Antibiotic resistance of Gardnerella vaginalis in recurrent bacterial vaginosis

Fifty strains of Gardnerella vaginalis isolated from 321 high vaginal swabs over a period of five months were tested for their antibiotic sensitivity. Sixty eight per cent of all isolates were resistant to metronidazole while 76% were sensitive to clindamycin. All the strains isolated from cases with recurrence of infection were resistant to metronidazole. Clindamycin therapy has a better clinical efficacy than metronidazole in cases of recurrent bacterial vaginosis.     

Correlation of local interleukin-1beta levels with specific IgA response against Gardnerella vaginalis cytolysin in women with bacterial vaginosis

PROBLEM: Mucosal immune system activation may represent a critical determinant of adverse sequelae correlated with bacterial vaginosis, as HIV sexual transmission, upper genital tract infections, cervicitis, endometritis, postsurgical infections, and adverse pregnancy outcomes as preterm delivery (PTD), low birth weight (LBW). METHOD OF STUDY: Levels of interleukin-1beta (IL-1beta), anti-Gardnerella vaginalis hemolysin (Gvh) IgA, pH, Nugent score, and number of leukocytes were measured in vaginal fluids of 60 fertile women with bacterial vaginosis and of 64 healthy controls. RESULTS: Vaginal IL-1beta levels were nearly 13-fold higher in women with bacterial vaginosis (BV) and were associated with anti-Gvh IgA response. IL-1beta was positively correlated with leukocyte counts in the smear both in healthy and bacterial vaginosis positive women. CONCLUSIONS: Induction of the proinflammatory cytokine IL-1beta may be a necessary event to elicit an innate immune response to control anaerobic genital tract infections. High levels of vaginal IL-1beta are associated with mounting of an antigen-specific mucosal immune response in women with bacterial vaginosis. Parallel induction of innate and adaptive immune response may be associated with protection from ascent of micro-organisms to the upper genital tract, and from acquiring viral infection through the vaginal tract.     

妊娠合并细菌性阴道病对妊娠结局影响的临床研究

目的探讨妊娠期细菌性阴道病对胎膜早破、早产、产褥感染和新生儿感染等不良妊娠结局的关系。方法在定期产检的孕妇410例进行细菌性阴道病的筛查,进行孕期及围产期随访,分析妊娠期细菌性阴道病与分娩方式、胎膜早破、早产、产褥感染和新生儿感染等的相关性。结果410例孕妇中检出妊娠合并细菌性阴道病52例,检出率为12.68%,妊娠合并细菌性阴道病者的胎膜早破、早产、产褥感染和新生儿感染发病率均明显高于未感染的孕妇。结论妊娠合并细菌性阴道病明显增加孕妇胎膜早破、早产、产褥感染和新生儿感染的机会。加强产前筛查,早期诊断,早期治疗是十分必要的。     

Trimethylamine content in vaginal secretion and its relation to bacterial vaginosis

The presence of a fishy odor emanating from women who present with a malodorous vaginal discharge is well known. The odor is due to bacterial reduction of trimethylamine oxide to trimethylamine (TMA) in vaginal secretion. The release of TMA from specimens of vaginal fluid following the addition of alkali is often used in making a clinical diagnosis of bacterial vaginosis (BV). We now report a sensitive gas chromatographic method for analysis and quantification of TMA in vaginal fluid in which weighed samples were used. In addition, a proper diagnosis of BV was obtained using Gram-stained smears of the vaginal fluid according to the method of Nugent et al. (R. P. Nugent et al., J Clin Microbiol 1991;29:297-301). We also diagnosed BV according to Hallén et al. (A. Hallén et al. Genitourin Med 1987;63:386-9). TMA was present in all women with a Nugent score between 7 and 10 and in almost all women diagnosed with BV according to the method of Hallén et al. TMA was not found or was only found in very low concentrations in vaginal fluid from women with Nugent scores of 0 to 3. TMA was also found in four women with a negative sniff test. It seems that high levels of TMA in samples of vaginal fluid are typical for BV regardless of the scoring method used for diagnosis. However, low levels of TMA, <5 microg/g vaginal fluid, do not always correlate with BV.     

