Isolation and characterization of bacteriophages against IMP-6-producing<i>Klebsiella pneumoniae</i>isolated from clinical settings in Japan
Isolation and characterization of bacteriophages against IMP-6-producing<i>Klebsiella pneumoniae</i>isolated from clinical settings in Japan
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从日本临床环境中分离出的针对产生 IMP-6 肺炎克雷伯菌的噬菌体的分离和表征
DOI：
10.1101/2022.11.05.515272
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Kawano Mitsuoki
中科院分区：
文献类型：
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作者：
Kondo Kohei;Nakano Satoshi;Hisatsune Junzo;Sugawara Yo;Kataoka Michiyo;Kayama Shizuo;Sugai Motoyuki;Kawano Mitsuoki
产碳青霉烯酶肠杆菌科（CPE）是世界范围内最有害的耐药菌之一。噬菌体治疗已成为治疗CPE病原体引起的感染的有效策略。在日本西部，携带pKPI-6质粒（编码blaIMP-6）的肺炎克雷伯氏菌的出现越来越多，这一问题日益受到关注。为了管理这些主要的耐药病原体，我们从日本的污水中分离出29种新的细菌，目标是31株K。pKPI-6质粒的大肠埃希菌1株。电镜分析表明，在分离的29株病毒中，肌尾病毒科（Myoviridae）21株（72.4%），管尾病毒科（Siphoviridae）5株（17.2%），短尾病毒科（Podoviridae）3株（10.3%）。宿主范围分析表明，20个肌尾病毒科成员感染25-26株克雷伯氏菌。pneumoniae，表明大多数分离的大肠杆菌具有广泛的宿主范围。K。肺炎克雷伯氏菌Kp 21只能被噬菌体øKp_21感染，而Kp 22可以被超过20 μ M的噬菌体感染。我们应用由10 μ g组成的噬菌体混合物对抗Kp 21和Kp 22，发现噬菌体混合物延迟了Kp 21菌株而不是Kp 22菌株的噬菌体抗性细菌的出现。此外，噬菌体抗性Kp 21（Kp 21 r）变得易于被其他噬菌体感染，作为对噬菌体øKp_21的抗性的“权衡”。我们提出的噬菌体集有足够数量的噬菌体来对抗K。日本分离的肺炎病毒株。值得注意的是，我们的工作证明了一个合适的噬菌体鸡尾酒如何减少噬菌体耐药bacteries.ImportanceKlebsiella pneumoniaeharboring质粒carryingblaIMP-6的发生正在成为一个越来越危险的物种在日本。我们收集并鉴定了29种感染K的新型噬菌体。携带pKPI-6质粒的肺炎，在日本西部的临床环境中分离。我们的寄生虫表现出广泛的寄主范围。我们应用了由10 μ g/ml构建的噬菌体鸡尾酒处理两种宿主菌株Kp 21和Kp 22，它们彼此显示出不同的噬菌体易感性模式。尽管噬菌体混合物延迟了抗噬菌体Kp 21的出现，但不能延迟抗噬菌体Kp 22的出现。此外，抗噬菌体的Kp 21变得对其他病原体敏感，这些病原体最初并不感染野生型Kp 21。我们的研究证明了合适的噬菌体鸡尾酒可以减少噬菌体抗性细菌的发生。
Carbapenemase-producingEnterobacteriaceae(CPE) are one of the most detrimental species of antibiotic-resistant bacteria worldwide. Phage therapy has emerged as an effective strategy for the treatment of infections caused by CPE pathogens. In west Japan, the increasing occurrence ofKlebsiella pneumoniaeharboring the pKPI-6 plasmid, which encodesblaIMP-6, is a growing concern. To manage such major antimicrobial-resistant pathogens, we isolated 29 novel phages from sewage in Japan, targeting 31 strains ofK. pneumoniaeand one strain ofEscherichia coliharboring the pKPI-6 plasmid. Electron microscopy analysis indicated that of the 29 isolated phages, 21 (72.4%), 5 (17.2%), and 3 (10.3%) belonged toMyoviridae, Siphoviridae, andPodoviridae, respectively. Host range analysis revealed that 20Myoviridaemembers in isolated phages infected 25–26 strains ofK. pneumoniae, indicating that most of the isolated phages have a broad host range. TheK. pneumoniaeKp21 can only be infected by phage øKp_21, while Kp22 can be infected by more than 20 phages. We applied a phage cocktail, which consists of 10 phages, against Kp21 and Kp22 and found that the phage cocktail delayed the emergence of phage-resistant bacteria for Kp21 strain but not for the Kp22 strain. Furthermore, phage-resistant Kp21 (Kp21r) became prone to be infected from other bacteriophages as a “trade-off” of resistance to phage øKp_21. Our proposed phage set has an adequate number of phages to combat theK. pneumoniaestrain isolated in Japan. Notably, our work demonstrates how a suitable phage cocktail diminishes the occurrence of phage-resistant bacteria.ImportanceKlebsiella pneumoniaeharboring the plasmid carryingblaIMP-6is becoming an increasingly hazardous species in Japan. We collected and characterized 29 novel bacteriophages that infectK. pneumoniaecarrying the pKPI-6 plasmid, isolated in clinical settings of west Japan. Our phages showed broad host ranges. We applied a phage cocktail treatment constructed from 10 phages against two host strains, Kp21 and Kp22, which show different phage susceptibility patterns each other. Although the phage cocktail delayed phage-resistant Kp21 emergence, the emergence of phage-resistant Kp22 could not be delayed. Moreover, phage-resistant Kp21 became sensitive to other phages, which did not originally infect wild-type Kp21. Our study demonstrates how a suitable phage cocktail can diminish the occurrence of phage-resistant bacteria.