraciones epiteliales. Desde 2007 al presente en los talleres BACOVA ERIGE (tinción de Giemsa) se evaluó la sensibilidad de la detección de células anormales exfoliadas. Un 99% de los participantes coincidió con la detección de koi-locitos. BACOVA/ERIGE no reemplaza al Papanicolaou de ninguna manera, pero puede y debe realizarse en laboratorios periféricos, con lo que además del diagnóstico de vaginosis/vaginitis con 100% de valor predictivo, aumentan la cobertura preventiva de estados proliferativos.

Vaginal dysfunction (DV) (vaginosis/vaginitis) is the generic syndrome of major prevalence, reaching 50% of all women in fertile age (symptomatic and asymptomatic). The Human Papillomavirus (HPV) is detected in 30-40% of women in fertile age (symptomatic and asymptomatic) and is associated to pre-neoplastic lesions and invading carcinoma of the uterine cervix. The diagnosis for the symptoms of DV and the alterations induced by HPV are inefficient and at present, the morphology (macroscopic and microscopic) is the standard gold, but it needs better classification. The Study of the Vaginal Content is the test of major request after urocultives in bacteriological laboratories. BACOVA establishes the procedure for the diagnosis of vaginosis/vaginitis and ERIGE increases the predictive value of cells that give the alarm on epithelial alterations. From 2007 to the present sensitivity in the detection of abnormal exfoliated cells from vagina and uterine cervix was evaluated during the BACOVA - ERIGE, (Giemsa's stain) workshops, 99% of the participants coincided with the detection of koilocytes. BACOVA/ERIGE does not replace the Papanicolaou by any means, but it can and must be performed in peripheral laboratories, where apart from the diagnosis of vaginosis/vaginitis with 100% of predictive value, it is possible to increase the detection of precocious proliferative changes of the squamous epithelium.

Disfungào vaginal (DV) (vaginose / vaginite) é a síndrome mais prevalente genérica, atingindo 50% de todas as mulheres em idade fértil (sintomáticas e assintomáticas). O Papilomavírus Humano (HPV) é detectado em 30-40% das mulheres em idade fértil (sintomáticas e assintomáticas) e está associado a alteragòes pré-neoplásicas e a carcinoma invasivo do colo do útero. O diagnóstico sindrómico de DV e alteragòes induzidas pelo HPV é ineficiente e atualmente a morfologia (macroscópica e microscópica) é o padrào ouro, mas precisa de ordenamento. O Estudo do Conteúdo Vaginal é o exame de laboratòrio bacteriológico mais solicitado, seguido da urocultura. BACOVA normatiza o diagnòstico de vaginose/ vaginite e ERIGE aumenta o valor preditivo de células que alertam a respeito de alteragòes epiteliais. Desde 2007 até hoje, nos workshops BACOVA/ERIGE (coloragào de Giemsa), foi avaliada a sensibili-dade da detecgào de células anormais esfoliadas. 99% dos participantes coincidiram com a detecgào de coilócitos. Bacova/Erige nào substitui o Papanicolaou de forma alguma, mas pode e deve ser feito em laboratórios periféricos, com o qual além do diagnóstico de vaginose / vaginite com 100% de valor preditivo, aumentam a cobertura preventiva de estados proliferativos.

Respuesta inflamatoria genital en la detección de alteraciones por virus del papiloma humano / The inflammatory genital reaction in the detection of alterations by human papilloma virus / ReagOo inflamatoria genital na detecgOo de alteragòes por Papilomavírus Humano

