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Phylogenetic analysis of aerobic chemoorganotrophic bacteria of the two extreme regions (Dead Sea and West Antarctic) was performed on the basis of the nucleotide sequences of the 16S rRNA gene. Thermotolerant and halotolerant spore-forming bacteria 7t1 and 7t3 of terrestrial ecosystems Dead Sea identified as Bacillus licheniformis and B. subtilis subsp. subtilis, respectively. Taking into account remote location of thermotolerant strain 6t1 from closely related strains in the cluster Staphylococcus, 6t1 strain can be regarded as Staphylococcus sp. In terrestrial ecosystems, Galindez Island (Antarctic) detected taxonomically diverse psychrotolerant bacteria. From ornithogenic soil were isolated Micrococcus luteus O-1 and Microbacterium trichothecenolyticum O-3. Strains 4r5, 5r5 and 40r5, isolated from grass and lichens, can be referred to the genus Frondihabitans. These strains are taxonomically and ecologically isolated and on the tree diagram form the joint cluster with three isolates Frondihabitans sp., isolated from the lichen Austrian Alps, and psychrotolerant associated with plants F. cladoniiphilus CafT13(T). Isolates from black lichen in the different stationary observation points on the south side of a vertical cliff identified as: Rhodococcus fascians 181n3, Sporosarcina aquimarina O-7, Staphylococcus sp. 0-10. From orange biofilm of fouling on top of the vertical cliff isolated Arthrobacter sp. 28r5g1, from the moss-- Serratia sp. 6r1g. According to the results, Frondihabitans strains most frequently encountered among chemoorganotrophic aerobic bacteria in the Antarctic phytocenoses.

The aerobic chemoorganotrophic bacteria, dominating in soils and phytocenosis of the Antarctic Region, on combination of morphological and biochemical properties belong to several taxons of Bacteria domain. Gram-negative strains 3189, 3415 (fam. Halomonadaceae, Halomonas sp.) and 3088, 3468, 3469 (fam. Moraxellaceae, Psychrobacter sp.) belong to phylum Proteobacteria, to class Gammaproteobacteria. Gram-negative strains 3294 3392 (Rhizobiales, fam. Methylobacteriaceae, Methylobacterium sp.) relate to class Alphaproteobacteria of this phylum. Gram-positive strains 3179, 3275, 3470, 3471 (fam. Microbacteriaceae, Cryobacterium sp.), 3054, 3058, 3411 (fam. Corynebacteriaceae, Corynebacterium sp.) and 3194, 3398 (fam. Micrococcaceae, Micrococcus sp.) relate to phylum Actinobacteria, class Actinobacteria. Thus, the psychrophilic and psychrotolerant Antarctic bacteria (aerobic chemoorganotrophic) isolated from phytocenosis and soils of polar region are characterized by wide taxonomic variety.

Such extreme factors as UV radiation, high temperature and salinity, and also the small amount of accessible water have an influence on microorganisms of coastal ecosystems of the Dead Sea. Resistance to these factors of the microorganisms isolated from ecosystems of this region (vertical steep gorge around the Dead Sea, clay-salt plain and black highly mineralized muds) is studied. Aerobic, chemoorganotrophic, thermotolerant, moderately halophilic bacteria which, according to their morphological and physiological properties, are similar to species Gracilibacillus halotolerans, Salimicrobium album and genus Caryophanon have been isolated from these ecosystems. All strains grew at 0-10% of NaCl in the medium (one strain--at 15% of NaCl), in the range of 30-50 degrees C. Resistance to UV radiation has been revealed in all the investigated bacteria Lethal doses of UV (LD90 and LD99.99) for spore-forming strains of genus Gracilibacillus were 100-170 and 1100-1500 J/m2, respectively; for strain Salimicrobium 6t1 (does not form spores)--70 and 400 J/m2; for the strain lt4 (genus Caryophanon), forming filamentous (or trychomes)--150 and 1400 J/m2. Some strains of genus Gracilibacillus had strong antagonistic effect on conditionally pathogenic test cultures Staphylococcus aureus 209p and Candida albicans UCM Y-690. It is conceivable that resistance of microorganisms of coastal ecosystems of the Dead Sea to extreme factors was generated under the influence of abiotic (physical and chemical) factors typical of this region.

Five strains of the black yeast similar to Exophiala nigra (Nadsoniela nigra), which we have isolated from the Antarctic biotopes, are studied. At cultivation in a periodic operation the maximum level of absolutely dry biomass in five tested strains constituted 3.2-7.8 g/l of medium, melanin pigment yield being 6-9% of absolutely dry mass of cells. Two highly productive strains have been selected. Pigments of the studied black yeast are water-insoluble, however dissolve in alkali and concentrated acids. The maximum absorption of the yeast pigments was in the range of 220 nm. The above-stated properties of pigments of the investigated yeast correspond to the description of melanin fractions of Nadsoniela nigra and some microscopic mushrooms. The water-soluble melanin-pigments have been obtained after the dialysis of alkaline solution of the pigment. UV-spectra and visible absorption spectra of water solution of melanin-pigments are almost identical to those of initial alkaline solutions. It is shown that the studied yeast are resistant to high concentrations of toxic metals (Hg2+, Cu2+, Co2+, Cr(VI) and Ni2+), and introduction of Co2+ into the cultivation medium leads to the increase of pigments synthesis.