Probiotics for the treatment of women with bacterial vaginosis

This review considers whether probiotics are effective agents for the treatment and/or prevention of bacterial vaginosis (BV). There seems to be an association between the absence of, or low concentrations of, vaginal lactobacilli and the development of BV. Many studies have suggested that the presence of H2O2-producing vaginal lactobacilli may protect against BV, although some studies do not support this hypothesis. In-vitro studies have suggested that certain specific strains of lactobacilli are able to inhibit the adherence of Gardnerella vaginalis to the vaginal epithelium and/or produce H2O2, lactic acid and/or bacteriocins, which inhibit the growth of bacteria causing BV. Clinical trials showed that intra-vaginal administration of Lactobacillus acidophilus for 6-12 days, or oral administration of L. acidophilus or Lactobacillus rhamnosus GR-1 and Lactobacillus fermentum RC-14 for 2 months, resulted in the cure of BV (defined as a 0-1 positive score according to Amsel's criteria), and/or reduced the recurrences of BV, and/or caused an increase in vaginal lactobacilli and restoration of a normal vaginal microbiota, significantly more frequently than did a placebo, acetic acid or no treatment. However, several trials have found no significant difference in the cure rate of BV and in the number of vaginal lactobacilli after intra-vaginal instillation of lactobacilli when compared with the effect of a placebo or oestrogen. Thus, although the available results concerning the effectiveness of the administration of lactobacilli for the treatment of BV are mostly positive, it cannot yet be concluded definitively that probiotics are useful for this purpose.     

Heterogeneity in restriction patterns of Gardnerella vaginalis isolates from individuals with bacterial vaginosis.

This study was undertaken to resolve the genetic make up of Gardnerella vaginalis present in bacterial vaginosis (BV). DNA from several G. vaginalis isolates from within and between individual BV patients were compared by BamHI, ClaI and EcoRI restriction endonuclease analysis (REA) followed by a restriction fragment length polymorphism (RFLP) study, utilizing a 5.7-kb BamHI G. vaginalis ATCC14018 DNA probe. Four G. vaginalis isolates from one patient (GVP-062) were composed of 3 different biotypes (biotypes 3, 5 and 8), and while the REA mirrored the biotype, in RFLP studies at least 3 isolates had DNA fragments in common. All of the isolates from 2 other patients (GVP-063 and GVP-072) represented a single biotype (biotype 2), but under REA and in RFLP studies, the isolates GVP-063 differed from GVP-072. An opposite case existed with the isolates GVP-072 (biotype 2) and GVP-065 (biotype 5), which appeared similar under REA and in RFLP studies. Finally, reisolates after 8 weeks (GVP-080) from a BV patient (isolates GVP-065) representing the same biotype (biotype 5) differed under REA and in RFLP studies. Thus, lacking any unique DNA fingerprint, G. vaginalis occurring in BV represents a (genetically) mixed population.     

乳杆菌活菌胶囊治疗妊娠期细菌性阴道病的临床观察

目的 调查妊娠期细菌性阴道病（BV）的发病情况，观察乳杆菌活菌胶囊治疗细菌性阴道病的疗效。方法对不同孕周的926名孕妇进行BV筛查，确定妊娠期细菌性阴道病的发病率。取BV患者164例，乳杆菌活菌胶囊治疗82例，孚舒达栓治疗82例，用药10d，停药1w后复查。结果妊娠期BV的检出率为17．71％。乳杆菌活菌胶囊治疗BV有效率89．02％，孚舒达栓治疗BV有效率87．80％，二者差异无显著性（P〉0．5）。结论妊娠期BV有较高的发病率，乳杆菌活菌胶囊是治疗BV的安全有效药物。     

Reciprocal Interference between Lactobacillus spp. and Gardnerella vaginalis on Initial Adherence to Epithelial Cells

Bacterial vaginosis (BV) is the most common vaginal disorder in women of child-bearing age. It is widely accepted that the microbial switch from normal microflora to the flora commonly associated with BV is characterized by a decrease in vaginal colonization by specific Lactobacillus species together with an increase of G. vaginalis and other anaerobes. However, the order of events leading to the development of BV remains poorly characterized and it is unclear whether the decrease in lactobacilli is a cause or a consequence of the increase in the population density of anaerobes. Our goal was to characterize the interaction between two Gardnerella vaginalis strains, one of which was isolated from a healthy woman (strain 5-1) and the other from a woman diagnosed with BV (strain 101), and vaginal lactobacilli on the adherence to cervical epithelial cells. In order to simulate the transition from vaginal health to BV, the lactobacilli were cultured with the epithelial cells first, and then the G. vaginalis strain was introduced. We quantified the inhibition of G. vaginalis adherence by the lactobacilli and displacement of adherent lactobacilli by G. vaginalis. Our results confirmed that pathogenic G vaginalis 101 had a higher capacity for adhesion to the cervical epithelial cells than strain 5-1. Interestingly, strain 101 displaced L. crispatus but not L. iners whereas strain 5-1 had less of an effect and did not affect the two species differently. Furthermore, L. iners actually enhanced adhesion of strain 101 but not of strain 5-1. These results suggest that BV-causing G. vaginalis and L. iners do not interfere with one another, which may help to explain previous reports that women who are colonized with L. iners are more likely to develop BV.     

