Background

Chloroform kills vegetative bacterial cells but not spores and thus is a useful treatment for enriching Clostridia, the dominant spore-forming group in the mammalian intestine. Experimental procedures for chloroform treatment of mouse feces exist (Momose et al., 2009; Yano et al., 2015). However, they utilize specialized equipment including an anaerobic chamber. We realized that several brief exposures to oxygen occur during experimental manipulation of intestinal contents in preparation for and after chloroform treatment. Therefore, we reasoned the sufficient recover of murine spore-forming bacteria could be obtained without the use of an anaerobic chamber. Since spore-forming Clostridia are a dominant species in the mammalian intestine, this protocol could potentially be used for isolation of spores from the intestines of other mammalian organisms, including larger rodents, primates, and humans.