Study of expression of cytokine genes in the process of cultivation of healthy donor leukocytes.RNA isolated before, after 3 and 24 hours of cultivation of leukocytes of 15 healthy donors aged 19 - 32 years was the object of the study. Study of the expression of genes of 8 cytokines by the level of their mRNA was performed by using polymerase chain reactionwith reverse transcription.In healthy donors among with individual differences, general regularities were traced: higher levels of IFNalpha and anti-inflammatory cytokine gene expression, as well as predominantly low expression of pro-inflammatory cytokines.In most of the cases in the process of cultivation of leukocytes isolated from blood donors the highest level of gene expression of pro-inflammatory cytokines is noted after 3 hours of cultivation, and expression of genes of anti-inflammatory cytokines remains constant for 24 hours of cultivation.

Resistance to toxic metals ( Hg2+, Cu2+, Ni2+, Co2+, Cr(VI)) of bacteria isolated from rock lichen samples of vertical cliffs located on the biogeographic polygon of Ukrainian Antarctic Station Akademik Vernadsky (island Galindez) is studied. Among the Antarctic rock microorganisms isolated on nonselective medium (without toxic metals), bacteria able to grow at toxic metal concentrations lethal for the majority of microorganisms (Hg2+, Cu2+, Ni2+, Co2+, Cr(VI)) are found out. The studied bacteria are most resistant to Cr(VI) possessing oxidative properties in concentration range 1.25-20.0 g/l depending on the strain. Maximal metal concentrations, at which the growth of bacteria was possible, was: Ni2+ to toxic metals - 2.0 g/l, Co(2+) - 0.1 g/l. In the presence of metal ions possessing both replacing and oxidative properties the strains grew in a concentration range: Hg(2+) - 0.005-0.05 g/l, Cu(2+) - 0.1-1.25 g/l. The highest toxic effect was shown for mercury ions. One of the isolated Antarctic strains which was superresistant to high toxic metal concentrations (g/l): Cr(VI) - 20.0, Ni(2+) - 2.0, Cu(2+) - 1.25, Co(2+) - 0.1, Hg(2+) - 0.05. Thus, for the Antarctic bacteria isolated from rock damaging or bactericidal toxic metal concentrations are by 2-3 orders higher than for the majority of microorganisms.

It is shown that the total number of chemoorganotrophic aerobic microorganisms in the Antarctic Region revealed at 1 degree C and 5 degrees C made from 10(4) up to 10(6) cells/g of plant-soil sample of biotopes: grass Deschampcia antarctica, grass Colobanthus, green mosses, crustose black lichens and a biofilm of accretion on vertical rocks. From 10(6) up to 10(8) cells/g of samples were revealed in the same Antarctic samples at 30 degrees C. At 42 degrees C thermotolerant bacteria were either absent, or their quantity was less than 10(4) cells/g of samples. Thus the fraction (part) of the Antarctic microorganisms, which grow at different temperatures, varied: at 1-5 degrees C their part made from 5 to 15%, and at 30 degrees C--from 10 to 45%. At 15-20 degrees C the growth of both psychrophilic/psychrotolerant, and mesophilic microorganisms was observed. When comparing the results of plating of samples from different climatic zones (the Antarctic Region and Ukraine), it is shown that in the Antarctic biotopes in comparison with biotopes of the zone with temperate climate: (1) the total number of microorganisms is lower, (2) quantity of psychrophilic/psychrotolerant bacteria is higher, (3) quantity of mesophilic microorganisms is less, (4) as a result the part psychrophilic/psychrotolerant microorganisms in the total number of microorganisms is much higher. It is evident, that low temperatures, and also daily cycles of freezing and thawing are factors which limit microbial colonization ofAntarctic biotopes.

To determine a possibility of application of phylogenetic criteria for estimating the taxa rank, the intra- and interspecies, as well as intergeneric relatedness of methanotrophs on the basis of 16S rRNA gene sequences was estimated. We used sequences of 16S rRNA genes of the studied isolates of obligate methanotrophs which have been deposited in UCM (Ukrainian Collection of Microorganisms), and of type strains of other obligate methanotrophs species (from GenBank database). It is shown, that the levels of interspecies and intergeneric relatedness in different families of methanotrophs are not identical, and therefore they can be used for differentiation of taxa only within one family. The carried out analysis has shown, that it is necessary to reconsider taxonomic position: (1) of two phenotypically similar species of Methylomonas (M. aurantiaca and M. fodinarum), similarity of 16S rRNA genes which is 99.4%, similarity of their total DNA--up to 80% that rather testifies to strain differences, than to species differences; (2) of species Methylomicrobium agile and M album which are phylogenetically more related to genus Methylobacter (97% of affinity), than Methylomicrobium (94% of affinity); (3) of genera of the family Beijerinckiaceae (Methylocella and Methylocapsa), and also genera of the family Methylocystaceae (Methylosinus and Methylocystis), whereas high level of relatedness (97% and more) of these bacteria with other methanotrophic genera (within one family) practically corresponds to a range of relatedness of species (within some genera) in the family Methylococcaceae. When determining phylogenetic criteria which can characterize the ranks of taxa, it was revealed, that the levels of interspecies relatedness of methanotrophic genera of the families Methylocystaceae and Beijerinckiaceae (97.8-99.1% and 97.8%, accordingly) considerably exceed the level of genera formation in the family Methylococcaceae (94.0-98.2%) and, moreover, approach the value of intraspecies relatedness of the family Methylococcaceae (97.5-99.3%). Coefficients of intraspecies relatedness of methanotrophs of the families Methylocystaceae and Beijerinckiaceae are sometimes equal to interspecies relatedness. Hence, taxa of various rank can have the identical level of genes divergence. Thus, methanotrophic taxons of the families Methylocystaceae and Beijerinckiaceae have not demonstrated precise phylogenetic criteria which could correspond both to the species rank, or the genus rank. At the same time, the criteria, being adequate to the rank of certain taxa, are revealed in methanotrophs of the family Methylococcaceae. The level of genotypic relatedness of strains of the same species is in the range of 97.5-99.3%, species of the same genus--94-98%, the highest levels of relatedness between genera of this family are 90-96%.

