Effect of metronidazole on the growth of vaginal lactobacilli in vitro

OBJECTIVE: To determine whether metronidazole has an adverse effect on the growth of Lactobacillus. METHODS: Hydrogen peroxide- and bacteriocin-producing strains of Lactobacillus were used as test strains. Concentrations of metronidazole used ranged from 128 to 7000 microg/ml. Susceptibility to metronidazole was conducted by the broth microdilution method recommended by the National Committee for Clinical Laboratory Standards. RESULTS: Growth of Lactobacillus was partially inhibited at concentrations between 1000 and 4000 microg/ml (p = 0.014). Concentrations > or = 5000 microg/ml completely inhibited growth of Lactobacillus. Concentrations between 128 and 256 microg/ml stimulated growth of Lactobacillus (p = 0.025 and 0.005, respectively). Concentrations of metronidazole between 64 and 128 microg/ml or > or = 512 microg/ml did not have an inhibitory or a stimulatory effect on the growth of Lactobacillus compared to the control. CONCLUSIONS: High concentration of metronidazole, i.e. between 1000 and 4000 microg/ml, partially inhibited the growth of Lactobacillus. Concentrations > or = 5000 microg/ml completely suppressed the growth of Lactobacillus. Concentrations between > or = 128 and < or = 256 microg/ml stimulated the growth of Lactobacillus. Further investigation to determine the ideal concentration of metronidazole is needed in order to use the antimicrobial agent effectively in the treatment of bacterial vaginosis.     

Defense factors of vaginal lactobacilli

OBJECTIVE: To determine the antagonistic relationship between vaginal lactobacilli and endogenous vaginal microflora. STUDY DESIGN: Twenty-two Lactobacillus strains were studied for the production of lactic acid, hydrogen peroxide, and bacteriocin. RESULTS: Under standardized growth conditions, most strains increased their biomass by more than 4 times. Lactobacillus species grew best at a pH > or = 4.5, and growth was retarded at a pH < 4.5. Lactic acid levels were 0.68 to 2.518 mg/mL and were not related to the number of cells or the pH of media. The pH of the media was caused by the secretion of lactic and other organic acids. Approximately 80% of the strains produced H(2)O(2) and were graded as 2+ in one third of the strains and 1+ in others. No statistical correlation was found between H(2)O(2) lactic acid and bacteriocin production. Bacteriocin activity was tested on 4 strains of Gardnerella vaginalis. Approximately 80% of the lactobacilli tested produced bacteriocin that inhibited growth of G vaginalis. Six of the strains did not produce bacteriocin. Thirteen strains produced all 3 defense factors, whereas the others lacked 1 or 2 properties. CONCLUSIONS: Lactobacillus species grow best at a pH > 4.5. The pH of the media is dependent on the cell mass and on all organic acids produced by Lactobacillus species. Although all species produce organic acids, not all produce H(2)O(2) and bacteriocin. Not all strains of G vaginalis can be inhibited by lactobacilli-producing bacteriocin.     

Hydrogen peroxide producing lactobacilli in women with vaginal infections

OBJECTIVE: Hydrogen peroxide producing lactobacilli are isolated from the vaginas of a majority of healthy reproductive age women. Their toxic and inhibitory effect against the overgrowth of pathogens in the vagina is documented by in vitro studies. Clinical studies concerning the role of hydrogen peroxide producing lactobacilli in vaginal infections are controversial. STUDY DESIGN: The aim of this study was to isolate lactobacilli from women with major vaginal infections: bacterial vaginosis (BV), vulvovaginal candidiasis (VVC) and trichomoniasis (TV), and from women with normal flora (NF), to test their ability to produce hydrogen peroxide, to quantitate lactobacilli and to evaluate factors related to the colonization of vagina with hydrogen peroxide producing lactobacilli. The study group comprised 108 women: 27 with VVC, 26 with BV, 20 with TV and 35 with NF. Data was analysed with Fisher exact test, chi2-test, and ANOVA. RESULTS: Lactobacilli were isolated from 81.5% of women with VVC, 58% of women with BV (p=0019), 55% of women with TV (p=0.015) and from 86% of women with NF. The majority of isolates of lactobacilli from women with VVC, TV and NF produced hydrogen peroxide (77%, 63.5% and 80%, respectively), compared with women with BV where only 26.5% of isolates produced hydrogen peroxide (p=0.01). Quantitative analysis showed that in an overwhelming majority of women colonized with lactobacilli, their count was >10(7) CFU/ml. Upon evaluation of behavioural and demographic characteristics, we found that smokers had a lower incidence of isolation of hydrogen peroxide producing lactobacilli (p=0.018). Our results showed that women with BV lack lactobacilli, especially hydrogen peroxide producing ones. Women with TV had a lower rate of lactobacilli. In women with VVC, there was neither significant difference in the isolation of lactobacilli, nor in their ability to produce hydrogen peroxide, compared to women with NF. CONCLUSION: Our results point out that hydrogen peroxide producing lactobacilli could protect against the development of BV, but not against VVC and TV.     

