Methods to control microbial contamination in confined livestock facilities are important to the health of both animals and workers. In addition, bacterial contamination is also a food safety issue. The most common disinfection technique employed in livestock facilities is the application of oxidizing agents (e.g., potassium peroxymonosulphate, chlorine, hydrogen peroxide, ozone). However, these techniques are associated with a number of limitations (e.g., toxicity, high cost, corrosiveness). Recently, engineered water nanostructures ( EWNS ) generated using an electrospray system was found effective in inactivating foodborne bacteria. Thus, this study investigated the efficacy of EWNS generated using a laboratory-scale electrospray system in inactivating bacteria found in poultry facilities. The effects of various operating conditions (distance between the injector and grounded electrode of the electrospray system, applied voltage, liquid pH and conductivity, liquid flow rate, and treatment time) on the efficacy were also assessed. In these various experiments, airborne bacterial samples were collected from a pullet room using tryptic soy agar plates and then exposed to EWNS under varying conditions. After treatment, the plates were incubated at 37°C prior to colony counting. Reductions in bacterial concentrations up to 1.26 logs were obtained. The results indicate that the EWNS generated by the electrospray system can be a potential chemical-free alternative to conventional disinfection methods. Future tests will focus on scaling up the system for larger scale trials.