A disinfectant was created for aerosol disinfection of premises in the presence of poultry, which will help reduce microbial contamination of premises, increase survival, weight of poultry and economic efficiency of meat production in general. The preparation based on essential oils can be used for disinfection in the presence of poultry and at the same time exhibits a therapeutic and prophylactic effect on respiratory infections. This disinfectant has a colloidal solution of silver (Ag), benzalkonium chloride and essential oils of thyme, fir and eucalyptus. The preparation based on essential oils contains (per 100 g): benzalkonium chloride – 16.0 g; thyme oil – up to 2.0 g; eucalyptus oil – up to 2.0 g; fir oil – up to 2.0 g; colloidal solution of silver (Ag) – 20–30 mg; distilled water - up to 100 cm3. Aerosol sanitation of indoor air was carried out with 0.3% solution of preparation in the period before housing poultry and once a day from the 20th to the 35th day of growing broilers with aerosol cold mist generator Dyna-Fog Tornado (model 2897, construction type – ULV-electric spray gnerator, manufacturer – Curtis Dyna-Fog, Ltd., USA) at a dose of 50.0 cm3 per 1 m3 at an exposure of 60 minutes. The size of the aerosol particles is 20 μm. On days 1, 4, 8, 11, 15, 28, 37, and 42, the chickens were weighed, and the blood was taken for examination. Blood was examined to study the number of red blood cells, hemoglobin content, the bactericidal activity of blood serum, phagocyte activity of leukocytes, lysozyme activity of blood serum. According to the results of the research, the technological modes of air disinfection of poultry premises in the presence of broiler chickens were substantiated during the use of preparation, which contains nanoparticles (NP) of silver, benzalkonium chloride and essential oils. The optimal mode of aerosol treatment of poultry houses using a 0.3% solution preparation based on essential oils is 50 mL/m3 of a room with a 60-minute exposure. The use of air disinfection in the presence of chickens during broiler rearing and one treatment per day from 20 to 35 days of the chickens’ growth reduced the microbial pollution of indoor air. Thus, the concentration of microbial cells in the room where the chickens were kept was 230.2 ± 15.6 thousand microbial cell/m3. Sixty minutes after disinfection, the concentration decreased to 1.4 ± 0.4 thousand microbial cell/m3. In addition, the bodyweight of chickens at 6 weeks increased by 449.4 ± 16.3 g (15.9%) compared with the controls. The method and mode of air treatment did not adversely affect the development of the internal organs of the poultry and their physiological state, which is confirmed by studies of the morphological parameters of the chicken blood. The data obtained indicate a positive effect of the developed methods and modes of aerosol air treatment with the preparation based on essential oils on the growth and development of broilers.