Response and Formation Mechanism of Highly Antibiotic-Resistant Dormant Subpopulations in Bioaerosol during Aerosolizing from Aquatic Environments

The characteristics and responses of bacteria aerosolized from aquatic to atmospheric environments are poorly understood. In this study, three antibiotic-resistant bacteria (cefotaxime (CTX)-resistant, polymyxin B (MCR)-resistant, and gentamycin (GEN)-resistant Escherichia coli DH5α) were used to explore microbial aerosolization responses and mechanisms. E. coli bioaerosols had improved resistance to aerosolizing stress through carrying antibiotic resistance genes, developing different phenotype distributions, including wild-type, small colony variant (SCV), and viable but nonculturable (VBNC) subpopulations. E. coli DH5α (CTX) bioaerosols showed the highest percentage of VBNCs (15.1%), while those E. coli DH5α (GEN) bioaerosols showed the highest percentage of SCVs (13.4%). A consistent variation of reduced growth and metabolic activity but increased ATP accumulation and ROS content was observed across SCVs of all strains. Notably, the SCVs of GEN-resistant E. coli exhibited the most pronounced ATP accumulation and a significant upregulation of protein synthesis genes. Collectively, oxidative stress responses were activated to defend against stress in E. coli bioaerosols and promoted the formation of dormant subpopulations (SCV and VBNC). However, a divergent mechanistic was observed: up-regulation of cell wall synthesis genes promoted VBNC formation, while up-regulation of protein synthesis genes promoted SCV formation. These findings highlight that antibiotic resistance in aquatic bacteria contributes to the formation of different dormant subpopulations in bioaerosols that pose different risks to ecosystems and human health.