Microbiological analysis of terrestrial biotopes of the Antarctic Region has shown, that vertical rocks of the Antarctic islands open for the Sun were characterized by special microcenoses. The wide distribution of pigmented microorganisms in the rock Antarctic samples, a higher frequency of their occurrence, the total number and biologic diversity, than in other Antarctic biotopes, has been demonstrated. For the first time the presence of bacteria and yeast, resistant to high doses of UV radiation on the vertical rocks in the Antarctic Region was shown. The lethal doze of UV radiation for the Antarctic pink pigmented Methylobacterium strains exceeded 200-300 J/m2, for coal-black yeast--500-800 J/m2, for red yeast--1200-1500 J/m2. The distinctions in lethal UV effect against strains of Methylobacterium isolated from the regions with different climate have not been found. Probably, adaptation of the rock microcenosis to extreme factors of the environment proceeds by natural selection of microorganisms, which resistance to this factor is genetically determined.

Microbiological analysis has been made of 120 samples from biotopes of the western coast of the Antarctic peninsula (Rasmussen cope, Tuxen cope, Waugh mountain), Argentine archipelago islands (Galindez, Skua, Corner, Barchans, Irizar, Uruguay, Cluls, Three Little Pigs, King-George), as well as neighbouring islands (Petermann--on the north, a group of Jalour islands--on the east, Berthelot--on the south-east); and more remote islands (Darboux, Lippmann, Booth). It was found out that the total number of chemoorganotrophic aerobic microorganisms was 10(6) - 10(8) cells/g of soil, that was by 2-3 orders lower than in the regions with temperate climate. One can observe a tendency of decreasing the quantity of chemoorganotrophic microorganisms in the Antartic biotopes (cells/g of a sample) in the following order: soil (1 x 10(7) - 8 x 10(8)), underground part of moss (1 x 10(6) - 5 x 10(7)), grass Deschampsia antarctica (10(6) - 10(8), slit of fresh-water reservoir (10(5) - 10(7)), ground part of moss (10(3) - 10(6)), lichens (10(3) - 10(6)). Representatives of several phylogenetic lines: Proteobacteria (genera Pseudomonas, Methylobacterium, Enterobacter), Firmicutes (genera Bacillus, Staphylococcus), Actinobacteria (genera Brevibacterium, Actinomyces, Streptomyces) have been found in the Antarctic samples. As a rule, genera of bacteria found in the Antarctic Region are widely distributed in different regions of the Earth with temperate climate. Microorganisms similar to the species Exophiala nigra (Issatsch.) Haats et de Hoog 1999, which was first detected 100 years ago by Academician B.L. Isachenko in the Arctic region water, were also isolated from biofilms on vertical rocks of the Galindez Island as well as from the soil of the Irizar Island.

Methylotrophs distribution has been studied in the terrestrial biotopes (moss, lichen, grass, soil, sludge of lakes) on the islands of Galindez, Barkhans, Irizar, Uruguay, Jalour, Petermann, Berthelot, Cruls, King George, Corner, Skua located in the Pacific sector of the Antarctic Region, as well as in analogous biotopes on the western shore of the Antarctic peninsula Basing on a complex of diagnosis features the isolated pink-pigmented strains, which facultatively use methanol and realize the serine cycle of assimilation of one-carbon compounds, are attributed to Methylobacterium genus. Methylobacterium strains occur more often in mosses, grass Deschampsia antarctica and lichens, than in the soil and lake sludge. Some regions ofAntarctica are comparable by the number of Methylobacterium cells with the same in the regions with moderate climate. An analysis of gene sequences 16S rRNA of the Antarctic methylobacteria with those of GenBank has shown a high extent of similarity with Methylobacterium extorquens (99.4-99.7%). Notwithstanding that the strains of Methylobacterium are resistant to the broad range of extreme factors (gamma-irradiation, UV-irradiation, dehydration), the Antarctic and collection strains of the genus were sensitive to the ions of such heavy metals as Cu, Hg, Cd, Cr (10 mg/l).

Now it is considered, that melanin is promising for application in medicine and pharmacology. Since black yeast were found in polar regions before, we have carried out screening of yeast-producers of melanin in the Antarctic biotopes. Dark pigmented microorganisms are revealed in 30% of samples from terrestrial biotopes at the western coast of the Antarctic peninsula, on islands of the Argentina archipelago (Galindez, Skua, Corner, Barchans, Irizar, Uruguay, Cruls, Three little pigs, King-George), and also on the neighbouring islands Petermann, Jalour, Berthelot, Darboux and Lippmann. In the Antarctic lichens the occurrence frequency of dark pigmented microorganisms, their total number and biodiversity are significantly higher, than in other Antarctic biotopes. Coal-black yeast are found on the crustose and bushy lichens on vertical rocks, they occur less often in soil, their quantity made 1 x 10(2) - 6 x 10(3)/g of a sample. Coal-black pigments were isolated from two Antarctic strains of yeast. These pigments are identical to melanin according to a complex of specific chemical tests, that is also confirmed by the character of UV-spectra (220-230 nm) and absorption spectra in the visible area (400-800 nm). The output of synthesized pigment/g in strain 36 made more than 10% of biomass amount. So, the yeast synthesizing intensively melanin are revealed in the Antarctic terrestrial biotopes.

