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The paper presents the idea of transparent evolution through the long-term reaction of the planet Earth on the external flow of radiant energy from the Sun. Due to limitations of matter on Earth, as well as on any other planet, the continuous pumping flow of radiant energy was shown to lead to cyclization and transport of substance on emerging gradients. The evolution of energy-matter interaction follows the path of capturing and transferring more energy by the fewer matter, i.e., the path of growth of the amount of energy used by each unit mass. For this indicator, the least effective mass transfer is a simple mass transfer as vortices of gases, in the gradients of temperature and pressure, which occurred on the primary surface of the planet. A long-term natural selection related to the accumulation of water on the planet has played a special role in developing the interaction of energy and matter. Phase transformations (ice, water, vapor) and mechanical transfers are the most common energy-matter processes. Based on water cycles, cyclic transports and transformations, chemical transformation of substances became possible developing over time into a biological transformation. This kind of the interaction of energy and matter is most efficient. In particular, during photosynthesis the energy of our star "is captured and utilized" in the most active part of the spectrum of its radiation. In the process of biological evolution of heterotrophs, a rise (by a factor of hundreds) in the coefficient that characterizes the intensity of energy exchange from protozoa to mammals is most illustratory. The development and the current dominance of humans as the most energy-using active species in capturing the energy and meaningful organization of its new flows especially on the basis of organic debris of former biospheres is admirable, but quite natural from the energy positions. In the course of technological evolution of humankind, the measure of the intensity of energy for homoeothermic (warm-blooded) animals has increased 20 times, based on the process energy used by the "average" inhabitant of the world. Thus, the victory of our species in planetary evolution is easy to fit into the mainstream of evolution through energy-matter interactions: multiple growth of star energy was used to transform the matter on the surface of the irradiated planet.

In this paper, the influence of the extent of openness of ecosystem that is defined by the dilution rate, which characterizes the extent of flowage of the pond, on the intensity of the biotic circulation in ecosystems with different regulation types, number of trophic links and extent of closing has been investigated. We considered open systems, we took into account the return of the limiting substances, such as nitrogen and phosphorous, into the cycle by degradation of detritus and products of vital functions of consumers. It was shown by the numerical calculations that the increase of the dilution rate in without recycle ecosystems leads to increase of the net primary production up to the maximum value corresponding to the two-link trophic chain (biogenic substance and producer) and then, to gradually decrease. The residual concentration of biogenic limiting substances monotone increases. Net primary production and residual concentration of biogenic limiting substances in systems with recycle with even number of links behaves similarly to that in without recycle ecosystems. In the systems with recycle with the odd number of links that values lies on the stable level. We showed that in wide range of the dilution rate the recycling of the ecosystem can highly increase the net primary production and reduce residual concentration of biogenic limiting substances. The influence of the dilution rate on numbers of links that may exist in the system was analyzed., (c2005 COSPAR. Published by Elsevier Ltd. All rights reserved.)

The problem of interaction between man and microorganisms in closed habitats is an inextricable part of the whole problem of co-existence between macro- and microorganisms. Concerning the support of human life in closed habitat, we can, conventionally, divide microorganisms, acting in life support system (LSS) into three groups: useful, neutral and harmful. The tasks, for human beings for optimal coexistence with microhabitants seem to be trivial: (1) to increase the activity of useful forms, (2) decrease the activity harmful forms, (3) not allow the neutral forms to become the harmful ones and even to help them to gain useful activity. The task of efficient management and control of microbial population's development in LSS highly depends on mission duration. As for short-term missions without recycling, the proper hygienic procedures are developed. For longer missions, the probability of transformation of the neutral forms into the harmful ones is becoming more dangerous. The LSS for long-term missions are to use cycling-recycling systems, including system with biological recycling. In these systems, microbial populations as regenerative link should be useful and active agents. Some problems of microbial populations control and management are discussed in the paper., (c2005 COSPAR. Published by Elsevier Ltd. All rights reserved.)