Disfunción vaginal (DV) (vaginosis/vaginitis) es el síndrome genérico de mayor prevalencia, alcanzando el 50% de todas las mujeres en edad fértil (sintomáticas y asintomáticas). El virus del Papiloma Humano (HPV) se detecta en 30 a 40% de mujeres en edad fértil (sintomáticas y asintomáticas) y se asocia a alteraciones pre-neoplásicas y a carcinoma invasor del cuello uterino. El diagnóstico sindrómico de DV y alteraciones inducidas por HPV es ineficiente y en la actualidad la morfología (macro y microscópica) es el gold standard, pero requiere ordenamiento. El Estudio del Contenido Vaginal es la prueba de laboratorio bacteriológico de mayor solicitud luego del urocultivo. BACOVA normatiza el diagnóstico de vaginosis/vaginitis y ERIGE aumenta el valor predictivo de células que alertan sobre alteraciones epiteliales. Desde 2007 al presente en los talleres BACOVA ERIGE (tinción de Giemsa) se evaluó la sensibilidad de la detección de células anormales exfoliadas. Un 99% de los participantes coincidió con la detección de koi-locitos. BACOVA/ERIGE no reemplaza al Papanicolaou de ninguna manera, pero puede y debe realizarse en laboratorios periféricos, con lo que además del diagnóstico de vaginosis/vaginitis con 100% de valor predictivo, aumentan la cobertura preventiva de estados proliferativos.(AU)

Vaginal dysfunction (DV) (vaginosis/vaginitis) is the generic syndrome of major prevalence, reaching 50% of all women in fertile age (symptomatic and asymptomatic). The Human Papillomavirus (HPV) is detected in 30-40% of women in fertile age (symptomatic and asymptomatic) and is associated to pre-neoplastic lesions and invading carcinoma of the uterine cervix. The diagnosis for the symptoms of DV and the alterations induced by HPV are inefficient and at present, the morphology (macroscopic and microscopic) is the standard gold, but it needs better classification. The Study of the Vaginal Content is the test of major request after urocultives in bacteriological laboratories. BACOVA establishes the procedure for the diagnosis of vaginosis/vaginitis and ERIGE increases the predictive value of cells that give the alarm on epithelial alterations. From 2007 to the present sensitivity in the detection of abnormal exfoliated cells from vagina and uterine cervix was evaluated during the BACOVA - ERIGE, (Giemsas stain) workshops, 99% of the participants coincided with the detection of koilocytes. BACOVA/ERIGE does not replace the Papanicolaou by any means, but it can and must be performed in peripheral laboratories, where apart from the diagnosis of vaginosis/vaginitis with 100% of predictive value, it is possible to increase the detection of precocious proliferative changes of the squamous epithelium.(AU)

DisfungOo vaginal (DV) (vaginose / vaginite) é a síndrome mais prevalente genérica, atingindo 50% de todas as mulheres em idade fértil (sintomáticas e assintomáticas). O Papilomavírus Humano (HPV) é detectado em 30-40% das mulheres em idade fértil (sintomáticas e assintomáticas) e está associado a alteragòes pré-neoplásicas e a carcinoma invasivo do colo do útero. O diagnóstico sindrómico de DV e alteragòes induzidas pelo HPV é ineficiente e atualmente a morfologia (macroscópica e microscópica) é o padrOo ouro, mas precisa de ordenamento. O Estudo do Conteúdo Vaginal é o exame de laboratòrio bacteriológico mais solicitado, seguido da urocultura. BACOVA normatiza o diagnòstico de vaginose/ vaginite e ERIGE aumenta o valor preditivo de células que alertam a respeito de alteragòes epiteliais. Desde 2007 até hoje, nos workshops BACOVA/ERIGE (coloragOo de Giemsa), foi avaliada a sensibili-dade da detecgOo de células anormais esfoliadas. 99% dos participantes coincidiram com a detecgOo de coilócitos. Bacova/Erige nOo substitui o Papanicolaou de forma alguma, mas pode e deve ser feito em laboratórios periféricos, com o qual além do diagnóstico de vaginose / vaginite com 100% de valor preditivo, aumentam a cobertura preventiva de estados proliferativos.(AU)