Sequence-analysis of genes 16S rRNA has demonstrated the high-level relationship (99%) of the strains Methylobacter ucrainicus UCM B-3159, and Methylobacter marinus A45(T). The strain UCM B-3159 has lower coefficients of similarity (97.4-96.4%) for other species of that genus. These strains are similar as to their phenotypical properties and form one branch on the dendrogram which demonstrates species relations of Methylococcaceae family, that permitted reclassifying M. ucrainicus as M. marinus. Phylogenetic analysis has confirmed the belonging of strains UCM B-3002 and UCM B-3494 to Methylococcus capsulatus species. Those strains were earlier related to this species on the basis of phenotype features.

The study was concerned with the effects of brown seaweeds on hemostasis and sarcoma M-1 growth in rats. Laminaria japonica and focus were shown to exert anticoagulant action to inhibit carcinogenesis. They may be used for adjuvant therapy of tumors.

In the course of photodynamic treatment with supernatants of Laminaria japonica and Fucus sp the former was found to significantly inhibit the growth of experimental tumor M-1.

Levels of expression of hBD-1 gene (beta-defensin 1) and Toll-like receptors (TLR1, TLR2, TLR6) in cells of cervical mucosa in healthy nonpregnant and healthy pregnant women as well as in pregnant women with urogenital infection was measured by developed RT-PCR systems. During normal pregnancy compared with nonpregnant women, increase of TLRs genes expression which was correlated with increase of hBD-1 gene expression was observed. During urogenital infection in pregnant women compared with healthy pregnant, 10- fold and 50-fold increase of TLR1 and TLR2 genes expression respectively was associated with 2.5-fold decrease of hBD-1 gene expression in cervical mucosa. In group of women with untrauterine infection more marked increase of TLRs genes expression was observed. Thus significant changes (TLRs, antimicrobial peptides, cytokines etc.) in cells of cervical mucosa can be used as prognostic criteria for development of intrauterine infection.

Noninbred rat experiments with M-1 tumor have found that combination of fenylin with photodynamic therapy produced a 1.5-fold slowing down in the growth of the tumor compared to the control animals on day 15 and 21 after the treatment.

High extracellular concentration of K+ stimulated methane oxygenation with Methylomonas rubra 15 [Russian character: see text], Methylococcus thermophilus 111 [Russian character: see text] and Methylococcus capsulatus 494 at neutral value of pH. That was determined by K+ arrival to the cells at neutral medium pH that resulted in the increase of pH difference between the exterior and interior sides of the membrane (ApH) and, respectively, in the increase of the methane oxygenation rate. Thus, methane monooxygenation depends on the availability of ion gradients on a membrane. Ionophores valinomycin and monensin inhibited methane oxygenation by the cells of Methylomonas rubra 15 [Russian character: see text] that evidenced for the methane oxygenation dependence on the protone-motive force which could be formed as the result both of protons displacement with oxygenation of methane monooxygenation products and of the gradient of potassium and sodium ions. Protonophore FCCP suppressed completely methane oxygenation in Methylococcus capsulatus 494 and M. thermophilus 111 [Russian character: see text] at neutral pH, and took no effect at the alkaline values of pH. This suggests that FCCP dissipates the proton-motive force and does not inhibit methane monooxygenase activity. The results obtained indicate that the process of methane oxygenation should be combined with energy generation in a form of the transmembrane electric charge (delta psi) and proton gradient (deltapH).

Strain Methylomonas rubra 15sh(T), deposited in several collections of microorganisms (NCIMB 11913(T) = UCM B-3075(T) = ACM 3303(T)), is the subject of numerous studies. However, the name of this strain is not valid, primarily due to the phenotypic similarity of the species M. rubra to the species M. methanica. The results of the present study and data available in the literature indicate that M. rubra deserves the status of a separate species. Strains of M. rubra differ from strains of M. methanica in a number of properties, such as the ability to reduce nitrates to nitrites, the structure of intracytoplasmic membranes, and the presence of a new coenzyme Q. The distinctions between the species M. rubra and M. methanica were confirmed by comparison of electrophoretic patterns of their cellular proteins, by results of DNA-DNA hybridization, and by the data from 16S rRNA gene sequencing (the level of phylogenetic homology between these two species was below 95%). Phylogenetic and phenotypic analyses showed that strains of M. rubra cannot be assigned to any species of the genus Methylomonas. Results of polyphasic analysis suggest an independent taxonomic status of strain Methylomonas rubra 15sh(T). This paper contains description of Methylomonas rubra sp. nov. with the type strain 15sh(T) = NCIMB 11913T = UCM B-3075(T) = ACM 3303(T). The nucleotide sequence of the 16S rRNA gene of strain 15sh(T) has been deposited with the GenBank database under the accession number AY995198.