Using biotic turnover of substances in trophic chains, natural and artificial ecosystems are similar in functioning, but different in structure. It is necessary to have quantitative criteria to evaluate the efficiency of artificial ecosystems (AES). These criteria are dependent on the specific objectives for which the AES are designed. For example, if AES is considered for use in space, important criteria are efficiency in use of mass, power, volume (size) and human labor and reliability. Another task involves the determination of quantitative criteria for the functioning of natural ecosystems. To solve the problem, it is fruitful to use a hierarchical approach suitable for both individual links and the ecosystem as a whole. Energy flux criteria (principles) were developed to estimate the functional activities of biosystems at the population, community and ecosystem levels. A major feature of ecosystems as a whole is their biotic turnover of matter the rate of which is restricted by the lack of limiting substances. Obviously, the most generalized criterion is to take into account the energy flux used by the biosystem and the quantity of limiting substance included in its turnover. The use of energy flux by ecosystem, E(USED)--is determined from the photoassimilation of CO2 by plants (per time unit). It can be approximately estimated as the net primary production of photosynthesis (NPP). So, the ratio of CO2 photoassimilation rate (sometimes, measured as NPP) to the total mass of limiting substrate can serve as a main universal criterion (MUC). This MUC characterizes the specific cycling rate of limiting chemical elements in the system and effectiveness of every ecosystem including the global Biosphere. Comparative analysis and elaboration of quantitative criteria for estimation of natural and artificial ecosystems activities is of high importance both for theoretical considerations and for real applications., (c2005 COSPAR. Published by Elsevier Ltd. All rights reserved.)

Studying material transformations and biotic cycling in artificial ecosystems (AES), we need to know the principles of biological adaptation of active organisms to change in the environment. Microorganisms in AES for water purification are the most active transforming organisms and consumers of the organic substances contained in wastes. Utilization of organic substances is directly connected with the energy fluxes used by AES. According to energy criteria, the energy fluxes used by a biological system tend to reach maximum values under stable conditions. Unutilized substrate concentration decreases as a result of biological adaptations. After a dramatic change in environmental factors, for example, after a sharp increase in the flow rate of organic substances, the biological system is not able to react quickly. The concentration of unutilized substrate increases and the energy flux used by the biological system decreases. The structure of the microbial community also changes, with a decrease in biological diversity. The efficiency of energy use by simple terrestrial ecosystems depends on the energetic intensity and interactions between plants and rhizospheric microorganisms., (c2005 COSPAR. Published by Elsevier Ltd. All rights reserved.)

Populations of Escherichia coli Z905/pPHL7, a transgenic microorganism, were heterogenic in the expression of plasmid genes when adapting to the conditions of water microcosms of various mineralization levels and structure of microbial community. This TM has formed two subpopulations (ampicillin-resistant and ampicillin-sensitive) in every microcosm. Irrespective of mineralization level of a microcosm, when E. coli Z905/pPHL7 alone was introduced, the ampicillin-resistant subpopulation prevailed, while introduction of the TM together with indigenous bacteria led to the dominance of the ampicillin-sensitive subpopulation. A high level of lux gene expression maintained longer in the freshwater microcosms than in sterile saline lake water microcosms. A horizontal gene transfer has been revealed between the jointly introduced TM and Micrococcus sp. 9/pSH1 in microcosms with the Lake Shira sterile water., (c2005 COSPAR. Published by Elsevier Ltd. All rights reserved.)

The processes resulting from the introduction of the tranagenic microorganism (TM) E. coli Z905/pPHL7 into aquatic microcosms have been modeled experimentally. It has been shown that the TM E. coli is able to adapt to a long co-existence with indigenous heterotrophic microflora in variously structured microcosms. In more complex microcosms the numerical dynamics of the introduced E. coli Z905/pPHL7 population is more stable. In the TM populations staying in the microcosms for a prolonged time, changes are recorded in the phenotypic expression of plasmid genes (ampicillin resistance and the luminescence level) and chromosome genes (morphological and physiological traits). However, in our study microcosms, the recombinant plasmid persisted in the TM cells for 6 years after the introduction, and as the population adapts to the conditions of the microcosms, the efficiency of the cloned gene expression in the cells is restored. In the microcosms with high microalgal counts (10(7) cells/ml), cells with a high threshold of sensitivity to ampicillin dominate in the population of the TM E. coli Z905/pPHL7., (c2003 COSPAR. Published by Elsevier Science Ltd. All rights reserved.)

