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Isolation and Characterization of Pseudomonas aeruginosa Phages with a Broad Host Spectrum from Hospital Sewage
Fangfang Dai1, Gengxia Yang2, Jinli Lou1
1Department of Clinical Laboratory, Beijing Youan Hospital, Capital Medical University, Beijing, People's Republic of China.
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.
Insights
Bacteriophages offer a promising alternative for treating drug-resistant Pseudomonas aeruginosa infections. Phage I demonstrated significant efficacy in an animal model, effectively reducing bacterial load and preventing mortality.
Area of Science:
- Microbiology
- Virology
- Infectious Diseases
- Bacteriophage Therapy
Background:
- Rising antimicrobial resistance in Pseudomonas aeruginosa poses a significant clinical challenge.
- Alternative therapeutic strategies are urgently needed to combat multidrug- and pan-drug-resistant bacterial infections.
Purpose of the Study:
- To isolate and characterize bacteriophages (phages) for treating drug-resistant Pseudomonas aeruginosa.
- To evaluate the therapeutic potential of isolated phages, particularly phage I, in an in vivo model.
Main Methods:
- Phage isolation from filtered sewage water using a double-layered agar spot test.
- Host spectrum screening of 14 isolated phages against 58 Pseudomonas aeruginosa strains.
- Genomic analysis (Random Amplification of Polymorphic DNA-typing PCR) and Transmission Electron Microscopy (TEM) for phage characterization.
- In vivo efficacy assessment using a mouse model of intraabdominal Pseudomonas aeruginosa infection.
Main Results:
- Four virulent bacteriophages with broad host spectrum against Pseudomonas aeruginosa were isolated.
- These phages were identified as double-stranded DNA viruses belonging to four distinct genotypes.
- Phage I exhibited superior characteristics, including high adsorption rate, short latent period, and large burst size.
- In vivo studies showed that phage I effectively prevented mortality in infected mice, with phage presence correlating to bacterial clearance.
Conclusions:
- Bacteriophage therapy presents a viable alternative for treating multidrug-resistant Pseudomonas aeruginosa.
- Phage I demonstrated significant therapeutic potential and efficacy in preclinical models.
- Further development of phage I as a treatment for drug-resistant Pseudomonas aeruginosa infections is warranted.
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