Phylogenetic Aspects of Antibiotic Resistance and Biofilm Formation of P. aeruginosa Isolated from Clinical Samples

Maryam Motevasel, Masoud Haghkhah, Negar Azimzadeh
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The results showed that 35.6% and 16.94% of all the samples were isolated from urine and wounds, 81.33% of the isolates were biofilm producers, 27.11% were multidrug-resistant (MDR), and 100% of the main biofilm former genes belonged to <i>pslA</i>. 94.91% of the isolates possessed <i>brlR</i> and <i>mexA</i>, and 91.5% of them expressed <i>pslA</i>. It was also indicated that neither ciprofloxacin nor imipenem could eradicate the formed biofilms. Moreover, we could identify 81.4% distinctive restriction profiles among the isolates, using an 80% similarity cutoff point; <i>brlR</i> and <i>pel</i> genes were significantly (<span><svg height=\"8.8423pt\" style=\"vertical-align:-0.2064009pt\" version=\"1.1\" viewbox=\"-0.0498162 -8.6359 19.289 8.8423\" width=\"19.289pt\" xmlns=\"http://www.w3.org/2000/svg\" xmlns:xlink=\"http://www.w3.org/1999/xlink\"><g transform=\"matrix(.013,0,0,-0.013,0,0)\"></path></g><g transform=\"matrix(.013,0,0,-0.013,11.658,0)\"></path></g></svg><span></span><span><svg height=\"8.8423pt\" style=\"vertical-align:-0.2064009pt\" version=\"1.1\" viewbox=\"22.8711838 -8.6359 28.182 8.8423\" width=\"28.182pt\" xmlns=\"http://www.w3.org/2000/svg\" xmlns:xlink=\"http://www.w3.org/1999/xlink\"><g transform=\"matrix(.013,0,0,-0.013,22.921,0)\"></path></g><g transform=\"matrix(.013,0,0,-0.013,29.161,0)\"></path></g><g transform=\"matrix(.013,0,0,-0.013,32.125,0)\"><use xlink:href=\"#g113-49\"></use></g><g transform=\"matrix(.013,0,0,-0.013,38.365,0)\"></path></g><g transform=\"matrix(.013,0,0,-0.013,44.605,0)\"></path></g></svg>;</span></span> <span><svg height=\"8.8423pt\" style=\"vertical-align:-0.2064009pt\" version=\"1.1\" viewbox=\"-0.0498162 -8.6359 19.289 8.8423\" width=\"19.289pt\" xmlns=\"http://www.w3.org/2000/svg\" xmlns:xlink=\"http://www.w3.org/1999/xlink\"><g transform=\"matrix(.013,0,0,-0.013,0,0)\"><use xlink:href=\"#g113-81\"></use></g><g transform=\"matrix(.013,0,0,-0.013,11.658,0)\"><use xlink:href=\"#g117-34\"></use></g></svg><span></span><span><svg height=\"8.8423pt\" style=\"vertical-align:-0.2064009pt\" version=\"1.1\" viewbox=\"22.8711838 -8.6359 28.182 8.8423\" width=\"28.182pt\" xmlns=\"http://www.w3.org/2000/svg\" xmlns:xlink=\"http://www.w3.org/1999/xlink\"><g transform=\"matrix(.013,0,0,-0.013,22.921,0)\"><use xlink:href=\"#g113-49\"></use></g><g transform=\"matrix(.013,0,0,-0.013,29.161,0)\"><use xlink:href=\"#g113-47\"></use></g><g transform=\"matrix(.013,0,0,-0.013,32.125,0)\"><use xlink:href=\"#g113-49\"></use></g><g transform=\"matrix(.013,0,0,-0.013,38.365,0)\"></path></g><g transform=\"matrix(.013,0,0,-0.013,44.605,0)\"><use xlink:href=\"#g113-53\"></use></g></svg>)</span></span> related to phylogenetic pulsotypes. Comparison of the dendrogram in the isolates revealed that the detected isolates from urine were present in 12 different pulsotypes. <i>Conclusion</i>. It was found that there was a relationship between MDR, biofilm production, and <i>brlR</i> and <i>pel genes</i> among the isolates. It is distinguished there were similar genetic patterns between detected isolates from urine and could be concluded that the urinary tract played a critical role in maintaining and transferring biofilm drug-resistant genes of <i>P. aeruginosa</i> in clinical sites. 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Abstract

