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Sewage as a rich source of phage study against Pseudomonas aeruginosa PAO
Reza Azizian1, Ahmad Nasser1, Hasan Askari1
1Department of Medical Microbiology, Faculty of Medicine, Ilam University of Medical Sciences, Ilam, Iran; Student Research Committee, Ilam University of Medical Sciences, Ilam, Iran.
Abstract:
Pseudomonas aeruginosa is a ubiquitous organism which has emerged as a major public health threat in hospital environments. Overuse of antibiotics has significantly exacerbated the emergence of multi-drug resistant bacteria such as P. aeruginosa. Phages are currently being utilized successfully for aquaculture, agriculture and veterinary applications. The aim of this study was to isolate and characterize of lytic P. aeruginosa phage from sewage of Ilam, Iran. Phage was isolated from sewage that was added to the enrichment along with the host and subsequently filtered. Plaque assay was done by using an overlay method (also called the double agar layer method). Purified plaques were then amplified for characterization. Finally, RAPD-PCR method was conducted for genotyping and Transition electron micrograph (TEM) recruited to determine the morphology and phage family. The phage had high concentration and tremendous effects against a variety of clinical and general laboratory strains (ATCC15693) of P. aeruginosa. Among a set of primers in RAPD panel, only P2 and RAPD5 primers, were useful in differentiating the phages. TEM images revealed that the isolated phages were members of the Siphoviridae family. The phage effectiveness and specificity towards target bacteria and potential to control biofilm formations will be investigate in our further studies.
Insights
Researchers isolated a lytic bacteriophage effective against multi-drug resistant Pseudomonas aeruginosa from sewage. This phage, belonging to the Siphoviridae family, shows potential for combating hospital-acquired infections caused by this resilient pathogen.
Area of Science:
- Microbiology
- Bacteriology
- Virology
Background:
- Pseudomonas aeruginosa is a significant hospital-acquired infection threat.
- Antibiotic overuse has led to multi-drug resistant strains of P. aeruginosa.
- Bacteriophages offer a promising alternative for combating resistant bacteria.
Purpose of the Study:
- To isolate and characterize a lytic bacteriophage targeting P. aeruginosa from sewage in Ilam, Iran.
- To evaluate the phage's efficacy against clinical and laboratory strains of P. aeruginosa.
- To determine the phage's morphology and family classification.
Main Methods:
- Phage isolation from sewage using enrichment and filtration.
- Plaque assay (double agar layer method) for purification and amplification.
- Random Amplified Polymorphic DNA-PCR (RAPD-PCR) for genotyping.
- Transmission Electron Microscopy (TEM) for morphology and family determination.
Main Results:
- A high-concentration lytic phage effective against various P. aeruginosa strains (including ATCC15693) was isolated.
- RAPD-PCR with primers P2 and RAPD5 differentiated the phages.
- TEM analysis identified the isolated phages as members of the Siphoviridae family.
Conclusions:
- The isolated Siphoviridae phage demonstrates significant lytic activity against P. aeruginosa.
- This phage represents a potential therapeutic agent against multi-drug resistant P. aeruginosa infections.
- Further studies will investigate phage efficacy and biofilm control capabilities.
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