Influence of some environmental variables and addition of r-lysozyme on efficacy of Vibrio harveyi phage for therapy

Bacteriophage therapy is a viable proposition for controlling luminous vibriosis caused by Vibrio harveyi in shrimp aquaculture. However, environmental factors influence the growth and activity of phage and affect its efficiency in controlling bacterial diseases. An essential problem in the use of vibrio phage as a therapeutic agent was the development of resistance to phage attachment, rendering them resistant to the lytic action of phage. This problem could be overcome by applying a cocktail of phages. This study aimed to evaluate the effect of salinity and pH on the phage activity and also to study the role of recombinant shrimp lysozyme on the performance of the V. harveyi phage. Out of three different levels of salinity (20, 25 and 30 ppt) and pH (6, 7 and 8) tested, optimum phage activity was observed at a salinity of 25 ppt and at neutral pH. Application of recombinant shrimp lysozyme in combination with V. harveyi phage significantly improved the activity of phage in in vitro assay as well as in microcosm study using seawater. The application of phage along with lysozyme can be a useful approach to overcome the inability of phage to enter the bacteria and thus eliminate or reduce fish/shrimp pathogenic bacteria in aquaculture.