Microbiological predication of using of vaginal suppositories "melanizol" for treatment nonspecific vaginitis

The article considers the vaginal suppositories “Melanizol” based on metronidazole, and tea tree oil influence on the museum and clinical microorganisms strains, that may be potential agents of vaginal infections. Established antimicrobial activity of vaginal suppositories “Melanizol” against the following strains of micro-organisms Staphylococcus aureus, Basillus subtilis, Klebsiella, and Escherichia coli, and also found their antifungal activity against Candida albicans.


INTRODUCTION
Non-specific (aerobic) vaginitis are arising from changes in normal vaginal microflora, causing signs of inflammation. The cause of vaginitis is mainly aerobic microorganisms microorganisms from intestinal commensals or other aerobic pathogenic aerobic bacteria (Escherichia, Enterococci, Streptococci, Staphylococci etc.). Clinical symptoms depend on the type and intensity, and have a high tendency to relapse and chronic leaks. Inflammation and ulcers in the non-specific (aerobic) vaginitis may increase the risk of infections, sexually transmitted infections [10].
The causes of recurrent infections of the genitourinary system in women are similar, including in healthy women with normal structure of the genitourinary system. According to statistics, 30 % to 44 % of women have infections relapse within six months. Relapses tend to cause the same kinds of pathogens that caused previous infection. Risk factors include: In premenopausal women, sexual intercourse three or more times per week, spermicide use, new or multiple sex partners. In postmenopausal women, risk is primarily increased by sequelae of lower estrogen levels. Patient-initiated treatment lowers the cost of diagnosis, number of physician visits, and number of symptomatic days compared with physician-initiated treatment. It also reduces antibiotic exposure compared with antibiotic prophylaxis. Antibiotic prophylaxis effectively limits the recurrence of urinary tract infection but increases the risk of antibiotic resistance and adverse effects [8].
Over the past decade streptococci lost their original meaning in the etiology of vaginitis. This is due to their lack of resistance to antibiotics. Staphylococci unlike strepto-cocci have the ability to form antibiotic-resistant strains that have high virulence, which contributes to their leading role in the etiology of inflammation of the female genital organs for a long time. But under the influence of antibiotics II and III generation in the first place were anaerobic gram-negative (Escherichia coli, Proteus vulgaris, Enterobacteria, Klebsiella and others). Increasingly began to meet microbial associations, causing severe course of inefficiency non-specific (aerobic) vaginitis and its treatment. According to the literature Escherichia coli is present in allocations of 80-85 % women, Klebsiella and Proteus -10-20 %, group B Streptococci, Enterococci and Staphylococci -5 %. Microorganisms are involved in the inflammatory process are excessive dangerous by reason of activation of inflammatory mediators [3,7].
Due to polyetiologic of non-specific vaginitis can be weighed both anaerobic and aerobic microorganisms and candida infection complicated, and requires a comprehensive approach to treatment. As such means tea tree oil can be recommended for using, which is very interesting materials for pharmaceutical use [1], contains over one hundred terms and their spirits from which terpinen-4-ol, α-pinene, linalool and α-terpineol considered essential components of the antimicrobial activity and fungicidal activity. Streptococcus mutans and Prevotela intermedia were sensitive to tea tree oil [5], and tea tree oil has bactericidal effect on Bacillus cereus, Corynebacterium sp., Entero-  [9].
On the base of National University of Pharmacy of TL department by staff under the supervision T. H. Yarnykh new vaginal suppositories "Melanizol" developed [4], suppositories contain compositions: metronidazole, tea tree oil and base (polyethylene oxide, PEO-1500: PEO-400) are promising for the treatment of nonspecific vaginitis and bacterial vaginosis.
The aim of our study was to investigate the antibacterial properties of new vaginal suppositories "Melanizol" to museum test-strains and clinical strains. We determined the reactivity of antibiotics on two layers of dense nutrient medium, poured into Petri dishes. Evaluation of antimicrobial activity carried out by the diameter of the zone delayed the growth of microorganisms. Differentiation susceptible, moderately resistant strains was performed according to the recommendations of the National Committee for clinical laboratory standards (NCCLS, USA). During the evaluation of new antimicrobial agents and strains against antibiotics in the study used the following criteria: no delays microbial growth zones around wells and delay zones of 10 mm indicate that the organism is not sensitive to the hole made by the drug or antibiotic concentration; stunted growth zone diameter of 10-15 mm indicate low sensitivity to the cultures studied concentrations of antibiotic substances; stunted growth zone diameter of 15-25 mm are measured as an indicator of the sensitivity of the microorganism to the IMP; stunted growth zone whose diameter exceeds 25 mm, indicate a high sensitivity of microorganisms to studied agents.

MATERIALS AND METHODS
Statistical analysis of experimental data was performed by conventional methods with statistical software Microsoft Excel 2007 and Statistic 6.0.

RESULTS AND DISCUSSION
The level of antimicrobial activity of experimental samples evaluated in diameter zone stunted growth of microorganisms. The experimental results are presented in Fig. 1.
According to the results of microbiological studies (Fig. 1), suppositories, vaginal "Melanizol" exhibit activity against strains Basillus subtilis level comparator suppositories "Hravahin" and significantly surpass it in terms of activity, inhibiting the growth of Staphylococcus aureus, Pseudomonas aeruginosa, and as Escherichia coli and Candida albicans, which activity caused by the presence of vaginal suppositories of tea tree oil, which suppositories "Melanizol" inferior to its activity. Given the preliminary results of microbiological testing in relation to the museum strains, it would conduct additional study of the effect of vaginal suppositories "Melanizol" and comparing drug suppositories "Hravahin" in clinical strains of bacteria. The data are presented in Fig. 2.
Data obtained from studies (Fig. 2) show that the suppositories vaginal "Melanizol" exhibit antibacterial activity against clinical strains of Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Klebsilla pneumoniae and Candida aibicans, significantly outperforming the comparator suppositories "Hravahin" and not yielding tea tree oil, and their effect on Enterococcus faecalis, Klebsilla pneumoniae significantly surpassing it.
Thus, based on the findings, we can conclude that vaginal suppositories "Melanizol" exhibit antimicrobial activity against clinical strains of microorganisms which are, at present, the most common agents of vaginal infections.  Note: * -significantly relative to comparator suppositories "Hravahin"; ** -significantly in relation to tea tree oil.