| Dog cafés have grown in popularity but lack standardized hygiene regulations, raising concerns about airborne microbial exposure. A promising disinfection system has been developed, but its effectiveness in indoor spaces shared by humans and animals remains unclear. In this study, we evaluated the efficacy of a portable highpressure spray device (Vi-Killer®) utilizing neutral electrolyzed water containing hypochlorous acid. Bacterial concentrations were measured in air from three dog cafés in South Korea before and after disinfection. ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) were identified and tested for antibiotic resistance. The average airborne bacterial concentration before disinfection was 193.2 CFU/m3, which significantly decreased by 57.3% to 80.9 CFU/m3 after disinfection (p < 0.05). A total of 466 bacterial isolates were obtained, of which 315 (67.6%) were identified. Staphylococcus spp., Bacillus spp., and Micrococcus luteus were dominant with minimal changes in community composition after disinfection. Six (1.3%) isolates were identified as ESKAPE pathogens; two exhibited multidrug resistance. The Vi-Killer® system effectively reduces microbial load in dog cafés and may mitigate the risk of airborne transmission of opportunistic and resistant bacteria. Regular air disinfection in animal–human shared facilities could improve public hygiene strategies. |