Isolation and Characterization of Pseudomonas aeruginosa Phages with a Broad Host Spectrum from Hospital Sewage Systems and Their Therapeutic Effect in a Mouse Model

Fangfang Dai Department of Clinical Laboratory, Beijing Youan Hospital, Capital Medical University, Beijing, People’s Republic of China;

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Gengxia Yang General Surgical Center, Beijing Youan Hospital, Capital Medical University, Beijing, People’s Republic of China;

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Jinli Lou Department of Clinical Laboratory, Beijing Youan Hospital, Capital Medical University, Beijing, People’s Republic of China;

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Xiuying Zhao Department of Clinical Laboratory, Beijing Tsinghua Changgung Hospital, School of Clinical Medicine, Tsinghua University, Beijing, People’s Republic of China

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Ming Chen Department of Clinical Laboratory, Beijing Youan Hospital, Capital Medical University, Beijing, People’s Republic of China;

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Guizhen Sun Department of Clinical Laboratory, Beijing Youan Hospital, Capital Medical University, Beijing, People’s Republic of China;

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Yanhua Yu Department of Clinical Laboratory, Beijing Youan Hospital, Capital Medical University, Beijing, People’s Republic of China;

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ABSTRACT.

This study aimed to isolate and characterize phages as an alternative treatment of multidrug- or pan-drug-resistant Pseudomonas aeruginosa. Phage titers and bacterial densities correlated, with the phages disappearing after bacteria were eliminated. We isolated phages in filtered sewage water by a double-layered agar spot test. Fifty-eight P. aeruginosa strains were used to screen the host spectrum of the 14 phages isolated. Random amplification of polymorphic DNA-typing polymerase chain reaction was used to analyze the genomic homologies of the 58 host bacteria strains and four phages with a broad host spectrum. Transmission electron microscopy was used to observe the morphology of the four phages with a broad host spectrum. Mice with intraabdominal P. aeruginosa infection were used as an in vivo animal model to investigate the therapeutic effect of the selected phage. Four virulent phages with a broad host spectrum specific to P. aeruginosa strains were isolated. They were all double-stranded DNA viruses and belonged to four different genotypes. The test curve showed that phage I had the highest adsorption rate, the shortest latent period, and the largest burst size. The infected mouse model indicated that small doses of phage I could prevent the death of infected mice. Phage titers and bacterial densities correlated, with phages disappearing after bacteria were eliminated. Phage I was the most effective and promising treatment of drug-resistant P. aeruginosa.

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Author Notes

Address correspondence to Jinli Lou, Department of Clinical Laboratory, Beijing Youan Hospital, Capital Medical University, Beijing 100069, People’s Republic of China, E-mail: loujinli@163.com or Xiuying Zhao, Department of Clinical Laboratory, Beijing Tsinghua Changgung Hospital, School of Clinical Medicine, Tsinghua University, Beijing 102218, People’s Republic of China, E-mail: zxya00340@btch.edu.cn or Yanhua Yu, Department of Clinical Laboratory, Beijing Youan Hospital, Capital Medical University, Beijing 100069, People’s Republic of China, E-mail: yyhs2005186@163.com

These authors contributed equally to this work.

Financial support: This study was supported by the China Primary Health Care Foundation-Youan Foundation of Liver Disease and AIDS and the Key Medical Professional Development Plan of the Beijing Hospital Authority (Grant no. ZYLX202124) and the Special Foundation for National Science and Technology Basic Research Program of China (Grant no. 2019FY101200).

Authors’ addresses: Fangfang Dai, Jinli Lou, Ming Chen, Guizhen Sun, and Yanhua Yu, Department of Clinical Laboratory, Beijing Youan Hospital, Capital Medical University, Beijing, People’s Republic of China, E-mails: daifangfang@126.com, loujinli@163.com, chenming@126.com, sunguizhen@126.com, and yyhs2005186@163.com. Gengxia Yang, General Surgical Center, Beijing Youan Hospital, Capital Medical University, Beijing, People’s Republic of China, E-mail: yanggengxia@126.com. Xiuying Zhao, Department of Clinical Laboratory, Beijing Tsinghua Changgung Hospital, School of Clinical Medicine, Tsinghua University, Beijing, People’s Republic of China, E-mail: zxya00340@btch.edu.cn.

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