Combined Biocidal Effect of Gaseous Ozone and Citric Acid on Acinetobacter baumannii Biofilm Formed on Ceramic Tiles and Polystyrene as a Novel Approach for Infection Prevention and Control

Acinetobacter baumannii is a prominent emerging pathogen responsible for a variety of hospital-acquired infections. It can contaminate inanimate surfaces and survive in harsh environmental conditions for prolonged periods of time in the form of biofilm. Biofilm is difficult to remove with only one method of disinfection, so combined disinfection methods and biocidal active substances are needed for biofilm eradication. Additionally, having in mind ecological demands, legislators are more prone using fewer toxic substances for disinfection that produce less solid waste and hazardous disinfection byproducts. Gaseous ozone and citric acid are natural biocidal compounds, and the purpose of this study was to determine their combined biocidal effects on A. baumannii biofilm formed on ceramics and polystyrene. Twenty-four-hour A. baumannii biofilm formed on ceramic tiles and polystyrene was exposed to different combinations of disinfection protocols with 25 ppm of gaseous ozone for 1 h exposure time and 15% citric acid for 10 min exposure. The total number of bacteria was counted afterwards and expressed as CFU/cm2. The determined disinfection protocols of A. baumannii biofilm with combined citric acid and gaseous ozone caused reduction of 2.8 to 5.89 log10 CFU (99.99% inhibition rate) of total viable bacteria for each method, with the citric acid–ozone–citric acid disinfection protocol being most successful in eradication of viable bacteria on both ceramics and polystyrene. In conclusion, gaseous ozone and citric acid showed good combined biocidal effects on A. baumannii biofilm and successfully reduced early A. baumannii biofilm from ceramic and polystyrene surfaces. The given combination of active substances can be a good option for eco-friendly disinfection of hospital inanimate surfaces from A. baumannii biofilm contamination with prior mechanical cleaning