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Published August 15, 2009 | public
Journal Article

Rapid endospore viability assay of Clostridium sporogenes spores

Abstract

A rapid Endospore Viability Assay (EVA), previously developed for Bacillus spores, was modified for enumeration of germinable Clostridium sporogenes spores. The EVA is based on the detection of dipicolinic acid (DPA), which is released during stage I germination and quantified by terbium (III) ion Tb-DPA luminescence. Germination of C. sporogenes spores in aqueous suspension was induced by L-alanine and NaHCO_3 addition, and germinable endospore numbers were determined by reference to a standard curve. Determination of the fractions of germinable C. sporogenes spores by EVA and phase-contrast microscopy yielded comparable results of 54.0% ± 2.9% and 59.3% ± 2.6%, respectively, while only 32.3% ± 5.3% of spores produced colonies on reinforced clostridial medium (RCM). Rates of germination were measured as a function of temperature (30 °C–60 °C) using EVA, yielding a linear relationship between the square root of the rate constant and inverse temperature.

Additional Information

Copyright © 2009 Elsevier. Received 12 September 2008; revised 20 April 2009; accepted 21 April 2009. Available online 4 May 2009. The authors would like to thank Dr. Stephanie A. Connon for editing and Dr. Christina N. Dock for insightful discussions. The research described in this paper was carried out at the Jet Propulsion Laboratory, California Institute of Technology, under a contract with the National Aeronautics and Space Administration. Supplementary data associated with this article can be found, in the online version, at doi:10.1016/j.ijfoodmicro.2009.04.024.

Additional details

Created:
August 21, 2023
Modified:
October 19, 2023