Wieczorek Piotr, Sacha Paweł, Ojdana Dominika, Milewski Robert, Jurczak Anna, Kaczyńska Katarzyna, Tryniszewska Elzbieta
Zakład Diagnostyki Mikrobiologicznej i Immunologii Infekcyjnej, Uniwersytet Medyczny w Białymstoku.
Med Dosw Mikrobiol. 2012;64(1):55-62.
Majority of nosocomial Acinetobacter baumannii strains are highly resistant to many available groups of antibiotics, causing therapy of infections the clinical challenge. The aim of study was to estimate of resistance development to sulbactam, cefoperazone and cefoperazone/sulbactam in Acinetobacter baumannii clinical strains.
Five Acinetobacter baumannii strains (Acb1, Acb2, Acb4, Acb13 and Acb25) were identified by the VITEK 2 GN card and the automatic system VITEK 2 according to the procedure and following the producer's instructions. Additionaly, the belonging of the strains to the species was confirmed by the presence of the bla(OXA-51-like) gene. Initial and after antibiotic exposure MIC values of sulbactam, cefoperazone and cefoperazone/sulbactam were determined by using a broth microdilution method. Antibiotic pressure of examined strains was performed in Mueller-Hinton broth containing 0,5x, 0,9x and 2x initial MIC of individual compounds during six-day passages and next six-day passages without antibiotic presence. The Mann-Whitney U test and Kruskal-Wallis non-prarametric Anova test were used to statistical analysis.
Serial passaging of Acinetobacter baumannii strains in the presence of antibiotics caused permanent increasing MIC value independently of used concentrations in the majority of examined strains. The highest MIC value increase of sulbactam was found in Acb4 strain. Even after two passages this isolate changed MIC from 0.5 microg/ml to 4 microg/ml (increase about four levels of concentration). Moreover, after incubation in 0.9x MIC concentration similar observation was noted. No normalization of MIC value of sulbactam after incubation during next six passages without sulbactam was observed. In case of cefoperazone the highest levels of induction were noted in Acb1, Acb13 and Acb25 strains. In these strains, after two passages in presence of cefoperazone (2xMIC) the exceedance of minimal of growth concentration over the highest examined concentration was observed. Similar effects were observed in Acbl strain after stimulation with 0.9x and 0.5x MIC cefoperazone. Return of initial MIC values was received only after induction with 0.5 x MIC cefoperazone. In some cases, no opportunities for evaluation of resistance development was noted, because during stimulation with 2x MIC of used antibiotics concentarations, bactericidal effect was found.
Sulbactam, cefoperazone and cefoperazone/sulbactam rapidly induce increasing of resistance in Acinetobacter baumannii clinical isolates. Statistically essential MIC increase after using higher concentration than lower was showed. This effect was particularly visible in the case of stimulation of cefoperazone/sulbactam combination.
大多数医院获得性鲍曼不动杆菌菌株对许多现有抗生素类别具有高度耐药性,这给感染治疗带来了临床挑战。本研究的目的是评估鲍曼不动杆菌临床菌株对舒巴坦、头孢哌酮和头孢哌酮/舒巴坦的耐药性发展情况。
使用VITEK 2 GN卡和自动系统VITEK 2,按照程序并遵循生产商说明,鉴定了五株鲍曼不动杆菌菌株(Acb1、Acb2、Acb4、Acb13和Acb25)。此外,通过bla(OXA - 51样)基因的存在确认菌株属于该物种。采用肉汤微量稀释法测定舒巴坦、头孢哌酮和头孢哌酮/舒巴坦的初始及抗生素暴露后的MIC值。在含各化合物初始MIC的0.5倍、0.9倍和2倍浓度的 Mueller - Hinton肉汤中,对受试菌株进行为期六天的传代培养,然后在无抗生素的情况下进行接下来的六天传代培养,以施加抗生素压力。采用Mann - Whitney U检验和Kruskal - Wallis非参数方差分析进行统计分析。
在抗生素存在的情况下,鲍曼不动杆菌菌株的连续传代导致大多数受试菌株的MIC值持续升高,与所用浓度无关。舒巴坦的MIC值升高幅度最大的是Acb4菌株。即使经过两次传代,该菌株的MIC值也从0.5μg/ml变为4μg/ml(浓度增加约四个水平)。此外,在0.9倍MIC浓度下培养后也观察到了类似现象。在接下来无舒巴坦的六次传代培养后,未观察到舒巴坦MIC值恢复正常。对于头孢哌酮,Acb1、Acb13和Acb25菌株的诱导水平最高。在这些菌株中,在头孢哌酮(2倍MIC)存在下传代两次后,观察到最低生长浓度超过最高检测浓度。在用0.9倍和0.5倍MIC头孢哌酮刺激后,Acbl菌株也观察到了类似效果。仅在用0.5倍MIC头孢哌酮诱导后才恢复初始MIC值。在某些情况下,未观察到评估耐药性发展的机会,因为在用2倍MIC的所用抗生素浓度刺激时,发现了杀菌作用。
舒巴坦、头孢哌酮和头孢哌酮/舒巴坦可迅速诱导鲍曼不动杆菌临床分离株的耐药性增加。使用较高浓度比使用较低浓度后,MIC在统计学上有显著增加。这种效应在头孢哌酮/舒巴坦联合刺激的情况下尤为明显。