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Документ Вплив детергенту, хімотрипсину та фагів на утворення біоплівок стафілококами(Українська медична стоматологічна академія, 2017) Семенчук, П. О.; Соколова, І. Є.; Воробєй, Є. С.; Вінніков, А. І.; Семенчук, П. А.; Соколова, И. Е.; Воробей, Е. С.; Винников, А. И.; Semenchuk, P. A.; Sokolova, I. E.; Vorobey, E. S.; Vinnikov, A. I.У статті представлено результати досліджень впливу детергенту (додецилсульфату натрію), хімотрипсину, і стафілококового фагу на утворення біоплівки культурою Staphylococcus aureus. Встановлено, що всі перелічені чинники виявляють відносно стафілококової біоплівки руйнівну дію, ступінь якої корелює з підвищенням концентрації означених речовин; В статье представлены результаты исследований влияния детергента (додецилсульфата натрия), химотрипсина, и стафилококкового фага на образование биоплёнки культурой Staphylococcus aureus. Установлено, что все перечисленные факторы проявляют относительно стафилококковой биоплёнки деструктивное действие, степень которого коррелирует с повышением концентрации указанных веществ; The aim of this work was investigation of influence of sodium dodecyl sulfate, chymotrypsin and phages on formation of biofilms by staphylococcus. Biological objects of study were able to film formation of clinical strain of Staphylococcus aureus isolated from patients with disorders of the gastrointestinal tract and bacteriophages from commercial preparation «Bacteriophage staphylococcal liquid» (JSC «Biopharma», Ukraine). To establish a dedicated clinical strain belonging to the species S. aureus were checked its physiological and biochemical properties, such as hemolytic, lipase and lecithinase activities in accordance seeding bacteria on blood and milk-yolk-salt agar, and also plasmocoagulase activity was determined using citrate rabbit plasma. Staphylococcus are capable to strike almost any organs and tissues of the human body. One of the important factors of pathogenic S. aureus is formation of biofilms, which is part of the life cycle and protect of bacteria from adverse environmental factors. The forming film staphylococcus rapidly acquire antibiotic resistance, and in microbial associations with other agents resistance is even more pronounced. The factors of internal environment such as hormones, enzymes, antibodies, endogenous antibiotics and other metabolites influence essentially on biofilmforming ability of opportunistic microorganisms. The substances whose effects on biofilm studied, chosen not by chance. Thus, there is no doubt that the process of film-forming on the mucosa of the intestine depend on digestive enzymes, especially proteases (trypsin and chymotrypsin) that can break down the protein components in the cell walls and membranes of bacteria and in matrix of biofilms. Determination of detergent effects on film-forming ability is important to develop means of disinfection. The study of bacteriophages action is need for creation of effective drugs for treatment of staphylococcal infections caused by film-forming strains. For biofilms growing 0.4 ml suspension of S. aureus, 1.5 ml nourishing broth and 0.1 ml examined substances in according concentration carried in each well of laboratory plate. The samples were incubating at 37°С during 72 h. After cultivation the medium rest removed from wells, the sediment dissolved in 2 ml physiological solution and tested the optical density at 540 nm. Definition of sodium dodecyl sulfate (SDS) influence on formation of biofilms by staphylococcus shown that optical density of bacteria in biofilms decreases with increasing concentrations of SDS. At the minimal concentration of SDS (0.05 mg/ml) the number of bacteria in the biofilm was 58.7% of control, increasing detergent concentrations (0.5 and 5 mg/ml) some more reduced biofilm density, but not so much as expected, and amounted to 52.3 and 42.2% of controls. Based on the findings, we can suggest that sodium dodecyl sulfate, as detergent, causing destructive effect on the cells of themselves staphylococcus and the elements of the matrix, so that part of bacteria fixed in the biofilm can come in free planktonic phase, and the density of the film as a result significantly reduce. In the study chymotrypsin impact on the biofilm-formation the enzyme concentrations selected so that they would be equivalent to the one that really exist in the body. The chymotrypsin concentration in healthy human intestinal is approximately equal to 0.025% of the total volume of secretions of the slim intestine. So the experimental final concentrations of enzyme in the wells of plate were 0.05, 0.025 and 0.0125 mg/ml. The optical density figures of staphylococcus suspensions in biofilms for increasing concentrations of chymotrypsin (0.0125, 0.025 and 0.05 mg/ml) were reduced after 3-day incubation with the enzyme to 63.3, 51.4 and 36.7% compared to control. Therefore destructive action of chymotrypsin relatively biofilms raise with increasing concentration of the enzyme. To study the influence of phage on the formation of staphylococcus biofilms used commercial preparation Bacteriophagum staphylococcus fluidum. The phage activity is first checked by Gracia method in their ability to form negative colonies on the medium with investigational clinical strain of S. aureus. The initial titre of bacteriophage was equal to 8,0 × 108 BFU/ml (bacteriophage forming units) and its final concentration in the wells was 4,0 × 107 BFU/ ml. The obtained experimental data had demonstrated reducing of staphylococcus biomass in biofilm almost twice after three days incubation with the bacteriophage, namely the optical density of microbial suspensions obtained from biofilm decreased to 51% compared to the control of native biofilm.