Hydrophobicity test in mutans Streptococci

Kinetic hydrophobic measurements were performed by confronting 40 mutans streptococci from thirty 10- to 20-year-old patients with 200 ml hexadecane (Sigma). Fourteen patients had high dental caries risk (Group A), dmft + DMFT >5 with 3 or more active caries, and 16 had low dental caries risk (Group B), dmft + DMFT <3 without active caries. Twenty bacteria from Group A and 20 bacteria from Group B were typed using De La Higuera's procedure and confirmed by API strip (bio- Merieux). From the 14 patients in Group A we obtained 12 S. mutans (8 hydrophobic/ 4 non-hydrophobic), 5 S. sobrinus (4 hydrophobic/ 1 non-hydrophobic) and 3 S. rattus (hydrophobic). From the 16 patients in Group B we obtained 11 Streptococcus mutans (10 non-hydrophobic/ 1 hydrophobic), 7 Streptococcus sobrinus (6 non-hydrophobic/ 1 hydrophobic) and 2 Streptococcus rattus (hydrophobic). Patients with high dental caries risk have a higher prevalence of hydrophobic bacteria than patients with low dental caries risk (p=0.0003). All typed S. rattus were hydrophobic.

Con el objeto de evaluar una posible relacion entre hidrofobicidad y caries dental, se estudiaron 40 cepas de Streptococcus grupo mutans provenientes de 30 pacientes de entre 10 y 20 anos, 14 pacientes con tres o mas caries activas e indice ceod mas CPOD > 5 (Grupo A) y 16 pacientes sin caries activas, con ceod mas CPOD < 3 (Grupo B). Las cepas fueron aisladas a partir de muestras de saliva en AMS-BT y tipificadas por pruebas bioquimicas y API-strep realizandose la medicion cinetica de hidrofobicidad, enfrentandolas con 200 ml de hexadecano (Sigma). En el grupo A se caracterizaron 12 cepas Streptococcus mutans: 8 hidrofobicas y 4 no hidrofobicas, 5 cepas de Streptococcus sobrinus: 4 hidrofobicas y 1 no hidrofobica y 3 cepas Streptococcus rattus hidrofobicas. En el grupo B, se caracterizaron 11 cepas de S. mutans, 10 no hidrofobicas y 1 hidrofobica, 7 cepas de S. sobrinus: 6 no hidrofobicas y 1 hidrofobica y 2 cepas de S. rattus hidrofobicas. Todos los pacientes del grupo A presentaron al menos 1 cepa hidrofobica. En las cepas aisladas de estos pacientes se demostro la existencia de una alta prevalencia de Streptococcus grupo mutans con caracteristicas hidrofobicas p=0,0003. Estos resultados indicarian la relacion entre la capacidad de adherencia a hexadecano y caries activa. Todas las cepas tipificadas como S. rattus fueron hidrofobicas independientemente del grupo de origen.

Prevalence and comparison of diagnostic methods for Trichomonas vaginalis infection in pregnant women in Argentina.

The objectives of this study were to conduct a prevalence survey of trichomoniasis in pregnant women and to evaluate the utility of different methods for its diagnosis. A total of 597 vaginal exudates from pregnant women who were examined at the Hospital de Clinicas in Buenos Aires, Argentina from 1 August 2005 to 31 January 2007, were prospectively and consecutively evaluated. The investigation of Trichomonas vaginalis was made by different microscopic examinations, and culture on liquid medium. The sensitivity and specificity of the microscopic examinations were assessed considering culture on liquid medium as the "gold standard". The prevalence of T. vaginalis obtained by culture on liquid medium was 4.0% (24/597). The prevalence of T. vaginalis obtained by direct wet smear, prolonged May-Grunwald Giemsa staining, and sodium acetate-formalin (SAF)/methylene blue staining-fixing technique was 1.8%, 2.3% and 2.5%, respectively. The sensitivity of the direct wet smear was 45.8%, that of the prolonged May-Grunwald Giemsa staining was 58.3%, and that of the SAF/methylene blue method was 62.5%. Considering the 3 microscopic examinations altogether, the sensitivity rose to 66.7% and the specificity was 100% for all of them. This is the first time that the prevalence data of T. vaginalis by culture in pregnant women are published in Argentina. Due to the low sensitivity obtained by microscopy in asymptomatic pregnant women, the use of the liquid medium is recommended during pregnancy, in order to provide an early diagnosis and treatment.

