[Reliable microbiological diagnosis of vulvovaginal candidiasis].

INTRODUCTION: Vulvovaginal candidiasis is common infection among those affecting the vulva and vagina. Is caused by the perpesentatives from the genus Candida, in most cases C. albicans (85-90%). An increase in the percentage of the so-called non-albicans agents is seen and these pathgogens are often resistant to the most commonly used in the practice antifungals. Faulty diagnosis, incorrect use of azoles, and self-treatment lead to selection of resistant strains and recurrent infections. AIM: Identification of Candida species associated with vulvovaginal candidiasis by conventional and PCR techniques. MATERIALS AND METHODS: For six months a total number of 213 vaginal secretions were tested applying Gram stain and cultivation on ChromAgar. API Candida fermentation tests and API 20CAUX assimilation tests were performed for the identification of the bacteria. Extraction of DNA of all the smears with subsequent PCR detection of different Candida species were done. RESULTS: 80.7% materials showed presence of blastospores and/or hyphae. Positive culture results were detected in 60 (28.2%) samples. The species specific identification revealed presence of C. albicans in 51 (85%) smears, C. glabrata--in 8 (13.3%), C. krusei--in 2 (3.3%), and S. cervisie--in 1 (2.1%). The PCR technique confirmed the results of the conventional methods. It is worth to mention that 51 of the tested smears were positive for G. vaginalis using additional PCR. CONCLUSIONS: The correct diagnosis of the cause of vulvovaginal candidiasis helps in the correct choice of appropriate antifungal therapy and prevents development of recurrent infections and consequences. The PCR based method is rapid, specific and sensitive. It perfectly correlates with the results from the conventional diagnostic tests so it could be selected as a method of choice for the diagnosis of vulvovaginal candidiasis.

May Chlamydia trachomatis be an aetiological agent of chronic prostatic infection?

Chlamydia trachomatis infection is the most common sexually transmitted bacterial disease. The objective of this study was to establish the presence/absence of C. trachomatis in 98 patients with chronic complaints about the prostate and to evaluate the role of this bacterium in the inflammation of the gland. We performed culture and microscopical examination of pre-massage/post-massage urine and expressed prostatic secretions (EPS). In all cases, culture on McCoy cells and polymerase chain reaction (PCR) of the EPS was performed. Based on laboratory findings in 53 cases (54.08%), Escherichia coli, Klebsiella, Enterobacter, Proteus, Pseudomonas and Staphylococcus were isolated and accepted as causative agents of chronic bacterial prostatitis. Forty-five patients were categorised as patients with chronic pelvic pain syndrome. The results from the PCR and the cell culture for detection of C. trachomatis were as follows - two positive probes detected at the same time by applying PCR and cultivation and 1 positive only by PCR but not by cultivation on the cell line. Based on these results, it is concluded that C. trachomatis is not so frequently detected in our patients. C. trachomatis may be accepted as one of the aetiological agents of chronic prostatitis and testing for this infection is highly recommended when presumption for chronic prostatitis is apparent.

Chlamydia trachomatis infection as a problem among male partners of infertile couples.

Chlamydia trachomatis infection is the most common bacterial sexually transmitted disease supposed to cause urethritis, epididymitis, prostatitis and infertility in men. The objective of this study was to assess the frequency of C. trachomatis infection in male partners of infertile couples at childbearing age. Sixty infertile couples and a control group of 40 healthy volunteers were included in the study. Urethral swabs were taken from all the male participants and cervical swabs from the female partners of the infertile couples. Culturing on McCoy cell line and PCR were the methods used for detection of the infection. C. trachomatis was found in five out of the 60 male urethral samples. Three of the female partners of these five positive males were diagnosed with C. trachomatis infection, too. We registered a woman with C. trachomatis infection whose partner's samples were negative for the bacterium. The control group showed one specimen positive for C. trachomatis. The frequency of C. trachomatis infection was 8.3% in the male partners of infertile couples at childbearing age when compared with 2.5% in the control group. It is most likely that infertility in the couples with chlamydial infection was due to the pathogen studied.

Prevalence and macrolide resistance phenotypes and genotypes among clinical isolates of Streptococcus pneumoniae collected in Sofia, Bulgaria from 2001 to 2005.

