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This paper studies the number of bacteria in typical chernozem and mountain-meadow soil by the traditional method and the cascade filtration technique. The total number of bacteria in these soils, which was obtained in filters of different diameters during filtering the suspension of a certain amount, is 1.5-5 times higher than that obtained by the traditional method. In the structure of the bacterial biomass in both soils, the biomass of bacterial cells with a diameter of 0.38-0.43 microm was dominating by 8-90%. In the typical chernozem, the biomass of cells with a diameter of 0.17 microm was slightly more than 1%; in the mountain-meadow soil, the percentage of the biomass of cells with a diameter of 0.17 microm increased by 5%. The average volume and diameter of the bacteria in the studied soils were calculated. In typical chernozem, the average volume of bacterial cells was equal to 0.0046 microm3 and the diameter was 0.206 microm. In the mountain-meadow soils, these values were slightly lower, 0.0038 microm3 and 0.194 microm, respectively. The biomass of the bacterial cells, which is usually calculated based on the cell volume of 0.1 microm3, is overestimated by about five times when counting the number on the filters. The percentage of the real biomass of soil bacteria is traditionally much lower than that estimated.

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A comprehensive study of chitinolytic microbial complexes of the phylloplane from cultured and forest plants has been conducted. An increase of the number and biomass of metabolically active cells of the representatives of the domain Bacteria and a decrease in fungal biomass in the experimental microcosms have been shown to occur after the introduction of chitin. The characteristic features of the taxonomic structure of metabolically active chitinolytic complexes of the phylloplane of the plants studied have been elucidated. Representatives of the phyla Proteobacteria, Bacteroidetes, and Verrucomicrobia have been shown to play important roles in the chitinolytic complexes of green leaf samples, while mycelial actinobacteria of the phylum Actinobacyteria played a similar role in needles of coniferous trees. A collection of chitinolytic microorganism cultures isolated from the phylloplane of different plant species has been created.

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It has been demonstrated that complexes of mycelial bacteria (actinomycetes), in which the amount of psychrotolerant actinomycetes reaches hundreds of thousands of CFU/g of the soil (frequently exceeding the portion of mesophilic forms), are developed in peat and podzolic soils of the tundra and taiga at low temperatures. As actinomycetes grow and develop in cold soils, their mycelium increases in length. Use of the molecular in situ hybridization method (fluorescent in situ hybridization, FISH) demonstrated that the portion of metabolically active mycelial actinobacteria exceeds the portion of unicellular actinobacteria in the Actinobacteria phylum. Specific peculiarities of psychrotolerant populations in relation to the spectrum of consumed substrates (histidine, mannitol, saccharose) were established by the method of multirespirometric testing.

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The method proposed in this study was used to isolate fungi grown under anaerobic conditions and to reveal distinctions in their abundance and species composition in different habitats. The ability of micromycetes of different taxa to grow under anaerobic conditions and ensure alcohol fermentation was determined for a representative sample (344 strains belonging to more than 60 species). The group of fungi growing under anaerobic conditions included species with high, moderate, and low fermentation activity. The ability for anaerobic growth and fermentation depended on the taxonomic affiliation of fungi. In some cases, the expression of these characteristics depended on the habitat from which the strain was isolated. The maximum level of ethanol accumulation in culture liquid (1.2-4.7%) was detected for Absidia spinosa, Aspergillus sp. of group flavus, Aspergillus terreus, Acremonium sp., Mucor circinelloides, Mucor sp., Fusarium oxysporum, F. solani, F. sambucinum, Rhizopus arrhizus var. Arrhizus, Trichoderma atroviride, and Trichoderma sp.

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The carbon content in microbial biomass (Cmic-BM) was determined in various horizons of the soil profile (sod-podzo, gray, podzol, and rzhavozem) of various forests (oak, spruce Yellow Archangel, spruce moss, aspen, spruce broadleaf) in the southern taiga of European Russia (Moscow and Kaluga oblasts) by the substrate induced respiration (SIR) and direct microscopy (DM) methods. The fungi-to-bacteria ratio was measured by the selective inhibition technique and DM. A quantitative differentiation of the fungal mycelium was suggested. The Cmic-DM/Cmic-SIR in various horizons of the soil profile was about 98%. The fungal contribution to MB was 52-74% and 92-99% according to the SIR and DM data, respectively. The microbial parameters were associated with the CO2 and N2O production by the soils. The contradictory data about the fungi proportion in the MB of soils of various ecosystems were discussed.

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By means of dye L7012, the number and physiological condition (damage rate of membranes) of bacterial cells is defined. The results testify to considerable physiological heterogeneity of bacteria cells in soils. In fresh samples of soil, the percentage of intact cells reached 60-70%. Damaged membranes occurred in 30-40% of cells. The number of damaged cells dramatically increased downwards through the soil profile. Drying and freezing of soil samples considerably reduced the quantity of intact cells and increased the percentage of cells with damaged membranes; the number of intact cells was 10-20%. Treatment with biocide agents resulted in lysis of the majority of cells and cells injuries. However, some of the cells kept an intact cellular membrane, which testifies to the high stability of bacteria in soil. These data allow us to offer a method of staining a soil suspension with the use of luminescent dye L7012 with the quality of an express method that gives the chance to monitor the number and physiological condition of the bacterial complex of soils.

