Prevalência e caracterização de espécies de lactobacilos vaginais em mulheres em idade reprodutiva sem vulvovaginites
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Data
2009-04-01
Autores
Brolazo, Eliane Melo
Simões, Jose Antonio
Nader, Maria Elena Fátima
Tomás, Maria Silvina Juárez
Gregoracci, Gustavo Bueno
Marconi, Camila [UNESP]
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Editor
Federação Brasileira das Sociedades de Ginecologia e Obstetrícia
Resumo
OBJETIVO: identificar espécies de lactobacilos isolados do conteúdo vaginal de mulheres saudáveis e assintomáticas; determinar as espécies mais prevalentes e caracterizá-las fenotipicamente. MÉTODOS: lactobacilos foram isolados em meio seletivo a partir de amostras de conteúdo vaginal de 135 mulheres, sem queixa de corrimento e com diagnóstico laboratorial negativo para infecções vaginais, acompanhadas em um ambulatório de Planejamento Familiar. Os isolados foram identificados por PCR multiplex e, quando necessário, submetidos ao sequenciamento do gene RNAr 16S. Foram também avaliados quanto à acidificação do meio de cultura, à produção de ácido láctico, de H2O2, bacteriocinas e a capacidade de adesão às células epiteliais. RESULTADOS: oitenta e três cepas de lactobacilos foram isoladas e identificadas, sendo as espécies predominantes L. crispatus (30,1%), L. jensenii (26,5%), L. gasseri (22,9%) e L. vaginalis (8,4%). Apenas 20 destes isolados não produziram H2O2 em quantidades detectáveis. Das 37 linhagens selecionadas para teste de adesão a células epiteliais, 12 apresentaram adesão entre 50 a 69%, 10 apresentaram 70% ou mais, e as restantes pouca ou nenhuma adesão. Nenhum dos isolados produziu bacteriocinas. CONCLUSÕES: as espécies de lactobacilos mais prevalentes em mulheres sem vulvovaginites, isoladas em meio de cultura seletivo e identificadas por métodos moleculares, foram L. crispatus, L. jensenii e L. gasseri. Além de mais frequentes, tais linhagens também apresentaram melhor produção de H2O2 e atingiram menores valores de pH em meio de cultura.
PURPOSE: to identify species of lactobacillus isolated from the vaginal contents of healthy and asymptomatic women, determining the most prevalent species and characterizing them phenotypically. METHODS: lactobacillus have been isolated in selective milieu from samples of the vaginal contents of 135 women without complaints of vaginal secretion, and with negative laboratorial diagnosis of vaginal infection, followed up at an outpatient clinic. After being identified by multiplex PCR, the isolates have been submitted to RNAr 16S gene sequencing, when necessary. They have also been evaluated concerning the production of lactic acid, H2O2, bacteriocins and the ability to adhere to epithelial cells. RESULTS: eight-three lactobacillus strains were isolated and identified, L. crispatus (30.1%), L. jensenii (26.5%), L. gasseri (22.9%) e L. vaginalis (8.4%), being the prevalent species. Only 20 of those isolates did not present H2O2 production, in detectable amounts. From the 37 strains selected for the test of adhesion to the epithelial cells, 12 presented 50 to 69% of adhesion, 10 presented 70% or more, and the remaining, little or no adhesion at all. None of the tested strains produced bacteriocins. CONCLUSIONS: the lactobacillus species more prevalent in women without vulvovaginitis, isolated in selective culture milieu and identified by molecular methods were L. crispatus, L. jensenii and L. gasseri. Besides the fact of being more prevalent, these strains also presented better production of H2O2, and reached lower pH values in the culture milieu.
PURPOSE: to identify species of lactobacillus isolated from the vaginal contents of healthy and asymptomatic women, determining the most prevalent species and characterizing them phenotypically. METHODS: lactobacillus have been isolated in selective milieu from samples of the vaginal contents of 135 women without complaints of vaginal secretion, and with negative laboratorial diagnosis of vaginal infection, followed up at an outpatient clinic. After being identified by multiplex PCR, the isolates have been submitted to RNAr 16S gene sequencing, when necessary. They have also been evaluated concerning the production of lactic acid, H2O2, bacteriocins and the ability to adhere to epithelial cells. RESULTS: eight-three lactobacillus strains were isolated and identified, L. crispatus (30.1%), L. jensenii (26.5%), L. gasseri (22.9%) e L. vaginalis (8.4%), being the prevalent species. Only 20 of those isolates did not present H2O2 production, in detectable amounts. From the 37 strains selected for the test of adhesion to the epithelial cells, 12 presented 50 to 69% of adhesion, 10 presented 70% or more, and the remaining, little or no adhesion at all. None of the tested strains produced bacteriocins. CONCLUSIONS: the lactobacillus species more prevalent in women without vulvovaginitis, isolated in selective culture milieu and identified by molecular methods were L. crispatus, L. jensenii and L. gasseri. Besides the fact of being more prevalent, these strains also presented better production of H2O2, and reached lower pH values in the culture milieu.
Descrição
Palavras-chave
Lactobacillus, Lactobacillus, Vagina, Reação em cadeia da polimerase, Vulvovaginite, Lactobacillus, Lactobacillus, Polymerase chain reaction, Vagina, Vulvovaginitis
Como citar
Revista Brasileira de Ginecologia e Obstetrícia. Federação Brasileira das Sociedades de Ginecologia e Obstetrícia, v. 31, n. 4, p. 189-195, 2009.