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Comparison of different sampling techniques and of different culture methods for detection of group B streptococcus carriage in pregnant women

BackgroundStreptococcus agalactiae (group B streptococcus; GBS) is a significant cause of perinatal and neonatal infections worldwide. To detect GBS colonization in pregnant women, the CDC recommends isolation of the bacterium from vaginal and anorectal swab samples by growth in a selective enrichment medium, such as Lim broth (Todd-Hewitt broth supplemented with selective antibiotics), followed by subculture on sheep blood agar. However, this procedure may require 48 h to complete. We compared different sampling and culture techniques for the detection of GBS.MethodsA total of 300 swabs was taken from 100 pregnant women at 35-37 weeks of gestation. For each subject, one rectovaginal, one vaginal and one rectal ESwab were collected. Plating onto Columbia CNA agar (CNA), group B streptococcus differential agar (GBSDA) (Granada Medium) and chromID Strepto B agar (CA), with and without Lim broth enrichment, were compared. The isolates were confirmed as S. agalactiae using the CAMP test on blood agar and by molecular identification with tDNA-PCR or by 16S rRNA gene sequence determination.ResultsThe overall GBS colonization rate was 22%. GBS positivity for rectovaginal sampling (100%) was significantly higher than detection on the basis of vaginal sampling (50%), but not significantly higher than for rectal sampling (82%). Direct plating of the rectovaginal swab on CNA, GBSDA and CA resulted in detection of 59, 91 and 95% of the carriers, respectively, whereas subculturing of Lim broth yielded 77, 95 and 100% positivity, respectively. Lim broth enrichment enabled the detection of only one additional GBS positive subject. There was no significant difference between GBSDA and CA, whereas both were more sensitive than CNA. Direct culture onto GBSDA or CA (91 and 95%) detected more carriers than Lim broth enrichment and subculture onto CNA (77%). One false negative isolate was observed on GBSDA, and three false positives on CA.ConclusionsIn conclusion, rectovaginal sampling increased the number GBS positive women detected, compared to vaginal and/or rectal sampling. Direct plating on CA and/or GBSDA provided rapid detection of GBS that was at least as sensitive and specific as the CDC recommended method of Lim broth subcultured onto non chromogenic agar.

Identification and genotyping of bacteria from paired vaginal and rectal samples from pregnant women indicates similarity between vaginal and rectal microflora

BackgroundThe vaginal microflora is important for maintaining vaginal health and preventing infections of the reproductive tract. The rectum has been suggested as the major source for the colonisation of the vaginal econiche.MethodsTo establish whether the rectum can serve as a possible bacterial reservoir for colonisation of the vaginal econiche, we cultured vaginal and rectal specimens from pregnant women at 35-37 weeks of gestation, identified the isolates to the species level with tRNA intergenic length polymorphism analysis (tDNA-PCR) and genotyped the isolates for those subjects from which the same species was isolated simultaneously vaginally and rectally, by RAPD-analysis.One vaginal and one rectal swab were collected from a total of each of 132 pregnant women at 35-37 weeks of gestation. Swabs were cultured on Columbia CNA agar and MRS agar. For each subject 4 colonies were selected for each of both sites, i.e. 8 colonies in total.ResultsAmong the 844 isolates that could be identified by tDNA-PCR, a total of 63 bacterial species were present, 9 (14%) only vaginally, 26 (41%) only rectally, and 28 (44%) in both vagina and rectum. A total of 121 (91.6%) of 132 vaginal samples and 51 (38.6%) of 132 rectal samples were positive for lactobacilli. L. crispatus was the most frequently isolated Lactobacillus species from the vagina (40% of the subjects were positive), followed by L. jensenii (32%), L. gasseri (30%) and L. iners (11%). L. gasseri was the most frequently isolated Lactobacillus species from the rectum (15%), followed by L. jensenii (12%), L. crispatus (11%) and L. iners (2%).A total of 47 pregnant women carried the same species vaginally and rectally. This resulted in 50 vaginal/rectal pairs of the same species, for a total of eight different species. For 34 of the 50 species pairs (68%), isolates with the same genotype were present vaginally and rectally and a high level of genotypic diversity within species per subject was also established.ConclusionIt can be concluded that there is a certain degree of correspondence between the vaginal and rectal microflora, not only with regard to species composition but also with regard to strain identity between vaginal and rectal isolates.These results support the hypothesis that the rectal microflora serves as a reservoir for colonisation of the vaginal econiche.

