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Rev Bras Ginecol Obstet. 2009;31(4):189-195
DOI 10.1590/S0100-72032009000400006
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.
Summary
Rev Bras Ginecol Obstet. 2009;31(4):189-195
DOI 10.1590/S0100-72032009000400006
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.
Summary
Rev Bras Ginecol Obstet. 2005;27(11):672-676
DOI 10.1590/S0100-72032005001100007
PURPOSE: to evaluate the influence of vaginal environment of pregnant women on group B streptococcus (GBS) survival after 8, 24 e 48 h in Amies and Stuart transport media. METHODS: Three vaginal samples were collected from 30 pregnant women attending the Prenatal Care Outpatient Clinic of the Centro de Atenção Integral à Saúde da Mulher (CAISM), Universidade Estadual de Campinas (UNICAMP). The first sample was placed directly onto Todd-Hewitt selective medium; the second was used to perform a gram-stained microscopy, and the third swab was placed in 2 mL physiological saline to which 200 µL of a suspension with 1-2 x 10(8) colony-forming units of GBS was added. After homogenization, six swabs were collected from this suspension (3 from Amie medium and 3 from Stuart medium). These six swabs were kept at room temperature for 8, 24 and 48 h and then incubated on blood agar. Bacterial growth at 37ºC was observed after a 24-h incubation period and it was semiquantitatively graded (0-3+) according to the number of colonies. Statistical analysis was performed by the exact Fisher test and the level of significance was set at 0.05. RESULTS: the recovery of GBS after 48-h storage in Amie and Stuart media was 97 e 87%, respectively. In one of the four cases where no GBS recovery was possible after 48 h of storage, vaginal pH was higher than 4.5, and in two of those cases cytolytic vaginosis was found. CONCLUSIONS: both transport media showed to be appropriate for GBS recovery up to 48 h after sampling. Characteristics of the vaginal enviroment did not influence GBS recovery as observed in this study.
Summary
Rev Bras Ginecol Obstet. 2005;27(11):672-676
DOI 10.1590/S0100-72032005001100007
PURPOSE: to evaluate the influence of vaginal environment of pregnant women on group B streptococcus (GBS) survival after 8, 24 e 48 h in Amies and Stuart transport media. METHODS: Three vaginal samples were collected from 30 pregnant women attending the Prenatal Care Outpatient Clinic of the Centro de Atenção Integral à Saúde da Mulher (CAISM), Universidade Estadual de Campinas (UNICAMP). The first sample was placed directly onto Todd-Hewitt selective medium; the second was used to perform a gram-stained microscopy, and the third swab was placed in 2 mL physiological saline to which 200 µL of a suspension with 1-2 x 10(8) colony-forming units of GBS was added. After homogenization, six swabs were collected from this suspension (3 from Amie medium and 3 from Stuart medium). These six swabs were kept at room temperature for 8, 24 and 48 h and then incubated on blood agar. Bacterial growth at 37ºC was observed after a 24-h incubation period and it was semiquantitatively graded (0-3+) according to the number of colonies. Statistical analysis was performed by the exact Fisher test and the level of significance was set at 0.05. RESULTS: the recovery of GBS after 48-h storage in Amie and Stuart media was 97 e 87%, respectively. In one of the four cases where no GBS recovery was possible after 48 h of storage, vaginal pH was higher than 4.5, and in two of those cases cytolytic vaginosis was found. CONCLUSIONS: both transport media showed to be appropriate for GBS recovery up to 48 h after sampling. Characteristics of the vaginal enviroment did not influence GBS recovery as observed in this study.