Eliane Melo Brolazo

Clinical diagnosis of bacterial vaginosis

Objective: To evaluate the accuracy of Amsels criteria individually or in combinations of two for the clinical diagnosis of bacterial vaginosis. Methods: This cross‐sectional study enrolled 135 women at UNICAMP, and evaluated Amsels four clinical criteria. Sensitivity, specificity, positive and negative predictive values were calculated for each criterion individually, in combinations of two, and for the classic test with at least three criteria present. The Nugent method was used as gold standard. Results: The most sensitive individual criteria were pH and vaginal discharge (97%), the latter having lowest specificity (26%). The criterion with highest specificity was the presence of clue cells (86%). The combination of two criteria showed sensitivity of 83% to 93%, and specificity of 82% to 94%. Conclusion: The accuracy of Amsels clinical criteria individually or in combinations of two was as accurate as the presence of at least three criteria for the diagnosis of bacterial vaginosis.

Phenotypical characteristics of group B streptococcus in parturients

Colonization by Group B Streptococcus (GBS) is highly prevalent among pregnant women, with prevalence rates ranging between 4% and 30%. The infection may be transmitted vertically and may result in serious neonatal consequences. In the period from November 2003 to May 2004, a cross-sectional study was carried out among 316 parturients at the Jundiaí Teaching Hospital to establish the prevalence of genital GBS colonization, to identify the factors associated with colonization and the characteristic phenotypes of these streptococci. Samples from rectal and vaginal areas were collected for selective culture in Todd-Hewitt broth. Susceptibility to 7 antimicrobial agents was tested using the antibiotic diffusion disk technique, and the isolated strains were classified using specific antisera. The prevalence of GBS colonization was 14.6%. No strain was resistant to penicillin, ampicillin, erythromycin or nitrofurantoin. The majority of strains were sensitive to cephalothin. Greatest resistance was to gentamicin (76.1%), followed by clindamycin (17.4%). The most frequent serotype was Ib (23.9%), followed by serotypes II and Ia (19.6% and 17.4%, respectively). There was no correlation between serotype and greater antimicrobial resistance. In conclusion, the prevalence of GBS in parturients was high and penicillin continues to be the drug of choice for intrapartum prophylaxis. The most frequent serotype (Ib) found in this study differs from those found in the majority of studies carried out in other countries, revealing the need to identify prevalent serotypes in each region so that specific vaccines can be designed.

Microbiological characteristics and inflammatory cytokines associated with preterm labor

PurposeTo evaluate vaginal microflora and interleukin-1β (IL-β), interleukin-6 (IL-6), interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-α) concentrations in the cervicovaginal fluid of a group of pregnant women in preterm labor when compared with a group of full-term pregnant women not yet in labor.MethodCase–control study performed in a University tertiary referral maternity in Campinas, Brazil with 45 pregnant women in preterm labor and 45 full-term pregnant women not in labor. All patients underwent speculum examination for the collection of cervicovaginal fluid. Bacterial vaginosis (BV) was diagnosed according to the criteria of Amsel and Nugent. Culture was performed for group B streptococcus (GBS) and lactobacilli, and hybrid capture assay for screening for chlamydial and gonococcal infection. Cytokine concentrations were measured using ELISA technique. Statistical analysis was performed using χ2, Fisher’s exact, and crude and adjusted odds ratios. Significance level was defined at 5%. The main outcome measures were cervicovaginal cytokines in preterm labor.ResultsIL-6 and IL-8 were significantly associated with preterm labor. The changes in vaginal microflora, as well as BV and GBS, were more frequent in women in preterm labor, although BV and GBS showed no statistical significance. The presence of Candida sp., absence of lactobacilli, positive screening for chlamydial and gonococcal infection and the presence of IL-1β and TNF-α were not associated with preterm labor.ConclusionsIL-6 and IL-8 and the presence of any type of vaginal infection were the factors that were significantly associated with preterm labor.

