Temperature and pH stability

The temperature sensitivity of the phage was studied by incubating phage at different temperatures from 4 °C, 16 °C, 25 °C, 37 °C, 45 °C, 65 °C and 95 °C for 1 h⁸⁶. Post incubation, phage were made to interact with the host and and the phage–host mixture was added to 5 ml of soft agar (0.7% Luria Bertani Agar) and overlaid on Nutrient Agar plates. The plates were allowed to solidify, and were incubated at 37 °C for 18–24 h subsequently, plaque titers were determined. Similarly, pH sensitivity of the phage was analysed by incubating the phage at different pH 3.0, 5.0, 7.0, 9.0 and 11.0 for 1 h. After incubation, the phage lysates were made to adsorb with the host and plated by agar overlay method as mentioned above to determine phage titers. After 18–24 h of incubation, plaque titers were determined and were expressed as PFU/ml³⁴. All experiments were performed in triplicates and represented as the percent survival rate.