An international study of the interobserver variation between interpretations of vaginal smear criteria of bacterial vaginosis

An international workshop on vaginal smear-based diagnosis of bacterial vaginosis was organized where 13 investigators scoring 258 slides with smears from vaginal fluid. Interobserver reproducibility of interpretations of Nugent scores, Hay/Ison scores and wet smear scores for the diagnosis of bacterial vaginosis was shown to be high. Detailed analysis of individual scoring results however indicated that basic standards of quality control to ensure robust individual readings of slides must be adhered to.     

Analysis of virulence factors in cases of enterococcal endocarditis

Eleven isolates of Enterococcus faecalis causing endocarditis were screened for possible virulence factors with PCR and phenotypic assays. The gene coding for the enterococcal surface protein (esp) was detected in one isolate only, and haemolysin was produced by two isolates. Aggregation substance, biofilm formation and gelatinase were present in seven, nine and eight isolates, respectively. Predisposing factors, particularly hospitalisation and multiple antibiotic therapy, appeared to be more relevant to the development of enterococcal endocarditis following bloodstream infections than the pattern of virulence factors.     

Normal flora and bacterial vaginosis in pregnancy: an overview

Abstract

The female genital tract is an intricate, yet balanced ecosystem that hosts a variety of different residential microflora. The physiological changes that occur during pregnancy may disrupt this balanced ecosystem and predispose women to a potentially pathogenic microbiota. Bacteria that are associated with bacterial vaginosis (BV) are opportunistic pathogens that frequently form part of this microbiota. The overgrowth of and infections with these bacteria are linked to poor obstetric outcomes and increased transmission of other reproductive tract infections (RTIs). These infections increase women's susceptibility of acquiring HIV, the rates of HIV shedding and the development of acquired immune deficiency syndrome (AIDS) in HIV-infected patients. It is unknown how the plethora of bacterial species associated with BV contributes to the dynamics of this condition. The use of high-throughput methods have led to the in-depth investigation of different BV-related bacterial species and the functional capabilities of these species. However, the pathogenesis of BV is still poorly defined and the role of individual BV-related bacterial species in specific pregnancy complications is unclear and controversial. The majority of BV infections are asymptomatic and successful diagnosis is complicated by the lack of reliable and standardized diagnostic tests.     

Genomic DNA fingerprint analysis of biotype 1 Gardnerella vaginalis from patients with and without bacterial vaginosis.

Of the 20 biotype 1 Gardnerella vaginalis isolates analyzed, 10 from patients with bacterial vaginosis and 10 from patients without bacterial vaginosis, none shared the same DNA fingerprint. However, a 1.18-kb HindIII fragment was common among 18 of the 20 biotype 1 isolates in a restriction fragment length polymorphism analysis with a 7.9-kb G. vaginalis DNA probe.     

定君生胶囊用于妊娠期细菌性阴道病的临床观察

目的调查妊娠期细菌性阴道病(BV)的发病情况,观察定君生胶囊治疗细菌性阴道病的有效性。方法采用珠海市银科生物技术应用研究所生产的细菌性阴道病快速检测卡,对不同孕周的1500名孕妇进行BV筛查,确定妊娠期细菌性阴道病的发病率。对确诊为BV的孕妇266例,随机分为两组。定君生组:136例,给予定君生胶囊(乳杆菌活菌胶囊)阴道给药治疗;孚舒达组:130例,给予孚舒达栓(复方甲硝唑栓)阴道给药治疗。两组均连续用药10d,停药7d后复查。结果妊娠期BV的检出率为17.73%。定君生胶囊治疗BV的总有效率为92.65%,孚舒达栓治疗BV的总有效率为91.53%,二者差异无显著性(P>0.5)。结论妊娠期BV的发病率较高。定君生胶囊能有效改善阴道内生态环境,治疗妊娠期细菌性阴道病安全、有效,易为孕妇所接受。     

灭滴灵阴道用药对细菌性阴道病晚孕妇女不良妊娠结局的预防效果

目的：探讨灭滴灵治疗细菌性阴道病对不良妊娠结局是否有改善作用。方法：在产科门诊对用阴道分泌物涂片革兰氏染色Ｎｕｇｅｎｔ诊断标准孕２８－３２周妇女进行细菌性阴道病筛查，对检出的患者随机分为二组；观察组５２例，未给予治疗；治疗组５０例，采用灭滴灵阴道用药治疗，必要时再次治疗。     

Effectiveness of Lactobacillus-containing vaginal tablets in the treatment of symptomatic bacterial vaginosis

