DOI: 10.36871 / vet.san.hyg.ecol.201902011
UDC 619: 579

Authors

KONONENKO A. B., BANNIKOVA D.A., BRITOVA S.V., PAVLOVA I.B., NABIULLINA D.N., SAVINOVA E.P.
Institute of Veterinary Sanitation, Hygiene and Ecology - BRANCH of FSBI Federal Research Center of the Russian Academy of Science and Technology

Abstract

Purpose of work: to monitor the formation of biofilms of opportunistic and pathogenic microorganisms. We used cultures of the genera Salmonella, Escherichia, Yersinia, Proteus, Citrobacter, Enterobacter, Providencia, Morganella, Klebsiella, Cronobacter, Pseudomonas, Bacillus, Staphylococcus. Materials and methods. The studied microorganisms were cultured in polystyrene 96-well plates. For this purpose, a daily culture of microorganisms was introduced into the wells with meat-peptone broth, having previously established a concentration of 104 mc / ml, and incubated for 24 ... 96 hours at a temperature of 37 ° C. Then the medium with plankton cells was removed from the wells. 200 μl of a filtered 0,1% crystalline violet solution were added to the plate wells and the plates were held for 10-15 minutes at room temperature. The dye was then removed from the wells. Unbound dye was thoroughly washed with saline or distilled water. The plates were turned onto filter paper and dried. The presence and density of the biomatrix (biofilm) was determined visually by the intensity of staining the surface of the tablet. Then, to extract paint from the film, 200 μl of 96% ethanol were added to the wells and the optical density was measured on a KFK-3KM spectrophotometer at a wavelength of 590 nm. Research results. The results of the experiments made it possible to argue that within 48 hours of cultivation, microorganisms form a mature biofilm, which can serve as a model for studying the biofilm formation process. The biofilm of microorganisms of different taxonomic groups varies in density. In addition, even bacteria belonging to the same genus, under the same conditions, can form a biofilm, the density of which varies by 30 ... 60%.

Keywords

microorganisms, opportunistic bacteria, bacterial biofilms, optical density.