Size Distribution and Pathogenic Potential of Culturable Airborne <i>Clostridium</i> spp. in a Suburb of Toyama City, Japan

Clostridium spp. are anaerobic, Gram-positive, spore-forming bacteria comprising more than 150 species, some of which are important pathogens of humans and animals. Members of this genus have been isolated from a number of environments, but are rarely found in the atmosphere. In the present study, we exami-ned culturable airborne Clostridium spp. and clarified their pathogenicity. We obtained 19 culturable Clostridium isolates from size-fractionated samples collected at a suburban site in Toyama, central Japan. Culturable Clostridium spp. were detected in particles larger than 1.1‍ ‍μm, and the size distribution peaked at 2.1-3.3‍ ‍μm, corresponding to the spore size of Clostridium spp. More Clostridium spp. were detected in coarse particles >2.1‍ ‍μm not only by culture methods, but also by 16S rRNA gene amplicon sequencing. Whole-genome sequencing (WGS) identified seven Clostridium species, among which Clostridium perfringens was predominant. Moreover, WGS revealed that C. perfringens isolates harbored many virulence and antibiotic resistance genes with the potential to cause gas gangrene. The detection and characterization of potential airborne pathogens are crucial for preventing the spread of diseases caused by these pathogens. To the best of our knowledge, this is the first study to demonstrate that anaerobic Clostridium spp. may be transported under aerobic conditions in the atmosphere and pose potential risks to human health.