Use of flow cytometry for analysis of phage-mediated killing of Enterobacter aerogenes

Verthé, Kristof and Verstraete, Willy (2006) Use of flow cytometry for analysis of phage-mediated killing of Enterobacter aerogenes. Research in Microbiology, 157 7: 613-618. doi:10.1016/j.resmic.2006.02.007


Author Verthé, Kristof
Verstraete, Willy
Title Use of flow cytometry for analysis of phage-mediated killing of Enterobacter aerogenes
Formatted title
Use of flow cytometry for analysis of phage-mediated killing of Enterobacter aerogenes
Journal name Research in Microbiology   Check publisher's open access policy
ISSN 0923-2508
1769-7123
Publication date 2006
Sub-type Article (original research)
DOI 10.1016/j.resmic.2006.02.007
Volume 157
Issue 7
Start page 613
End page 618
Total pages 6
Place of publication Cedex, France
Publisher Elsevier Masson
Language eng
Subject 100204 Environmental Biotechnology Diagnostics (incl. Biosensors)
Formatted abstract
In this study, the use of flow cytometry to analyze phage-mediated killing of Enterobacter aerogenes under varying conditions of temperature and nutrient availability was assessed. Bacteriophage UZ1, specific for an E. aerogenes strain, was applied at a multiplicity of infection (MOI) of 1 and 1000 to a Teflon surface, artificially infected with its host at a level of 4.5 log cells. After incubation for 20 h, bacteriophages were quantified using the soft agar layer method. For the quantification of bacterial cells, plate counting and flow cytometric analysis of live/dead stained cells were performed in parallel. At an MOI of 1, phage treatment was successful only after incubation under nutrient-rich conditions at 37 ◦C: E. aerogenes cells were not detected and a tenfold increase in phage UZ1 was observed. At a MOI of 1000, no E. aerogenes cells could be cultured after incubation at 37 and 4 ◦C. However, flow cytometric analysis revealed that lysis did not occur at 4 ◦C but was achieved during subsequent plate culture. In conclusion, the use of flow cytometry enabled identification of culture-based bias during plate culture. The flow cytometric assay used in this study proved to be rapid, as this culture-independent method does not require lengthy incubation periods post-sampling. The bacteriophagemediated killing of E. aerogenes cells on Teflon surfaces indicated that disinfection of E. aerogenes with bacteriophage UZ1 can be successful when high MOIs are achieved, while at low multiplicities of infection conditions favorable for phage replication are required.
Keyword Bacteriophage
Enterobacter aerogenes
Phage therapy
Surface disinfection
Flow cytometry
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
Advanced Water Management Centre Publications
 
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