The authors have performed sequence-analysis of 16S rRNA genes of thermophylic methane-oxidizing bacteria UCM B-3026, UCM B-3032, UCM B-3109, UCM B-3014 which were isolated from sludge pond of different regions in Ukraine and deposited at Ukrainian Collection of Microorganisms (UCM) as Methylococcus thermophilus and "M. gracilis". A comparative analysis of 16S rRNA gene sequences of the studied bacteria with those sequences of various strains of bacteria in the GenBank databases has shown that the level of similarity of the strains UCM B-3026, UCM B-3032, UCM B-3109 with Methylocaldum szegediense was 98.0-98.7%, strain UCM B-3014 with Methylocaldum gracile--99.1%, that allows to relate them to these species. As to other species of the genus Methylocaldum, these strains had a lower level of similarity (94.8-97.2 %) and with Methylococcus thermophilus ACM 3585T (= IMV-B-3037T = UCM B-3037T)--less than 90%. Thus, the strains UCM B-3026, UCM B-3032, UCM-3109 were reclassified as Methylocaldum szegediense, strain UCM B-3014--as Methylocaldum gracile. When constructing phylogenetic tree based on the comparison of 16S rRNA genes with the use of various algorithms realized in the packages of programs Tree View (version 1.5.2) and ClastalX (version 1.81) certain clusters have formed genera Methylocaldum, Methylococcus and Methylomonas. Genera Methylobacter and Methylomicrobium have formed three clusters, one of the clusters contained two species of Methylobacter and two species of Methylomicrobium. Apparently, the taxonomy of genera Methylobacter and Methylomicrobium is to be specified.

Psychrotolerant bacteria which use obligately methane were found in the moss samples and in soil-vegetation samples in the island part of the Antarctica during the VII expedition (2003) at the station "Akademik Vernadsky". The number of methane-oxidizing bacteria in the samples from the Antarctica (101- 10(3)/g of the sample) was lower than in the samples from the regions with moderate climate (10(2)- 106/g of the sample). Psychrotolerant strains of Methylobacteriium genus which use facultatively methanol were found in the bottom sediments of the fresh-water and Krasnoye lakes, as well as in the soil-plant samples. The psychrophilic strain which is probably a new species of the genus Methylobacterium has been isolated from one soil-plant sample from the Antarctica at 10 degrees C. It is established that most collection mesophilic strains of Methylobacterium, which have been isolated from the soil and plant phyllosphere in Ukraine, also could grow at 10 degrees C.

Phylogenetic analysis of Methylobacterium genus species using analysis of nucleotide sequence of 16S rRNA genes has shown that Methylobacterium zatmanii and M. rhodesianum possess high-level similarity with M. extorquens (99.4-98.8%), and M. fujisawaense and M. radiotolerans--with M. mesophilicum (98.9-97.8%). These species are also similar as to their phenotypical properties and their total DNAs have also close similarity. Thus, the right of M. zatmanii, M. rhodesianum, M. fujisawaense and M. radiotolerans species for existence needs confirmation. Some phenotypical properties which were assummed as a basis for formation of certain species of the genus Methylobacterium (radioresistance, e.g.) cannot serve as the differentiating characteristic of this genus species. High resistance to gamma- and UV-radiation is a specific sign of representatives of Methylobacterium genus.

The drying of soil samples reduced the abundance (especially of predominant species) and the diversity of bacteria isolated from these samples, making easier the isolation of rare bacterial species. Some bacterial species that were minor before soil drying became dominant in dried soil samples. In general, soil drying allowed the diversity of soil bacteria to be determined more adequately. The bacteria that were isolated from dried soil samples turned out to be resistant to gamma radiation (with LD90 = 2.8-4.6 kGy) and desiccation. It is concluded that soil drying may serve as a model for the action of stress factors on natural bacterial populations. The hypothesis that periodic desiccation was the primary cause of formation of bacterial radioresistance in nature is discussed.

Historical aspects of development of phylogenetic bacteria taxonomy and essence of species in systematics of bacteria are considered. Comparative analysis of phenotypic and phylogenic classification of bacteria is conducted. Basic results of 16S rRNA gene sequence for bacteria are adduced. Comparative estimation of molecular biological methods in microbiologic researches and also estimation of potentialities and shortcomings of 16S rRNA gene sequence-analysis in systematics of bacteria are conducted.

The basic trends of the department scientific activity are presented. The priority results of investigations of methaneoxidizing bacteria biology are given. The biothechnological development of the department (obtaining of products of microbial synthesis: protein, polysaccharides, antialcoholic and antinarcotic preparations) are considered in detail. Possible areas of their application are presented.

In experiments on mice F1 total exposed to gamma-irradiation at the 5.0 Gy, showed that a intraperitoneal injection neogen at the dose 10 micrograms/kg oue time at the day and night during the 8 days after irradiation results in recovery of the number blood platelets and time issue of blood from standard wound to normal level on the week by compare with irradiated control animals.

