Quantitative assessment of vaginal microflora during use of tampons of various compositions.

Although the effect of vaginal tampons on the microbial flora during menstruation has recently been studied by several investigators, quantitative effects attributable to particular tampon fibers have received less attention. The purposes of the present study were (i) to determine and then to compare the effects of polyacrylate rayon tampons and viscose rayon tampons on the normal vaginal flora, (ii) to compare quantitative bacterial counts obtained from these tampons with those obtained from concomitant vaginal swabs, and (iii) to determine whether either of these tampon types alters the vaginal microflora when compared with the microflora in the same women using all-cotton tampons or external catamenial pads. Tampon and swab samples were obtained at predetermined times from 18 women for an average of seven menstrual cycles. Samples consisting of swabs from women wearing menstrual pads were compared with swab and tampon samples taken at predetermined times during the menstrual cycle from women using cotton, polyacrylate rayon, or viscose rayon tampons. Samples were analyzed for total aerobic, facultative, and anaerobic bacterial counts. Statistical evaluation of the results indicated that, on the whole, total bacterial counts decreased during menstruation and that the numbers of bacteria in tampons tended to be lower than those in swab samples taken at the same time. The tampon type had little effect on the vaginal microflora.     

Methods for quantitative and qualitative evaluation of vaginal microflora during menstruation.

The quantitative and qualitative changes in the bacterial flora of the vagina during menstruation have received inadequate study. Similarly, the effect of vaginal tampons on the microbial flora as well as the relationship between the microbial flora of the vagina and that of the tampon has not been adequately evaluated. The purposes of the present study were (i) to develop quantitative methods for studying the vaginal flora and the flora of tampons obtained during menstruation and (ii) to determine whether there were differences between the microflora of the tampon and that of the vaginal vault. Tampon and swab samples were obtained at various times from eight young healthy volunteers for 8 to 10 menstrual cycles. Samples consisted of swabs from women wearing menstrual pads compared with swab and tampon samples taken at various times during the menstrual cycle. Samples were analyzed for total facultative and anaerobic bacterial counts, and the six dominant bacterial species in each culture were identified. Statistical evaluation of the results indicates that total bacterial counts decreased during menstruation and that swab and tampon samples yielded similar total counts per unit weight of sample. The numbers of bacteria in tampons tended to be lower than in swabs taken at the same time. Overall, during menstruation, the concentrations of lactobacilli declined, but otherwise there was little difference among the species found during menstruation compared with those found in intermenstrual samples. Cotton tampons had little discernible effect on the microbial flora.     

Methods for quantitative and qualitative evaluation of vaginal microflora during menstruation

The quantitative and qualitative changes in the bacterial flora of the vagina during menstruation have received inadequate study. Similarly, the effect of vaginal tampons on the microbial flora as well as the relationship between the microbial flora of the vagina and that of the tampon has not been adequately evaluated. The purposes of the present study were (i) to develop quantitative methods for studying the vaginal flora and the flora of tampons obtained during menstruation and (ii) to determine whether there were differences between the microflora of the tampon and that of the vaginal vault. Tampon and swab samples were obtained at various times from eight young healthy volunteers for 8 to 10 menstrual cycles. Samples consisted of swabs from women wearing menstrual pads compared with swab and tampon samples taken at various times during the menstrual cycle. Samples were analyzed for total facultative and anaerobic bacterial counts, and the six dominant bacterial species in each culture were identified. Statistical evaluation of the results indicates that total bacterial counts decreased during menstruation and that swab and tampon samples yielded similar total counts per unit weight of sample. The numbers of bacteria in tampons tended to be lower than in swabs taken at the same time. Overall, during menstruation, the concentrations of lactobacilli declined, but otherwise there was little difference among the species found during menstruation compared with those found in intermenstrual samples. Cotton tampons had little discernible effect on the microbial flora.     

Qualitative assessment of vaginal microflora during use of tampons of various compositions

The effect of vaginal tampons on the microbial flora during menstruation has recently been studied by several investigators. However, little information regarding the qualitative effects attributable to particular tampon fibers is available. The purpose of the present study was to compare the effects of polyacrylate rayon tampons and cotton-viscose rayon blend tampons on the qualitative bacterial counts obtained from tampons and concomitant vaginal swabs and to determine whether either of these tampon types alters the qualitative makeup of the vaginal microflora when compared with the microflora in the same women using all-cotton tampons or external catamenial pads. Tampon and swab samples were obtained as described previously (A. B. Onderdonk, G. R. Zamarchi, M. L. Rodriguez, M. L. Hirsch, A. Mu?oz, and E. H. Kass, Appl. Environ. Microbiol. 53:2774-2778). The genus and species of the six dominant bacterial species in each sample were identified, if possible. A statistical evaluation of the qualitative makeup of the microflora revealed that the same numerically dominant phenotypes were present regardless of sample type, sample time, or catamenial product. Predictable changes in total numbers among the dominant species were also noted when the data were evaluated by day of menstrual cycle. The correlation between the total numbers of each dominant species present was evaluated by day of cycle, and the findings are discussed.     