A mathematical model was used to investigate the effect of cannibalism intensity on the net primary production and the dynamics of trophic links in an aquatic ecosystem characterized by cannibalism at the upper trophic level. A mathematical model of an aquatic ecosystem has been constructed, with the following principal trophic links: limiting nutrient concentration, producers (phytoplankton), nonpredatory and predatory zooplankton. The model takes into account the age structure of the predator and includes two age groups (the young and adults). The adult predators are cannibals feeding on both nonpredatory zooplankton and their own young, which consume phytoplankton. It has been found that when cannibalism intensity increases above a certain level, the concentrations of both adults and the young of the predators decrease. At the same time, the concentrations of the nonpredatory zooplankton and of nutrients increase, while the biomass of producers decreases. When the cannibalism intensity is low, the net primary production of the system increases to a certain level correlated with the increase in cannibalism intensity and drops sharply when the level of consumption of young is high. There is an optimal intensity of cannibalism, at which the productivity in the photosynthesis link is maximal., (c2003 COSPAR. Published by Elsevier Science Ltd. All rights reserved.)

The anthropogenic impact on the Earth's ecosystems are leading to dramatic changes in ecosystem functioning and even to destruction of them. System analysis and the use of heuristic modeling can be an effective means to determine the main biological interactions and key factors that are of high importance for understanding the development of ecosystems. Cycling of limiting substances, induced by the external free energy flux, and trophic links interaction is the basis of the mathematical modeling studies presented in this paper. Mathematical models describe the dynamics of simplified ecosystems having different characteristics: 1) different degrees of biotic turnover closure (from open to completely closed); 2) different numbers of trophic links (including both "top-down", "bottom-up" regulation types); 3) different intensities of input-output flows of the limiting nutrient and its total amount in the system. Adaptive values of the changes of lower hierarchical levels (populational, trophic chain level) are to be estimated by integrity indices for total system functioning (e.g. NPP, total photosynthesis). The approach developed can be used for evaluating the contributions of lower hierarchical levels to the functioning of the higher hierarchical levels of the system. This approach may have value for determining biomanipulation management and their assessment., (c2003 COSPAR. Published by Elsevier Science Ltd. All rights reserved.)

Simple models of terrestrial ecosystems with a limited number of components are an efficient tool to study the main laws of functioning of populations, including microbial ones, and their communities, as components of natural ecosystems, under variable environmental conditions. Among other factors are the increase of carbon dioxide in the atmosphere and limitation of plants' growth by biogenic elements. The main types of ecosystems' responses to changes in environmental conditions (a change in CO2 concentration) have been demonstrated in a "plants-rhizospheric microorganisms-artificial soil" simple experimental system. The mathematical model of interactions between plants and microorganisms under normal and elevated atmospheric CO2 and limitation by nutrients (nitrogen and phosphorus) yielded a qualitative agreement between calculated and experimental values of limiting substances concentrations and release rates of exudates., (c2003 COSPAR. Published by Elsevier Science Ltd. All rights reserved.)

It has been demonstrated that the transgenic microorganism Escherichia coli Z905/pPHL7 (AprLux+) can exist for a long time at an elevated concentration of mineral salts. The microorganism was introduced into microcosms with sterile brackish water (salinity variable from 21 to 22 g l-1) taken from Lake Shira (Khakasia, Russia). The survival of the microorganism was estimated both by measuring the growth of the colonies on solid nutrient media and by the bioluminescence exhibited by the transgenic strain in samples from the microcosms and in the enrichment culture with the added selective factor-ampicillin (50 micrograms/ml). In the enrichment culture, the bioluminescent signal was registered through the 160-day experiment. It has been shown that in the closed microcosms with brackish water the E. coli strain becomes heterogeneous in its ampicillin resistance. The populations of the transgenic strain were mainly represented by isolates able to persist in the medium containing 50 micrograms/ml, but there were also the cells (about 10%) with the threshold of ampicillin resistance not more than 0.05 micrograms/ml. Thus, it was shown that in the microcosms with brackish water and in the absence of the selective factor the transgenic strain survives and retails the recombinant plasmid., (c2003 COSPAR. Published by Elsevier Science Ltd. All rights reserved.)