Methylobacterium extorquens, M. mesophilicum, and Bacillus subtilis strains were found to be resistant to gamma-radiation, irrespective of whether they were isolated from the alienated zone around the Chernobyl Nuclear Power Plant or outside this zone. The LD90 of Methylobacterium and B. subtilis strains with respect to gamma-radiation was 2.0-3.4 and 3.7-4.4 kGy, respectively, whereas their LD99.99 values were 4.5-6.9 and more than 10 kGy, respectively. The high threshold levels of gamma-radiation for Methylobacterium and B. subtilis imply the efficient functioning of DNA repair systems in these bacteria. Unlike Bacillus polymyxa cells, the cells of M. extorquens, M. mesophilicum, and B. subtilis were also resistant to desiccation. Pseudomonas sp., Nocardia sp., and nocardioform actinomycetes were sensitive to both gamma-radiation and desiccation. Similar results were obtained when the bacteria studied were exposed to hydrogen peroxide and ultraviolet radiation. The results obtained indicate that the bacteria that are resistant to gamma-radiation are also resistant to desiccation, UV radiation, and hydrogen peroxide. The possibility of using simple laboratory tests (such as the determination of bacterial resistance to UV light and desiccation) for the evaluation of bacterial resistance to gamma-radiation is discussed.

A number of Methylococcus thermophilus 111p clones have been obtained which have acquired resistance to tetracycline. The stability of maintenance of marker resistance in these clones and also in already designed Methylomonas rubra 15sh mutants has been investigated. Chromosomal markers resistance to antibiotics or formaldehyde were maintained in the marked strains Methylococcus thermophilus 111p and Methylomonas rubra 15sh after storage in nonselective conditions. The markers of resistance to antibiotics, which were coded by plasmids (pAS8-121 and pULB113), were not always preserved in Methylomonas rubra and Methylococcus thermophilus. The stability of maintenance of chromosomal markers in the investigated methane oxidizing bacteria testifies to the fact that they can be used in laboratory and industrial practice for testing the marked bacteria on selective media. The collection of the marked bacteria-mutants Methylomonas rubra 15sh and Methylococcus thermophilus 111p has been created. These strains stably support the marker resistance to various antibiotics or formaldehyde in unselective conditions.

It was established, that EDTA (1.0 mM) and formamide (100 mM) are inhibitors of methanol dehydrogenase in Methylobacter luteus 12b, Methylomonas rubra 15sh and Methylococcus thermophilus 111p. The investigated strains co-metabolised ethane with the use of formate as the co-substrate. The application of formamide (or EDTA) as inhibitors of methanol dehydrogenase prevented from further transformation of ethanol and resulted in accumulation of extracellular ethanol. It was shown, that M. rubra 15sh accumulated extracellular ethanol under cultivation in a chemostate. The carried out researches have shown a regulation path of co-metabolism process of hydrocarbons by methane utilizing bacteria. Using the specific inhibitors of methanol dehydrogenase and a source of reducing agent (energy) for methane monooxygenase with the help of the cells of methane-oxidizing bacteria it is possible to obtain from ethane or other hydrocarbons the products of their monooxygenation--alcohols.

The pink-pigmented facultative methylotrophic bacteria (PPFMB) of the genus Methylobacterium are indespensible inhabitants of the plant phyllosphere. Using maize Zea mays as a model, the ways of plant colonization by PPFMB and some properties of the latter that might be beneficial to plants were studied. A marked strain, Methylobacterium mesophilicum APR-8 (pULB113), was generated to facilitate the detection of the methylotrophic bacteria inoculated into the soil or applied to the maize leaves. Colonization of maize leaves by M. mesophilicum APR-8 (pULB113) occurred only after the bacteria were applied onto the leaf surface. In this case, the number of PPFMB cells on inoculated leaves increased with plant growth. During seed germination, no colonization of maize leaves with M. mesophilicum cells occurred immediately from the soil inoculated with the marked strain. Thus, under natural conditions, colonization of plant leaves with PPFMB seems to occur via soil particle transfer to the leaves by air. PPFMB monocultures were not antagonistic to phytopathogenic bacteria. However, mixed cultures of epiphytic bacteria containing Methylobacterium mesophilicum or M. extorquens did exhibit an antagonistic effect against the phytopathogenic bacteria studied (Xanthomonas camprestris, Pseudomonas syringae, Erwinia carotovora, Clavibacter michiganense, and Agrobacterium tumifaciens). Neither epiphytic and soil strains of Methylobacterium extorquens, M. organophillum, M. mesophilicum, and M. fujisawaense catalyzed ice nucleation. Hence, they cause no frost injury to plants. Thus, the results indicate that the strains of the genus Methylobacterium can protect plants against adverse environmental factors.

In connection with the Chernobyl Nuclear Power Plant (ChNPP) accident and the negative ecological after-effects for biota in this zone the interest has arisen to radioresistant bacteria, as to the most dynamic model of the given ecosystem, and to mechanisms which provide resistance of bacteria to ionizing radiation. The analysis of published data has shown that the radioresistant bacteria are not interrelated taxonomically and phylogenetically. The extreme radioresistant bacteria are represented by the Deinococcus species, which form a group phylogenetically close to the line Thermus-Meiothermus. Other radioresistant bacteria are the representatives of the genera Rubrobacter, Methylobacterium, Kocuria, Bacillus and some archebacteria. Data on natural habitats, of radioresistant bacteria are not numerous. In a number of cases it is difficult to distinguish their natural habitats, as they were isolated from the samples which were previously exposed to X-ray or gamma-irradiation, or from the ecosystems with the naturally raised radioactivity. To understand the strategy of survival of radioresistant bacteria, we briefly reviewed the mechanism of action of various species of radiation on cells and macromolecules; physiological signs of the cell damage caused by radiation; mechanisms eliminating (repairing) these damages. More details on mechanisms of the DNA repair in D. radiodurans are described. The extreme resistance of D. radiodurans to the DNA damaging factors is defined by 1) repair mechanisms which fundamentally differ from those in other procaryotes; 2) ability to increase the efficiency of a standard set of the DNA repairing proteins. Literary and own data on the effect of radiation on survival of various groups of bacteria in natural ecosystems are summarized. The ecological consequences of the ChNPP accident for soil bacteria in this region were estimated. The reduction of the number of soil bacteria and recession of microbial diversity under the effect of anthropogenic radiation was shown.