Longitudinal Study of the Dynamics of Vaginal Microflora during Two Consecutive Menstrual Cycles

Background Although the vaginal microflora (VMF) has been well studied, information on the fluctuation of the different bacterial species throughout the menstrual cycle and the information on events preceding the presence of disturbed VMF is still very limited. Documenting the dynamics of the VMF during the menstrual cycle might provide better insights. In this study, we assessed the presence of different Lactobacillus species in relation to the BV associated species during the menstrual cycle, assessed the influence of the menstrual cycle on the different categories of vaginal microflora and assessed possible causes, such as menstruation and sexual intercourse, of VMF disturbance. To our knowledge, this is the first longitudinal study in which swabs and Gram stains were available for each day of two consecutive menstrual cycles, whereby 8 grades of VMF were distinguished by Gram stain analysis, and whereby the swabs were cultured every 7th day and identification of the bacterial isolates was carried out with a molecular technique. Methods Self-collected vaginal swabs were obtained daily from 17 non pregnant, menarchal volunteers, and used for daily Gram staining and weekly culture. Bacterial isolates were identified with tDNA-PCR and 16 S rRNA gene sequencing. Results Nine women presented with predominantly normal VMF and the 8 others had predominantly disturbed VMF. The overall VMF of each volunteer was characteristic and rather stable. Menses and antimicrobials were the major disturbing factors of the VMF. Disturbances were always accompanied by a rise in Gram positive cocci, which also appeared to be a significant group within the VMF in general. Conclusions We observed a huge interindividual variability of predominantly stable VMF types. The importance of Gram positive cocci in VMF is underestimated. L. crispatus was the species that was most negatively affected by the menses, whereas the presence of the other lactobacilli was less variable.

Longitudinal qPCR Study of the Dynamics of L. crispatus, L. iners, A. vaginae, (Sialidase Positive) G. vaginalis, and P. bivia in the Vagina

Background To obtain more detailed understanding of the causes of disturbance of the vaginal microflora (VMF), a longitudinal study was carried out for 17 women during two menstrual cycles. Methods Vaginal swabs were obtained daily from 17 non-pregnant, menarchal volunteers. For each woman, Gram stains were scored, the quantitative changes of 5 key vaginal species, i.e. Atopobium vaginae, Lactobacillus crispatus, L. iners, (sialidase positive) Gardnerella vaginalis and Prevotella bivia were quantified with qPCR and hydrogen-peroxide production was assessed on TMB+ agar. Results Women could be divided in 9 subjects with predominantly normal VMF (grades Ia, Ib and Iab, group N) and 8 with predominantly disturbed VMF (grades I-like, II, III and IV, group D). VMF was variable between women, but overall stable for most of the women. Menses were the strongest disturbing factor of the VMF. L. crispatus was present at log7–9 cells/ml in grade Ia, Iab and II VMF, but concentrations declined 100-fold during menses. L. crispatus below log7 cells/ml corresponded with poor H2O2-production. L. iners was present at log 10 cells/ml in grade Ib, II and III VMF. Sialidase negative G. vaginalis strains (average log5 cells/ml) were detected in grade I, I-like and IV VMF. In grade II VMF, predominantly a mixture of both sialidase negative and positive G. vaginalis strains (average log9 cells/ml) were present, and predominantly sialidase positive strains in grade III VMF. The presence of A. vaginae (average log9 cells/ml) coincided with grade II and III VMF. P. bivia (log4–8 cells/ml) was mostly present in grade III vaginal microflora. L. iners, G. vaginalis, A. vaginae and P. bivia all increased around menses for group N women, and as such L. iners was considered a member of disturbed VMF. Conclusions This qPCR-based study confirms largely the results of previous culture-based, microscopy-based and pyrosequencing-based studies.