Qualitative assessment of vaginal microflora during use of tampons of various compositions.

The effect of vaginal tampons on the microbial flora during menstruation has recently been studied by several investigators. However, little information regarding the qualitative effects attributable to particular tampon fibers is available. The purpose of the present study was to compare the effects of polyacrylate rayon tampons and cotton-viscose rayon blend tampons on the qualitative bacterial counts obtained from tampons and concomitant vaginal swabs and to determine whether either of these tampon types alters the qualitative makeup of the vaginal microflora when compared with the microflora in the same women using all-cotton tampons or external catamenial pads. Tampon and swab samples were obtained as described previously (A. B. Onderdonk, G. R. Zamarchi, M. L. Rodriguez, M. L. Hirsch, A. Muñoz, and E. H. Kass, Appl. Environ. Microbiol. 53:2774-2778). The genus and species of the six dominant bacterial species in each sample were identified, if possible. A statistical evaluation of the qualitative makeup of the microflora revealed that the same numerically dominant phenotypes were present regardless of sample type, sample time, or catamenial product. Predictable changes in total numbers among the dominant species were also noted when the data were evaluated by day of menstrual cycle. The correlation between the total numbers of each dominant species present was evaluated by day of cycle, and the findings are discussed.     

In vivo assessment of human vaginal oxygen and carbon dioxide levels during and post menses.

Previous in vitro and in vivo animal studies showed that O(2) and CO(2) concentrations can affect virulence of pathogenic bacteria such as Staphylococcus aureus. The objective of this work was to measure O(2) and CO(2) levels in the vaginal environment during tampon wear using newly available sensor technology. Measurements by two vaginal sensors showed a decrease in vaginal O(2) levels after tampon insertion. These decreases were independent of the type of tampons used and the time of measurement (mid-cycle or during menstruation). These results are not in agreement with a previous study that concluded that oxygenation of the vaginal environment during tampon use occurred via delivery of a bolus of O(2) during the insertion process. Our measurements of gas levels in menses showed the presence of both O(2) and CO(2) in menses. The tampons inserted into the vagina contained O(2) and CO(2) levels consistent with atmospheric conditions. Over time during tampon use, levels of O(2) in the tampon decreased and levels of CO(2) increased. Tampon absorbent capacity, menses loading, and wear time influenced the kinetics of these changes. Colonization with S. aureus had no effect on the gas profiles during menstruation. Taken collectively, these findings have important implications on the current understanding of gaseous changes in the vaginal environment during menstruation and the potential role(s) they may play in affecting bacterial virulence factor production.     

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 7^th 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.     

Toxic shock syndrome in Canada.

Since 1976, 53 confirmed or suspected cases of toxic shock syndrome (TSS) have been reported in Canada. Twenty-two cases occurred in 1980, and by October 1981 another 21 had been reported. In Canada, like the United States, where nearly 1200 cases have been recorded, TSS appears to be associated with tampon use, although a few cases have occurred in males and in nonmenstruating women. Of the 53 patients 3 died. The enterotoxin produced by Staphylococcus aureus is probably responsible for TSS. Manufacturers of tampons have now placed warning labels on tampon boxes and information on TSS in the instruction inserts. Women should select tampons of appropriate absorbency for the various stages of menstruation.     

The correction of dysbiotic disorders of the vaginal microflora by using a preparation made from highly adhesive lactobacteria

The effectiveness of a new bacterial preparation obtained from highly adhesive lactobacteria and intended for the correction of dysbiotic disturbances of vaginal microflora was studied in the treatment of 60 pregnant women with dysbacteriosis of the maternal passages. 30 pregnant women were simultaneously treated by the vaginal application of Lactobacterin. The study showed that the use of the preparation of highly adhesive lactobacteria caused the pronounced and stable correction of the microflora of the maternal passages. This correction was manifested by the domination of lactic acid bacterial flora and a decrease in the number of opportunistic microorganisms.     

