Related Experiment Video
Updated: Sep 22, 2025
Time-Lapse Epifluorescence Microscopy Imaging of Pseudomonas aeruginosa and Staphylococcus aureus Heterogeneous Phenotypes
Published on: February 14, 2025
Isolation and Characterization of Lytic Bacteriophage Against Multi-drug Resistant Pseudomonas aeruginosa
Archana Maharjan1, Roshan Nepal1, Gunaraj Dhungana1
1Central Department of Biotechnology, Institute of Science and Technology, Tribhuvan University, Kirtipur, Nepal.
Background:
Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen frequently causing healthcare-associated infections. The apocalyptic rise of antimicrobial resistance has rekindled interest in age-old phage therapy that uses phages (viruses that infect bacteria) to kill the targeted pathogenic bacteria. Because of its specificity, phages are often considered as potential personalized therapeutic candidate for treating bacterial infections.
Methods:
In this study, we isolated and purified lytic phages against multi-drug resistant P. aeruginosa using soft agar overlay technique. Phage characteristics like thermal and pH stability, latent period and burst size were determined using one-step growth assay while multiple host range spectrum was determined by spot assay. The phages were further characterized using protein profiling.
Results:
Three Pseudomonas phages (øCDBT-PA31, øCDBT-PA56 and øCDBT-PA58) were isolated from the holy rivers of Kathmandu valley. Among 3 phages, øCDBT-PA31 demonstrated multiple host range and could lyse multi-drug resistant strain of P. aeruginosa. Further, øCDBT-PA31 showed latent period of 30 minutes with corresponding burst sizes of 423-525 PFU/cell. Interestingly, øCDBT-PA31 also tolerated a wide range of adverse conditions, such as high temperature (50°C) and pH 3-11. Further, protein profiling revealed that øCDBT-PA31 has 4 and øCDBT-PA11 had 3 distinct bands in the gradient gel ranging from approximately 3.5-29 kilodaltons (kDa) suggesting them to be morphologically distinct from each other.
Conclusions:
As multi-drug resistant bacteria are emerging as a global problem, lytic phages can be an alternative treatment strategy when all available antibiotics fail.
Insights
Lytic phages offer a promising alternative to antibiotics for combating multi-drug resistant Pseudomonas aeruginosa. Researchers isolated phage øCDBT-PA31, effective against resistant strains and stable under harsh conditions.
Area of Science:
- Microbiology
- Virology
- Biotechnology
Background:
- Pseudomonas aeruginosa is a significant cause of healthcare-associated infections.
- Rising antimicrobial resistance necessitates novel therapeutic strategies.
- Phage therapy, using viruses to target bacteria, is a potential personalized treatment.
Purpose of the Study:
- To isolate and characterize lytic phages effective against multi-drug resistant Pseudomonas aeruginosa.
- To evaluate the stability and host range of isolated phages.
- To explore phage therapy as an alternative to antibiotics.
Main Methods:
- Isolation of lytic phages using the soft agar overlay technique.
- Determination of phage characteristics (thermal/pH stability, latent period, burst size) via one-step growth assay.
- Host range determination by spot assay and protein profiling for characterization.
Main Results:
- Three Pseudomonas phages (øCDBT-PA31, øCDBT-PA56, øCDBT-PA58) were isolated.
- Phage øCDBT-PA31 exhibited a broad host range, lysing multi-drug resistant P. aeruginosa.
- øCDBT-PA31 demonstrated stability across a wide temperature (50°C) and pH (3-11) range, with a latent period of 30 minutes and burst size of 423-525 PFU/cell.
Conclusions:
- Lytic phages represent a viable alternative treatment for multi-drug resistant bacterial infections.
- Phage therapy provides a potential solution when conventional antibiotics are ineffective.
- Further research into specific phages like øCDBT-PA31 is warranted for clinical application.
More Related Videos
Related Concept Videos
01:30Lytic Cycle of Bacteriophages
01:28Gene Regulation in Microbial Communities: Quorum Sensing