[Electron microscopic detection and in situ characterization of bacterial nanoforms in extreme biotopes].

The morphology, ultrastructure, and quantity of bacterial nanoforms were studied in extreme biotopes: East Siberia permafrost soil (1-3 Ma old), petroleum-containing slimes (35 years old), and biofilms from subsurface oil pipelines. The morphology and ultrastructure of microbial cells in natural biotopes in situ were investigated by high-resolution transmission electron microscopy and various methods of sample preparation: ultrathin sectioning, cell replicas, and cryofractography. It was shown that the biotopes under study contained high numbers of bacterial nanoforms (29-43% of the total number of microorganisms) that could be assigned to ultramicrobacteria due to their size (diameter of < or =0.3 microm and volume of < or =0.014 microm3) and structural characteristics (the presence of the outer and cytoplasmic membranes, nucleoid, and cell wall, as well as their division patterns). Seven different morphostructural types of nanoforms of vegetative cells, as well as nanospores and cyst-like cells were described, potentially representing new species of ultramicrobacteria. In petroleum-containing slimes, a peculiar type of nanocells was discovered, gram-negative cells mostly 0.18-0.20 x 0.20-0.30 microm in size, forming spherical aggregates (microcolonies) of dividing cells in situ. The data obtained promoted the isolation of pure cultures of ultramicrobacteria from petroleum-containing slimes; they resembled the ultramicrobacterium observed in situ in their morphology and ultrastructure.

[Microbiological degradation of asymmetrical dimethylhydrazine--a toxic component of rocket fuel]. / Mikdrobiologicheskaia degradatsiia nesimmetrichnogo dimetilgidrazina--toksichnogo komponenta raketnogo topliva.

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.

[Study of the integral toxicity of aqueous media, polluted by petroleum and petroleum products using bacterial tests]. / Issledovanie integral'noi toksichnosti vodnoi sredy, zagriaznennoi neft'iu i nefteproduktami, s ispol'zovaniem bakterial'nykh testov.

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.

[Plant biotests of soil and water, polluted with petroleum and petroleum products]. / Biotestirovanie pochvy i vody, zagriaznennykh neft'iu i nefteproduktami, s pomoshch'iu rastenii.

Reactiona of higher plants (mustard, oat, rye, salad, dill and barley) and microalgae (Euglena gracilis) on the contamination of soil and water with petroleum and oil products was studied. The germination of seeds was analyzed. The length of sprouts, dry biomass and length of plant roots, as well as the optical density of micro-algal broth culture were determined. Negative effects of soil and water contamination with petroleum and oil products on plant and microalgal parameters examined was shown. After biological destruction of contaminants by an association of destructor strains (Acinetobacter sp., Mycobacterium flavescens and Rhodoccocus sp.), the toxicity of contaminated mediums decreased. The data suggest that the integral toxicity of soil and water contaminated with petroleum and oil products and toxicity change during biodestruction of these pollutants can be analyzed by using plant test organisms.

[Development and testing of "Ekosorb" biosorbent based on association of petroleum-oxidizing bacteria for purifying petroleum-polluted soils]. / Razrabotka i ispytaniia biosorbenta "Ekosorb" na osnove assotsiatsii nefteokisliaiushchikh baketerii dlia ochistki neftezagriaznennykh pochv.

A biosorbent containing an association of oil-oxidizing bacteria as a main constituent was developed, in which Lessorb, a product of moss and wood thermal processing, was used as a carrier. Xeroprotectors preserving the cell viability and oil-oxidizing activity in the biosorbent on drying and after long-term storage were selected. The use of this biosorbent for cleaning oil-polluted sod-podzol soils showed a two-threefold cleanup rate acceleration at different pollution levels (8 and 24 l/m2), especially in the presence of a nitrogen-phosphate fertilizer. The biosorbent increased the populations of certain groups of soil microorganisms and the total soil biological activity.

[Creation and use of a liquid substance based on association of petroleum-oxidizing bacteria]. / Sozdanie i primenenie zhidkogo preparata na osnove assotsiatsii nefteokisliaiushchikh bakterii.

An association of four bacterial strains with high oil-oxidizing and bioemulsifying activities, psychrophilicity, resistance to chemical pollutants, and lack of pathogenicity was selected from a collection of natural oil-oxidizing microorganisms. A new liquid preparation containing stabilizers and preservatives that maintain the cell viability and oil-oxidizing activity during long-term storage was developed. A field experiment in oil-polluted sod-podzol and clay sand soils demonstrated that this preparation accelerated the biodegradation of oil and its individual fractions, especially in the presence of mineral and organic fertilizers. Treatment of oil-polluted soil with this preparation and additives decreased the oil-induced suppression of certain groups of soil microflora.

[Effect of water pollution by oil and oil products on barrier functions of bacterial cell cytoplasmic membranes]. / Vliianie zagriazneniia vodnoi sredy neft'iu i nefteproduktami na bar'ernye svoistva tsitoplazmaticheskikh membran bakterial'nykh kletok.

The effects of oil, diesel fuel, and kerosene on the electroorientational spectra and osmo-optical characteristics of bacterial cells were studied. Electroorientational spectra were found to be affected over the entire frequency range studied; changes in low-frequency (< 100 kHz) electroorientation were related to alterations in the cell surface, and those in high-frequency electroorientation, to the impairment of the barrier function of the plasma membrane. The membranotropic activity of petroleum products was also demonstrated by the osmo-optical method. Of nine bacterial species studied, Pseudomonas fluorescens VKM B-894, P. oleovorans VKM B-1522, and P. stutzeri VKM B-903 were most susceptible to the membranotropic action of kerosene; P. putida VKM B-1292 was the most resistant. Other bacterial strains studied were moderately sensitive.