Correlation between API 50 CH and multiplex polymerase chain reaction for the identification of vaginal lactobacilli in isolates

Identification of Lactobacillus sp. strains by phenotypic methods may lead to doubtful results possibly interfering in the reliability of the epidemiological and probiotics studies. Therefore this study aimed to determine the best methodology for the identification of the large diversity of lactobacilli species found in the vagina by comparing two techniques, one based on their biochemical profile and other employing molecular biology. A carbohydrate fermentation test (API 50 CH) was compared with multiplex polymerase chain reaction (PCR) for the identification of species of vaginal lactobacilli from 135 healthy women. The kappa index was used to evaluate agreement between the methods. Using the molecular technique, L. crispatus (32.6%), L. jensenii (25%) and L. gasseri (20.6%) were the most frequent species. However, using the biochemical technique, the most frequent species were: L. acidophilus (34.8%), L. crispatus (27.2%) and L. fermentum (13%). Although L. acidophilus was the most frequent specie found by biochemical tests, no strain of this microorganism was detected by PCR. Agreement between the methods was low for identification of all the most common species. Although rates of L. crispatus detected were similar using both methods (32.6% and 27.2%), agreement between them was relatively low (kappa = 0.52). Conclusions: Our results confirmed the limitation of the biochemical method and the applicability of a previously published molecular method (Multiplex PCR) for the identification of lactobacilli in the vaginal tract, focusing on further necessity of its improvement for also targeting L. vaginalis and L. iners.

The effect of "breathable" panty liners on the female lower genital tract.

To evaluate whether the use of “breathable” panty liners (BPLs) alters the normal vaginal flora, increases the incidence of bacterial vaginosis and/or vaginal candidiasis, or causes vulvar irritation.

Prevalência e caracterização de espécies de lactobacilos vaginais em mulheres em idade reprodutiva sem vulvovaginites

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.

Influência do conteúdo vaginal de gestantes sobre a recuperação do estreptococo do grupo B nos meios de transporte Stuart e Amies

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 Atencao Integral a Saude 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 108 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.

Tinidazole versus cefazolin in antibiotic prophylaxis of vaginal and abdominal hysterectomy

PURPOSE to compare the efficacy of tinidazole and cephazolin on the febrile and infectious morbidity of post vaginal and abdominal hysterectomy antibiotic prophylaxis. METHODS randomized clinical study, where women admitted to hospital for hysterectomy were randomly allocated in one of the following antibiotic prophylaxis groups: Group C (2 g of IV cephazolin in the anesthetic induction); Group T (2 g of tinidazole orally, 12 hours before the surgery); or Group C+T (2 g of tinidazole orally 12 hours before the surgery and 2g of IV cephazolin in the anesthetic induction). Cervicovaginal smears were collected for specific cultures and the diagnosis of bacterial vaginosis (BV) was based in Amsel and Nugents criteria. The patients were reevaluated 7 and 30 days after the surgery for signs of febrile and/or infectious morbidity. The chi2 or the Fishers exact test was used to assess differences among the three groups, with a significance level of 5%. The sample power (1-beta) was calculated through the SAS program. RESULTS seven days after the hysterectomy, infectious morbidity was diagnosed in 6.6% of the women, but with no significant difference among the three groups studied (p=0.12). There was no febrile or infectious morbidity at the immediate post-surgical period or after 30 days from the surgery. BV ratio at the pre-surgical period was significantly higher among the women submitted to vaginal hysterectomy, rather than among the ones submitted to abdominal hysterectomy (27 versus 7%, p=0.02). BV ratio was also higher after 30 days, among the women submitted to vaginal hysterectomy (20 versus 8%), though without statistical significance (p=0.19). CONCLUSIONS the use of tinidazole, isolated or associated with cephazolin has not presented higher efficacy, than the use of cephazolin, alone to prevent febrile or infectious morbidity post hysterectomy. The high ratio of BV at the immediate pre-surgery period among the women submitted to vaginal hysterectomy suggests that this infection must be better investigated and properly treated before the surgery.