Genotyping of Streptococcus agalactiae(group B streptococci) isolated from vaginal and rectal swabs of women at 35-37 weeks of pregnancy

BackgroundGroup B streptococci (GBS), or Streptococcus agalactiae, are the leading bacterial cause of meningitis and bacterial sepsis in newborns. Here we compared different culture media for GBS detection and we compared the occurrence of different genotypes and serotypes of GBS isolates from the vagina and rectum.MethodsStreptococcus agalactiae was cultured separately from both rectum and vagina, for a total of 150 pregnant women, i) directly onto Columbia CNA agar, or indirectly onto ii) Granada agar resp. iii) Columbia CNA agar, after overnight incubation in Lim broth.ResultsThirty six women (24%) were colonized by GBS. Of these, 19 harbored GBS in both rectum and vagina, 9 only in the vagina and 8 exclusively in the rectum. The combination of Lim broth and subculture on Granada agar was the only culture method that detected all GBS positive women. Using RAPD-analysis, a total of 66 genotypes could be established among the 118 isolates from 32 women for which fingerprinting was carried out. Up to 4 different genotypes in total (rectal + vaginal) were found for 4 women, one woman carried 3 different genotypes vaginally and 14 women carried two 2 different genotypes vaginally. Only two subjects were found to carry strains with the same genotype, although the serotype of both of these strains was different.Eighteen of the 19 subjects with GBS at both sites had at least one vaginal and one rectal isolate with the same genotype.We report the presence of two to four different genotypes in 22 (61%) of the 36 GBS positive women and the presence of identical genotypes in both sites for all women but one.ConclusionThe combination of Lim broth and subculture on Granada medium provide high sensitivity for GBS detection from vaginal and rectal swabs from pregnant women. We established a higher genotypic diversity per individual than other studies, with up to four different genotypes among a maximum of 6 isolates per individual picked. Still, 18 of the 19 women with GBS from both rectum and vagina had at least one isolate from each sampling site with the same genotype.

Imbalances between matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases (TIMPs) in maternal serum during preterm labor

Background Matrix metalloproteinases (MMPs) are involved in remodeling of the extracellular matrix (ECM) during pregnancy and parturition. Aberrant ECM degradation by MMPs or an imbalance between MMPs and their tissue inhibitors (TIMPs) have been implicated in the pathogenesis of preterm labor, however few studies have investigated MMPs or TIMPs in maternal serum. Therefore, the purpose of this study was to determine serum concentrations of MMP-3, MMP-9 and all four TIMPs as well as MMP:TIMP ratios during term and preterm labor. Methods A case control study with 166 singleton pregnancies, divided into four groups: (1) women with preterm birth, delivering before 34 weeks (PTB); (2) gestational age (GA) matched controls, not in preterm labor; (3) women at term in labor and (4) at term not in labor. MMP and TIMP concentrations were measured using Luminex technology. Results MMP-9 and TIMP-4 concentrations were higher in women with PTB vs. GA matched controls (resp. p = 0.01 and p<0.001). An increase in MMP-9:TIMP-1 and MMP-9:TIMP-2 ratio was observed in women with PTB compared to GA matched controls (resp. p = 0.02 and p<0.001) as well as compared to women at term in labor (resp. p = 0.006 and p<0.001). Multiple regression results with groups recoded as three key covariates showed significantly higher MMP-9 concentrations, higher MMP-9:TIMP-1 and MMP-9:TIMP-2 ratios and lower TIMP-1 and -2 concentrations for preterm labor. Significantly higher MMP-9 and TIMP-4 concentrations and MMP-9:TIMP-2 ratios were observed for labor. Conclusions Serum MMP-9:TIMP-1 and MMP-9:TIMP-2 balances are tilting in favor of gelatinolysis during preterm labor. TIMP-1 and -2 concentrations were lower in preterm gestation, irrespective of labor, while TIMP-4 concentrations were raised in labor. These observations suggest that aberrant serum expression of MMP:TIMP ratios and TIMPs reflect pregnancy and labor status, providing a far less invasive method to determine enzymes essential in ECM remodeling during pregnancy and parturition.

