Annotation. P. aeruginosa, A. baumannii are the causative agents of severe wound infections, ventilator-associated pneumonia, catheter-associated septic conditions and other severe pathological processes. The ability of bacteria to form biofilms in affected tissues plays an important role in the pathogenesis of these diseases. Therefore, the purpose of our work is to study the influence of certain external factors on the ability of bacteria to form biofilms. The ability of bacteria to form biofilms in affected tissues plays an important role in the pathogenesis of these diseases. 5 clinical strains of P. aeruginosa and 5 strains of A. baumannii were used in this research. The ability to form biofilms was studied in 96-well polystyrene plates by the spectrophotometric method, which is based on the ability of the crystal violet dye to bind to the cells and matrix of the biofilm. Statistical processing of the obtained results was carried out using Excel Microsoft Office tables. The effect of the presence of certain sugars and nitrogen-containing compounds in the cultivation medium on the processes of biofilm formation by pseudomonads and acinetobacteria was investigated. As a result of the research, it was established that acinetobacteria show the ability to form films, unlike pseudomonads, under the condition of their cultivation in sterile distilled water. The introduction of certain carbohydrates or urea into the cultivation medium intensified the biofilm-forming properties of pseudomonads to a much greater extent, compared to acinetobacteria. According to the research results, it was established that the most active process of biofilm formation occurs in both types of non-fermenting Gram-negative bacteria when there are animal blood serum proteins in the culture medium. The formation of biofilms is one of the main strategies that increases the survival of bacteria in the environment, as well as in the human body. The ability of bacteria to form biofilms is, among other things, a factor in the pathogenicity and virulence of bacteria. So, in recent years, special attention has been paid to the ability of microorganisms to form biofilms, and to the study of factors affecting this process.