Документ Етіологічні особливості негоспітальної пневмонії у хворих різних вікових груп(Українська медична стоматологічна академія, 2017) Декун, Т. Ю.; Соколова, І. Є.; Ювко, А. В.; Братусь, О. В.; Вінніков, А. І.; Декун, Т. Ю.; Соколова, И. Е.; Ювко, А. В.; Братусь, Е. В.; Винников, А. И.; Dekun, T. Y.; Sokolova, I. E.; Yvko, A. V.; Bratys, E. V.; Vinnikov, A. I.У роботі представлені результати мікробіологічного дослідження мокротиння 273 пацієнтів, що мали підозру на негоспітальну пневмонію, за період з січня по вересень 2016 року з метою виявлення етіологічних факторів захворювання в залежності від віку. В результаті було встановлено, що домінуючими збудниками пневмонії були Haemophilus influenzae і Haemophilus parainfluenzaе (47,31 і 33,33%,), рідше виділялись Streptococcus pneumoniae і Moraxella catarrhalis (10,75 і 8,60%). У осіб середнього (36-60 р.) та похилого віку (61-90 р.) відсоток усіх виявлених патогенів був найвищим (40,93 і 24,58% відповідно), у осіб молодого (20-35 р.) та юного (14-19 р.) віку він склав 19,51 і 6,75%, у молодших вікових групах даний параметр був найнижчим – 4,74 (8-13 р.) і 3,29 (0-7 р.) %; В работе представлены результаты микробиологического исследования мокроты 273 пациентов, что имели подозрение на внебольничную пневмонию, за период с января по сентябрь 2016 года з целью выявления этиологических факторов заболевания в зависимости от возраста. В результате было установлено, что доминирующими возбудителями пневмонии были Haemophilus influenza и Haemophilus parainfluenzaе (47,31 и 33,33%,), реже выделялись Streptococcus pneumoniae и Moraxella catarrhalis (10,75 и 8,60%). У пациентов среднего (36-60 лет) и преклонного возраста 61-90 лет) процент всех выявленных патогенов был наивысшим (40,93 и 24,58% соответственно), у пациентов молодого (20-35 лет) и юного (14-19 лет) возраста он составил 19,51 и 6,75%, в младших возрастных группах данный параметр был самым низким – 4,74 (8-13 лет) и 3,29 (0-7 лет) %; A community-acquired pneumonia (CAP) is one of the most widespread and life-threatening infectious disease. A quantity of lung inflammation incidences in 2011 was 494.3 per 100 thousands of population. A certain progress in diagnostics and treatment of CAP was reached, despite this mortality is increasing, in consequence of complications and pathogenicity modification of microorganisms, formation of antibiotic resistance and also decreased immune status of population. Considering relevance of this problem, the aim of this work was identification of CAP pathogens from sputum of patients and also analysis of their etiologic structure in patients of different ages. The samples were grown in different media for cultivation – 5% sheep blood agar, chocolate agar, Sabouraud agar and EMB agar. We have investigated the sputum samples of 273 patients with CAP, who were treated during 2016 year at the laboratory of Diagnostic center of LLP Pharmacies of medical academy of Dnipro. The bacteria were isolated in 93 (34%) of 273 clinical samples, from which the leading microorganisms were Haemophilus influenzae (47.31%), H. parainfluenzae (33.33%,), S. pneumoniae (10.75%) and M. catarrhalis (8,60% of all isolates). The monoinfections (59.62%) in the etiological structure of CAP pathogens were dominate, the associations of two or three infectious agents were found according in 29.81% and 10.52% causes. The largest number of detected CAP were observed when the concentration of bacteria H. influenza, S. pneumoniae and M. catarrhalis were ≥ 106 and ≥ 108 CFU/ml (Colonies formation units per ml), while frequency of detection rate of H. parainfluenzae was highest in low titer ≤ 104 CFU/ml. Frequency of definition of certain CAP pathogens widely varied in depends of year season. Thus, the maximum number of pneumonia caused by H. influenzae (38%) and H. parainfluenzae (27%) were detected in the spring, but the degree of S. pneumoniae detection was low (9%) at this period. In summer the number of identified pathogens decreased in all organisms, and the growth of infection began with arrival of autumn. The presented data about etiological structure of community-acquired pneumonia can be used in clinical practice and underscores the needs of further study of the role of significant and atypical pathogens in development of community-acquired pneumonia.