The role of lactobacilli and probiotics in maintaining vaginal health

Background

The vaginal microbiota of healthy women consists typically of a diversity of anaerobic and aerobic microorganisms. Lactobacilli are the most prevalent and often numerically dominant microorganisms and are relevant as a barrier to infection. The capacity of lactobacilli to adhere and compete for adhesion sites in the vaginal epithelium and the capacity to produce antimicrobial compounds (hydrogen peroxide, lactic acid, bacteriocin-like substances), are important in the impairment of colonization by pathogens.

Objective

This review summarizes the role of lactic acid bacteria in preventing illness of the host, including bacterial vaginosis, yeast vaginitis, urinary tract infection and sexually transmitted diseases.

Conclusions

The administration of probiotics that colonize the vaginal tract can be important in maintaining a normal urogenital health and also to prevent or treat infections.     

Study on the effects of an oral lactobacilli and lactoferrin complex in women with intermediate vaginal microbiota

Purpose

The aim of our meta-analysis was to explore whether pre-pregnancy obesity is regarded as an important risk factor for predicting macrosomia or not.

Methods

Three databases were systematically reviewed and reference lists of relevant articles were checked. Meta-analysis of published cohort studies comparing whether pre-pregnancy obesity was associated with macrosomia and adjusting for potential confounding factors. Calculations of pooled estimates were conducted in random-effect model. Heterogeneity was tested by using Chi-square test and I ² statistics. Publication bias was estimated from Egger’s test (linear regression method) and Begg’s test (rank correlation method).

Results

Sixteen cohort studies met the inclusion criteria. The meta-analysis showed that pre-pregnancy obesity was associated with macrosomia as an important risk factor. The adjusted odds ratio was 1.93, 95% CI (1.65, 2.27) in random-effect model, stratified analyses showed no differences regarding different quality grade, definition of macrosomia, location of study and number of confounding factors adjusted for. There was no indication of a publication bias either from the result of Egger’s test or Begg’s test.

Conclusion

Our findings indicated that pre-pregnancy obesity should be considered as an important risk factor for macrosomia. The effect of pre-pregnancy obesity on macrosomia need to be carefully assessed and monitored.

     

The effects of vaginal lubricants on sperm function: an in vitro analysis

Background

Despite being marketed as “sperm friendly”, some vaginal lubricants are known to be detrimental to sperm function and therefore could negatively affect fertility. Many others have not yet been assessed in regards to their effect on sperm function. This issue may concern couples trying to conceive both naturally and via artificial reproductive technologies (ART).

Aim

The aim of this research was to analyse the effects that lubricants, commonly used in the setting of natural conception and ART, have on sperm function in an in vitro setting. This was done by assessing sperm motility, vitality and DNA fragmentation following treatment with commercial lubricants or control preparations. We have attempted to mimic the conditions of the vaginal environment in our clinical trial, and so have compiled a list of lubricants that are likely to have minimal negative effect on sperm function in vivo or are “sperm friendly”.

Methods

Ten samples were obtained for the study from patients attending a fertility clinic. Once collected, the sperm samples were prepared by density gradient centrifugation and incubated with 11 different lubricants including positive and negative controls for 30 min at 37 °C to mimic the temperature inside the female reproductive tract. Sperm motility, vitality and DNA fragmentation were assessed to determine the effects of the lubricants on sperm function and DNA integrity.

Results

Nine lubricants were investigated including Sylk™, Conceive Plus®, glycerol, Johnson’s® Baby Oil, SAGE® Culture Oil, Yes®, Forelife™, MaybeBaby® and Pre-seed®. The lubricant which had the best results in terms of vitality, at 92 %, was Pre-seed® and the worst was Forelife™ with 28 % vitality. In terms of motility, Pre-seed® resulted in the highest percentage of spermatozoa with progressive motility at 86 % and Sylk™ resulted in the lowest percentage of progressively motile cells in the sample with 31 % of sperm progressively motile. There were no significant effects on DNA integrity.