A mathematical model describing the interaction of plants and rhizospheric microorganisms on complete mineral medium at a higher CO2 level in the atmosphere was constructed. The positive effect of CO2-enrichment on the system plant--rhizospheric microorganisms was shown. The effect of rhizospheric microorganisms on plant growth at normal and high level of carbon dioxide was demonstrated. It was shown that the biomass of plant in the system is smaller than the biomass of plant growing without microorganisms. It was experimentally demonstrated that a simple ecosystem wheat--Pseudomonas putida--artificial soil develops and functions differently than its individual constituents in the case of a wheat-artificial soil system. With unlimited nutrition and a higher CO2 level (0.06%), plants with roots inoculated with microorganisms have a smaller biomass than plants that were not inoculated with microorganisms.

The copy number of R plasmids weakly depends on the selective pressure of the respective antibiotic but does depend on the physiology of the host species and the type of plasmids and cloned genes, whose expression leads to a further load on the biosynthetic apparatus of cells. The last factor is critical in the maintenance of recombinant plasmids in transgenic microorganisms.

The process of biotic turnover in a closed ecological system (CES) with an external energy flow was analyzed by mathematical modeling of the biotic cycle formation. The formation of hierarchical structure in model CESs is governed by energy criteria. Energy flow through the ecosystem increases when a predator is introduced into a "producer-reducer" system at steady state. Analysis of the model shows that under certain conditions the presence of the primary predator with its high mineralization ability accelerates the biotic turnover measured by primary production. We, therefore, conclude that for every system it is possible to find a suitable predator able to provide the system with a higher biotic turnover rate and energy consumption. Grant numbers: 99-04-96017/2000., (c 2001. COSPAR. Published by Elsevier Science Ltd. All rights reserved.)

A mathematical model was used to study the response of ecosystems of different structures to external impact. The response was measured as a sensitivity coefficient: the magnitude of the system's response vs. the change of the factor in the inflow. The formula has been obtained to calculate the sensitivity coefficient for ecosystems containing different numbers of trophic links. The derived sensitivity coefficients demonstrate that the degree of compensation for the external impact can differ depending on the type of system regulation and the length of the trophic chain. E. g. the sensitivity coefficient decreases with complexity of trophic links in an ecosystem for top-down controlled systems and impact of degree of openness on sensitivity e.g. closed ecosystems show higher sensitivity then fully open ecosystem to impacts also bottom-up control system show less sensitivity then top-down. Grant numbers: N99-04-96017, N25., (c 2001. COSPAR. Published by Elsevier Science Ltd. All rights reserved.)

Effect of the size of rhizospheric bacterial populations on germination of seeds and development of simple terrestrial "wheat plants--rhizospheric microorganisms--artificial soil" and "wheat plants-artificial soil" systems has been studied. Experiments demonstrated that within specify ranges in the inoculate, the rhizospheric bacteria are capable of increasing the yield of germinated seeds and stimulate the growth of plantlets. Germination of seeds inoculated with bacteria was either stimulated, or inhibited or remained at control levels depending on the amount of bacteria. Plant biomass growth and total photoassimilation has been found to depend on the amount of bacteria on the plant roots: the higher the amount of bacteria on plant roots, the smaller is the biomass of plants but the total photoassimilation is, higher. Thus, depending on the amount of bacteria on the roots of plants the system either increases the biomass of plants or increases the total photoassimilation, i.e. "pumps" carbon through itself involving bacteria. Grant numbers: N99-04-96017, N15., (c 2001. COSPAR. Published by Elsevier Science Ltd. All rights reserved.)

The role of key environmental factors in adaptation of spore-forming and non-spore-forming transgenic microorganisms (TM) have been studied in model ecosystems. Model TM Escherichia coli Z905 (bearing plasmid genes of bacterial luminescence Ap (r) Lux+) has been found to have a higher adaptation potential than TM Bacillus subtilis 2335/105 (bearing genes of human alpha 2-interferon Km (r) Inf+), planned for employment as a living vaccine under varying environmental conditions. Effects of abiotic factors on migration of natural and recombinant plasmids between microorganisms under model ecosystem conditions has been estimated. The transgenic microorganisms with low copy number survived better under introduction conditions in the microcosms studied. This trend has been shown to be independent of the microcosm type and its complexity. Grant numbers: 99-04-96017, 25, 00-07-9011., (c 2001. COSPAR. Published by Elsevier Science Ltd. All rights reserved.)

