Isolation identification and characterization of bacteriophage specific to antibiotics resistant bacteria

This study aimed to isolate and characterize bacteriophage specific to the multidrug-resistant (MDR) Pseudomonas aeruginosa strains. The result found that the bacteriophage named WJ9 (phageWJ9) could be isolated from the wastewater treatment plant of Sanprasitthiprasong Hospital, Ubon Ratchathani Province. The phageWJ9 was highly specific to bacteria MDR Pseudomonas aeruginosa. The strains were isolated from patients who were hospitalized at Sanpasitiprasong Hospital. Bacteriophage properties that were thoroughly investigated in this study include determination of bacteriophage’s host range, stability in ethanol and normal saline solution, bacteriophage genome analysis and bacteriophage morphology. The results showed that phageWJ9 was highly specific to P.aeruginosa at percentage of 70(7-10) of all strains tested. However, other tested bacteria which include Escherichia coli ATCC 25922, Enterobacter sp. Klebsiella pneumonia, Salmonella typhimurium DMSc5784, Shigella dysenteriae MSc2137, Staphylococcus aureus ATCC25923, Staphylococcus epidermidis and Bacillus subtilis were not killed by phageWJ9. Stability in ethanol and normal saline solution demonstrated that phageWJ9 was able to survive after incubating in 0.85% normal saline for 30 minutes but was not tolerated in the ethanol solution. Genome analysis revealed that phageWJ9’s genome consisted of a double-stranded alcohol because it was digested by restriction enzyme HindIII and NcoI and DNase enzyme, but it was not digested by RNase A. Study of phage morphology by transmission electron microscopy revealed that the phage had an icosahedral head (83nm in diameter) and a long contractile tail (200nm in length). Thus, based on phage genome and morphology, phageWJ9 can be categorized in the family Myoviridae of the order Caudovirales. In conclusion, the highly specific bacteriophage to MDR Pseudomonas aeruginosa had good properties and should be considered for further advanced study such as determination of the P.aeruginosa growth inhibition in cell line cukture, animal models, and application for wound healing of P.aeruginosa in patients with infectious diseases.