Sialidase Activity in the Cervicovaginal Fluid Is Associated With Changes in Bacterial Components of Lactobacillus-Deprived Microbiota.

INTRODUCTION: Sialidase activity in the cervicovaginal fluid (CVF) is associated with microscopic findings of bacterial vaginosis (BV). Sequencing of bacterial 16S rRNA gene in vaginal samples has revealed that the majority of microscopic BV cases fit into vaginal community-state type IV (CST IV), which was recently named "molecular-BV." Bacterial vaginosis-associated bacterial species, such as Gardnerella spp., may act as sources of CVF sialidases. These hydrolases lead to impairment of local immunity and enable bacterial adhesion to epithelial and biofilm formation. However, the impact of CVL sialidase on microbiota components and diversity remains unknown. OBJECTIVE: To assess if CVF sialidase activity is associated with changes in bacterial components of CST IV. METHODS: One hundred forty women were cross-sectionally enrolled. The presence of molecular-BV (CST IV) was assessed by V3-V4 16S rRNA sequencing (Illumina). Fluorometric assays were performed using 2-(4-methylumbelliferyl)-α-D-N-acetylneuraminic acid (MUAN) for measuring sialidase activity in CVF samples. Linear discriminant analysis effect size (LEfSe) was performed to identify the differently enriched bacterial taxa in molecular-BV according to the status of CVF sialidase activity. RESULTS: Forty-four participants (31.4%) had molecular-BV, of which 30 (68.2%) had sialidase activity at detectable levels. A total of 24 bacterial taxa were enriched in the presence of sialidase activity, while just two taxa were enriched in sialidase-negative samples. CONCLUSION: Sialidase activity in molecular-BV is associated with changes in bacterial components of the local microbiome. This association should be further investigated, since it may result in diminished local defenses against pathogens.

Characterization of the Vaginal Microbiome in Women of Reproductive Age From 5 Regions in Brazil.

BACKGROUND: Composition of the vaginal microbiome is strongly related to a woman's reproductive health and risk of sexually transmitted infections. Ethnoracial, behavioral, and environmental factors can influence microbiome. The Brazilian population is unique in terms of miscegenation of ethnic groups and behavioral characteristics across different regions. We aimed to characterize the vaginal microbiome of women from 5 geographical regions of Brazil. METHODS: We sequenced V3-V4 regions of 16S rRNA gene in vaginal samples of 609 reproductive-aged women. We performed logistic regression analyses to estimate odds ratios (OR) and 95% confidence intervals (CI) for the association between sociodemographic and behavioral factors with Lactobacillus-depleted microbiome (community state type [CST] IV). RESULTS: Vaginal samples were grouped into 5 CST: CST I (L. crispatus predominant, 30.5%), CST II (L. gasseri predominant, 4.4%), CST III (Lactobacillus iners predominant, 36.5%), CST IV (Lactobacillus-depleted, 27.4%), and CST V (L. jensenii predominant, 1.2%). Several factors were independently associated with CST IV, such as smoking (OR, 1.80; 95% CI, 1.02-3.18), number of partners (OR, 2.11; 95% CI, 1.20-3.70), and vaginal douching (OR, 2.24; 95% CI, 1.34-3.74). A protective effect was observed for milk/dairy intake (OR, 0.47; 95% CI, 0.27-0.82) and sitz bathing (OR, 0.43; 95% CI, 0.19-0.98). CONCLUSIONS: Nearly two thirds of Brazilian women may be at an increased risk for adverse outcomes associated with a vaginal microbiota characterized by the depletion of Lactobacillus or dominance by L. iners, whose protective role has been widely questioned. Several factors related to sexual behavior and intimate hygiene were associated with CST IV.

Cervicovaginal levels of proinflammatory cytokines are increased during chlamydial infection in bacterial vaginosis but not in lactobacilli-dominated flora.

OBJECTIVE: The purpose of this study was to assess the cervicovaginal levels of proinflammatory cytokines in women with Chlamydia trachomatis (CT) infection in the presence of bacterial vaginosis (BV) and normal flora and to compare with those negative for CT. MATERIALS AND METHODS: In this cross-sectional study, nonpregnant women were enrolled at 2 outpatient clinics and at 1 primary medical care unit in São Paulo State, Brazil. Cervicovaginal samples from 256 women with BV, of which 68 (26.6%) had concomitant CT infection and 188 (73.4%) were CT-negative, were measured for interleukin-1ß (IL-1ß), IL-6, and IL-8 by enzyme-linked immunosorbent assay. A matching number of samples from women with normal flora, CT-positive (n = 68) and negative (n = 188), were evaluated as control. Cytokine levels were compared by Mann-Whitney test and differences were considered significant at p < .05. RESULTS: In CT-negative women, IL-1ß was increased in BV (p < .001) when compared to normal flora, while the levels of IL-6 and IL8 were unchanged. The presence of CT infection was not associated with differences on cytokine levels in women with normal flora. However, women with BV had higher levels of IL-1ß (p = .02), IL-6 (p = .02), and IL-8 (p = .03) in the presence of CT when compared to those who tested negative for CT. CONCLUSIONS: Detection of endocervical CT is associated with increased cervicovaginal IL-1ß, IL-6, and IL-8 levels in women with concomitant BV but not in those with normal flora.

