Production and characterization of pure Clostridium spore suspensions
Article first published online: 15 DEC 2008
© 2008 The Authors. Journal compilation © 2008 The Society for Applied Microbiology
Journal of Applied Microbiology
Volume 106, Issue 1, pages 27–33, January 2009
How to Cite
Yang, W.-W., Crow-Willard, E.N. and Ponce, A. (2009), Production and characterization of pure Clostridium spore suspensions. Journal of Applied Microbiology, 106: 27–33. doi: 10.1111/j.1365-2672.2008.03931.x
- Issue published online: 12 JAN 2009
- Article first published online: 15 DEC 2008
- 2008/0745: received 1 May 2008, revised 19 June 2008 and accepted 24 June 2008
- quorum sensing;
- rapid techniques;
- spores (inc. endospores);
- waste water
Aims: A general protocol was derived for optimizing the production of pure, high concentration Clostridium endospore suspensions.
Methods and Results: Two sporulation methods were developed that yielded high concentrations of notably pure Clostridium sporogenes, C. hungatei and C. GSA-1 (Greenland ice core isolate) spore suspensions (10 ml of 109 spores ml−1 with >99% purity each). Each method was derived by evaluating combinations of three sporulation conditions, including freeze drying of inocula, heat shock treatment of cultures, and subsequent incubation at suboptimal temperatures that yielded the highest percentage of sporulation. Pure spore suspensions were characterized in terms of dipicolinic acid content, culturability, decimal reduction time (D) value for heat inactivation (100°C) and hydrophobicity.
Conclusions: While some Clostridium species produce a high percentage of spores with heat shock treatment and suboptimal temperature incubation, other species require the additional step of freeze drying the inocula to achieve a high percentage of sporulation.
Significance and Impact of the Study: Pure Clostridium spore suspensions are required for investigating species of medical and environmental importance. Defining the conditions for optimal spore production also provides insight into the underlying mechanisms of Clostridium sporulation.