妊娠妇女与无性生活女性阴道分泌物 微生物组学分析

目的探讨妊娠妇女与无性生活女性阴道分泌物中菌群结构差异,为临床女性泌尿生殖系统疾病研究提供可靠的依据。方法运用高通量二代测序技术检测20例妊娠妇女和29例无性生活女性阴道分泌物中微生物;采用秩和检验进行组间显著性差异分析,使用生物信息学软件进行数据处理。结果 49份样本共注释出14个门、23个纲、33个目、28个科、38个属和23个种水平物种。妊娠妇女与无性生活女性阴道分泌物中菌群结构差异显著。妊娠妇女在种水平仅有Atopobium vaginae、Campylobacter ureolyticus、Lactobacillus coleohominis、德氏乳杆菌、瑞氏乳杆菌、惰性乳杆菌、罗伊乳杆菌、白假丝酵母菌和近平滑假丝酵母菌共9个物种。结论妊娠能引起女性阴道分泌物菌群结构发生改变,且菌种数量明显减少。Lactobacillus coleohominis和德氏乳杆菌可能在维持妊娠妇女阴道菌群平衡和自净作用中起到关键性作用。     

Assessment of Lactobacillus species colonizing the vagina of apparently healthy Nigerian women, using PCR-DGGE and 16S rRNA gene sequencing

The identity of lactobacilli colonizing Africa-American women, have previously been based on culture-dependent methods. This led to some misleading speculations that black women lack lactobacilli in their vagina and are therefore highly susceptible to Bacterial vaginosis and STDs including HIV. In this study we used culture-independent procedures.About 241 vaginal swabs were obtained from ‘apparently’ healthy premenopausal women, between 18 and 48 years. Samples were Gram stained for the Nugent score evaluation. DNA was extracted from the bacteria on the vaginal swabs and amplified using polymerase chain reaction (PCR), with Lactobacillus primers. Samples with PCR products were separated with denaturing gradient gel electrophoresis (DGGE) and cut bands were re-amplified with Lactobacillus primers without the GC clamp. The re-amplified product was purified and sequenced with ABI PRISM Big-Dye Terminator. The sequence was identified with BLAST algorithm having the highest Gene Bank Accession number.Out of the 241 vaginal swab samples, 207 (85.8%) had PCR products, indicating the presence of lactobacilli, while 34 (14.2%) showed absence of lactobacilli and the Nugent scores were synonymous to either intermediate bacterial vaginosis or bacterial vaginosis (BV). Out of the 207 samples that had PCR product for Lactobacillus, 149 (72%) had sequence results as revealed by the BLAST algorithm. Most of the women (64%) were colonized by Lactobacillus iners as the predominant strain. Lactobacillus gasseri had 7.3%, followed by L. plantarum and L suntoryeus (6.0%) each. Others were colonized by Lactobacillus crispatus (3.0%), Lactobacillus rhamnosus (2.7%), Lactobacillus vaginalis (2.7%), Lactobacillus rennanqilfy (2.7%), followed by Lactobacillus fermentum (1.3%), Lactobacillus helveticus (1.3%), Lactobacillus johnsonii (1.3%) and Lactobacillus salivarus (1.3%). To our knowledge, this is the first report of Lactobacillus iners being the predominant species colonizing the vagina of the Nigerian women examined. This finding is in line with those from a recent study conducted among Canadian and Swedish women.     

An Evaluation of Risk Factors in Pregnant Women with <Emphasis Type="Italic">Candida</Emphasis> Vaginitis and the Diagnostic Value of Simultaneous Vaginal and Rectal Sampling

In this study, we investigated the epidemiological characteristics of VVC among pregnant women. We conducted a prospective survey among 372 pregnant women to investigate the prevalence, clinical forms, etiological agents, and predisposing factors of VVC. In addition, we determined the relationship between vaginal and rectal flora by simultaneously obtaining one high vaginal swab and one rectal swab from each patient using sterile cotton-tipped swabs. Furthermore, we compared the recovery and identification performances of chromID Candida agar to Sabouraud dextrose agar with gentamicin and chloramphenicol. Clinically and mycologically confirmed cases of VVC were detected in 139 (37.4%) and vaginal colonization described in 42 (11.3%) of 372 pregnant women. Rectal cultures were also positive in 98 of the 139 (70.5%) VVC cases. Candida albicans and C. glabrata were identified in vaginal samples in 58.0 versus 19.0% and from rectal samples in 49.0 versus 13.5%, respectively. Increases in gestational week and gravidae were identified to be statistically significant in patients with acute VVC (AVVC) and symptomatic recurrent VVC (RVVC), and asymptomatic RVVC (P = 0.04 and P = 0.03, respectively). In the laboratory diagnosis of VVC, specifically tailored chromogenic media are reliable tools for both the recovery and rapid identification of common Candida spp., particularly C. albicans, as well as for the detection of polyfungal populations in vaginal samples (P > 0.05). In addition, rectal colonization is a common finding in cases of AVVC and symptomatic-RVVC cases and corresponds well with the presence of the same yeast species in the vagina.     

