[Desulfovibrio hontreensis sp. nov., a Sulfate-Reducing Bacterium Isolated from Marine Biofoulings at the South Vietnam Coastal Area].

A Desulfovibrio strain physiologically similar to and phylogeneticall related to "D. caledoniensis" SEBR 7250, D. portus MSL79, and D. dechloracetivorans ATCC 700912 (96.9, 95.9, and 95.8% similarity of the 16S rRNA gen sequences, respectively) was isolated from marine biofouling in the coastal zone of the South China Sae (Nha Trang, South Vietnam). The cells of strain ME were gram-negative motile vibrios (0.4-0.6 x 1.3-2 µm) with a single flagellum. The strain grew at 20 to 39 degrees C (growth optimum at 34-37 degrees C), pH 5.8 to 8.5 (pH optimum at 6.8-7.5), and salinity from 0.08 to 1.1 M Na+ (optimum at 0.2-0.3 M Na+). In the presence of sulfate, the strain grew autotrophically with hydrogen or on lactate, formate, pyruvate, fumarate, and malate. Weak growth occurred on succinate, glycerol, and fructose. In the absence of sulfate, the strain was able to ferment pyruvate, malate (weakly), but not lactate. Sulfate, sulfite, thiosulfate, elemental sulfur, and dimethyl sulfoxide were used as electron acceptors. Vitamins and yeast extract were not required for growth. The G+C content was 52.4 mol %. Predominant fatty acids were C18:0 (13.9%), C16:0 (9.6%), iso-C16:0 (9.5%), C18: 1w7 (8.8%), anteiso-C15:0 (8.1%), and iso-C 17:1 (7.2%). The fatty acid composition was close to that of D. dechloracetivorans BO and has some similarity to that of D. portus. Based on its genotypic and phenotypic characteristics, strain ME maybe considered as a new species, for which the name Desulfovibrio hontrensis sp. nov. is proposed.

[Microbial potential for cleaning the oiled iron scale].

The possibility of using microorganisms to clean oiled iron scale of metallurgical production was investigated with the goal of recuperation. A stable microbial association growing on mineral oil as the sole carbon source was isolated from a sample from oiled iron scale taken directly from a metallurgical plant. For microbial cultures isolated from this association, the taxonomic position, as well as their morphological and cultural characteristics, were determined. The microorganisms belonged to the genera Luteimonas, Alcanivorax, Flavobacterium, and Pseudomonas. Microbial associations oxidizing mineral oil were found to contain some microorganisms incapable of its utilization, which stimulated the hydrocarbon-oxidizing microflora. Application of the isolates, as well as of the strains from microbial collections, resulted in a 58% decrease in residual oil content in treated samples of the oiled iron scale.

[Antimicrobic features of phenolic lipids].

The connection between the efficiency of phenolic lipids (PL) and their hydrophobic property (solubility) and hydrophobic property of microorganisms' cell structure is shown. The mixture of amphiphilic di(oxiphenil)-phenil-methanes, which act bacteriostatically under 15 mg/l, possesses maximal efficiency against Staphylococcus aureus. Against Mycobacterium smegmatis with hydrophobic cell wall, hydrophobic 2,4-dialkylocibenzol 70 mg/l was the most effective. Hexylresorcin (HR) stops the development of gram-positive bacteria in concentrations 20-50 mg/l, that of gram-negative bacteria in concentration 65 mg/l, that of M. smegmatis at 70 mg/l, and that of yeast and fungi at 300 mg/l. HR prevails bacteria spores germination in the concentration 25-100 mg/l. The dependence of antibacterial action of isomers and homologues of alkylresorcins on their structure--number, position, and length of alkyl substituents--is studied.

[Effect of a synthetic analogue of bacterial anabiosis autoinducers hexylresorcinol on the stability of membrane structures].

The effect of hexylresorcinol (HR), a chemical analogue of microbial anabiosis autoinducers of the alkylhydroxybenzene (AHB) group, on the stability of biological membranes and monolamellar liposomes formed of egg phosphatidylcholine (ePC) was studied. According to spectrophotometry and electron microscopy studying of HR-loaded liposomes in the presence of a surface-active agent Tween 20, the critical ratio between HR and ePC for liposome preservation was found to be close to equimolar. The trends in HR influence on membrane structural organization and stability confirmed in experiments on liposomes were also reproduced on intact bacterial cells explaining non-species-specific effect of AHBs. The demonstrated high efficiency of AHB biocides may be used in material and equipment protection against biocorrosion.

[Changes in physicochemical properties of proteins, caused by modification with alkylhydroxybenzenes].