Hydrophobicity test in mutans streptococci.

Kinetic hydrophobic measurements were performed by confronting 40 mutans streptococci from thirty 10- to 20-year-old patients with 200 ml hexadecane (Sigma). Fourteen patients had high dental caries risk (Group A), dmft + DMFT > 5 with 3 or more active caries, and 16 had low dental caries risk (Group B), dmft + DMFT < 3 without active caries. Twenty bacteria from Group A and 20 bacteria from Group B were typed using De La Higuera's procedure and confirmed by API strip (bio-Merieux). From the 14 patients in Group A we obtained 12 S. mutans (8 hydrophobic/4 non-hydrophobic), 5 S. sobrinus (4 hydrophobic/1 non-hydrophobic) and 3 S. rattus (hydrophobic). From the 16 patients in Group B we obtained 11 Streptococcus mutans (10 non-hydrophobic/1 hydrophobic), 7 Streptococcus sobrinus (6 non-hydrophobic/ 1 hydrophobic) and 2 Streptococcus rattus (hydrophobic). Patients with high dental caries risk have a higher prevalence of hydrophobic bacteria than patients with low dental caries risk (p = 0.0003). All typed S. rattus were hydrophobic.

Balance del contenido vaginal en el diagnóstico diferencial de vaginosis-vaginitis: Reacción inflamatoria vaginal en embarazadas sintomáticas / Balance of vaginal content in vaginosis-vaginitis differential diagnosis: Vaginal inflammatory reaction in symptomatic pregnant women

El objetivo del trabajo fue validar el alto valor predictivo de BACOVA (Balance del contenido vaginal, disponible en www.fba.org.ar/proeco) en el diagnóstico microscópico diferencial de vaginosis bacteriana (VB) y vaginitis microbiana inespecífica (VMI). Fueron estudiadas 299 embarazadas sintomáticas. Se determinó por microscopía el Valor Numérico (Nugent) (VN), células guía (CG), morfotipos extraños (Mex), leucocitos por campo (Lpc), tricomonas (TV) y levaduras (LE) con lectura a 400X. Los resultados globales fueron: VN de 7 a 10, 16,4% del total de casos. BACOVA permitió diferenciar 11,7% de los casos con VN de 7 a 10, con reacción inflamatoria vaginal (RIV) menor de 10 Lpc, como casos típicos de BV. Con igual VN se detectó 4,7% de casos con RIV con más de 10 Lpc, compatibles con VMI. En sólo 7,3% de los casos de LE detectadas por microscopía (20,4% en total), se confirmó una RIV significativa. Hubo 2,7% del total de casos con RIV significativa, pH<4,5 y resultado negativo para el resto de los criterios estudiados. La determinación de la RIV, fue imprescindible, junto al VN, en el diagnóstico diferencial de VB y VMI. La RIV también es necesaria para establecer la significación clínica de la presencia de levaduras. BACOVA detecta casos con RIV significativa (2,7%), como único marcador, con pH inferior a 4,5, compatibles con infección del tracto genital superior.

