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Membrane filtration immobilization technique—a simple and novel method for primary isolation and enrichment of bacteriophages

IR@CSIR-NEERI

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Title Membrane filtration immobilization technique—a simple
and novel method for primary isolation and enrichment of
bacteriophages
 
Creator Ghugare, G S
Nair, A
Nimkande, V
Sarode, P
Rangari, P
Khairnar, Krishna
 
Subject Microbiology
 
Description To develop a method for the isolation and enrichment of bacteriophages selectively against specific bacteria coupled with a 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 a specific host immobilized on a 0�45 lm membrane in a membrane filtration unit. The principle behind this method is the exploitation
of host-specific interaction of bacteriophages with their host and maximizing this interaction using a classic membrane filtration method. This provides a chance for each bacteriophage in the sample to interact with the specific host on the membrane filter fitted with a vacuum pump. Specific bacteriophages of the host are retained on the membrane along with its host cells due to the effect of adsorption and these adsorbed bacteriophages (along with their hosts) on the filter disc are then amplified and enriched in regular nutritive broth tryptose soya broth by incubation. With the help of the 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 fold using a lower quantity of an environmental water sample, or other phage suspensions. Enrichment of phages from single plaques was also achieved.
Significance and Impact of the Study: The isolation and detection of hostspecific bacteriophages from a low density bacteriophage water sample in a single step by the use of a simple and basic microbiological technique can be achieved. Enrichment of phages from low phage titre suspensions is also achieved very effectively
 
Publisher Wiley
 
Date 2017-02
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://neeri.csircentral.net/641/1/EVC_2016_Journal_of_Applied_Microbiology.pdf
Ghugare, G S and Nair, A and Nimkande, V and Sarode, P and Rangari, P and Khairnar, Krishna (2017) Membrane filtration immobilization technique—a simple and novel method for primary isolation and enrichment of bacteriophages. Journal of Applied Microbiology, 122 (2). pp. 531-539. ISSN 1364-5072, ESSN: 1365-2672
 
Relation http://onlinelibrary.wiley.com/journal/10.1111/(ISSN)1365-2672
http://neeri.csircentral.net/641/