Investigation of Ethidium Bromide Interaction with Yersinia Bacterial Cells in Lyophilized/Rehydrated Preparations
https://doi.org/10.21055/0370-1069-2012-1(111)-54-56
Abstract
Investigated was ethidium bromide interaction with the cells of avirulent Y. pestis and Y. pseudotuberculosis strains in lyophilized/rehydrated preparations. The population heterogeneity regarding particle size and concentration of nucleic acid was identified by flow cytofluorimetry and registration of total fluorescence intensity. Shown was the correlation between concentration of colony-forming units (CFU) and fluorescence increase after supplementing suspension with cetyltrimethylammoniumbromide. This approach was suggested as the express method to estimate viable cell count in lyophilized/rehydrated preparations and to control the process of preparing and storage of collection strains, dry live vaccine and probiotic preparations of gramnegative bacteria.
Keywords
616.91/98
About the Authors
V. A. Govorunova
State Research Center for Applied Microbiology and Biotechnology
Russian Federation
Russian Federation
I. G. Govorunov
State Research Center for Applied Microbiology and Biotechnology
Russian Federation
Russian Federation
V. V. Firstova
State Research Center for Applied Microbiology and Biotechnology
Russian Federation
Russian Federation
E. V. Zyrina
State Research Center for Applied Microbiology and Biotechnology
Russian Federation
Russian Federation
References
1. Govorunov I.G., Kosarev N.B., Evtodienko Yu.V., Puchkov E.O. [Permeability of the membranes of the Escherichia coli envelope for ethidium bromide]. Mikrobiologia. 1982; 51(5):731–4.
2. Irkhin A.I., Kondrashenko T.N., Puchkov E.O. [Sensitivity of Escherichia coli cell membranes to various classes of detergents]. Mikrobiologia. 1989; 58(2):217–21.
3. Kosarev N.V., Puchkov E.O. [Methods for the Measurement of Bacterial Concentration in Suspension]. SU Inventor Certificate 1231077.
4. Puchkov E.O. [Methods for the assessment of the presence and vi- ability of microorganisms]. Biotekhnologia. 1988; 4(1):132-42.
5. Puchkov E.O., Govorunov I.G., Evtodienko Yu.V., KosarevN.B. [Effect of low temperature shock and extracellular ice formation on the ex- ternal and cytoplasmic membranes of Escherichia coli cells]. Mikrobiologia. 1983; 52(1):136–9.
6. Sanitary Regulations “Safety work with microorganisms of patho- genicity (risk) groups I–II” SR 1.3.1285-03. Bull. Norm. Metod. Doc. Gossanepidnadzora. 2003; 3(13):61–144.
7. Tikhonravov V.V., Ageev V.A., Kovtun V.P., Kovtun A.L., Shvedov V.V. [The method for assessment of the number of bacteria in biological prepara- tion]. RF Patent 02117291.
8. Davey H.M., Kell D.B. Flow cytometry and cell sorting of heterogeneous microbial populations: the importance of single-cell analyses. Microbiol. Rev. 1996; 60:641-96.
9. Greenstocka C.L., Ruddocka G.W. Interaction of ethidium bromide with DNA as studied by kinetic spectrophotometry. Chemico-Biological Interactions. 1975; 11(5):441-7.
10. Martins M., Viveiros M., Couto I., Costa S.S., Pacheco T., Fanning S. et. al. Identification of Efflux Pump-mediated Multidrug-resistant Bacteria by the Ethidium Bromide-agar Cartwheel Method. In Vivo. 2011; 25:171-8.
11. Puchkov E., Melkozernov A. Fiuorimetric assessment of Pseudomonas fluorescens viability after freeze-thawing using ethidium bromide. Lett. Appl. Microbiol. 1995; 21:368-72.
12. Viveiros M., Martins A., Paixão L., Rodrigues L., Martins M., Couto I. et al. Demonstration of intrinsic efflux activity of Escherichia coli K-12 AG100 by an automated ethidium bromide method. International Journal of Antimicrobial Agents. 2008; 31:458-62.
Review
For citations:
Govorunova V.A., Govorunov I.G., Firstova V.V., Zyrina E.V. Investigation of Ethidium Bromide Interaction with Yersinia Bacterial Cells in Lyophilized/Rehydrated Preparations. Problems of Particularly Dangerous Infections. 2012;(1(111)):54-56. (In Russ.) https://doi.org/10.21055/0370-1069-2012-1(111)-54-56
Views:
606
Email this article 