Membrane filtration Immobilization technique ‐ A simple and novel method for primary isolation and enrichment of bacteriophages

Aim To develop a method for isolation and enrichment of bacteriophages selectively against specific bacteria coupled with membrane filtration technique. Methods and Results Rapid isolation and concentration of host specific bacteriophages was achieved by exposure of the sample suspected to contain bacteriophages to specific host immobilized on a 0.45μm membrane in a membrane filtration unit. The principle behind this method is exploitation of host specific interaction of bacteriophages with their host and maximizing this interaction using classic membrane filtration method. This provides a chance to every bacteriophage in the sample to interact with the specific host on membrane filter under the effect of vacuum with the help of vacuum pump. Specific bacteriophages of the host are retained onto the membrane along with its host cells due to effect of adsorption and these adsorbed bacteriophages (along with their hosts) on filter disc are then amplified and enriched in regular nutritive broth Tryptose Soya Broth (TSB) by incubation. With help of plaque assay method, host specific phages of various bacterial species were isolated, segregated and enriched. Conclusions The phage concentration method coupled with membrane filtration immobilization of host bacteria was able to isolate and enrich the host specific bacteriophages by several folds using less quantity of environmental water sample, or other phage suspensions. Enrichment of phages from single plaques too was achieved. Significance and impact of the study Isolation and detection of host specific bacteriophages from low bacteriophage density water sample in a single step by use of a simple and basic microbiological technique can be achieved. Enrichment of phages from low phage titre suspensions is also achieved very effectively. This article is protected by copyright. All rights reserved.