Evaluation of novel assays to assess the influence of different iron sources on the growth of Clostridium difficile

Ramona C. Cernat; Karen P. Scott

doi:10.1016/j.anaerobe.2012.04.007

Evaluation of novel assays to assess the influence of different iron sources on the growth of Clostridium difficile

Ramona C. Cernat, Karen P. Scott^*

^*Corresponding author for this work

The Rowett Institute of Nutrition and Health

University of Aberdeen

Research output: Contribution to journal › Article › peer-review

21 Citations (Scopus)

Abstract

The ability of four Clostridium difficile strains to utilize various exogenous organic and inorganic iron sources for growth under iron-depleted (250 μM DPP) and iron-limited (75 μM 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 FeSO ₄, FeCl ₃, 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.

Original language	English
Pages (from-to)	298-304
Number of pages	7
Journal	Anaerobe
Volume	18
Issue number	3
Early online date	25 Apr 2012
DOIs	https://doi.org/10.1016/j.anaerobe.2012.04.007
Publication status	Published - Jun 2012

Bibliographical note

Acknowledgements
We thank Professor Ian Poxton at University of Edinburgh for providing C. difficile VPI 10463 and NCTC 11223 strains, Dr. Peter Mullany at University College London for providing C. difficile strain 630 and Professor Harry Flint for his critical review of the manuscript.
This work was supported by The European Commission (grant number PIEF-GA-2008-220815). The Rowett Institute of Nutrition and Health receives support from the Scottish Government (Rural and Environment Science and Analytical Services; RESAS).

Keywords

Clostridium difficile
Iron requirements
Iron utilization bioassays
Microtiter growth assay

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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abstract = "The ability of four Clostridium difficile strains to utilize various exogenous organic and inorganic iron sources for growth under iron-depleted (250 μM DPP) and iron-limited (75 μM 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 FeSO 4, FeCl 3, 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.",

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note = "Acknowledgements We thank Professor Ian Poxton at University of Edinburgh for providing C. difficile VPI 10463 and NCTC 11223 strains, Dr. Peter Mullany at University College London for providing C. difficile strain 630 and Professor Harry Flint for his critical review of the manuscript. This work was supported by The European Commission (grant number PIEF-GA-2008-220815). The Rowett Institute of Nutrition and Health receives support from the Scottish Government (Rural and Environment Science and Analytical Services; RESAS).",

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AB - The ability of four Clostridium difficile strains to utilize various exogenous organic and inorganic iron sources for growth under iron-depleted (250 μM DPP) and iron-limited (75 μM 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 FeSO 4, FeCl 3, 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.

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Evaluation of novel assays to assess the influence of different iron sources on the growth of Clostridium difficile

Abstract

Bibliographical note

Keywords

UN SDGs

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