Introduction. Biofilm production and drug resistance phenomenon play a critical role in P. aeruginosa infections. Several genes, including psl, pel, brlR, and mex, are involved in the phenomenon. The aim of this study was to find the relationship between the mentioned genes and the sources of P. aeruginosa infections. Materials and Methods. Fifty-nine P. aeruginosa isolates detected from clinical specimens were used to determine antibiotic susceptibility patterns, prevalence of the genes using PCR, biofilm formation, biofilm eradication concentration assay (MBEC), and epidemiological characteristics using pulsed-field gel electrophoresis (PFGE). Results. The results showed that 35.6% and 16.94% of all the samples were isolated from urine and wounds, 81.33% of the isolates were biofilm producers, 27.11% were multidrug-resistant (MDR), and 100% of the main biofilm former genes belonged to pslA. 94.91% of the isolates possessed brlR and mexA, and 91.5% of them expressed pslA. It was also indicated that neither ciprofloxacin nor imipenem could eradicate the formed biofilms. Moreover, we could identify 81.4% distinctive restriction profiles among the isolates, using an 80% similarity cutoff point; brlR and pel genes were significantly (; ) related to phylogenetic pulsotypes. Comparison of the dendrogram in the isolates revealed that the detected isolates from urine were present in 12 different pulsotypes. Conclusion. It was found that there was a relationship between MDR, biofilm production, and brlR and pel genes among the isolates. It is distinguished there were similar genetic patterns between detected isolates from urine and could be concluded that the urinary tract played a critical role in maintaining and transferring biofilm drug-resistant genes of P. aeruginosa in clinical sites. The study highlights the importance of urine in distribution of clinical biofilm formation and drug-resistant P. aeruginosa isolates.
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从临床样本中分离出的铜绿微囊桿菌的抗生素耐药性和生物膜形成的系统发育特征
导言。生物膜的产生和耐药性现象在铜绿假单胞菌感染中起着至关重要的作用。包括 psl、pel、brlR 和 mex 在内的多个基因参与了这一现象。本研究旨在找出上述基因与铜绿假单胞菌感染源之间的关系。材料与方法。对从临床标本中检测到的 59 株铜绿假单胞菌进行抗生素敏感性模式测定、利用 PCR 测定基因的流行率、生物膜形成、生物膜根除浓度测定(MBEC)以及利用脉冲场凝胶电泳(PFGE)测定流行病学特征。结果显示结果显示,35.6%和 16.94%的样本分别从尿液和伤口中分离出来,81.33%的分离株是生物膜生产者,27.11%的分离株具有多重耐药性(MDR),100%的主要生物膜形成基因属于 pslA。94.91% 的分离物拥有 brlR 和 mexA,91.5% 的分离物表达 pslA。研究还表明,环丙沙星和亚胺培南都无法根除已形成的生物膜。此外,以 80% 的相似度为临界点,我们可以在分离株中识别出 81.4% 的独特限制性图谱;brlR 和 pel 基因与系统发育脉型有显著(;)关系。对分离物的树枝图进行比较后发现,从尿液中检测到的分离物存在于 12 个不同的脉型中。结论研究发现,分离物中的 MDR、生物膜生成、brlR 和 pel 基因之间存在关系。尿液中检测到的分离株之间存在相似的基因模式,因此可以得出结论,尿道在维持和转移铜绿假单胞菌生物膜耐药基因方面在临床中起着关键作用。这项研究强调了尿液在临床生物膜形成和耐药铜绿假单胞菌分离物分布中的重要性。
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