Isolation and characterisation of six novel mycobacteriophages and investigation of their antimicrobial potential in milk

Using Mycobacterium smegmatis as a rapidly growing host, six mycobacteriophages were isolated from a variety of environmental sources and characterised by restriction endonuclease digestion. All six phages had long, non-contractile tails and isometric heads characteristic of Siphoviridae and were relatively heat-stable up to 72 °C and generally retained infectivity at pH4–pH10 for 60 min. Each individual phage, when tested against M. smegmatis in both growth media and reconstituted milk, typically reduced host numbers by up to 7 logs. When all 6 phages were combined as a cocktail in milk, a strong bactericidal effect on M. smegmatis was observed over 96 h, evidenced by a 9-log reduction in host cell numbers. These results achieved against M. smegmatis, indicate that mycophages may have a potential application as anti-mycobacterial agents when used against more virulent pathogens such as Mycobacterium avium subsp. paratuberculosis, which has been detected in both raw and pasteurised milk.