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The content of microbial biomass (MB) was determined in samples of gray forest, chestnut, and tundra soils with different physicochemical properties (0.4-22.7% Corg; 8.4-26.8% silt particles; pH 4.3-8.4) by the methods of substrate-induced respiration (MB(SIR)) and direct microscopy (MB(M)). The samples of two upper soil layers, 0-5 and 5-10 cm (without plant litter), from different ecosystems (forest, forest shelter belt, meadow, fallow, and arable land) and elements of relief of interfluvial tundra (block/upper land plateau, depression between blocks) have been analyzed. The content of microbial biomass in the 0-5-cm soil layer was 216-8134 and 348-7513 microg C/g soil as measured by the methods of substrate-induced respiration and direct microscopy, respectively. The MB(SIR) and MB(M) values closely correlated with each other: r = 0.90 and 0.74 for 0-5 and 5-10 cm, respectively. The average MB(SIR)/MB(M) ratio was 90 and 60% for 0-5 and 5-10 cm, respectively. The portion of microbial carbon in total organic soil carbon was, on average, 4 and 3% (SIR) and 5 and 7% (direct microscopy) for 0-5 and 5-10 cm, respectively. Possible reasons for the differences between MB(SIR) and MB(M) values in the soils under study are discussed.

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The numbers of microscopic fungi isolated from soil samples after anaerobic incubation varied from tens to several hundreds of CFU per one gram of soil; a total of 30 species was found. This group is composed primarily of mitotic fungi of the ascomycete affinity belonging to the orders Hypocreales (Fusarium solani, F. oxysporum, Fusarium sp., Clonostachys grammicospora, C. rosea. Acremonium sp., Gliocladium penicilloides, Trichoderma aureoviride, T. harzianum, T. polysporum, T. viride. T. koningii, Lecanicillum lecanii, and Tolypocladium inflatum) and Eurotiales (Aspergillus terreus, A. niger, and Paecilomyces lilacimus), as well as to the phylum Zygomycota, to the order Mucorales (Actinomucor elegans, Absidia glauca, Mucor circinelloides, M. hiemalis, M. racemosus, Mucor sp., Rhizopus oryzae, Zygorrhynchus moelleri, Z. heterogamus, and Umbelopsis isabellina) and the order Mortierellales (Mortierella sp.). As much as 10-30% of the total amount of fungal mycelium remains viable for a long time (one month) under anaerobic conditions.

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Bacterial succession in soil was studied for two variants of initiation (moistening and moistening with addition of glucose). To determine the numbers of viable gram-negative bacteria, the modified nalidixic acid method was applied. The numbers of gram-negative bacteria revealed by this method were 2 to 3.5 times higher than those determined by the traditional method. In a developing community, the highest total bacterial numbers were observed on day 7; afterwards their numbers decreased and stabilized at a level exceeding four- to fivefold the initial one. In both experimental variants, the highest numbers of viable gram-negative bacteria were revealed on day 15 (75-85% of the total bacterial numbers). Morphology of these bacteria suggests their classification as cytophagas (chitinophagas) utilizing chitin from the dead fungal mycelium.

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The dynamics of assimilation of chitin by soil microorganisms (primarily prokaryotes) as a source of carbon and nitrogen has been determined by gas chromatography and fluorescence microscopy. The highest rates of chitin decomposition in chernozem were detected at humidity levels corresponding to the pressure of soil moisture (P) of -1.4 atm. The rate of microbial consumption of chitin is three times higher than that of the carbon of soil organic matter. Fluorescence microscopy revealed that an increase in the pressure of soil moisture from P = -10 atm to P = -0.7 atm resulted in a considerable increase in the proportion of the specific surface of mycelial bacteria (actinomycetes).

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Very large microbial biomass was revealed in peat bogs by means of fluorescence microscopy. In ombrotrophic peatlands, the pool of the dry-weight microbial biomass in the 1.5-m layer constituted 3-4 t/ha and was twice as high as in the minerotrophic peat bogs. Fungal biomass was predominant (55-99%) in ombrotrophic peatlands, while bacterial biomass predominated in minerotrophic peatlands (55-86%). In ombrotrophic peatlands, the microbial biomass was concentrated in the upper layers, while in minerotrophic peatlands, it was uniformly distributed in the bulk. After drainage, the microbial pool in the ombrotrophic peatlands increased twofold; that in the minerotrophic peatlands remained at the same level. The potential activity of nitrogen fixation and denitrification was revealed across the whole profile of the peatlands. The average values of these potential activities were five times higher in the minerotrophic peatlands, where bacterial biomass predominated.

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The abundance of actinomycetes isolated from the soils of Mongolian desert steppes varies from several thousand to hundreds of thousands of CFU/g soil, depending on soil type and isolation medium. Eight actinomycete genera have been found in these soils: Streptomyces, Micromonospora, Saccharopolyspora, Actinomadura, Microtetraspora, Thermomonospora, Nocardia, and Dactylosporangium. The streptomycete complexes of brown desert-steppe and gray-brown desert alkaline soils include halophilic, alkaliphilic, and haloalkaliphilic species that grow most successfully on the media with a salt concentration of 5% and pH 8-9.