This study was undertaken to demonstrate the high diagnostic predictive value of BACOVA (Balance of Vaginal Content, www.fba.org.ar/proeco) for bacterial vaginosis and microbial non specific vaginitis, in pregnant women primary health care. BACOVA including Nugent score (Gram 1000X) and leucocyte count (Wet mount and Giemsa, using 400X) was evaluated in 299 symptomatic pregnant women. Nugent score 7 to 10 was detected in 16.4%. Crossing Nugent value 7 to 10 with leucocytes counts shows that only 11.7% were below 10 leucocytes per field and 4.7% had a significant vaginal inflammatory response. Yeasts were detected in 20.4% but only in 7.2% of cases they show a significant association with vaginal inflammatory response. In 2.7% of the cases there was a significant vaginal inflammatory response, with pH below 4.5, VN from 0 to 3 and negative for TV, fishy odor, and exogenous microbial morphotypes. Simultaneous study of vaginal inflammatory response and Nugent score is mandatory in order to detect true cases of bacterial vaginosis (11.7%) from those of potential microbial non specific vaginitis (4.7%) (Donders' "aerobic vaginitis") Besides, vaginal inflammatory response became a strong criterion to define yeast vulvovaginitis (7.2%). Cases (2.7%) with score 0 to 3, negative for other criteria, with a high vaginal inflammatory response, are predictors of upper genital infections.

Enterocin CRL35 inhibits late stages of HSV-1 and HSV-2 replication in vitro.

The replication of herpes simplex virus (HSV) type 1 and 2 in Vero cells is inhibited in the presence of enterocin CRL35 (ECRL), a bacteriocin produced by Enterococcus faecium CRL35. Attempts to resolve the mode of action of ECRL indicate that virus adsorption and penetration are not affected. Instead, a late step of virus multiplication is hindered since the addition of 100 microg/ml of ECRL at 8h post infection still causes a 90% inhibition of virus release. The effect of ECRL on HSV antigen expression was studied by immunofluorescence using a polyclonal serum and a monoclonal antibody against glycoprotein D (gamma protein). These studies indicated that ECRL impeded the second round of infection, apparently as a consequence of the inhibition of glycoprotein D expression. The replication of syncytial mutants of HSV-1 was significantly inhibited at a ECRL concentration of 25 microg/ml. Both the percentage of fused cells and the polykaryocyte size were affected. Studies on the effect of ECRL on viral protein synthesis showed that in the presence of ECRL, HSV late gamma proteins were not synthesized. From these findings, it is concluded that inhibition of HSV spreading by ECRL is due to the prevention of mainly late glycoprotein synthesis.

[Prevalence of associated microorganisms in genital discharge, Argentina]. / Prevalencia de microorganismos asociados a secrecíon genital femenina, Argentina.

OBJECTIVE: There was a significant increase in the number of women demanding gynecological care in public hospital of the Great Buenos Aires, Argentina, between 1997 and 1998. It was necessary to update the prevalence of associated microorganisms in order to review the laboratory support and adjust prevention and control guidelines. METHODS: Samples from vaginal and endocervical discharge, from total cases: 84 adolescents (15 to 19 years) and 784 adults (20-60 years) attended in 1997-1998, were studied. Neisseria gonorrhoeae, Streptococcus agalactiae, Trichomonas vaginalis, Candida spp, and bacterial vaginosis, were diagnosed applying direct detection methods and specific culture isolation. Chlamydia trachomatis (antigen detection), Ureaplasma urealyticum and Mycoplasma hominis (culture) were also studied in part of the population. RESULTS: Patient care increased steadily from 1997 and there was an increase of 2.1 times from the first semester to the last one in 1998. Bacterial vaginosis was the most prevalent disease in the adult group, with 23.8%; followed by Candida spp 17.8%; S. agalactiae 5.6%; T.vaginalis 2.4%. In 50.3% of total adult cases neither bacterial vaginosis or presence of any sought microorganisms, was detected. In the adolescents group the most frequent detection was Candida spp with a 29.7%; bacterial vaginosis in 17.8%; followed by S. agalactiae 3.6%, T.vaginalis 2.4%. Also in this group on an important number of cases, 46.4%, none bacterial vaginosis or the presence of the sought microorganism were found. In some of the adult group, C. trachomatis (7/400) 1.76%, U. urealyticum (209/340) 61.4% and M. hominis (45/272) 16.5% were detected. CONCLUSIONS: Bacterial vaginosis and Candida spp prevalence is important in both groups. The absence of N. gonorrhoeae and lower prevalence of T. vaginalis and C. trachomatis is remarkable. A high prevalence of U. urealyticum and M. hominis were also detected, but the actual pathogenic role in adult women is still under discussion. The significant increase of attendance in the public system is due to social changes in the population and not due to changes in any particular pathology. Laboratory support has to be adjusted to the most cost-effective procedures to solve the most prevalent problems.

