Aerosolization Affects Bacillus globigii Vegetative Cell and Spore Behaviors

Antimicrobial resistance (AMR) in bacteria is a critical global health threat, yet the impact of environmental stressors such as aerosolization on resistance remains unclear. We previously showed that aerosolization can induce antibiotic resistance in Escherichia coli MG1655, a gram-negative pathogen simulant. Here, we investigated Bacillus globigii, a surrogate for the gram-positive pathogen Bacillus anthracis, to assess how aerosolization affects bacterial survival and antibiotic resistance. B. globigii vegetative cells and spores were aerosolized under varying conditions and durations (5, 10, 15, 30, and 45 min) into a sterile, airtight chamber and collected using the wetted wall cyclone (WWC) system. Samples were analyzed via antibiotic susceptibility testing, culture-based assay, and quantitative polymerase chain reaction (qPCR). Vegetative cells exhibited the lowest culturability after 5 and 30 min aerosolization, while spores showed reduced culturability at 15-45 min. Both vegetative cells and spores displayed lowest antibiotic susceptibility profiles after 15 min of aerosolization. Our findings suggest that aerosolization duration and bacterial state (vegetative vs. spores) can influence bacterial survival and development of antibiotic resistance. Understanding these dynamics is essential for designing strategies to mitigate the airborne spread of antibiotic-resistant bacteria.