A total of 328 clinical isolates of Streptococcus pneumoniae were analyzed to determine the rate of macrolide and penicillin resistance as well as macrolide resistance phenotypes and genotypes. Erythromycin resistance was found in 81 pneumococcal isolates (24.7%) and 10.7% of isolates were clindamycin resistant. The prevalence of penicillin G-intermediate (minimum inhibitory concentrations, MICs, 0.125 to 1 microg/ml) and penicillin-resistant (MICs, >or=2 microg/ml) S. pneumoniae isolates was 25.6% and 13.7%, respectively. The rate of ceftriaxone-intermediate and ceftriaxone-resistant strains was 2.7% and 1.2%, respectively. Among erythromycin-resistant S. pneumoniae isolates, strains harboring mef(A) genes (n=42; 51.8%) were found to be predominant over strains with erm(B) genes (n=34; 42.0%). One (1.2%) isolate carried both erm(B) and mef(A), while 4 (4.9%) isolates carried L4 protein mutations. By using the erythromycin, clindamycin and rokitamycin triple-disk test, 42 strains were assigned to the M phenotype of macrolide resistance, 31 isolates were assigned to the partially inducible (iMcLS) phenotype, 4 were assigned to the constitutive (cMLS) phenotype. Four strains with L4 gene showed a rare phenotype with the triple-disk test. Serotyping of S. pneumoniae isolates suggested that serotype (or serogroup) 14, 6 and 19 were predominant (81.5%) among erythromycin-resistant strains. Among mef(A) positive isolates serotype 14 was predominant, among erm(B) positive isolates serogroups 6 and 19 were the most prevalent.

Widespread detection of VEB-1-type extended-spectrum beta-lactamases among nosocomial ceftazidime-resistant Pseudomonas aeruginosa isolates in Sofia, Bulgaria.

A total of 132 ceftazidime-resistant clinical isolates of Pseudomonas aeruginosa were collected during 2001-2005 from 5 university hospitals in Sofia, Bulgaria to assess the current levels of antimicrobial susceptibility and to evaluate resistance mechanisms to beta-lactams. Antimicrobial susceptibilities were detected by a disk diffusion method and E-test. Polymerase chain reaction amplification and sequencing of bla(VEB-1 )and bla(PER-1 )were performed. The antibiotic resistance rates were: to piperacillin 90.2%, piperacillin/tazobactam 52.3%, ceftazidime 94.7%, cefepime 88.6%, cefpirome 98.5%, aztreonam 85.6%, imipenem 66.6%, meropenem 63.6%, amikacin 81.1%, gentamicin 84.8%, tobramycin 89.4%, netilmicin 57.6%, ciprofloxacin 83.4%. Structural genes for VEB-1 extended-spectrum beta -lactamases (ESBLs) were found in 75 (56.8%) of the isolates. PER-1 ESBLs were not detected. The VEB-1-producing strains were more resistant than VEB-1 non-producers to amikacin, gentamicin, tobramycin and ciprofloxacin ( P<0.001). VEB-1 appears to have a significant presence among ceftazidime-resistant P. aeruginosa isolates from Sofia.

Group B Streptococci and inducible nitric oxide synthase: modulation by nuclear factor kappa B and ibuprofen.

Group B Streptococci (GBS) neonatal infections cause a complex inflammatory process involving numerous biochemical mediators. Nitric Oxide (NO) is generated by many cell types in response to different inflammatory signals. Nuclear factor kappa B (NFkappaB) plays an important role in the inflammatory process and may induce a number of biochemical mediator genes, including the inducible nitric oxide synthase (iNOS). We tested the hypothesis that GBS induces iNOS gene expression through activation of NFkappaB. We also tested whether ibuprofen (IBU) will suppress iNOS expression by blocking NFkappaB activation. Cerebral microvascular endothelial cells isolated from newborn piglets were harvested for the determination of iNOS gene expression and activation of NFkappaB. GBS significantly induced iNOS mRNA expression (5- to 6-fold, P < .005) and iNOS protein (3- to 4-fold, P < .01) at 24 hours. DNA-NFkappaB binding activity was detected within 15 minutes of GBS treatment and reached a maximal effect at 3 hours. Treatment with IBU significantly suppressed GBS-induced iNOS mRNA expression at 24 hours, and NFkappaB activity at 3 hours, suggesting that suppression of GBS-induced iNOS mRNA expression by IBU occurs by blocking of NFkappaB activation. These data show that NFkappaB activation is an early step in the induction of iNOS gene expression by GBS and that this interaction may play a vital role in the pathogenesis of GBS neonatal infections.