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Actinomycete communities demonstrated a replacement of the generic composition in time as a function of soil moisture. Representatives of the genera Streptomyces, Micromonospora, Actinomadura, Saccharopolyspora, and Microbispora were repeatedly isolated from soil under different moisture conditions (field capacity, maximum molecular capacity, and maximum adsorption capacity). Representatives of some rare genera (Thermomonospora and Kibdelosporangium) were isolated from soil with low moisture levels inhibiting growth of more hydrophilic actinomycetes and bacteria. Spores of some actinomycetes could grow at low relative air humidity (RH) (50 and 67%). The complete growth cycle of all actinomycetes starting from spore germination to sporulation was observed only at RH of 98%.

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The first demonstration that samples of lignite at a depth of 10 m are considerably enriched in bacteria is reported. According to direct microscopy, the abundance of bacteria was about 10(7) cells/g. About 70% of cells had intact cell membranes and small size, which points to their anabiotic state. The fungal mycelium length was no more than 1 m. Lignite inoculation onto solid glucose-yeast-peptone medium allowed as to isolate bacteria of the genera Bacillus, Rhodococcus, Arthrobacter, Micrococcus, Spirillum, and Cytophaga. Representatives of the genera Penicillium and Trichoderma were identified on Czapek medium. Moistening of lignite powder increased the microbial respiration rate and microbial and fungal abundance but did not increase their generic diversity. This finding suggests that the studied microorganisms are autochthonous to lignite.

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The reaction of soil bacteria and fungi to the digestive fluid of the earthworm Aporrectodea caliginosa was studied. The fluid was obtained by centrifugation of the native enzymes of the digestive tract. The inhibition of growth of certain bacteria, spores, and fungal hyphae under the effect of extracts from the anterior and middle sections of the digestive tract of A. caliginosa was discovered for the first time. In bacteria, microcolony formation was inhibited as early as 20-30 s after the application of the gut extracts, which may indicate the nonenzymatic nature of the effect. The digestive fluid exhibited the same microbicidal activity whether the earthworms were feeding on soil or sterile sand. This indicates that the microbicidal agents are formed within the earthworm's body, rather than by soil microorganisms. The effect of the digestive fluid from the anterior and middle divisions is selective in relation to different microorganisms. Of 42 strains of soil bacteria, seven were susceptible to the microbicidal action of the fluid (Alcaligenes.faecalis 345-1, Microbacterium sp. 423-1, Arthrobacter sp. 430-1, Bacillus megaterium 401-1, B. megaterium 413-1, Kluyvera ascorbata 301-1, Pseudomonas reactans 387-2). The remaining bacteria did not die in the digestive fluid. Of 13 micromycetes, the digestive fluid inhibited spore germination in Aspergillus terreus and Paecilomyces lilacinus and the growth of hyphae in Trichoderma harzianum and Penicillium decumbens. The digestive fluid stimulated spore germination in Alternaria alternata and the growth of hyphae in Penicillium chrysogenum. The reaction of the remaining micromycetes was neutral. The gut fluid from the posterior division of the abdominal tract did not possess microbicidal activity. No relation was found between the reaction of microorganisms to the effects of the digestive fluid and the taxonomic position of the microorganisms. The effects revealed are similar to those shown earlier for millipedes and wood lice in the following parameters: quick action of the digestive fluid on microorganisms, and the selectivity of the action on microorganisms revealed at the strain level. The selective effect of the digestive gut fluid of the earthworms on soil microorganisms is important for animal feeding, maintaining the homeostasis of the gut microbial community, and the formation of microbial communities in soils.

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Anaerobic chitinolytic complex was studied in three soil types: chernozem, gray forest soil, and chestnut soil. The abundance and biomass of anaerobic chitinolytic microbial complex of fungi, bacteria, and actinomycetes were evaluated by luminescent microscopy. The dynamics of methane emission from soil during chitinolytic succession was studied by gas chromatography. All three studied microbial groups proved to participate in chitin transformation in soil under anaerobic conditions. The highest biomass growth was observed among prokaryotes, particularly actinomycetes, whose biomass doubled. The increase in methane emission during chitinolytic succession was most pronounced in soils with low organic matter content.

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The population density of bacteria in peat deposits along the landscape profile of the Vasyugan Marsh has been found to be as high as tens of millions of CFU/g peat. The abundance and diversity of bacteria increased with depth within the peat deposit, correlating with an increasing level of peat degradation. Variations in these parameters with depth and season were greater in peat deposits located in transaccumulative and transitional positions than in the sedge-sphagnum bogs located at the eluvial region of the profile. In the upper 1-m-thick layer of the peat deposits studied, bacilli, represented by five species, dominated, whereas, in the deeper layers, spirilla and myxobacteria prevailed. These bacteria were major degraders of plant polymers. Unlike the bacterial communities found in the peat deposits of European Russia, the dominant taxa in the studied peat deposits of Western Siberia are represented by bacteria resistant to extreme conditions.

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