Functional, regulatory and indicator features of microorganisms in development and functioning of the systems and sustaining stability of three man-made ecosystem types has been studied. 1) The functional (metabolic) feature was studied in aquatic ecosystems of biological treatment of sewage waters for the reducer component. 2) The regulatory feature of bacteria for plants (producer component) was studied in simple terrestrial systems "wheat plants-rhizospheric microorganisms-artificial soil" where the behavior of the system varied with activity of the microbial component. For example with atmospheric carbon dioxide content elevated microbes promote intensification of photosynthesis processes, without binding the carbon in the plant biomass. 3) The indicator feature for the humans (consumer component) was studied in Life Support Systems (LSS). High sensitivity of human microflora to system conditions allowed its use as an indicator of the state of both system components and the entire ecosystem. Grant numbers: N99-04-96017, N15., (c 2001. COSPAR. Published by Elsevier Science Ltd. All rights reserved.)

High reproduction rates make the bacterial component of ecosystems a good indicator of the state of the system on the whole. This determines the necessity to develop rapid monitoring of the functional state of the bacterial component of small ecosystems. Information about substrate concentration in the population is indicative of the state of the bacterial culture. Conventional methods of monitoring the concentration of integral substrate in the system take time much longer than the changes in the ecosystem. The paper presents theoretical foundations for the logical sequence "catalase activity--intracellular substrate concentration--estimate of substrate consumed by bacteria" for experimental verification and as a consequence of development of the integral method of monitoring the bacterial population on the basis of determining bacterial catalase activity. Grant numbers: 04-96017, N25., (c 2001. COSPAR. Published by Elsevier Science Ltd. All rights reserved.)

The work analyzes functioning of a "producer-consumer" closed aquatic system with spatially separated links, where each component consisted of two species. Producers in the system were the microalgae of Chlorella vulgaris and Scenedesmus sp., consumers--Paramecium caudatum infusoria and Brachionus sp. rotifers. In the experiment the competing predators were consuming on a mixed culture of algae, and the competition of algae was studied under nitrogen limitation. Under these conditions competitiveness of Scenedesmus was higher than that of Chlorella vulgaris. Metabolism products of Scenedesmus algae have been found to have negative effect on reproduction of Paramecium caudatum protozoa. Predator population dynamics in the "consumer" link demonstrated that the rotifers that consume two algal species are more competitive compared to protozoa feeding on chlorella only. Grant numbers: N99-04-96017, N25., (c 2001. COSPAR. Published by Elsevier Science Ltd. All rights reserved.)

Closed Artificial ecosystems (CAES) have good prospects for wide use as new means for quantitative studies of different types of both natural ecosystems and man-made ones. The paper deals with the discussion of three points of CAES applications. The first one is of importance for theoretical ecology development and is connected with bringing together "holistic" and "merological" approaches in ecosystems studies. Using CAES, we can combine both approaches, taking into account the biotic turnover of limiting substrates which few in number even for complicated natural ecosystems. The second CAES use concerns the development of "ecosystems health" concept and application of a key-factor-approach for the indication and measurement of healthy unhealthy state and functioning of ecosystems or their links. The third use is more of an applied nature, oriented to the intensification of bioremediation or biodepollution processes in different types of ecosystems, including the global biosphere. Grant numbers: N 99-04-96017, N25., (c 2001. COSPAR. Published by Elsevier Science Ltd. All rights reserved.)

A structural approach to studying the regularities of the population dynamics of unstable recombinant bacterial strains in a chemostat was elaborated. The approach is based on the mathematical modeling of cell distribution in a population with different numbers of plasmid copies. The effect of decreased selective preference of plasmidless variants of the recombinant strain in the chemostat, which is related to a decrease in the number of plasmid copies in cells upon long-term incubation was analyzed. It is shown that the time of half-elimination of plasmids from the bacterial population in the steady state in the chemostat T1/2 does not depend on the maximum number of plasmid copies in cells N but is determined only by the mean time of generation g and the probability of the loss of one plasmid copy tau. The dependence of the preference of bacterial plasmidless variants on the efficiency of expression of genes cloned into plasmids in chemostat was analyzed using the recombinant strain E. coli Z905, whose plasmids pPHL-7 contain cloned genes for the luminescence system of marine luminescing bacteria Photobacterium leiognathi.

