Влияние цефтриаксона и тетрациклина на формирование биопленки штаммами Staphylococcus epidermidis

O. I. Sidashenko, O. S. Voronkova, Y. A. Sirokvasha, A. I. Vinnikov
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Abstract

122 strains of staphylococci were identified. Among the examined 122 clinical strains of staphylococci, 67 strains belonged to coagulase-positive, and 55 strains to the coagulase-negative ones. According to the study of physiological and biochemical properties, it was found that 37 strains (30.3%) belonged to S. epidermidis species. One of the biological properties of many bacteria is the ability to film formation and these strains attract special attention, since it is known that the film antibiotic resistance is higher than in planktonic cultures. It was determined that 20 strains of those under study were film-forming, 17 strains – non-biofilm forming ones. The film was formed during three days, and settled to the bottom of the plate holes. The clinical (Cl) strain of S. epidermidis was sensitive to ceftriaxone and tetracicline. The control (C) strains of S. epidermidis were sensitive to ceftriaxone, tetracycline and sizomicine. The study of biofilm growth for 2, 3 and 4 days of incubation was carried out. The maximum rate of biofilm S. epidermidis C was observed during 2–3 days; there is the most intense increase of cells number from 5.2 × 10 8 CFU/ml, for S. epidermidis Cl to 5.6 × 10 8 CFU/ml. The effect of ceftriaxone and tetracycline on biofilm formation by 2 investigation strains of S. epidermidis was found. We determined differences in minimal inhibitory concentrations (MIC) for planktonic cultures and biofilm of strains under study. It was established that MIC antibiotics inhibited the growth of planktonic cultures on average 2 times lower compared to the MIC which inhibited the biofilm formation. MIC for planktonic culture of S. epidermidis Cl defined for ceftriaxone was equal to 10 mg/ml, and for tetracycline – 1 mg/ml. MIC of ceftriaxone for the control strain was equal to 12 mg/ml, MIC of tetracycline – 0.7 mg/ml. MIC values for dynamics biofilm formation of S. epidermidis Cl strain on the plater were as follows: to ceftriaxone – 20 mg/ml and for tetracycline – 2 mg/ml, MIC of ceftriaxone for S. epidermidis C strain – 24 mg/ml, MIC of tetracycline – 1.5 mg/ml. The effect of ceftriaxone and tetracycline was defined to the larger extent, than MIC for biofilm-forming on the plate (10, 50 and 100 times). More effective action of tetracycline was shown for 1- and 2-daily biofilm cultures of S. epidermidis clinical strain. Adding tetracycline concentration of 20 mg/ml in the culture medium of the 1-day biofilm of S. epidermidis Cl strain reduced the cell number of biofilm formation 590 times, increasing concentrations of tetracycline to 100 mg/ml and added to the 1-day biofilm of the clinical strain reduced the number of bacterial cell 4400 times compared with control.
头孢曲松和四环素对生物胶片形成的影响
共检出葡萄球菌122株。122株葡萄球菌临床检出凝固酶阳性67株,凝固酶阴性55株。经生理生化特性研究,发现37株(30.3%)属表皮葡萄球菌。许多细菌的生物学特性之一是能够成膜,这些菌株引起了特别的注意,因为众所周知,这种成膜的抗生素耐药性比浮游生物培养的要高。其中成膜菌株20株,非成膜菌株17株。胶片在三天内形成,并沉降到板孔的底部。临床(Cl)株表皮葡萄球菌对头孢曲松和四环素敏感。对照(C)株表皮葡萄球菌对头孢曲松、四环素和西佐米辛敏感。对2、3、4 d的生物膜生长情况进行了研究。生物膜表皮葡萄球菌C在2 ~ 3 d达到最大值;表皮葡萄球菌Cl的细胞数从5.2 × 10 8 CFU/ml增加到5.6 × 10 8 CFU/ml,增幅最大。研究了头孢曲松和四环素对2株表皮葡萄球菌生物膜形成的影响。我们确定了浮游生物培养和生物膜的最低抑制浓度(MIC)的差异。结果表明,MIC抗生素对浮游生物培养物生长的抑制作用平均比MIC低2倍,而MIC抑制生物膜的形成。浮游培养表皮葡萄球菌的MIC为头孢曲松为10 mg/ml,四环素为1 mg/ml。对照菌株头孢曲松的MIC为12 mg/ml,四环素的MIC为- 0.7 mg/ml。表皮葡萄球菌Cl菌株在平板上动态生物膜形成的MIC值为:头孢曲松- 20 mg/ml,四环素- 2 mg/ml,表皮葡萄球菌C菌株头孢曲松的MIC - 24 mg/ml,四环素的MIC - 1.5 mg/ml。头孢曲松和四环素对平板生物膜形成的影响比MIC更大(10倍、50倍和100倍)。四环素对表皮葡萄球菌临床菌株每日1次和2次生物膜培养的作用更有效。在表皮葡萄球菌(S. epidermidis) Cl菌株1 d生物膜的培养基中添加浓度为20 mg/ml的四环素,使生物膜形成的细胞数减少了590倍,将四环素浓度增加到100 mg/ml,使临床菌株1 d生物膜的细菌细胞数比对照减少了4400倍。
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