Journal
ANAEROBE
Volume 18, Issue 3, Pages 298-304Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.anaerobe.2012.04.007
Keywords
Clostridium difficile; Iron requirements; Iron utilization bioassays; Microtiter growth assay
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Funding
- European Commission [PIEF-GA-2008-220815]
- Scottish Government (Rural and Environment Science and Analytical Services
- RESAS)
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The ability of four Clostridium difficile strains to utilize various exogenous organic and inorganic iron sources for growth under iron-depleted (250 mu M DPP) and iron-limited (75 mu M DPP) conditions was analyzed in liquid broth cultures grown in tubes and in microtiter plates, and data compared with results from a bioassay developed on solid media. The growth profile of C difficile varied depending on the iron source and availability. Addition of FeSO4, FeCl3, Fe citrate and ferritin allowed growth in an iron-depleted environment whereas glycoproteins (iron-saturated and low-iron lactoferrin, apo- and holo-transferrin) and heme proteins (hemoglobin, hematin and hemin) did not. All iron sources, except lactoferrin, were able to restore bacterial growth under iron-limited conditions to varying extents. The results demonstrated that the broth microtiter assay developed here was reproducible, reliable and convenient for high-throughput analysis of the growth of C difficile compared to alternative traditional methods. (C) 2012 Elsevier Ltd. All rights reserved.
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