The population heterogeneity of recombinant and plasmid-free Bacillus subtilis strains introduced into aquatic microcosms was studied. After introduction, the population of the plasmid-free strain B. subtilis 2335 in microcosms has long been represented by both vegetative cells and spores, whereas, already ten days after introduction, the population of the recombinant strain B. subtilis 2335/105 (Km[symbol: see text]nf+) was represented only by spores. The number of plasmid copies in the spore isolates of the recombinant strain was the same as before introduction, but the plasmid abundance in the vegetative isolates of this strain decreased. The isolates of B. subtilis 2335/105 obtained from microcosms and the variants of this strain obtained by ten successive subcultures on M9 and 0.1 x M9 media with and without kanamycin (Km) differed in the number of plasmid copies, Km resistance, and maximum biomass yield during batch cultivation. Irrespective of the presence of Km, more than 50% of the variants subcultured on M9 medium showed reduced plasmid abundance. At the same time, about 70% of the variants subcultured on 0.1 x M9 medium with Km and 90% of the variants subcultured on the same medium without Km retained the initial number of plasmid copies. The variants subcultured on media with Km retained the initial biomass level. In more than 70% of the variants isolated from media without Km, the biomass yield increased.

Effect of different concentrations of salts on natural and recombinant strains of Bacillus subtilis and Escherichia coli was studied. The recombinant strain of B. subtilis was found to be more osmotolerant than the wild-type strain of this bacterium, whereas the opposite situation was observed for the recombinant and wild-type strains of E. coli. Some salts exerted a bacteriostatic effect on E. coli and B. subtilis. The adaptive capacity of recombinant strains depended on the number of plasmid copies in the cells. The introduction of recombinant bacteria into model ecosystems resulted in the generation of their variants with increased osmotolerance.

This article deals with some methodological aspects of biospherics connected with theoretical sciences development and prospective use for practical application. Properties of experimental objects, methods and goals of biospherics as synthesising science have been discussed. The problem of stability of incomplete (natural and artificial) ecosystems has been considered. The concept of the ecosystem health based on effective functioning of different types of ecosystems has been developed.

Experimental and theoretical models of closed "autotroph-heterotroph" (chlorella-yeast, chlorella-protozoa) ecosystems with spatially separated components have been created and studied. The chart of flows and interaction of components of gas-closed "chlorella-yeast" system have formed the basis describe mathematically the functioning of the given system, experimental results have been found to agree with computer solution of the problem in terms of quality. Investigation of the experimental model of the "producer-consumer" trophic chain demonstrated the role of protozoa in nitrogen turnover. "Production-decomposition" and "production-grazing-decomposition" cycle models has been theoretically analyzed and compared. The predator has been shown to be a more intensive mineralizer than the reducer component.

Experimental models of small biotic cycles of different degree of closure and complexity with spatially separated components based on unicellular organisms have been studied. Gas closure of components looped into "autotroph-heterotroph" (chlorella-yeast) system doubled the lifetime of the system (as opposed to individually cultivated components). Higher complexity of the heterotroph component consisting of two yeast species also increased the lifetime of the system through more complete utilization of the substrate by competing yeast species. The lifetime of gas and substrate closed "producer-consumer" trophic chain (chlorella-paramecia) increased to 7 months. In 60 days the components' numbers reached their steady state followed by more than 40 cycles of the medium. The role of a predator organism (protozoan) in nitrogen cycling was demonstrated; reproduction of protozoa correlated directly with their emission of nitrogen in the ammonia form that is most optimum for growth of chlorella.

A mathematical model concerning the interaction of plants and rhizospheric microorganisms on complete mineral medium and under nitrogen limitation has been constructed. The model takes into account the closeness of plants and microorganisms in terms of the matter released by the plant and consumed by the microorganisms. The effect of rhizospheric microorganisms on plant growth with normal carbon dioxide and complete mineral medium has been demonstrated. Plants interacting with microorganisms have a greater biomass than plants growing without microorganisms. Wheat growth stimulation by metabolites of rhizospheric microorganisms under laboratory conditions on artificial soil has been experimentally demonstrated (Pechurkin, 1997). Under nitrogen limitation, the biomass of plants, with or without microorganisms, is identical, and is substantially reduced as compared with the medium with standard nitrogen.

