Isolation and Characterization of Klebsiella Phages for Phage Therapy.

PHAGE (New Rochelle, N.Y.) Pub Date : 2021-03-01 Epub Date: 2021-03-17 DOI:10.1089/phage.2020.0046
Eleanor M Townsend, Lucy Kelly, Lucy Gannon, George Muscatt, Rhys Dunstan, Slawomir Michniewski, Hari Sapkota, Saija J Kiljunen, Anna Kolsi, Mikael Skurnik, Trevor Lithgow, Andrew D Millard, Eleanor Jameson
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Abstract

Introduction: Klebsiella is a clinically important pathogen causing a variety of antimicrobial resistant infections in both community and nosocomial settings, particularly pneumonia, urinary tract infection, and sepsis. Bacteriophage (phage) therapy is being considered a primary option for the treatment of drug-resistant infections of these types. Methods: We report the successful isolation and characterization of 30 novel, genetically diverse Klebsiella phages. Results: The isolated phages span six different phage families and nine genera, representing both lysogenic and lytic lifestyles. Individual Klebsiella phage isolates infected up to 11 of the 18 Klebsiella capsule types tested, and all 18 capsule-types were infected by at least one of the phages. Conclusions: Of the Klebsiella-infecting phages presented in this study, the lytic phages are most suitable for phage therapy, based on their broad host range, high virulence, short lysis period and given that they encode no known toxin or antimicrobial resistance genes. Phage isolates belonging to the Sugarlandvirus and Slopekvirus genera were deemed most suitable for phage therapy based on our characterization. Importantly, when applied alone, none of the characterized phages were able to suppress the growth of Klebsiella for more than 12 h, likely due to the inherent ease of Klebsiella to generate spontaneous phage-resistant mutants. This indicates that for successful phage therapy, a cocktail of multiple phages would be necessary to treat Klebsiella infections.

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用于噬菌体疗法的克雷伯氏菌噬菌体的分离和特征描述。
导言:克雷伯氏菌是一种重要的临床病原体,可在社区和医院内引起各种耐药性感染,尤其是肺炎、尿路感染和败血症。细菌噬菌体(噬菌体)疗法被认为是治疗此类耐药感染的主要选择。方法:我们成功分离并鉴定了 30 种新型、基因多样的克雷伯氏菌噬菌体。结果:分离出的噬菌体横跨六种不同的噬菌体:分离出的噬菌体跨越了 6 个不同的噬菌体家族和 9 个属,代表了溶菌性和溶解性两种生活方式。分离出的克雷伯氏菌噬菌体感染了所测试的 18 种克雷伯氏菌胶囊类型中的 11 种,所有 18 种胶囊类型都至少被其中一种噬菌体感染。结论在本研究介绍的克雷伯氏菌感染噬菌体中,溶菌噬菌体最适合用于噬菌体疗法,因为它们的宿主范围广、毒力强、溶解时间短,而且没有编码已知的毒素或抗菌药耐药性基因。根据我们的特征分析,认为属于 Sugarlandvirus 和 Slopekvirus 属的噬菌体分离物最适合噬菌体疗法。重要的是,当单独使用噬菌体时,没有一种噬菌体能够抑制克雷伯氏菌的生长超过 12 小时,这可能是由于克雷伯氏菌本身容易产生自发的噬菌体抗性突变体。这表明,要想成功进行噬菌体治疗,必须使用多种噬菌体的鸡尾酒来治疗克雷伯氏菌感染。
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Beyond Antibiotics: Exploring the Potential of Bacteriophages and Phage Therapy. Correction to: Addressing the Research and Development Gaps in Modern Phage Therapy, by Paul E. Turner et al., PHAGE: Ther Appl Res 2024;5(1):30-39; doi: 10.1089/phage.2023.0045. A New Insight into Phage Combination Therapeutic Approaches Against Drug-Resistant Mixed Bacterial Infections. Bacteriophage Dosing and Its Effect on Bacterial Growth Suppression in a Staphylococcus epidermidis Model: An In Vitro Study. Editorial.
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