Electrophysical analysis of Escherichia coli cell damage caused by silver ions]

The influence of Ag+ (0.5-10 microM) on Escherichia coli K-12 cells was studied by electrophoresis and electro-orientation spectroscopy methods. It was shown that the pH-dependency of the cell electrokinetic potential (phosphate-citrate buffer with ion strength 0.02) practically didn't changed after Ag+ treatment, but in low-conductive media electrophoretical mobility of intact and inactivated by heat (70 degrees, 15 min) cells gradually decreased as the Ag+ concentration increased. It was due to the Ag+ adsorption on the cell surface and could not be used for the definite characterization of the cell damage. The high-frequency decrease in the cell electro-orientation spectrum shifted to the region of lower frequencies, K+ was excreted by cells, slight raise of the medium pH occurred and significant changes of cell osmotic properties were observed as a result of Ag+ action. All these changes showed the disturbance of barrier properties of the cytoplasmic membrane. Besides the damaging action of Ag+ on cell membranes increased with the decrease of pH and decreased after the addition of Mg2+, Ca2+ and Sr2+ in low concentrations.     

Study of the integral toxicity of aqueous media, polluted by petroleum and petroleum products using bacterial tests

A biotest kit was used to assess the integral toxicity level of aquatic medium contamination with petroleum and petroleum-based products. The integral toxicity dynamics was also monitored during biodegradation of petroleum and petroleum-based products by an association of petroleum-degrading strains including Acinetobacter sp., Mycobacterium flavescens, and Rhodococcus sp. The following bacterial tests were used: the bioluminescence (BL) test based on Photobacterium leiognathi; electro-orientation (EO), optoosmotic (OO), and growth test; as well as the reducing activity (RA) test based on the Agrobacterium radiobacter culture. No significant increase in the integral toxicity level of aquatic medium was observed when diesel fuel and kerosene contamination had been subjected to biodegradation. Although express biotests (EO, OO, RA, and BL) detected a pronounced increase in the integral toxicity of aquatic medium, long-term growth biotest revealed no statistically significant increase in the toxicity level.     

Microbiological degradation of asymmetrical dimethylhydrazine--a toxic component of rocket fuel

A possibility of microbiological cleaning of water and soil polluted with asymmetric dimethylhydrazine (ADMH), a highly toxic rocket fuel ingredient (RFI), was studied. Several isolates (bacteria, yeast, and micromycetes) capable of utilizing ADMH as the only source of nitrogen, carbon, and energy were isolated from RFI-polluted tundra soil. Acceleration of RFI biodegradation was achieved using a biosorbent that involved cells of the degrader strain immobilized on granulated activated carbon. Biological testing in Escherichia coli and cereals (wheat and barley) demonstrated that biodegradation significantly decreased the integral toxicity of solutions containing ADMH, suggesting its utility for microbiological cleaning of polluted territories.     

Assessment of the Integral Toxicity of Aquatic Medium Contamination with Oil and Oil Products Using Bacterial Tests

A biotest kit was used to assess the integral toxicity level of aquatic environment contamination with oil and oil products. The integral toxicity dynamics were also monitored during biodegradation of oil and oil products by an association of oil-degrading strains, including Acinetobacter sp., Mycobacterium flavescens, and Rhodococcussp. The following bacterial tests were used: the bioluminescence (BL) test based on Photobacterium leiognathi; electroorientation (EO), optoosmotic (OO), and growth tests; and the reducing activity (RA) test based on an Agrobacterium radiobacter culture. No significant increase in the integral toxicity level of the aquatic medium was observed when diesel fuel and kerosene contamination had been subjected to biodegradation. Although rapid biotests (EO, OO, RA, and BL) detected a pronounced increase in the integral toxicity of the aquatic environment, long-term growth biotests revealed no statistically significant increase in the toxicity level.     

Rapid assay for the assessment of a potential of chemical biocides to microbial destructors of industrial materials

A colorimetric rapid assay for estimating the biocide potential of various chemicals towards metal biocorrosive and petroleum product degrading microbes was developed based on the reducing potential of live microbial cell. A water-soluble organic redox indicator, blue in the oxidized form and pink in the reduced form, was used as an indicator of the reducing potential of microbial cells. Once added to a suspension of vital microbial cells, it was reduced and changed in color. A good correlation between the results of this assay and viability control was obtained by employing surfactants and heavy metal ions.     