Do Atopobium vaginae, Megasphaera sp. and Leptotrichia sp. change the local innate immune response and sialidase activity in bacterial vaginosis?

OBJECTIVES: To investigate if the participation of Atopobium vaginae, Megasphaera sp. and Leptotrichia sp. in the bacterial community of bacterial vaginosis (BV) is associated with distinct patterns of this condition. METHODS: In this cross-sectional controlled study, 205 women with BV and 205 women with normal flora were included. Vaginal rinsing samples were obtained for measuring the levels of pro-inflammatory cytokines and bacterial sialidases. Real-time PCR was used to quantify the BV-associated bacteria and to estimate the total bacterial load using the 16S rRNA. Principal component analysis (PCA) using the measured parameters was performed to compare the BV samples with lower and higher loads of the species of interest. RESULTS: Higher bacterial load (p<0.001), levels of interleukin 1-ß (p<0.001) and sialidase activity (p<0.001) were associated with BV. Women with BV and higher relative loads of A vaginae, Megasphaera sp. and Leptotrichia sp. presented increased sialidase activity, but unchanged cytokine levels. PCA analysis did not indicate a different pattern of BV according to the loads of A vaginae, Megasphaera sp. and Leptotrichia sp. CONCLUSIONS: Greater participation of A vaginae, Megasphaera sp. and Leptotrichia sp. in vaginal bacterial community did not indicate a less severe form of BV; moreover, it was associated with increased sialidase activity.

Chlamydial infection in a high risk population: association with vaginal flora patterns.

PURPOSE: This study aimed to determine the frequency of Chlamydia trachomatis (CT) infection among high risk Brazilian women and evaluate its association with vaginal flora patterns. METHODS: This was a cross-sectional study, performed in an outpatient clinic of Bauru State Hospital, São Paulo, Brazil. A total of 142 women were included from 2006 to 2008. Inclusion criteria was dyspareunia, pain during bimanual exam, presence of excessive cervical mucus, cervical ectopy or with three or more episodes of abnormal vaginal flora (AVF) in the previous year before enrollment. Endocervical CT testing was performed by PCR. Vaginal swabs were collected for microscopic assessment of the microbial flora pattern. Gram-stained smears were classified in normal, intermediate or bacterial vaginosis (BV), and recognition of Candida sp. morphotypes. Wet mount smears were used for detection of Trichomonas vaginalis and aerobic vaginitis (AV). RESULTS: Thirty-four of 142 women (23.9%) tested positive for CT. AVF was found in 50 (35.2%) cases. The most frequent type of AVF was BV (17.6%). CT was strongly associated with the presence of AV (n = 7, 4.9%, P = 0.018), but not BV (n = 25, 17.6%, P = 0.80) or intermediate flora (n = 18, 12.7%, P = 0.28). CONCLUSIONS: A high rate of chlamydial infection was found in this population. Chlamydia infection is associated with aerobic vaginitis.

Diagnóstico da situaçäo do aleitamento materno em área urbana do Sudeste do Brasil: utilizaçäo de metodologia simplificada / Diagnosis of breastfeeding status in an urban area of Southeastern Brazil: use of a simplified methodology

O diagnóstico da situaçäo do aleitamento materno em populaçöes é necessário para a definiçäo de metas e avaliaçäo de programas de promoçäo e apoio a esta prática. Neste sentido, testou-se a viabilidade de realizar tal diagnóstico concomitante à Campanha Nacional de Multivacinaçäo, no Município de Botucatu, SP, Brasil. Utilizou-se um questionário simplificado (3 questöes tipo sim/näo) para estudar a alimentaçäo atual de 1.550 crianças menores de um ano (91,8 por cento de cobertura) que compareceram aos postos de vacinaçäo, em 19 de agosto de 1995. As medianas e freqüências das três categorias de aleitamento materno foram calculadas pela técnica de análise de probitos. As medianas obtidas foram: aleitamento materno exclusivo = 17 dias (IC: 4,6-28,7); aleitamento materno completo = 64 dias (IC: 53,0-74,5) e aleitamento materno = 167 dias (IC: 153,7-182,2). O bom ajuste dos três modelos foi evidenciado pelos valores de R2 e pelos testes de Kolmogorov-Smirnov (p<0,05). Os resultados confirmaram a necessidade do programa no município. A metodologia simplificada revelou-se viável. Recomenda-se sua utilizaçäo na monitorizaçäo da tendência do aleitamento materno e em estudos sobre o impacto de intervençöes