Female genital mutilation: Knowledge, attitude and practices of Flemish midwives

BACKGROUND health professionals in Belgium are confronted with female genital mutilation (FGM). To date, no survey to assess knowledge, attitudes and practices on FGM was conducted among midwives in the Northern region of Belgium. OBJECTIVE the objective of this study was to assess the knowledge, attitude and practices of Flemish midwives regarding female genital mutilation (FGM). DESIGN we used a quantitative design, using KAP study (semi-structured questionnaire). SETTING labour wards, maternity wards and maternal intensive care units (MIC) in 56 hospitals in Flemish region of Belgium. PARTICIPANTS 820 midwives, actively working in labour wards, maternity wards and maternal intensive care units (MIC). FINDINGS 820 valid questionnaires (40.9%) were returned. More than 15% of the respondents were recently confronted with FGM. They were mostly faced with the psychological and sexual complications caused by FGM. Few respondents were aware of existing guidelines regarding FGM in their hospitals (3.5%). The results also showed that only 20.2% was aware of the exact content of the law. The majority of midwives condemned the harmful traditional practice: FGM was experienced as a form of violence against women or a violation of human rights. Only 25.9% declared that FGM forms a part of their midwifery program. The vast majority of respondents (92.5%) indicated a need for more information on the subject. KEY CONCLUSIONS this study indicated that midwives in Flanders are confronted with FGM and its complications and highlighted the gaps in the knowledge of Flemish midwives regarding FGM. This may interfere with the provision of adequate care and prevention of FGM for the new-born daughter. IMPLICATIONS FOR PRACTICE there is an important need for appropriate training of (student)midwives concerning FGM as well as for the development and dissemination of clear guidelines in Flemish hospitals.

Skin hydration in nursing home residents using disposable bed baths

The objective of this study was to evaluate a new way for applying bed baths and reducing the risk for dry skin by comparing the effect of two washing methods on skin hydration. A cluster randomized trial was conducted. Skin hydration was measured before and after implementation of disposable wash gloves, using a MoistureMeter SC at three skin sites. Total skin hydration did not differ between residents at the start of the study in both groups. After implementation, the post minus pre hydration scores were higher for the intervention group than the control group at all skin sites. However, the difference was only significant at cheek site. The use of disposable wash gloves does not increase the risk for dry skin in comparison with traditional washing methods. These results may encourage the introduction of disposable wash gloves as an innovation in daily skin care practice.

Analysis of smoking cessation beliefs in pregnant smokers and ex-smokers using the Theory of Planned Behavior

AimTo analyze the association between smoking cessation beliefs and smoking status, and between smoking cessations beliefs and intention to quit, using the Theory of Planned Behavior (TPB).Subjects and methodsAn observational study using a questionnaire was performed; data were collected from 264 Flemish pregnant smokers and ex-smokers before week 16 of pregnancy.ResultsThere was a significant difference in the behavioral beliefs of the TPB between smokers and ex-smokers, after controlling for education and age. All respondents experienced more support from their non-smoking partners to maintain abstinence during pregnancy. We found no significant difference in the behavioral beliefs of the TPB between respondents with low and high intention to quit smoking.ConclusionOur results suggest that attitude, subjective norms, support especially from the partner, and perceived behavioral control are associated with actual smoking behavior in pregnant women and not with intention to quit smoking. It is important to engage the partner and/or significant others in smoking cessation counselling so that they can support the pregnant woman in an attempt to quit or to maintain abstinence.

The Modified Reasons for Smoking Scale: factorial structure, validity and reliability in pregnant smokers

RATIONALE, AIMS AND OBJECTIVES Smoking during pregnancy can cause several maternal and neonatal health risks, yet a considerable number of pregnant women continue to smoke. The objectives of this study were to test the factorial structure, validity and reliability of the Dutch version of the Modified Reasons for Smoking Scale (MRSS) in a sample of smoking pregnant women and to understand reasons for continued smoking during pregnancy. METHODS A longitudinal design was performed. Data of 97 pregnant smokers were collected during prenatal consultation. Structural equation modelling was performed to assess the construct validity of the MRSS: an exploratory factor analysis was conducted, followed by a confirmatory factor analysis.Test-retest reliability (<16 weeks and 32-34 weeks pregnancy) and internal consistency were assessed using the intraclass correlation coefficient and the Cronbachs alpha, respectively. To verify concurrent validity, Mann-Whitney U-tests were performed examining associations between the MRSS subscales and nicotine dependence, daily consumption, depressive symptoms and intention to quit. RESULTS We found a factorial structure for the MRSS of 11 items within five subscales in order of importance: tension reduction, addiction, pleasure, habit and social function. Results for internal consistency and test-retest reliability were good to acceptable. There were significant associations of nicotine dependence with tension reduction and addiction and of daily consumption with addiction and habit. CONCLUSIONS Validity and reliability of the MRSS were shown in a sample of pregnant smokers. Tension reduction was the most important reason for continued smoking, followed by pleasure and addiction. Although the score for nicotine dependence was low, addiction was an important reason for continued smoking during pregnancy; therefore, nicotine replacement therapy could be considered. Half of the respondents experienced depressive symptoms. Hence, it is important to identify those women who need more specialized care, which can include not only smoking cessation counselling but also treatment for depression.