Key factors of ecosystem functioning are of the same nature for artificial and natural types. An hierarchical approach gives the opportunity for estimation of the quantitative behavior of both individual links and the system as a whole. At the organismic level we can use interactions of studied macroorganisms (man, animal, higher plant) with selected microorganisms as key indicating factors of the organisms immune status. The most informative factor for the population/community level is an age structure of populations and relationships of domination/elimination. The integrated key factors of the ecosystems level are productivity and rates of cycling of the limiting substances. The key factors approach is of great value for growth regulations and monitoring the state of any ecosystem, including the life support system (LSS)-type.

An experimental approach for investigation of genetically modified microorganisms (GMMO) introduced into model ecosystems to evaluate potential risk of propagation of recombinant plasmids in surrounding medium has been developed. The object of modeling was Escherichia coli Z905 strain with a recombinant plasmid with bacterial luminescence genes, which was introduced into water microcosms of different structure. The approach involves comprehensive investigation of GMMO at four hierarchical levels: molecular (retaining the structure of the plasmid and expression of cloned genes); cellular (variation of metabolic activity); population (competitive power and metabolic interactions of GMMO with indigenous microflora, migration of recombinant and natural plasmids); ecosystem (effect of GMMO and cloned genes on ecosystem parameters). The experimental evidence and theoretical estimates are intended to form grounds to develop a basic version of an ecological certificate for different GMMO variants.

Response of "wheat plants--rhizospheric microorganisms--artificial soil"--a simple terrestrial ecosystem--to carbon dioxide increased in its atmosphere to 0.06% has been studied. It has been experimentally demonstrated that a simple ecosystem develops and functions different from its individual elements (components), in this case "plants-artificial soil" without microorganisms. With mineral nutrition unlimited and CO2 enrichment the system is capable of binding (involving into turnover) 40% more carbon than the system without microorganisms. With material balance as the basis, this article evaluates the contribution of a system's elements into its development, namely, the contribution of the photosynthesizing component and the contribution of microorganisms.

The possibility of introducing genetically engineered microorganisms (GEM) into simple biotic cycles of laboratory water microcosms was investigated. The survival of the recombinant strain Escherichia coli Z905 (Apr, Lux+) in microcosms depends on the type of model ecosystems. During the absence of algae blooming in the model ecosystem, the part of plasmid-containing cells E. coli decreased fast, and the structure of the plasmid was also modified. In conditions of algae blooming (Ankistrodesmus sp.) an almost total maintenance of plasmid-containing cells was observed in E. coli population. A mathematics model of GEM's behavior in water ecosystems with different level of complexity has been formulated. Mechanisms causing the difference in luminescent exhibition of different species are discussed, and attempts are made to forecast the GEM's behavior in water ecosystems.

We studied preservation of recombinant Escherichia coli strain Z905 (AprLux+) in liquid microecosystems (LME) after the introduction. E. coli cells were shown to remain viable and preserve the ability to express the cloned lux genes for a long time (more than a year) in LME. The majority of the clones have reduced efficiency of the expression due to either changed regulation of the lux operon or decreased number of copies of the plasmid. These mechanisms could be realized either independently or simultaneously depending on LME conditions. We have exposed the major factors affecting the metabolic activity of the E. coli strain Z905 (AprLux+) introduced into model ecosystems and the level of expression of the cloned genes.

Expression of the lux-genes cloned on the recombinant plasmid pPHL7 (AprLux+) in Escherichia coli z905 cells was studied in various environments, including model aquatic ecosystems. Expression of the lux-genes strongly depended on the nutritional status of the medium. In particular, the cultivation of cells in nutrient-rich medium favored the maintenance of the initial level of expression of the lux-operon, whereas nutrient limitation induced recombinant cell variants with an impaired control of the catabolite-dependent lux-operon. On the other hand, long-term laboratory cultivation of recombinant strain in nutrient-deficient media or its long-term life in model aquatic ecosystems led to the accumulation of cells with a stringent control on the cloned lux-genes in the bacterial population. The presence of the selective factor (ampicillin) in the medium had no significant effect on the expression of the lux-operon.

