A rapid staining technique for the detection of the initiation of germination of bacterial spores

Aims: We propose to apply the Wirtz-Conklin staining technique to evaluate spore germination. Methods and Results: Spores at different stages of germination were stained with modified spore stain (Wirtz-Conklin) and evaluated for staining properties. Bacillus spores suspended in deionized water, which does not support germination, stained greenish-blue. Spores suspended in germination enhancers that did not form bacilli stained pink, indicating the initiation of germination. Spores suspended in culture media, which promotes bacterial outgrowth, formed bacilli and were also stained pink. Conclusions: Modified spore stain (Wirtz-Conklin) was found to be useful to detect the initiation of spore germination as early as 30 min following incubation in a germination environment. Significance and Impact of the Study: This simple staining procedure is useful in detecting the initiation of germination of bacterial spores.