Electrophysical analysis of the damage of Escherichia coli outer membrane

The method of electro-orientational spectroscopy was used to study the damaging action of SDS and Triton X-100 on Escherichia coli cells in which the barrier properties of the outer membrane were impaired by treatment with Triton B (10(-2) M Tris-HCl buffer, pH 8.0) and a heat shock (47 degrees C, 15 min). When either SDS (10(-4)-2.10(-4) M) or Triton X-100 (10(-4)-10(-3) M) was added to such cells, the high-frequency region of their electroorientational spectrum was found to undergo considerable changes. The mode of these changes indicated that the barrier properties of cell cytoplasmic membranes were damaged. These changes were not detected in the case of intact cells. Changes in the low-frequency region of the spectra for intact and damaged cells stemmed from the adsorption of these surfactant molecules on the cell surface.     

Influence of heavy metal ions on the electrophysical properties of Anacystis nidulans and Escherichia coli cells]

The influence of heavy metal ions (Ag+, Cu2+, Cd2+, Pb2+, Mn2+, Zn2+, Gd3+, 1 microM-1 mM) on Anacystis nidulans and Escherichia coli cells has been studied by means of electrophoresis and electro-orientation spectroscopy methods. It has been shown that changes of cell electrophoretic mobility (EM) and low-frequency (20 Hz) electro-orientation effect (EOE) observed with the increase of metal cation concentration characterize the adsorption of these ions on surface layers of cell envelopes. The degree and the character of these changes depend on cation valency and the initial value of cell EM. At the same time different changes of EM and EOE as a result of the multivalent cation adsorption allows to conclude that in that case the anisotropy of the cell surface increases. Cell damages were determined by changes in high-frequency EOE of cells which indicated the disturbance of barrier properties of their cytoplasmic membrane. Toxic effects of Ag+, Cu2+, Cd2+ ions on cells of both species and of Pb2+ on E.coli cells were observed. By toxic effects on the cytoplasmic membrane these ions could be placed in the order: for A.nidulans cells--Ag+ greater than Cu2+ greater than Cd2+; for E.coli cells Ag+ greater than Cu2+ greater than Cd2+ greater than Pb2+. Higher toxicity of heavy metals on E.coli cells seems to be connected with the more negative charge of deep layers of the cell surface.     

Rapid Assay for Assessment of the Potential of Chemical Biocides against Microbial Destructors of Industrial Materials

A colorimetric rapid assay for estimating the biocide potential of various chemicals towards metal- biocorrosive and petroleum-product-degrading microbes was developed based on the reducing potential of live microbial cells. A water-soluble organic redox indicator, blue in the oxidized form and pink in the reduced form, was used as an indicator of the reducing potential of microbial cells. Once added to a suspension of vital microbial cells, it was reduced and changed in color. A good correlation between the results of this assay and viability control was obtained by employing surfactants and heavy metal ions.     

Microbiological Degradation of Asymmetric Dimethylhydrazine,a Toxic Rocket Fuel Ingredient

The possibility of microbiological cleaning of water and soil polluted with asymmetric dimethylhydrazine (ADMH), a highly toxic ingredient of rocket fuel (IRF), was studied. Several isolates (bacteria, yeast, and micromycetes) capable of utilizing ADMH as the only source of nitrogen, carbon, and energy were isolated from IRF-polluted tundra soil. Acceleration of IRF biodegradation was achieved using a biosorbent that involved cells of the degrader strain immobilized on granulated activated charcoal. Biological testing in Escherichia coli and cereals (wheat and barley) demonstrated that biodegradation significantly decreased the integral toxicity of solutions containing ADMH, suggesting its utility for microbiological cleaning of polluted territories.     

Effect of surface-active agents on the electrical properties of bacterial cells

The work was aimed at studying the effect of cationic, anionic and non-ionogenic surfactants on the frequency dependence of the electroorientation effect (EOE) and on the electrophoretic mobility (EPM) of rod-like bacteria. The character of concentration dependences was found to differ for EOE and EPM at a low frequency of the electric field (20 to 10(4) Hz). Analysis of EOE changes at a high frequency (4 X 10(5) to 3 X 10(7) Hz) showed that anionic and non-ionogenic surfactants at a concentration up to 10(-3) M did not damage Escherichia coli, a Gram-negative bacterium, in contrast to Bacillus cereus, a Gram-positive bacterium. Cationic surfactants affected the cells of the both species. The optical properties of bacterial cells were found to change under the action of cationic surfactants.