Determinación de la concentración inhibitoria mínima de azitromicina para Chlamydia trachomatis en distintas líneas celulares / Determination of the minimun inhibitory concentration of azythromicin for chlamydia trachomatis in different cells cultures

Las bacterias del orden Chlamydiales son parásitos intracelulares obligados responsables de diferentes patologías tanto en humanos como en animales. Esta característica biológica hace que la determinación de la Concentración Inhibitoria Mínima (CIM) presente dificultades en su realización e interpretación. Se carece de una metodología normatizada, lo que dificulta el análisis y homologación de los pocos resultados publicados. Entre las múltiples variables a tener en cuenta, es importante considerar la línea celular a utilizar. En éste trabajo, se ha determinado la CIM de azitromicina para la cepa de Chlamydia trachomatis L2/434/BU utilizando las líneas celulares LLCMK2, McCoy y Vero, detectándose diferencias destacables en cuánto a los valores obtenidos. Se sugiere como el sustrato celular de mayor reproducibilidad a la línea LLCMK2 y se propone la normatización de los otros parámetros necesarios para determinar la CIM de las bacterias de éste orden (AU)

Genotipos de Chlamydia trachomatis circulantes en el área de la Ciudad de Buenos Aires y su Conurbano / Circulating genotypes of Chlamydia trachomatis in Buenos Aires metropolitan area and suburbs (Argentina)

El objetivo de éste estudio es aportar información sobre los genotipos de Chlamydia trachomatis (Ct) que circulan en el área metropolitana de Buenos Aires y su Conurbano. Se estudiaron 38 muestras positivas para Ct, provenientes de distintos hospitales, obtenidas del tracto urogenital de mujeres y hombres, y de conjuntivas de neonatos. La confirmación de la presencia de Ct se realizó por amplificación génica de un segmento del plásmido críptico mediante PCR. Para la genotipificación de Ct se amplificó el gen omp-1, los productos de ésta amplificación fueron digeridos con el enzima AluI, y los fragmentos obtenidos fueron separados por electroforesis en geles de poliacrilamida al 13,5 por ciento. Los resultados obtenidos fueron confirmados por digestión con EcoRI. Cuando fue necesario se recurrió a la digestión con una segunda enzima. El genotipo E, detectado en 20 casos, fue el más frecuente y estuvo presente en muestras de origen cervical, uretral y conjuntival, lo que en distribución muestra un perfil similar a la mayoría de las regiones donde se han estudiado. Esta mayor frecuencia de detección podría deberse a una mejor adaptación de éste genotipo a las condiciones de transmisión e infección y a las islas de virulencia particulares del genotipo (AU)

Determinación de la concentración inhibitoria mínima de azitromicina para Chlamydia trachomatis en distintas líneas celulares / Determination of the minimun inhibitory concentration of azythromicin for chlamydia trachomatis in different cells cultures

