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A method, based on germination with 50 mm CaCl2 and 40 mm sodium dipicolinate (Na2 DPA), was developed for the determination of total viable counts of bacterial spores requiring heat activation. Incorporation of these germinating agents into Tryptone Glucose Extract Agar permitted plate-count enumeration of essentially 100% ofBacillus subtilis strain 5230 spores without a heat treatment. Other spore suspensions were surveyed for their response to CaCl2 and Na2 DPA, and for the subsequent removal of the heat-activation requirement for enumeration of maximal populations.

Use of Calcium Dipicolinate for Enumeration of Total Viable Endospore Populations Without Heat Activation