MICROSCOPIC CANDIDA GENUS FUNGI IN THE STRUCTURE OF MICROBIAL ASSOCIATIONS IN THE CONDITIONS OF GENERALIZED PERIODONTITIS AND THEIR SENSITIVITY TO ANTIBIOTICS AND ESSENTIAL OILS

Abstract

The significance of opportunistic pathogenic bacteria in the development of inflammatory diseases and complications is shown to be continuously growing over the past several decades. Of them, spread of antibioticresistant agents of opportunistic infections has become a most burning problem. Under such conditions, it becomes especially important to perform research aimed at the search for alternative anti-microbial materials. The plantbased materials, essential oils in particular, often demonstrate a high level of direct antimicrobial activity. Essential Oils (EOs) are complex natural mixtures of volatile secondary metabolites isolated from plant material (flowers, buds, seeds, leaves, twigs, bark, herbs, wood, fruits and roots) by hydro or steam distillation and by expression. The purpose of the present study was to determine Ñandida strains in microbial associations of the oral cavity in the conditions of generalized periodontitis, to clarify their sensitivity to antibiotics and essential oils. Object and methods. The isolates that caused periodontal inflammatory processes were isolated on the basis of the Dental Polyclinic at the Uzhhorod National University; the antimicrobial activity was studied at the Microbiology Laboratory at the Department of Genetics, Plant Physiology and Microbiology, Uzhhorod National University. The biological material was collected from the mucous membrane of the inflammatory site using a sterile transport system (a test tube with gel and an applicator for biological fluids produced by FLmedical (Italy). The material was plated according to Gold on nutrient media: Sabourund Dextrose Agar, HiCrome ™ Candida Differential Agar (Himedia) for the cultivation of microscopic fungi, hemolytic microflora, namely, the Streptococcus and Neisseria genera bacteria on the blood agar, Enterobacteriaceae genera bacteria – on Endo and Ploskirev agar (Farmaktiv, Ukraine), the Staphylococcus genus bacteria – on Mannitol Salt Agar (Biolif-Italia), Enterococci were identified with Bile esculin agar (Biolif-Italia ) The antibiotic sensitivity of bacteria and microscopic fungi was identified by the disc diffusion method according to the accepted procedure (Order No. 167 of the MOH of Ukraine dated 05/04/2007; EUCAST (European Committee on Antimicrobial Susceptibility Testing). The fungi isolates were screened for susceptibility to the following antibiotics: (in 6 mm diameter wells) with nystatin (50 µg), itroconazole (10 mkg), fluconazole (25 µg), ketoconazole (10 µg), voriconazole (1 µg), klotrimazole (10 µg), miconazole (50 µg). The microorganisms’ sensitivity to plant-based materials and disinfectants was determined by the standard agar diffusion test (with 8 mm diameter wells). Results. Isolation of 44 strains of microscopic fungi of the genus Candida were isolated from the oral cavity patients with generalized periodontitis. 15 isolates were characterized by resistance to antimycotic preparations, 13 belonged to C.albicans, 1 to C.glabrata and 1 to C.tropicalis. It is shown that microscopic fungi of the genus Candida isolated from the oral cavity of people with persistent inflammation, as a rule in the structure of associations of other conventionally pathogenic microorganisms: C.glabrata+S.saprophyticus+E.cloaceae; S.aureus+H.alvei+C. albicans; S.haemolyticus+C.albicans; S.aureus+C.albicans; K.pneumoniae+C.albicans; S.aureus+C.albicans+Ñ. freundii; Enterococcus faecalis+Candida albicans; C.albicans+ S.aureus+E.cloaceae. The low sensitivity of isolates to antibiotics and high sensitivity of microorganisms to essential oils (Lavandula angustifolia Mill., Origanum vulgare L., Anethum graveolis L., Thymus vulgaris L.) was installed and that substantiates the relevance of further research and development of oral hygiene products based on them. Conclusion. Establishment of antimicrobial properties of essential oils makes it possible to combine them in preparations applied for prevention of inflammatory processes of parodentium. Application of essential oils in preparations for mouth care and prevention of oral cavity infections looks promising due to their ability to inhibit the development of inflammatory processes at cost of their antimicrobial properties.

Keywords

• Ñandida strains
• essential oils
• antibiotic resistant microoganisms
• antimycotic activity.