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Objective. The study objective it to analyze the dynamics of microbial content of burn wounds and to assess the markers of hospital-acquired infection (HAI) in different periods of hospitalization.Methods: 617 microbiological samples from the surface of burn wounds of 515 patients in the period from the first to ninth days after the injury were analyzed. The presence of gramnegative bacteria Pseudomonas aeruginosa and Acinetobacter baumannii, multiresistant microorganisms and microbial associations was determined. Spectrophotometry was used to identify pathogens and plate method was used to assess microbial content of wounds.Results: During treatment, there was change of pathogens associated with duration of stay in the hospital. With an increase in hospitalization, the degree of microbial contamination of wounds, the rate of gram-negative bacteria, multiresistant microorganisms, and microbial associations increased.Conclusions: The greatest severity of all signs of nosocomial infection occurs on the 3–4th day of hospitalization in a burn care facility. This period can be considered ad a start of active contamination of burn wounds with nosocomial flora. To prevent this, it is necessary as early as possible, from 2–3 days after hospitalization to apply active strategy for treating burn wounds and closing burn surfaces.