Department disciplines
Scientific directions of the department:
1. Formation of highly productive pharmaceutical screening methods of new drugs based on bacterial bioluminescence.
2. Study of biological properties of nanoparticles
3. Synthesis, physical, chemical and biological properties in a series of condensed azaheterocyclic compounds.
The department has the capacity to conduct research in the field of:
Polyacrylamide gel electrophoresis;
Conductometry;
Spectrophotometry in the visible and ultraviolet regions;
Bio-and chemiluminescence;
Bioassay using luminescent bacteria;
Analytical high performance liquid chromatography (HPLC);
The department has its own collection of luminous bacteria isolated from the Black and Azov Seas, which includes 21 strains:
Two strains of Vibrio fischeri F1 and Photobacterium phosphoreum Sq3 were deposited at the Ukrainian Collection of Microorganisms (Institute of microbiology and virusology NAS of Ukraine) and awarded certificates of deposit: Vibrio fіscheri IMB B-7070, Photobacterium phosphoreum IMB B-7071, June 27, 2002.
Use of luminescent bacteria for bioassay
For more than 20 years luminous bacteria are used for analytical purposes. It happened due to a number of fundamental advantages they possess. Photobacteria are alive independent sources of visible radiation, that, under certain conditions, can remain stable for a long time. The level of bioluminescence in vivo reflects the total metabolic activity of cells depending on their amount. A great number of physical, chemical and biological factors that influence the growth and cell respiration, synthesis of proteins and lipids, the functioning of cell membranes and LUX family genes affect bacterial bioluminescence and can be measured by its registration.
Methods of acute toxicity determination: 0.8 ml of the test solution, 1 - 3% of solution NaCl, 100 mkl of buffer and 100 mkl of a suspension of luminescent bacteria are mixed in the luminometer cuvette. Under a certain temperatures the change in the intensity of bioluminescence for 30-120 minutes is recorded. The level of toxicity is expressed as the concentration of a substance (for solutions of individual compounds) or degree of dilution (for solutions of unknown composition), causing 20 %, 50 % or 80 % decrease in bacterial luminescence intensity relative to the control.
Methods of the chronic toxicity determination: liquid medium for photobacteria is diluted 10 times with a solution of sterile 2.5% NaCl. Overnight culture of luminescent bacteria in a volume ratio of 1:100 (bacteria : fresh medium) was inoculated to the solution. The resulting bacterial suspension was dispensed into 24 well plates (1 ml per well) together with the test solution. The plate was placed in an incubator for 16 - 18 hours at the optimal temperature, followed by analysis of the level of bioluminescence produced in each well. Chronic toxicity was evaluated by the EC50 - effective concentration which reduces bioluminescence by 50 % (20 %, 80 % ) compared with the control. (bacteria with no toxin) or minimal concentration effect which causes 20 % decrease in the level of bioluminescence as compared with the control (LEC - Lowest Effective Concentration).
