Author:
Bourdin Gilles,Schmitt Bertrand,Marvin Guy Laure,Germond Jacques-Edouard,Zuber Sophie,Michot Lise,Reuteler Gloria,Brüssow Harald
Abstract
ABSTRACTWe investigated the amplification and purification of phage preparations with respect to titer, contamination level, stability, and technical affordability. Using various production systems (wave bags, stirred-tank reactors, and Erlenmeyer flasks), we obtained peak titers of 109to 1010PFU/ml for T4-like coliphages. Phage lysates could be sterilized through 0.22-μm membrane filters without titer loss. Phages concentrated by differential centrifugation were not contaminated with cellular debris or bacterial proteins, as assessed by electron microscopy and mass spectrometry, respectively. Titer losses occurred by high-speed pelleting of phages but could be decreased by sedimentation through a sucrose cushion. Alternative phage concentration methods are prolonged medium-speed centrifugation, strong anion-exchange chromatography, and ultrafiltration, but the latter still allowed elevated lipopolysaccharide contamination. T4-like phages could not be pasteurized but maintained their infectivity titer in the cold chain. In the presence of 10 mM magnesium ions, phages showed no loss of titer over 1 month at 30°C.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Reference20 articles.
1. High rate of resistance to locally used antibiotics among enteric bacteria from children in Northern Ghana;Djie-Maletz;J. Antimicrob. Chemother.,2008
2. High frequency of antimicrobial drug resistance of diarrheagenic Escherichia coli in infants in Peru;Ochoa;Am. J. Trop. Med. Hyg.,2009
3. Enterotoxigenic Escherichia coli in developing countries: epidemiology, microbiology, clinical features, treatment, and prevention;Qadri;Clin. Microbiol. Rev.,2005
4. Emergence of a new antibiotic resistance mechanism in India, Pakistan, and the UK: a molecular, biological, and epidemiological study;Kumarasamy;Lancet Infect. Dis.,2010
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