UV irradiation is proposed for use in studying the effect of radioactive irradiation, since radioresistant bacteria are, as a rule, resistant to UV, and the mechanisms of repair of cell damage induced by UV and ionizing radiation are similar. It was found that the total number of bacteria and the number of dominant species in soil samples exposed to UV radiation decreased, indicating the unfavorable effect of UV radiation on bacterial diversity in soil ecosystems. The percentage of cells of bacteria belonging to dominant species varied significantly depending on the intensity of UV irradiation. It can be inferred that long-term irradiation of soils must impair the stability of soil ecosystems, a phenomenon that was indeed observed in the zone around the Chernobyl Nuclear Power Plant. At the same time, the UV irradiation of soil samples made it possible to reveal minor species, primarily UV-resistant pigmented bacteria. UV irradiation can probably be used as a selective factor for the isolation of radioresistant species.

Seventy strains of chemoorganotrophic bacteria isolated by our group in 1993-1994 from soil sampled in the zone around the Chernobyl Nuclear Power Plant (ChNPP) were studied with respect to their sensitivity to various stress factors damaging DNA. Bacillus subtilis, B. cereus (both spores and vegetative cells), Methylobacterium extorquens, M. mesophilicum, and unidentified pigmented bacteria were found to be the most resistant to ultraviolet (UV) radiation, exhibiting LD90 values of 40 to more than 211 J/m2. The same bacteria, as well as Bacillus polymyxa, were tolerant to hydrogen peroxide (lethal concentrations of H2O2 ranged from 0.3 to 1.0 M); i.e., UV-resistant strains were also tolerant to hydrogen peroxide and vice versa. Fluorescent pseudomonads were the most sensitive to both UV radiation and H2O2, showing LD90 from 6 to 18 J/m2 and a lethal concentration of H2O2 lower than 0.1 M. All of the soil samples collected in the alienated zone around the ChNPP, where the radioactivity of the soil had decreased from 1000 to 2 microCi/kg soil over the period from 1987 to 1995, contained not only resistant bacteria but also a small number of bacteria sensitive to UV radiation and H2O2.

The diversity of aerobic chemoorganotrophic (capable of growing on nutrient agar) bacteria in radioactive soil (0.3-17.0 microCi/kg soil) sampled in the 10-km zone around the Chernobyl Nuclear Power Plant (CNPP) was found to be lower than that observed in control, uncontaminated soil with a radioactivity of 0.002-0.006 microCi/kg soil. All the radioactive soil samples contained the bacteria Bacillus cereus and Methylobacterium extorquens or M. mesophillicum, which exhibited a high tolerance to 0.3-1.0 M hydrogen peroxide, whose action can to a certain extent simulate the effect of ionizing radiation. Some of the contaminated soil samples contained other species of chemoorganotrophic bacteria with a low tolerance to H2O2. The survival of bacteria in the Chernobyl accident zone is probably due to the functioning of mechanisms efficiently neutralizing peroxide compounds and repairing radiation-damaged DNA. The population of cellulolytic, nitrifying, and sulfate-reducing bacteria in contaminated soil was found to be 1-2 orders of magnitude less than in control soil, indicating the unfavorable effect of anthropogenic radiation on the abundance and diversity of soil bacteria.

The catastrophe at Chernobyl Atomic Power Station (ChAPS) of 1986 has created a natural model for studying the after-effects of prolonged action of radiation on the biota. Our work is aimed to estimate the variability of heterotrophic bacteria in the soil of 10-km zone of ChAPS which have formed as affected by prolonged action of radiation, as well as to create the corresponding collection of bacteria. Microbiological analysis of soils was carried out in 1993-1994 (in spring, summer, autumn), allowing for bacteria destroying different organic substance (in soil). It is shown that in the surface layer of soils (at the depth of 0-2 cm) the total number of cells of heterotrophic bacteria as well as the number of found species of bacteria is considerably less than in the control samples. Atypical distribution of bacteria in the soil profile was established. Indices of the species diversity of bacteria in these soils permit one to consider that microbe content of soil of the 10-km zone of the ChAPS has become considerably less after the disaster at the ChAPS in 1986 and has not recovered by 1993. A collection of various physiological groups of bacteria including representatives of Methylobacterium genus has been created. They have been isolated from the soils of 10-km zone of the ChAPS for following genetic investigations.

The effect of berberin sulfate (single and chronic administration during 30 days in doses of 0.14-80 mg/kg) on the functional state of hemostasis system was studied on gamma-irradiated (dose 5.0 Gy) and intact Wistar rats and mice (CBA x C57BL)F1. Administration of the drug increases the number of thrombocytes, decreases the activity of factor XIII and promote blood coagulation.

Exposure of hen's eggs to irradiation by doses 0.05-1.66 Gy (dose rate 2.56x10-4 Gy/s) in early embryonal period (1st day of incubation of eggs doesn't essentially affect on indices of functional state of haemostasis system of two-month aged individuals, except the activation of contact phase of blood coagulation, in which a hypercoagulation shift within normal fluctuations has been noted.

