Isolation and characterization of a lytic bacteriophage

In recent years, the use of bacteriophages (or ‘phages’) against multidrug-resistant (MDR) bacteria including Pseudomonas aeruginosa has drawn considerable attention, globally. In this work, we report the isolation and detailed characterization of a highly lytic Pseudomonasphage DRL-P1 isolated from wastewater. Under TEM, DRL-P1 appeared as a member of the phage family Myoviridae. DRL-P1 featured rapid adsorption (~ 5 min), short-latency (~ 30 min), and large burst size (~ 100 PFU per infected cell). DRL-P1 can withstand a wide temperature range (4 °C to 40 °C) and pH (5.0 to 10.0) conditions. The 66,243 bp DRL-P1 genome (MN564818) encodes at least 93 ORFs, of which 36 were functionally annotated based on homology with similar phage proteins available in the databases. Comparative analyses of related genomes suggest an independent evolutionary history and discrete taxonomic position of DRL-P1 within genus Pbunavirus.

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FIG: Plaques of Pseudomonasphage DRL-P1 on P. aeruginosa lawn. (a) A clear zone of lysis was observed in spot-test. (b) Clear plaques were observed after double layer agar plating of enriched phages (c) ‘Web-pattern’ plates for preparation of high titer phage lysate for different studies.

No toxin or antibiotic resistance genes was identified. DRL-P1 is tolerant to lyophilization and encapsulation techniques and retained lytic activity even after 18 months of storage. We also demonstrated decontaminating potentials of DRL-P1 in vitro, on an artificially contaminated cover-slip model. To the best of our knowledge, this is the first Pbunavirus to be reported from India. Our study suggests DRL-P1 as a potential candidate for various applications.

Sharma, S., Datta, S., Chatterjee, S. et al. Isolation and characterization of a lytic bacteriophage against Pseudomonas aeruginosaSci Rep 11,19393 (2021). https://doi.org/10.1038/s41598-021-98457-z

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