Anti-Infective Research Laboratory, Department of Pharmacy Practice, Eugene Applebaum College of Pharmacy and Health Sciences, Wayne State University, Detroit, Michigan, USA.
Department of Clinical Pharmacy & Outcomes Sciences, Medical University of South Carolina, College of Pharmacy, Charleston, South Carolina, USA.
J Appl Microbiol. 2022 Sep;133(3):1636-1649. doi: 10.1111/jam.15647. Epub 2022 Jun 28.
Here, we investigate the impact of phage-antibiotic combinations (PAC) on bacterial killing, resistance development and outer membrane vesicle (OMV) production in multidrug-resistant (MDR) P. aeruginosa.
After screening 10 well-characterized MDR P. aeruginosa strains against three P. aeruginosa phages, representative strains, R10266 and R9316, were selected for synergy testing based on high phage sensitivity and substantial antibiotic resistance patterns, while phage EM was chosen based on host range. To understand the impact of phage-antibiotic combinations (PAC) against MDR P. aeruginosa, time-kill analyses, OMV quantification and phage/antibiotic resistance testing were performed. Phage and meropenem demonstrated synergistic activity against both MDR strains. Triple combination regimens, phage-meropenem-colistin and phage-ciprofloxacin-colistin, resulted in the greatest CFU reduction for strains R9316 (3.50 log CFU ml ) and R10266 (4.50 log CFU ml ) respectively. PAC resulted in regained and improved antibiotic susceptibility to ciprofloxacin (MIC 2 to 0.0625) and meropenem (MIC 32 to 16), respectively, in R9316. Phage resistance was prevented or reduced in the presence of several classes of antibiotics and OMV production was reduced in the presence of phage for both strains, which was associated with significantly improved bacterial eradication.
These findings support the potential of phage-antibiotic synergy (PAS) to augment killing of MDR P. aeruginosa. Systematic in vitro and in vivo studies are needed to better understand phage interactions with antipseudomonal antibiotics, to define the role of OMV production in P. aeruginosa PAC therapy and to outline pharmacokinetic and pharmacodynamic parameters conducive to PAS.
This study identifies novel bactericidal phage-antibiotic combinations capable of thwarting resistance development in MDR and XDR P. aeruginosa strains. Furthermore, phage-mediated OMV reduction is identified as a potential mechanism through which PAC potentiates bacterial killing.
本研究旨在探讨噬菌体-抗生素联合治疗（PAC）对多重耐药（MDR）铜绿假单胞菌的杀菌效果、耐药性发展和外膜囊泡（OMV）产生的影响。
通过对 10 株经过充分表征的 MDR 铜绿假单胞菌菌株进行三种铜绿假单胞噬菌体的筛选，根据噬菌体敏感性和抗生素耐药模式，选择 R10266 和 R9316 两种代表性菌株进行协同作用测试，而噬菌体 EM 则基于宿主范围进行选择。为了了解噬菌体-抗生素联合治疗（PAC）对 MDR 铜绿假单胞菌的影响，进行了时间杀伤分析、OMV 定量和噬菌体/抗生素耐药性测试。噬菌体和美罗培南对两种 MDR 菌株均表现出协同作用。噬菌体-美罗培南-黏菌素和噬菌体-环丙沙星-黏菌素三联组合治疗方案分别使 R9316（减少 3.50 log CFU ml）和 R10266（减少 4.50 log CFU ml）的 CFU 减少量最大。PAC 使 R9316 对环丙沙星（MIC 2 至 0.0625）和美罗培南（MIC 32 至 16）的抗生素敏感性得到恢复和改善。在几种抗生素存在的情况下，噬菌体耐药性得到了预防或降低，而在两种菌株中，噬菌体的存在都降低了 OMV 的产生，这与细菌清除率的显著提高有关。
这些发现支持噬菌体-抗生素协同作用（PAS）增强对 MDR 铜绿假单胞菌的杀菌效果。需要进行系统的体外和体内研究，以更好地了解噬菌体与抗假单胞菌抗生素的相互作用，确定 OMV 产生在铜绿假单胞菌 PAC 治疗中的作用，并概述有利于 PAS 的药代动力学和药效学参数。
本研究确定了新型杀菌噬菌体-抗生素联合治疗方案，能够阻止 MDR 和 XDR 铜绿假单胞菌菌株的耐药性发展。此外，噬菌体介导的 OMV 减少被确定为 PAC 增强杀菌效果的潜在机制。