Serum medium-size molecules (MSM) and blood coagulation parameters have been studied before and after surgery in 60 patients randomized into 2 groups. Patients in group I were subjected to multicomponent endotracheal anesthesia with neuroleptanalgesia and cardiopulmonary bypass and to postoperative analgesia with non-narcotic and narcotic analgesics. Patients in group II were additionally subjected to epidural anesthesia into upper chest segments with trimecaine + adrenaline and low morphine doses. Prior to surgery many patients showed signs of the first-degree disseminated intravascular coagulation syndrome. Parallel changes in the postoperative clinical course, pathological shifts in coagulograms and changes in MSM levels (which were maximum on the fourth day postoperatively in both groups) have been established. Patients in group II were found to have a more favourable coagulation profile.

Ethane and propane were found to be competitive inhibitors of methane oxidation. Obligate methylotrophs can oxidize ethane only in the presence of methane, i.e. co-oxidation of ethane takes place. Studies on the kinetics of simultaneous oxidation of ethane and methane have shown that their oxidation is catalysed by one and the same enzyme system, within one and the same enzyme active site; therefore, the absolute substrate specificity is absent. The reactions of methane and ethane oxidation are coupled since ethane can be oxidized only after the reduced coenzyme had been provided by the reaction of methane oxidation.

Obligate methane oxidizing bacteria were classified within groups using numerical analysis and computer techniques. Employment of properties coinciding for all the species of the bacterial group under study was found to result in an insignificant (5--10%) change in the degree of similarity rather than in the redistribution of interrelationship; it had no effect on the formation of phenons. The Smirnov taxonomical analysis according to which the weight of a property is inversely proportional to its frequency has a number of advantages over other numerical methods where all the properties are assumed to be of equal value. Comparative evaluation of the data obtained by different methods which have been used by us to calculate the coefficients for coupled similarity of species (the coefficient of association, the coefficient of correlation, the index of distance, the coefficient of taxonomic ratio) shows that similar results are found in all the cases with respect to group division of the species belonging to the family Methylomonadaceae Leadbetter 1974. The species of methane oxidizing bacteria should be divided into the following genera: Methylosinus Whittenbury, Phillips a. Wilkinson 1970, Methylocystis Whittenbury, Phillips a. Wilkinson 1970, Methylococcus Foster a. Davis 1966, Methylomonas Leadbetter 1974.

Various numerical methods were used to estimate the coordination of taxa of gram-negative aerobic and facultative anaerobic organoheterotrophic and chemolithotrophic bacteria. Stable phena were found to be formed by cultures belonging to the families Rhizobiaceae, Halobacteriaceae, Enterobacteriaceae, Nitrobacteriaceae (except the genus Nitrobacter), and Methylomonadaceae (except the genus Methylococcus). The unstable position was found in the genera Thermus, Zoogloea, Xanthomonas, Sulfolobus, Methylococcus, Alcaligenes, Brucella, and Acetobacter. The greatest scatter among the objects being analysed was detected among genera belonging to the family Pseudomonadaceae. The taxonomic position of these genera must be defined more precisely. The family Methylomonadaceae is related to such physiologically unique groups of microorganisms as nitrifying, sulfate-reducing, extreme thermophilic and halophilic forms. All in all, the data reported in this work show that numerical analysis can be used to specify the classification structure of bacteria. In a number of cases, the results are consistent with those changes which are performed in the Bergey Manual 9 using logical analysis (for instance, concerning the position of the genera Gluconobacter, Acetobacter, Beijerinckia, and Derxia).

The diagnoses of taxons have been corrected for methane assimilating bacteria of the family Methylomonadaceae Leadbetter 1974, of the genera Methylomonas Leadbetter 1974. Methylococcus Foster a. Davis 1966, Methylosinus Whittenbury. Phillips a. Wilkinson 1970, Methylocytis Whittenbury, Phillips a. Wilkinson 1970, and of 16 species. The diagnosis of the species Methylomonas rubra Whittenbury, Phillips a. Wilkinson 1970 has been completed, and the new species Methylomonas gracilis sp. nov. and Methylococcus luteus sp. nov. have been described. Since information about the properties of certain strains is not complete, they should be transferred to the category incertae sedis. Diagnostic keys for identification of methane assimilating bacteria are presented in the paper.

Methane hydroxylase (MH) from the obligate methane assimilating culture of Methylococcus thermophilus catalyses oxygenation of both CH4+ and NH4+; therefore, we studied the specificity of enzyme systems catalysing the subsequent oxidation of compounds produced upon the oxygenation of these substrates (CH3OH and NH2OH). CH3OH and NH2OH were shown to be oxidized by different enzymes, viz. methanol dehydrogenase (MD) and hydroxylamine oxidase (HO), respectively. Similar to MH, MD is characterized by the absence of strict substrate specificity, and catalyses oxidation of primary alcohols other than methanol, rather than hydroxylamine. HO catalyses oxidation of hydroxylamine rather than methanol and possesses the activity of hydroxylamine:cytochrome c oxidoreductase. The constitutive character of HO from the methane assimilating bacteria and the substrate specificity of the enzyme suggest that a lithotrophic pathway for producing energy operates in these bacteria. The HO of Methylococcus thermophilus is similar in certain properties to the HO of the nitrifying bacterium Nitrosomonas europaea.