Diagnostic value of Gram stain for assessment of vaginal smears during pregnancy

The main aim of this study was to compare the diagnostic value of Giemsa stained method with Gram stained method for the evaluation of vaginal smears among pregnant women. A study population comprised 111 pregnant between 6 and 30 weeks of gestation. The vaginal smears from every subject was diagnosed according to Giemsa and Gram stained method and micro-organisms were isolated by culture. In 29.3% cases diagnosed as normal flora (2a) on the basis of Giemsa method bacterial vaginosis was detected in Gram stains according to Spiegel's criteria and pathological microflora in concentration > or = 10(5) CFU/ml was cultured among 75.9% of them. Among 31.7% women who had grade 3a (abnormal) in Giemsa stains method normal flora was diagnosed on the basis on Gram's method and from 17.1% pregnant women from this group we did not isolated any pathogens. For evaluation of vaginal smears during pregnancy the Giemsa method should be replaced by Gram stained method.     

阴道微生态失调与早产相关性研究进展

健康育龄妇女的阴道微生态环境呈现多样性,阴道内微生物菌群以乳杆菌为主。乳杆菌可维持妇女阴道微环境平衡和保持阴道酸性环境以抑制致病性病原体生长。当乳杆菌比例低至致病阈值以下或无乳杆菌存在时,阴道微环境的菌群组成转变成以兼性需氧及厌氧菌为主。越来越多的证据表明,怀孕期间阴道菌群的异常与早产、及因早产和围产期结局不佳导致的潜在新生儿后遗症有关。本文综述了妊娠期阴道菌群的变化、细菌性阴道炎与早产发生风险等方面的问题,目的是帮助临床医生确定怀孕期间有阴道炎症状女性是否可接受治疗,从而降低早产、早产胎膜早破、绒毛膜羊膜炎和母胎传染病的发病率。     

The evaluation of prevalence and the impact of pathological microflora of the lower genital tract among women at early pregnancy on the risk of preterm delivery

The main aim of this prospective study was to determine the prevalence and an association between pathological microflora of the lower genital tract diagnosed at early pregnancy and the risk of preterm delivery. The study group comprised 179 randomly selected pregnant women from Lodz region, between 8 and 16 week of pregnancy. For the qualitative and quantitative assessment of biocenosis of the lower genital tract vaginal and cervical swabs were collected from the pregnant women under study. The C. trachomatis antigen was detected by direct immunofluorescence assay. The vaginal swabs were tested for aerobic and anaerobic bacteria. Bacterial vaginosis was diagnosed by Gram stain according to Spiegel's criteria. To evaluate the risk factors odds ratios were calculated using EPI INFO software. 21 (11.7%) women delivered before 37th week of pregnancy. Bacterial vaginosis was diagnosed among 51 (28.5%) pregnant women while intermediate microflora was diagnosed by Gram stain in 62 (34.6%) women. The shortest mean gestational age at delivery was noted among women with BV. The rate of preterm delivery in BV group was 15.7% comparing to 9.1% among women with normal microflora. Among women with preterm delivery BV was diagnosed in 38.1% (OR = 1.86). Based on culture results only 84 (46.9%) women had normal microflora at early pregnancy. The pathological culture was associated with slightly increased preterm delivery rate (12.6%) as compare to 10.7% in control group. Positive culture for Bacteroides and Mobiluncus was connected with nonstatistical rise in the risk of preterm delivery. No association between C. trachomatis infection at early pregnancy and elevated risk of preterm delivery was found. Early pregnancy diagnosis of bacterial vaginosis and its treatment should lower the rate of prematurity in Poland.     

