Vašková S, Slobodníková L, Fajtl D, Blažíčková S, Botek R, Melicháčová V
Epidemiol Mikrobiol Imunol. 2020 Winter;69(1):3-9.
The presented study was to compare in vitro biofilm production by bacterial strains from chronic/recurrent and from acute non-complicated UTIs. The activity of gentamicin and colistin on biofilm form of these strains has also been detected, with goal to predict the gentamicin and colistin therapeutic efficacy in the antimicrobial treatment of patients with a suspected presence of biofilm in urinary tract.
The group of 40 bacterial strains repeatedly isolated from patients with chronic or recurrent UTIs was compared with the group of 40 strains from acute UTIs. Both groups contained comparable number of strains of Escherichia coli, Klebsiella spp., Proteus mirabilis and Pseudomonas aeruginosa. Biofilm production was assessed by method in polystyrene microtiter plate. The MIC and MBC values of gentamicin and colistin were detected by broth microdilution assay. The minimal biofilm inhibitory (MBIC) and biofilm eradication concentrations (MBEC) were tested by microdilution method. Non-inactivated biofilm-associated bacteria were detected after overnight incubation in broth medium free of antimicrobials. The statistical analysis of results was performed by Fisher's exact test and by Student's t-test.
Biofilm was produced by 90% strains from chronic UTIs, but only by 52% of strains from acute UTIs (p = 0.0004). In the biofilm producing strains, the MBIC values of gentamicin reached from four to 256 mg/L, the MBIC levels of colistin from two to 64 mg/L. The minimal biofilm eradicating concentrations were even higher: for gentamicin from eight to > 512 mg/L, and for colistin from 32 to > 512 mg/L. The differences between MIC and MBIC/MBEC levels were statistically highly significant (p < 0.0001). Presumably, the therapeutic success of parenterally applied gentamicin or colistin on biofilm-related urinary tract infections would be, without respect to the high concentration of gentamicin or colistin achievable in urine during parenteral application, rather unpredictable. Local intravesical instillation would allow for achieving higher gentamicin and colistin concentrations; however, there is need for interpretation criteria for MBEC values concerning therapy, as well as for clinical studies allowing for application of those values to predict clinical success of therapy.
Laboratory detection of biofilm production and evaluation of the MBIC/MBEC values of antimicrobials for strains producing biofilm might be a valuable complement to the microbiologic diagnostics of chronic and recurrent UTIs. It might provide valuable information for more reliable individualised therapy and so decrease the risk of emergence and selection of multiresistant strains during repeated and non-eradicating therapy of chronic and recurrent UTIs.
本研究旨在比较慢性/复发性尿路感染和急性非复杂性尿路感染患者分离出的细菌菌株的体外生物膜形成情况。同时检测庆大霉素和黏菌素对这些菌株生物膜形式的活性,以预测庆大霉素和黏菌素在疑似尿路存在生物膜患者抗菌治疗中的疗效。
将从慢性或复发性尿路感染患者中反复分离出的40株细菌菌株与从急性尿路感染患者中分离出的40株细菌菌株进行比较。两组中大肠埃希菌、克雷伯菌属、奇异变形杆菌和铜绿假单胞菌的菌株数量相当。采用聚苯乙烯微量滴定板法评估生物膜形成情况。通过肉汤微量稀释法检测庆大霉素和黏菌素的MIC和MBC值。采用微量稀释法检测最小生物膜抑制浓度(MBIC)和生物膜根除浓度(MBEC)。在无抗菌药物的肉汤培养基中过夜培养后,检测未灭活的生物膜相关细菌。结果采用Fisher精确检验和学生t检验进行统计学分析。
90%的慢性尿路感染菌株可产生生物膜,而急性尿路感染菌株中只有52%能产生生物膜(p = 0.0004)。在产生生物膜的菌株中,庆大霉素的MBIC值为4至256 mg/L,黏菌素的MBIC水平为2至64 mg/L。最小生物膜根除浓度更高:庆大霉素为8至>512 mg/L,黏菌素为32至>512 mg/L。MIC与MBIC/MBEC水平之间的差异具有高度统计学意义(p < 0.0001)。推测,静脉应用庆大霉素或黏菌素治疗生物膜相关尿路感染的疗效,无论静脉应用期间尿液中可达到的庆大霉素或黏菌素浓度多高,都可能难以预测。膀胱内局部灌注可使庆大霉素和黏菌素达到更高浓度;然而,需要有关治疗的MBEC值的解读标准,以及允许应用这些值来预测治疗临床成功的临床研究。
实验室检测生物膜形成情况以及评估产生生物膜菌株的抗菌药物MBIC/MBEC值,可能是慢性和复发性尿路感染微生物诊断的有价值补充。它可能为更可靠的个体化治疗提供有价值的信息,从而降低慢性和复发性尿路感染反复及未根除治疗期间多重耐药菌株出现和选择的风险。