Las bacterias del orden Chlamydiales son parásitos intracelulares obligados responsables de diferentes patologías tanto en humanos como en animales. Esta característica biológica hace que la determinación de la Concentración Inhibitoria Mínima (CIM) presente dificultades en su realización e interpretación. Se carece de una metodología normatizada, lo que dificulta el análisis y homologación de los pocos resultados publicados. Entre las múltiples variables a tener en cuenta, es importante considerar la línea celular a utilizar. En éste trabajo, se ha determinado la CIM de azitromicina para la cepa de Chlamydia trachomatis L2/434/BU utilizando las líneas celulares LLCMK2, McCoy y Vero, detectándose diferencias destacables en cuánto a los valores obtenidos. Se sugiere como el sustrato celular de mayor reproducibilidad a la línea LLCMK2 y se propone la normatización de los otros parámetros necesarios para determinar la CIM de las bacterias de éste orden

Genotipos de Chlamydia trachomatis circulantes en el área de la Ciudad de Buenos Aires y su Conurbano / Circulating genotypes of Chlamydia trachomatis in Buenos Aires metropolitan area and suburbs (Argentina)

El objetivo de éste estudio es aportar información sobre los genotipos de Chlamydia trachomatis (Ct) que circulan en el área metropolitana de Buenos Aires y su Conurbano. Se estudiaron 38 muestras positivas para Ct, provenientes de distintos hospitales, obtenidas del tracto urogenital de mujeres y hombres, y de conjuntivas de neonatos. La confirmación de la presencia de Ct se realizó por amplificación génica de un segmento del plásmido críptico mediante PCR. Para la genotipificación de Ct se amplificó el gen omp-1, los productos de ésta amplificación fueron digeridos con el enzima AluI, y los fragmentos obtenidos fueron separados por electroforesis en geles de poliacrilamida al 13,5 por ciento. Los resultados obtenidos fueron confirmados por digestión con EcoRI. Cuando fue necesario se recurrió a la digestión con una segunda enzima. El genotipo E, detectado en 20 casos, fue el más frecuente y estuvo presente en muestras de origen cervical, uretral y conjuntival, lo que en distribución muestra un perfil similar a la mayoría de las regiones donde se han estudiado. Esta mayor frecuencia de detección podría deberse a una mejor adaptación de éste genotipo a las condiciones de transmisión e infección y a las islas de virulencia particulares del genotipo

Characterization of intermediate phenotypes induced by chemically undefined laboratory media in virulent Bordetella bronchiseptica strains.

The expression of many virulence factors of Bordetella bronchiseptica is regulated by the bvgAS locus and reduced in response to environmental signals called modulators. Virulent strains can alternate between virulent (Bvg(+)), intermediate (Bvg(i)), and modulated (Bvg(+)mod) phenotypes. Potential vaccine antigens can be expressed by Bvg1 strains grown only in the absence of modulators. In the present study we evaluated filamentous hemagglutinin (FHA) and outer membrane protein (OMP) expression in Bvg(+) B. bronchiseptica strains grown in chemically undefined media: nutrient agar (NA), tryptic soy agar (TSA), tryptose phosphate broth (TPB), and brain-heart infusion (BHI). Our results suggest that TSA and TPB usually induce semimodulation, since Bvg(+) strains cultured in these media retained the expression of FHA and virulence-associated OMPs in the 30 kDa region, but failed to express other virulence markers such as OMPs in the regions of 90 and 200 kDa, though they expressed flagellin (avirulence marker). On the other hand, NA and BHI usually induce modulation. Thus the assayed chemically undefined media should not be used in vaccine production. Semimodulation induced by TSA and TPB can be accurately detected by SDS-PAGE Sarkosyl-insoluble OMP-enriched profiles. The reduction or absence of OMPs in the regions of 90 and 200 kDa is the most sensitive marker, and in some cases the presence of flagellin in intermediate profiles is another trait of the Bvg(i) phenotypes. Therefore these markers could be useful for selecting media for vaccine production. We also characterized the phenotype of Bvg(+) strains grown in Stainer-Scholte broth, an expensive medium, with and without glutathione, and we have detected no differences; this is the first attempt to reduce the cost of a Bordetella growth medium for veterinary vaccine production.