Conclusions

Pre-seed® was the lubricant which had the least negative effect on sperm function, with Conceive Plus® a close second, due to the significantly higher sperm motility and vitality parameters measured following lubricant exposure.     

Collagen-coated polypropylene mesh in vaginal prolapse surgery: an observational study

Objective

To determine the incidence and factors associated with the development of bowel, urinary and sexual symptoms following obstetric anal sphincter injury (OASIS).

Study design

A prospective cohort study involving 435 women who sustained OASIS, over a five-year period, in a large UK teaching hospital. Details of bowel, urinary and sexual function were documented using a structured questionnaire. The outcome measures included the incidence of symptoms following OASIS and factors which modify the risk of developing symptoms.

Results

The majority (96%) of women were faecally continent three months after primary OASIS repair. Nevertheless, 34.2% reported faecal urgency, 25% suffered poor flatal control, and nearly 30% reported pain and bleeding on defaecation. Sixteen percent of women reported stress urinary incontinence, 15% experienced urgency and 20% reported urinary frequency. Fifty-seven percent of women had resumed intercourse but 32% of those women reported dyspareunia. Women who developed faecal symptoms were significantly more likely to develop urinary symptoms. Advancing maternal age and the use of forceps, in particular rotational forceps, significantly increase the risk of developing faecal and urinary symptoms.

Conclusions

Obstetric anal sphincter injuries continue to be responsible for significant morbidity, with approximately 30% of women reporting faecal, urinary or sexual symptoms, three months postpartum. This large prospective UK study provides up-to-date information relating to factors which increase the likelihood of such symptoms occurring. These data are useful for counselling and targeting more intensive follow up to women at higher risk of developing symptoms.     

Randomized, double-blind, placebo-controlled study of oral lactobacilli to improve the vaginal flora of postmenopausal women

Objective

The purpose of this randomized, double-blind, placebo-controlled study was to evaluate the influence of the orally administered probiotic strains Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 on the quality of the vaginal flora in postmenopausal women.

Study design

Postmenopausal women with Nugent scores between 4 and 6 in initial vaginal swab, were randomized into two groups. Women in the intervention group received probiotic capsules containing 2.5 × 10⁹ CFU (colony forming units) each of lyophilized L. rhamnosus GR-1 and L. reuteri RC-14 and women in the control group received an oral placebo once daily, in both groups for 14 days. Final vaginal swabs were taken 1 day after the last administration of the medication. The primary efficacy variable was a change in the Nugent score between baseline and the end of the study of at least two grades in each individual patient.

Results

Seventy two women were recruited in the study, 35 assigned to the intervention group and 37 to the control group. Twenty-one of the 35 subjects (60%) in the intervention group and 6 of the 37 subjects (16%) in the control group showed a reduction in the Nugent score by at least two grades. The difference in the number of patients with improvement was highly significant (p = 0.0001). The median difference in Nugent scores between baseline and the end of the study was 3 in the intervention group and 0 in the control group (p = 0.0001).

Conclusion

Our results provide evidence for an alternative modality to restore the normal vaginal flora using specific probiotic strains administered orally.     

Analysis of lactobacillus products for phages and bacteriocins that inhibit vaginal lactobacilli

Objective: Bacterial vaginosis is associated with an unexplained loss of vaginal lactobacilli. Previously, we have identified certain vaginal lactobacilli-released phages that can inhibit in vitro other vaginal lactobacilli. However, there is no apparent route for phages to be transmitted among women. The purpose of this study was to identify whether certain Lactobacillus products commonly used by women release phages or bacteriocins that can inhibit vaginal lactobacilli.Methods: From 26 Lactobacillus products (2 acidophilus milks, 20 yogurts, 3 Lactobacillus pills, and 1 vaginal douche mix), lactobacilli were isolated with Rogosa SL agar (Difco, Detroit, MI). From these lactobacilli, phages and bacteriocins were induced with mitomycin C and tested against a collection of vaginal Lactobacillus strains.Results: From the 26 products, 43 Lactobacillus strains were isolated. Strains from 11 yogurts released phages, among which 7 inhibited vaginal lactobacilli. Eleven strains released bacteriocins that inhibited vaginal lactobacilli. While about one-half of the vaginal strains were lysed by bacteriocins, less than 20% were lysed by phages.Conclusions: Some vaginal lactobacilli were inhibited in vitro by phages or bacteriocins released from Lactobacillus products used by women, implying that vaginal lactobacilli may be reduced naturally due to phages or bacteriocins from the environment.     

