Application of New Instrumental Methods for Evaluation of the Functional State of Cells of Francisella Tularensis in a Stressful Environment
The structure of the cell surface reflects the functional state of the bacterium as a whole. The presence of specific features of cells affects the biophysical characteristics of their surface: charge, free energy and hydrophobicity. The interaction of bacteria with biomolecules and particles is also related to these surface characteristics. The aim of our work was to study the function of F. tularensis 15 NIIEG cells under cultivation under stress using electro-optical (EO) monitoring. It was shown that under the influence of elevated temperature in combination with peroxide stress, the cell viability indices, as well as the surface of the cell wall, change. At the same time, stress stimulates expression of immunoreactive stress proteins in bacteria. The effect of small doses of antibiotic on the biophysical parameters of F. tularensis cells was studied for a short time – 1 hour. In the experiment, antibiotics used in the treatment of tularemia – streptomycin, kanamycin and ampicillin at a concentration of 0.5 g/l were used. It was found that the greatest damaging effect on the cell in this concentration was by ampicillin. When exposed to kanamycin and streptomycin, changes in the viability of F. tularensis cells were also observed, but to a lesser extent. Thus, the conditions of cultivation F. tularensis affect the vital signs of cells and the level of antigen expression. Electro-optical analysis successfully registers various types of exposure to microbial cells and is a promising method of control in the development of preventive and diagnostic medicaments.
- DunaevaT. N., Emelyanova O. S., May I. N., Myasni- kov Y. A., Olsufiev N. G. , Rudnev G. P., Saveliev R. A., Silchenko V. S. Tularemia. Eds. G. Olsufyev, G. P. Rudnev. Moscow, Medgiz Publ., 1960. 460 p.
- Alberts B., Johnson A., Lewis J., Raff M., Roberts K., Walter P. Molecular Biology of the Cell. 5th ed. New York, Garland Science, 2007. 1392 p.
- Erisson E., Ericsson M., Tarnvik A., Kuoppa K., Sand- strom G., Sjostedt A. Increased Synthesis of DnaK, GroEL, and GroES Homologs by Francisella tularensis LVS in Response to Heat and Hydrogen Peroxide. Infection and Immunity, 1994, Jan, pp. 178–183.
- Laemmli U. K. Cleavage of Structural Proteins During the Assembly of the Head of Bacteriophage T4. Nature, 1970, vol. 277, pp. 680–685.
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA, 1979, vol. 76, iss. 9, pp. 4350–4354.