Kinetic characteristics of model enzymes and physicochemical properties of globular proteins modified by chemical analogues of low-molecular-weight microbial autoregulators (alkylhydroxybenzenes, AHBs) have been studied. C7 and C12 AHB homologues were used, differing in the length of the alkyl radical and the capacity for weak physicochemical interactions. Both homologues affected the degree of protein swelling, viscosity, and the degree of hydrophobicity. The effects depended on the structure of AHBs, their concentration, and pH of the solution, which likely reflects changes in the charge of the protein globule and its solvate cover. Variations of hydrophobicity indices of AHB-modified enzymes (trypsin and lysozyme) were coupled to changes in the catalytic activity. The values of K(M), measured for the enzymes within both AHB complexes, did not change, whereas V(max) increased (in the case of C7 complexes) or decreased (C12 complexes). Possible molecular mechanisms of changes in the physicochemical and catalytic parameters of enzymatically active proteins, induced by modification with structurally distinct AHBs, are described, with emphasis on targeted regulation of functional activity.

[A microbiological study of an underground gas storage in the process of gas extraction].

The numbers of microorganisms belonging to ecologically significant groups and the rates of terminal microbial processes of sulfate reduction and methanogenesis were determined in the liquid phase of an underground gas storage (UGS) in the period of gas extraction. The total number of microorganisms in water samples from the operation and injection wells reached 2.1 x 10(6) cells/ml. Aerobic organotrophs (including hydrocarbon- and oil-oxidizing ones) and various anaerobic microorganisms (fermenting bacteria, methanogens, acetogens, sulfate-, nitrate-, and iron-reducing bacteria) were constituent parts of the community. The radioisotopic method showed that, in all the UGS units, the terminal stages of organic matter decomposition included sulfate reduction and methanogenesis, with the maximal rate of these processes recorded in the aqueous phase of above-ground technological equipment which the gas enters from the operation wells. A comparative analysis by these parameters of different anaerobic ecotopes, including natural hydrocarbon fields, allows us to assess the rate of these processes in the UGS as high throughout the annual cycle of its operation. The data obtained indicate the existence in the UGS of a bacterial community that is unique in its diversity and metabolic capacities and able to make a certain contribution to the geochemistry of organic and inorganic compounds in the natural and technogenic ecosystem of the UGS and thus influence the industrial gas composition.

[A microbiological study of an underground gas storage in the process of gas injection].

The liquid phase of different units of an underground gas storage (UGS) in the period of gas injection was studied with respect to its hydrochemical composition and characterized microbiologically. The presence of viable aerobic and anaerobic bacteria was revealed in the UGS stratal and associated waters. An important source of microorganisms and biogenic elements in the ecosystem studied is water and various technogenic admixtures contained in trace amounts in the gas entering from the gas main in the period of gas injection into the storage. Owing to this fact, the bacterial functional diversity, number, and activity are maximal in the system of gas treatment and purification and considerably lower in the observation well zone. At the terminal stages, the anaerobic transformation of organic matter in the UGS aqueous media occurs via sulfate reduction and methanogenesis; exceptionally high rates of these processes (up to 4.9 x 10(5) ng S(2-)l(-1) day(-1) and 2.8 x 10(6) nl CH4 l(-1) day(-1), respectively) were recorded for above-ground technological equipment.

[The new facultatively chemolithoautotrophic, moderately halophilic, sulfate-reducing bacterium Desulfovermiculus halophilus gen. nov., sp. nov., isolated from an oil field].

The new mesophilic, chemolithoautotrophic, moderately halophilic, sulfate-reducing bacterium strain 11-6 could grow at a NaCl concentration in the medium of 30-230 g/l, with an optimum at 80-100 g/l. Cells were vibrios motile at the early stages of growth. Lactate, pyruvate, malate, fumarate, succinate, propionate, butyrate, crotonate, ethanol, alanine, formate, and H2 + CO2 were used in sulfate reduction. Butyrate was degraded completely, without acetate accumulation. In butyrate-grown cells, a high activity of CO dehydrogenase was detected. Additional growth factors were not required. Autotrophic growth occurred, in the presence of sulfate, on H2 + CO2 or formate without other electron donors. Fermentation of pyruvate and fumarate was possible in the absence of sulfate. Apart from sulfate, sulfite, thiosulfate, and elemental sulfur were able to serve as electron acceptors. The optimal growth temperature was 37 degrees C; the optimum pH was 7.2. Desulfoviridin was not detected. Menaquinone MK-7 was present. The DNA G+C content was 55.2 mol %. Phylogenetically, the bacterium represented a separate branch within the cluster formed by representatives of the family Desulfohalobiaceae in the subclass Deltaproteobacteria. The bacterium was assigned to a new genus and species, Desulfovermiculus halophilus gen. nov., sp. nov. The type strain is 11-6T (= VKM B-2364), isolated from the highly mineralized formation water of an oil field.

[The properties of hydrocarbon-oxidizing bacteria isolated from the oilfields of Tatarstan, Western Siberia, and Vietnam].