Rho kinase-related proteins in human vaginal arteries: an immunohistochemical and functional study

IntroductionThe calcium‐sensitizing Rho A/Rho kinase pathway has been suggested to play a role in the control of nongenital vascular smooth muscle. Rho‐associated kinases (ROKs) cause calcium‐independent modulation of smooth muscle contraction, and have been demonstrated in the bladder, prostate, and corpus cavernosum. Until now, it is not known whether ROKs and related proteins play a role in the control of vaginal blood flow.AimTo investigate by means of functional studies and immunohistochemistry the significance of the Rho pathway in human vaginal arteries.MethodsVaginal tissue was obtained from five postmenopausal women. Specimens were processed for immunohistochemistry for ROK1, ROK2, RhoA, and RhoGDI. Segments of sub‐epithelial vaginal arteries were mounted in a tissue bath. Effects of Y27632 on the concentration–response curves to phenylephrine (Phe) or Phe‐precontracted preparations were investigated.Main Outcome MeasureThe expression of Rho kinases ROK1, ROK2, and the Rho‐associated protein RhoGDI in human vaginal arteries was investigated by means of immunohistochemistry. Tissue bath studies were conducted in order to characterize the effects of the ROK inhibitor Y27632 on isolated vaginal arteries.ResultsA meshwork of α‐actin immunoreactive arterioles was located in the sub‐epithelium of human vaginal specimens. Immunoreactivities for ROK1, ROK2, RhoA, and RhoGDI were expressed in the smooth musculature of these arteries. At 0.1 and 1 µM Y27632, the contraction to Phe (10 µM) was 99 ± 17% and 28 ± 12% that of 124 mM K⁺. In Phe‐contracted preparations, Y27632 produced relaxant responses.ConclusionsThe activation of alpha₁‐adrenoceptors contracts sub‐epithelial human vaginal arteries via ROK‐sensitive mechanisms. A role for these signals in the regulation of vaginal blood flow might be considered. Ückert S, Waldkirch ES, Kauffels W, Kuczyk MA, and Hedlund P. Rho kinase‐related proteins in human vaginal arteries: An immunohistochemical and functional study. J Sex Med 2011;8:2739–2745.     

223例健康育龄妇女阴道内乳酸杆菌菌群的鉴定

目的检测健康育龄妇女阴道内乳酸杆菌的主要菌群种类及其产生过氧化氢的能力。方法采集223例健康育龄妇女阴道侧壁的分泌物,经过乳酸杆菌选择性培养基（Rogosa）与肉汤培养基（MRS）分离培养后,提取细菌染色体DNA,PCR扩增细菌的16SrRNA基因,测定16SrRNA核苷酸序列,然后与基因库（GenBank）中的数据进行比对,确定细菌的种类。并对乳酸杆菌产生过氧化氢的情况进行分析。结果从223例健康育龄妇女阴道中分离到8种乳酸杆菌种属,较常见的种属为卷曲乳杆菌（53．7％）,詹氏乳杆菌（27．9％）与格氏乳杆菌（10．3％）。其他少见的细菌类型包括阴道乳杆菌（2．2％）,L．coleohominis（1．5％）,约氏乳杆菌（1．5％）,嗜酸乳杆菌（1．5％）及发酵乳杆菌（1．5％）;大多数乳酸杆菌均能产生过氧化氢,其中卷曲乳杆菌和詹氏乳杆菌产生过氧化氢的能力较强。结论卷曲乳杆菌、詹氏乳杆菌和格氏乳杆菌是健康育龄妇女阴道内乳酸杆菌的主要种属。卷曲乳杆菌和詹氏乳杆菌有较强的产生过氧化氢的能力。     

Effectiveness of infection prevention with latamoxef in vaginal hysterectomy. A prospective study

A prospective clinical trial was designed to examine the influence of a single-dose prophylaxis with 2 g Latamoxef on postoperative infectious morbidity in women undergoing vaginal hysterectomy. 91 vaginal hysterectomies with latamoxef prophylaxis were compared with a similar group of 99 patients without prophylaxis. Patients in the latamoxef group had significantly less postoperative infectious morbidity (23.1%) than those in the non prophylaxis group (45.1%). Prophylactic latamoxef reduced postoperative additional antibiotic therapy in the prophylaxis group significantly. The postoperative infectious morbidity (urinary tract infections, pelvic infections, febrile morbidity) was significantly reduced by a single-dose prophylaxis with 2 g Latamoxef.     