The purpose of this study was to determine the effectiveness of Lactobacillus -containing vaginal tablets in the treatment of bacterial vaginosis (BV) and in the restoration of a healthy vaginal flora. Thirty-nine women with BV were enrolled in a double-blind, placebo-controlled clinical trial. Patients received either one Lactobacillus -containing tablet or placebo daily for 7 days. Clinical criteria, vaginal Gram stain scores and symptoms were compared with those at the initial visit and those at completion of therapy and 2 weeks later. After completion of therapy, all of the patients in the Lactobacillus -treated group ( n  = 18) were free of BV, showing a normal (83%) or intermediate (17%) vaginal flora, as compared with only two patients free of BV with intermediate flora (12%) from among the 16 placebo-treated women (p   <0.001). Two weeks after completion of therapy, treatment was successful (score <7) in 61% of Lactobacillus -treated patients as compared with 19% of those in the placebo group (p   <0.05). In the treatment group, the total number of symptomatic patients and the intensity of their symptoms, in particular vaginal malodour, were significantly reduced at both follow-up visits. The data indicate that intravaginal administration of exogenous selected strains of lactobacilli can restore a normal vaginal microbiota and be used in treating bacterial vaginosis.     

Validation of the use of Pap-stained vaginal smears for diagnosis of bacterial vaginosis

Papanicolaou-stained cervicovaginal smears (Pap smears) are used to screen for cervical cancer. Since there is a lack of consensus in published reports respecting the efficacy of Pap-stained smears in BV diagnostics, there is a need to validate their use for diagnosis of BV. Slides from the international BV00 workshop were Pap stained and independently analyzed by four investigators under a phase-contrast microscope. All workshop slides--whether Pap-stained, Gram-stained or rehydrated air-dried smears--were scored according to the same Nugent classification. The diagnostic accuracy of Pap smears for diagnosis of BV had a sensitivity of 0.85 and a specificity of 0.92, with a positive and negative predictive value of 0.84 and 0.93, respectively. The interobserver weighted kappa index was 0.86 for Pap-stained smears compared to 0.81 for Gram-stained smears, and 0.70 for rehydrated air-dried smears using the mean Nugent score as the criterion standard. Provided that the samples are taken from equivalent locations (the vaginal fornix) and analyzed according to the same scoring criteria, there is no discernable difference in the diagnostic accuracy of the three smear-staining methods. The Pap-stained vaginal smears can be used as a wholly adequate alternative to Gram-stained smears for BV diagnosis.     

Use of DNA hybridization test for diagnosing bacterial vaginosis in women with symptoms suggestive of infection

The purpose of this study was to evaluate a DNA hybridization test (Affirm VPIII) as an alternative to Gram stain for the rapid diagnosis of bacterial vaginosis in women with clinical signs of vaginal infection. Vaginal specimens were collected from 321 symptomatic women, and analyzed for bacterial vaginosis by both Gram stain using Nugent criteria and DNA hybridization test. Sensitivity, specificity, positive predictive value, and negative predictive value of the DNA hybridization test were determined using the Gram staining as the standard for diagnosis of bacterial vaginosis. Of the 321 patients, 115 (35.8%) were Gram positive for bacterial vaginosis and 126 (39.2%) were negative. 80 patients (25.0%) demonstrated intermediate Gram staining that was also considered negative. The Affirm system detected G. vaginalis in 107 (93.0%) of 115 vaginal specimens positive for bacterial vaginosis diagnosed by Gram stain. Compared to the Gram stain, DNA hybridization test had a sensitivity of 87.7% and a specificity of 96.0%. Positive and negative predictive values of the DNA hybridization test were 93.0% and 92.7%, respectively. In conclusion, Affirm VPIII hybridization test correlated well with Gram stain and may be used as a rapid diagnostic tool to exclude bacterial vaginosis in women with genital complaints.     

Vaginal nitric oxide in pregnant women with bacterial vaginosis

PROBLEM: To evaluate vaginal nitric oxide (NO) production in response to alterations in the vaginal microbial flora. METHOD OF STUDY: Cervicovaginal lavage samples from 206 women at 18-22 weeks of gestation were tested for NO, interleukin-1beta (IL-1beta), IL-1 receptor antagonist (IL-1ra), tumor necrosis factor-alpha and the inducible 70 kDa heat shock protein (hsp70). Bacterial vaginosis (BV) was diagnosed based on gram staining of vaginal smears. RESULTS AND CONCLUSIONS: Elevated NO (>2.14 mmol/L) was associated with a diagnosis of BV (38% versus 11%, P < 0.008) as well as an increased median vaginal IL-1ra concentration (72.5 ng/ml versus 36.6 ng/ml, P = 0.041). Elevated vaginal NO was also associated with vaginal hsp70 and this relationship was independent of BV status or IL-1ra concentrations (P < 0.026). We conclude that vaginal hsp70 release in response to abnormal vaginal microflora may trigger NO production in an attempt to minimize the pathological consequences of this altered milieu.