Eleven strains of hydrocarbon-oxidizing bacteria, isolated from oilfields, representing the genera Rhodococcus, Gordonia, Dietzia, and Pseudomonas, were characterized as mesophiles and neutrophiles. Rhodococci were halotolerant microorganisms growing in a media containing up to 15% NaCl. All the strains oxidized n-alkanes of crude oil. An influence of the cultivation temperatures (28 or 45 degrees C) and organic supplements on the degradation of C12-C30 n-alkanes in oxidized oil by two bacterial strains of the genus Pseudomonas was shown. The introduction of acetate, propionate, butyrate, ethanol, and sucrose led mainly to the decreased oxidation of petroleum paraffins. At certain cultivation temperatures, the addition of volatile fatty acid salts increased the content of individual n-alkanes in oxidized vs. crude oil.

[Utilization of H2O2 as the oxygen source by bacteria of the genera Pseudomonas and Rhodococcus]. / Ispol'zovanie H2O2 v kachestve istochnika kisloroda bakteriiami rodov Pseudomonas i Rhodococcus.

The growth of bacteria of the genera Pseudomonas and Rhodococcus in the presence of hydrogen peroxide as the sole source of oxygen was studied. The toxic effect of H2O2 in the concentration range of 100-200 microg/ml was shown to extend the lag phase by 2 to 3 days. Apart from the peroxide toxicity, the bacterial growth was inhibited by the toxic effect of dissolved oxygen in concentrations over 100 microg O2/ml; in the presence of a liquid hydrocarbon phase, this effect was alleviated. Under decreased partial pressure of oxygen in the presence of hydrocarbons (12-15 vol %), the culture growth was initiated at high initial concentrations of H2O2 (300 microg/ml). When hydrogen peroxide concentrations exceeded 320 microg/ml, no growth occurred, no matter how much hydrocarbon was added.

[Dynamics of microbial processes in the stratal waters of the Romashkinskoe oil field]. / Dinamika mikrobnykh protsessov v plastovykh vodakh Romashkinskogo neftianogo mestorozhdeniia.

Dynamics of the microbial processes developing in parallel with the exploitation of the Romashkinskoe oil field (Tatarstan) was studied in two areas differing in the degree of stratal water freshening. Flooding the strata in conjunction with purposeful measures on stratal microflora activation was shown to increase the microbial population density and activate both methanogenesis and sulfate-reduction; the latter process was limited by the low sulfate concentration. Development of anaerobic processes correlated with changes in acetate concentration in the stratal water. High mineralization (over 200 g/l) inhibited the stratal water microflora even if other conditions were favorable. Isotopic analysis of the carbonate carbon showed that the bicarbonate concentration increased in the stratal water due to microbial degradation of oil hydrocarbons and further participation of the biogenic carbon dioxide in dissolution of the carbonate cement of the oil-bearing strata. In strongly desalinated stratal water, the proportion of the newly formed bicarbonate was as high as 80%.

[Microbiological processes in a high-temperature oil field]. / Mikrobiologicheskie protsessy v vysokotemperaturnom neftianom mestorozhdenii.

Thermophilic sulfate-reducing bacteria (SRB) oxidizing lactate, butyrate, and C12-C16 n-alkanes of oil at a temperature of 90 degrees C were isolated from samples of water and oil originating from oil reservoirs of the White Tiger high-temperature oil field (Vietnam). At the same time, no thermophiles were detected in the injected seawater, which contained mesophilic microorganisms and was the site of low-temperature processes of sulfate reduction and methanogenesis. Thermophilic SRB were also found in samples of liquid taken from various engineering reservoirs used for oil storage, treatment, and transportation. These samples also contained mesophilic SRB, methanogens, aerobic oil-oxidizing bacteria, and heterotrophs. Rates of bacterial production of hydrogen sulfide varied from 0.11-2069.63 at 30 degrees C and from 1.18-173.86 at 70 degrees C micrograms S/(1 day); and those of methane production, varied from 58.4-100 629.8 nl CH4/(1 day) (at 30 degrees C). The sulfur isotopic compositions of sulfates contained in reservoir waters and of hydrogen sulfide of the accompanying gas indicate that bacterial sulfate reduction might be effective in the depth of the oil field.

[Halophilic archaebacteria from the Kalamkass oilfield]. / Galfil'nye arkhebakterii neftianogo mestorozhdeniia Kalamkass.

Two strains of halophilic archebacteria, growing in the range from 10 to 25% NaCl, were obtained from the brines of the Kalamkass (Mangyshlak) oilfield. Both strains are extremely halophilic archaebacteria according to their total phenotypic properties. Strain M-11 was identified as Haloferax mediterranei on the basis of the composition of polar lipids and DNA-DNA homology. The composition of polar lipids and 16S rRNA sequences of M-18 strain permitted to include it in Haloferax genus. This strain differs from the affirmed species of Haloferax genus--H. volcanii and H. mediterranei. However, the additional investigations are necessary for its relation to new species.