Epidemiology of vaginal Candida infection: significance of numbers of vaginal yeasts and their biotypes

The prevalence, quantity and biotypes of vaginal yeasts were determined for 1082 randomly selected nonpregnant patients attending two genitourinary medicine clinics. The overall yeast prevalence was 22.1% with a geometric mean of 40 and a median of 75 yeast colonies per positive isolate (on 5-cm Petri dishes). There was no statistically significant variation in prevalence, quantity or biotype of yeasts with the patients' age, season of the year, stage of the menstrual cycle, recent antibiotic history, contraceptive use or main diagnosis (excluding candidosis). A clinical score for Candida infection, based on symptoms of pruritus and signs of Candida vulvovaginitis, showed significant variation with the prevalence and quantity of yeast isolates. Unequivocal clinical evidence of candidosis was strongly associated with high concentrations of vaginal yeasts. There was also some association between certain groups of Candida albicans biotypes and the clinical score. These observations reemphasize the need for consideration of both clinical and mycological factors in establishing a diagnosis of vulvovaginal Candida infection. It is suggested that isolation of fewer than 10 yeast colonies from a vaginal swab is usually unlikely to indicate an infection requiring treatment.     

The effectiveness of live lactobacilli in combination with low dose oestriol (Gynoflor) to restore the vaginal flora after treatment of vaginal infections

OBJECTIVE: To evaluate the effectiveness of live lactobacilli in combination with low dose oestriol for restoration of the vaginal flora after anti-infective treatment. DESIGN: The study was designed as a single centre, randomised, placebo-controlled, double-blind clinical trial. SETTING: University Hospital. SAMPLE: Three hundred and sixty women out of 1750 were randomised. METHODS: Three hundred and sixty women with the complaints of vaginal infections (bacterial vaginosis, candidiasis, trichomoniasis or fluor vaginalis) were randomly assigned two to seven days after the end of the anti-infective therapy, to therapy with live lactobacilli in combination with low dose oestriol (study group, n= 240) or placebo (n= 120). The follow up visits occurred three to seven days and four to six weeks after the end of the restoration therapy. MAIN OUTCOME MEASURES: The Normal Flora Index (NFI), which consists of numbers of lactobacilli, pathogenic microorganisms, leucocytes and vaginal pH, was used as the primary outcome of the study. Secondary outcomes included the total symptoms score, the degree of purity of the vaginal flora and the global assessment of the treatment by the investigator and the women. RESULTS: During restoration therapy, the NFI increased significantly more in the study group than in the control group in both first and second control visits (P= 0.002 and P= 0.006, respectively). The degree of purity of the vaginal flora also increased significantly more in the study group compared with the control group (P < 0.0001 and P= 0.001, respectively). No serious adverse event was reported during restoration therapy. CONCLUSION: Restoration of the vaginal flora can be significantly enhanced by the administration of live lactobacilli in combination with low dose oestriol.     

Operative vaginal delivery and post-partum infection

During the past decade, there has been an increase in the awareness of infections associated with pregnancy and delivery. The most significant cause of post-partum infection is caesarean section; 20–25% of operations are followed by wound infections, endometritis or urinary tract infections. Approximately 13% of women in the UK undergo operative vaginal delivery (OVD) with forceps or vacuum, which is also associated with an increased risk of infection, estimated at 0.7%–16% of these deliveries. Despite this, previous reviews have identified only one small trial of antibiotic prophylaxis in 393 women and concluded that there was insufficient evidence to support the routine use of prophylactic antibiotics after OVD. The ANODE trial, a multicentre, blinded, placebo-controlled trial from the UK, is due to report findings from more than 3400 women in 2019 and will be the largest study to date of antibiotic prophylaxis following OVD.     

Perinatal infection and vaginal flora.

Vaginal cultures were obtained from 1, 083 mothers on admission to the labor floor. The additional bacteriologic information gained from this survey was not sufficient to warrant continued use of the admission vaginal culture as a screening procedure.