Rely and Toxic Shock Syndrome: a technological health crisis

This essay examines factors leading to the identification of Toxic Shock Syndrome with the bacteria Staphylococcus aureus in 1978 and the specific role of Rely tampons in generating a technologically rooted health crisis. The concept biologically incompatible technology is offered to explain the relationship between constituent bacteria, women's menstrual cycles, and a reactive technology that converged to create the ideal environment for the S. aureus bacteria to live and flourish in some women. The complicated and reactive relationship of the Rely tampon to emergent disease, corporate interests, public health, and injury law reveals the dangers of naturalizing technologies.     

Studying the Effects of Reproductive Hormones and Bacterial Vaginosis on the Glycome of Lavage Samples from the Cervicovaginal Cavity

The cervicovaginal fluid (CVF) coating the vaginal epithelium is an important immunological mediator, providing a barrier to infection. Glycosylation of CVF proteins, such as mucins, IgG and S-IgA, plays a critical role in their immunological functions. Although multiple factors, such as hormones and microflora, may influence glycosylation of the CVF, few studies have examined their impact on this important immunological fluid. Herein we analyzed the glycosylation of cervicovaginal lavage (CVL) samples collected from 165 women under different hormonal conditions including: (1) no contraceptive, post-menopausal, (2) no contraceptive, days 1-14 of the menstrual cycle, (3) no contraceptive, days 15-28 of the menstrual cycle, (4) combined-oral contraceptive pills for at least 6 months, (5) depo-medroxyprogesterone acetate (Depo-Provera) injections for at least 6 months, (6) levonorgestrel IUD for at least 1 month. Glycomic profiling was obtained using our lectin microarray system, a rapid method to analyze carbohydrate composition. Although some small effects were observed due to hormone levels, the major influence on the glycome was the presence of an altered bacterial cohort due to bacterial vaginosis (BV). Compared to normal women, samples from women with BV contained lower levels of sialic acid and high-mannose glycans in their CVL. The change in high mannose levels was unexpected and may be related to the increased risk of HIV-infection observed in women with BV, as high mannose receptors are a viral entry pathway. Changes in the glycome were also observed with hormonal contraceptive use, in a contraceptive-dependent manner. Overall, microflora had a greater impact on the glycome than hormonal levels, and both of these effects should be more closely examined in future studies given the importance of glycans in the innate immune system.     

Evaluation of rayon swab surface sample collection method for Bacillus spores from nonporous surfaces

AIM: To evaluate US Centers for Disease Control and Prevention recommended swab surface sample collection method for recovery efficiency and limit of detection for powdered Bacillus spores from nonporous surfaces. METHODS AND RESULTS: Stainless steel and painted wallboard surface coupons were seeded with dry aerosolized Bacillus atrophaeus spores and surface concentrations determined. The observed mean rayon swab recovery efficiency from stainless steel was 0.41 with a standard deviation (SD) of +/-0.17 and for painted wallboard was 0.41 with an SD of +/-0.23. Evaluation of a sonication extraction method for the rayon swabs produced a mean extraction efficiency of 0.76 with an SD of +/-0.12. Swab recovery quantitative limits of detection were estimated at 25 colony forming units (CFU) per sample area for both stainless steel and painted wallboard. CONCLUSIONS: The swab sample collection method may be appropriate for small area sampling (10 -25 cm2) with a high agent concentration, but has limited value for large surface areas with a low agent concentration. The results of this study provide information necessary for the interpretation of swab environmental sample collection data, that is, positive swab samples are indicative of high surface concentrations and may imply a potential for exposure, whereas negative swab samples do not assure that organisms are absent from the surfaces sampled and may not assure the absence of the potential for exposure. SIGNIFICANCE AND IMPACT OF THE STUDY: It is critical from a public health perspective that the information obtained is accurate and reproducible. The consequence of an inappropriate public health response founded on information gathered using an ineffective or unreliable sample collection method has the potential for undesired social and economic impact.     

Microbiological Quality of Protein Feed Supplements Produced by Rendering Plants

Samples of protein feed supplements produced by rendering plants were examined for salmonellae, total aerobic bacterial counts, coliform counts, and enterococci. Isolations of salmonellae were more frequent from products with high counts; however, 6% of the samples with total counts of less than 1,000 per gram and 14% of the samples with coliform counts of less than 1 per gram contained salmonellae. Serotypes of Escherichia coli which have been associated with disease in domestic animals and poultry were also isolated from products. Although the distribution of serotypes of salmonellae isolated from environmental swabs and flies was similar to that isolated from products, the isolation of several serotypes from flies which had not been isolated in plants suggested that flies may be a potential source of contamination.