Порівняльна активність антисиньогнійних антибіотиків та бактеріофагів відносно позалікарняних штамів Pseudomonas aeruginosa

Abstract

Проведено in vitro дослідження чутливості позалікарняних штамів P. aeruginosa до антибіотиків з антисиньогнійною активністю та специфічних бактеріофагів («Синьогнійний бактеріофаг» та «Піобактеріофаг»). Найбільшою активністю по відношенню до досліджених позалікарняних штамів P. aeruginosa володіли іміпенем, ципрофлоксацин, цефтазидим, частота чутливості до яких слала 64,8%, 64,8%, 53,7% відповідно. Чутливість штамів Pseudomonas aeruginosa до комерційних препаратів – бактеріофагів склала близько 50% і залежала від біотопу вилучення. 24,0% штамів виявлялися стійкими як до антибіотиків, так і до бактеріофагів, а 13,0% полірезистентних псевдомонад були фагочутливими.

Проведено in vitro исследование чувствительности внебольничных штаммов P. aeruginosa к антибиотикам с антисинегнойной активностью и специфическим бактериофагам («Синегнийний бактериофаг» и «Пиобактериофаг»). Наибольшей активностью по отношению к исследованным штаммам P. aeruginosa владели имипенем, ципрофлоксацин, цефтазидим, частота чувствительности к которым составляла 64,8%, 64,8% и 53,7% соответственно. Чувствительность к коммерческим препаратам – бактериофагам составила около 50% и зависела от биотопа выделения. 24,0% штаммов оказались стойкими как к антибиотикам, так и к бак- териофагам, а 13,0% полирезистентных псевдомонад были фагочувствительными.

The article is sanctified to the study of activity of antibiotics and bacteriophages in relation to community- made strains of Pseudomonas aeruginosa. Pseudomonas aeruginosa, as a typical species of the genus Pseudomonas, is known in the territories of all continents, since, on the one hand, this stick is an ordinary inhabitant of reservoirs, various plant substrates, and on the other hand, it exhibits pathogenic properties in relation to various objects of nature, people, warm blooded animals, arthropods, and the like. Due to the peculiarities of its ecology, the widespread use of the Pseudomonas spider was due to the lack of requirements for organic matter for life, the ability to persistence in a non-culturally active state and in the composition of bacterial biofilms, a wide range of resistance to various environmental factors: antibiotics, bacteriophages, disinfectants, and antiseptics. In spite of the achievement of microbiology and practical medicine, the problems of Pseudomonas aerobic infection, both in clinical practice and in veterinary medicine, remain relevant and diverse, include issues of diagnosis, determination of the pathogenic potential of the pathogen, choice of effective treatment tactics, advanced preventive measures, etc. In modern conditions, when the monopoly position of antibiotics in the complex of antimicrobials is called into question, scientific researches aimed at increasing the efficiency of bacteriophagotherapy become of increasing importance. For experimental studies, we used commercial bacteriophages: «Pseudomonas bacteriophage» (FGUP NPO «Microgen», Perm.) and «Piobacteriophage» (FSUE NPO «Microgen» MH RF, Nizhny Novgorod). The sensitivity to specific bacteriophages was determined by the drop method. For the experiment, a suspension of daily agar culture was prepared in an isotonic sodium chloride solution, which was applied to the nutrient agar according rule, after the phage addition. Most strains of P. aeruginosa, which were taken at a concentration of 109 to 107 CFU / ml for the determination of their sensitivity to the bacteriophages, were assigned to a persistent or weakly sensitive (“-” or “+”, according to the generally accepted evaluation scheme) up to 106 – 104 CFU / ml – became medium-sensitive (on “++” – “+++”). Particular importance for adsorption of phage to a bacterial cell (the initial and most important stage of interaction), has the pH value of the medium. It should be in parameters 7-10. Adsorption occurs only on a negatively charged shell of a bacterium of a positively charged phage. In acidic environment, phage acquires a negative charge and repels from target cells. This should be remembered and during phagotherapy, trying to neutralize the acidic environment of the stomach in the oral application of the drug. An aim of these researches is an increase of efficiency of fight against Pseudomonas aeruginosa infection by determination of perspective of application of modern antibiotics and bacteriophages. Research object: antibiotic and phagosensitivity of 54 community-made strains of Pseudomonas aeruginosa. Pseudomonas aeruginosa, as a typical species of Pseudomonas aeruginosa, withdrawn from the different biotopes of patients. Research methods: the sensitivity to specific bacteriophages was determined by the drop method. For the experiment, a suspension of daily agar culture was prepared in an isotonic sodium chloride solution, which was applied to the nutrient agar according rule, after the phage addition. Antibiotic susceptibility studied by disc diffusion method to 8 antibiotics: to the gentamicin, amicacin, cephtriacson, cephipim, cephtazidim, norphloxacin, ciprophloxacin, imipenem and by determination of them minimum repressing concentration. Results it was registered by a statistical method with the use of indexes of p and х2. Results and discussions. It is conducted in of vitro research of sensitiveness of community-made strains of P. aeruginosa to the antibiotics with activity against pseudomonads and specific bacteriophages («Pseudomonas bacteriophage » and «Piobacteriophage»). By most activity in relation to the investigational of P. aeruginosa owned imipenem, ciprophloxacin, cephtazidim frequency of sensitiveness to that was 64,8%, 64,8%, 53,7% accordingly. Activity of amicacin and gentamicin was below – sensible to them were 33,3% and 38,9% accordingly. The smallest activity was owned by cephtriacson, sensible to that were only 18,5% strains ( p<0,05). The sensitiveness of community-made strains of Pseudomonas aeruginosa to commercial preparations – bacteriophages laid down about 50% and depended on the biotope of exception. 24,0% strains appeared proof both to the antibiotics and to the bacteriophages, and 13,0% polyresistant pseudomonads there were phagosensitive. No significant difference in the activity of the investigated phages was detected (х2>0,05). The antibiotics of choice for starting therapy are imipenem, ciprophloxacin, cephtazidim. For the increase of efficiency of phagotherapy it is desirable to create collection of regional strains – bacteriophages for producing of home preparations.