The effect has been studied of inoculation of seeds of wheat with two species of rhizospheric microorganisms--Pseudomonas fluorescens and Pseudomonas putida--on young plant growth with complete and with nitrogen deficit mineral nutrition. With complete mineral medium, plants grown from seeds inoculated with bacteria of Pseudomonas genus (experiment plants) have been found to have better growth over plants not inoculated with these bacteria (control plants). The experiment plants had increased transpiration and their biomass had higher organic nitrogen content. With nitrogen deficit medium, the plants inoculated with bacteria and those without them, have not revealed changes in growth. Neither case demonstrated competition of microorganisms with plants for nitrogen sources.

The behavior of Escherichia coli Z905, carrying a recombinant plasmid pPHL7 with genes determining ampicillin resistance and bacterial luminescence, and the efficiency of expression of cloned genes were studied after introduction of the strain into model aqueous ecosystems with different trophic chain lengths. The E. coli Z905 variants isolated from ecosystems after different periods of time were found to vary in their resistance to ampicillin (from 50 to 0.05 micrograms/ml) and in the intensity of bioluminescence. An increase in the concentration of the selective factor (ampicillin) or in the extent of the aqueous microcosm blooming restored the expression of the recombinant plasmid genes in some clones.

The populational structure of the Escherichia coli strain Z905 containing the recombinant plasmid with the phenotype AprLux+ was studied in chemostat. It was shown that the stability of the ratio of plasmid containing cells and cells without plasmids depends in the first place on the presence of the selective factor (ampicillin) in the medium and on the sources of carbon and energy limiting growth.

The effect of phenol on the growth rate and respiration was studied with the yeast Candida mycoderma cultivated in the pH-static conditions with continuous recording of the principal kinetic parameters of the population. The kinetics of growth inhibition with phenol was studied. Adaptation of the culture in terms of the growth rate and the rate of oxygen uptake was detected within 10--15 hours of cultivation. A new strain of C. mycoderma Phen. R. isolated using the technique of autoselection upon continuous cultivation for a long period of time, at a phenol concentration of 1.5 g/l in the growth medium, had an elevated growth rate (2.2 times higher) in these conditions as compared with the parent culture and required less oxygen.

The growth rate of the phage for Brevibacterium was studied as a function of the population of the host bacterium and its growth rate. The interaction was not additive when the phage-bacterium system was modeled in chemostat and in periodic regime. This fact was taken into account to describe more precisely and completely the qualitative characteristics of such a system as compared to models of the predator-victim type.

The physicochemical properties of a model medium consisting of two substances, sodium sulfite and albumin, were studied. It was shown that the presence of a SAS in the sulfite solution significantly lowered the rate of oxygen sorption by the model medium. The higher the SAS concentration, the lower the sorption rate. When yeasts were grown on sulfite liquor under conditions of the reduced medium the rate of their growth and respiration did not significantly depend on aeration within wide ranges of its variation. The experiments with the model medium demonstrated that the aeration independence was due to indifference to oxygen not of the cells but of the medium as the result of its physicochemical properties. The effect of sorption hysteresis was also studied on the model medium. It was revealed that the rate of oxygen desorption from the medium was lower than that of oxygen sorption by the same medium. The effect was not observed when a nonionic SAS was added to the medium and the isoelectric point was almost reached. The effect was connected with impairment of oxygen diffusion through the medium surface film. Interaction of the substances included in the model medium was of physical nature.

The models of evolution of Eigen's hypercycles and microbial populations were compared. It has been proved that both above mentioned model descriptions of evolution completely coincide for the case of support constant organization (1). It has been shown that in the case of constant flows (2) for the Eigen's model it is necessary to take into account the dynamics of concentration of energy monomers. The experimental results of microevolution of microbial populations for conditions 1 and 2 are presented.

The aim of this work was to investigate phenol decomposition by Candida tropicalis 29-10 adapted to phenol as a sole source of carbon and energy, and to elucidate the nature of resistance of continuous cultures decomposing phenol in processes with two types of control, viz. in chemostat and in pH-stat. The results suggest the resistance of the pH-stat regime of microbial cultivation during substrate inhibition. A region of instability in the chemostat cultivation has been determined when the yeast growth was inhibited with phenol.