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The study describes a small closed ecosystem used to test technologies to be further employed in full-scale manned closed ecosystems. The experimental ecosystem is designed to use a certain portion of human metabolism, which is included in the gas, water, and organic waste loops of the system. In this experimental ecosystem, gas and water loops are fully closed, and the model enables processing of human waste and plant inedible biomass. A physicochemical method is used to remove pollutants from the air in the system. A human takes part in the gas exchange of the system through its respiration loop. This experimental ecosystem can be used for testing and improving new technologies to be further used in the future space stations., (Copyright © 2018 The Committee on Space Research (COSPAR). Published by Elsevier Ltd. All rights reserved.)

The flow of long-chain polyunsaturated fatty acids (PUFAs) of the omega-3 family, namely, eicosapentaenoic acid (20:5n-3, EPA) and docosahexaenoic acid (22:6n-3, DHA), exported by amphibian metamorphs from water to terrestrial ecosystems in the Medveditsa River floodplain, was quantified for the first time. The total biomass export by three amphibian species (Pelobates fuscus, Bombina bombina, and Pelophylax ridibundus) per unit area of the lake surface was 0.594 g/m 2 per year (as a mean for 2 years). The biomass flow per unit area of land was 0.726 g/ha per year (0.302 g/ha per year for organic carbon) in 2015-2016. The average annual total removal of EPA + DHA by amphibians from the floodplain lake was 1.47 mg/m 2 of water surface area. Due to the high content of EPA and DHA in biomass, amphibians are potentially a valuable food for terrestrial predators having no access to other sources of essential PUFAs.

The study of the composition of fatty acid markers of tadpoles of cohabiting amphibian species for the first time revealed differences in their diets: the moor frog Rana arvalis prefers bacteria not associated with plant detritus, whereas the diet of the common spadefoot Pelobates fuscus is based on cyanobacteria, green algae, diatoms, and possibly higher plants. Major differences in the fatty acid composition are determined by the difference in the percentage of eicosapentaenoic and myristic acids.

Methods of physicochemical further oxidation of hardly soluble sediment obtained from "wet combustion" of human exometabolites applied to space-purpose Bio Technological Life Support Systems (BTLLS) were studied. Most hardly dissoluble sediment containing Ca, P, Mg, and other essential plant nutrition elements were shown to dissolve in H 2 O 2 and HNO 3 aqueous media activated by alternating electric current. Dissolved additional mineral elements allowed (as demonstrated for lettuce) to increase the productivity of BTLLS phototrophic unit plants more than twice, which is comparable to their productivity on standard Knop solution with balanced chemical composition. Thus, dissolved mineral elements can be involved into BTLLS turnover process and increase its closure degree.

Synthesis of polyhydroxyalkanoates (PHAs) by a new strain of Cupriavidus - Cupriavidus eutrophus B-10646 - was investigated under autotrophic growth conditions. Under chemostat, at the specific flow rate D=0.1h(-1), on sole carbon substrate (CO2), with nitrogen, sulfur, phosphorus, potassium, and manganese used as growth limiting elements, the highest poly(3-hydroxybutyrate) [P(3HB)] yields were obtained under nitrogen deficiency. In batch autotrophic culture, in the fermenter with oxygen mass transfer coefficient 0.460 h(-1), P(3HB) yields reached 85% of dry cell weight (DCW) and DCW reached 50 g/l. Concentrations of supplementary PHA precursor substrates (valerate, hexanoate, γ-butyrolactone) and culture conditions were varied to produce, for the first time under autotrophic growth conditions, PHA ter- and tetra-polymers with widely varying major fractions of 3-hydroxybutyrate, 4-hydroxybutyrate, 3-hydroxyvalerate, and 3-hydroxyhexanoate monomer units. Investigation of the high-purity PHA specimens showed significant differences in their physicochemical and physicomechanical properties., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Microbial polyhydroxyalkanoates (PHAs), because of their well studied complex physiology and commercial potential, are vehicles for carbon and potential storage reduction for many microbial species. Even with the wealth of studies about microbial PHAs in the scientific literature, polymer accumulation and degradation are still not comprehensively understood. Poly(3-hydroxybutyrate) (P3HB) granule formation and polymer mobility were studied here in the bacterium Ralstonia eutropha strain B5786 in autotrophic cultures. Electron microscopy studies revealed decreasing cell size concomitant with enlargement of size and number of intracellular granules, and inhibition of cell division during intracellular polymer production. Activities of key P3HB biosynthetic enzymes demonstrated correlations with each other during polymer accumulation, suggesting an intricately regulated P3HB cycle in autotrophically grown R. eutropha cells., (Copyright © 2012. Published by Elsevier Masson SAS.)

Synthesis of 3- and 4-hydroxybutyrate copolymer (3HB-CO-4HB), the most promising member of the biodegradable polyhydroxyalcanoate (PHA) family, has been studied. Cultivation conditions of naturally occurring strains of hydrogen-oxidizing bacteria Ralstonia eutropha B5786 and Cupriavidus eutrophus B10646 have been optimized to ensure efficient synthesis of the 3HB-CO-4HB copolymer. A set of highly pure samples of the 3HB-CO-4HB copolymer with 4HB content varying from 8.7 to 24.3 mol % has been obtained. Incorporation of 4-HB into the copolymer was shown to cause a more pronounced decrease in polymer crystallinity than the incorporation of 3-hydroxyvalerate or 3-hydroxyhexanoate; samples with a degree of crystallinity below 30% have been obtained. The weight average molecular mass of the 3HB-CO-4HB copolymers was shown to be independent on the monomer ratio and to vary broadly (from 540 to 1110 kDa).

Microparticles obtained by using (14)C-labeled resorbable hydroxyaminobutyric acid polymer were injected into the caudal vein of laboratory animals without negative aftereffects for their growth and development and without changes in the macro- and microstructure of organs and tissues. The distribution of microparticles in the viscera and the dynamics of accumulation of carbon-containing polymer degradation products in the viscera were studied. The main targets for the particles are liver tissues, as well as renal and splenic tissues. The polymeric matrix of the microparticles is most actively destroyed in the spleen and liver. The presence of high-molecular-weight polymeric matrix in organs indicates the integrity of microparticles and the possibility of long-term (up to 12 weeks) functioning of polymeric particles in vivo.

The possibility of use of polyhydroxyalkanoates (PHAs), biodegradable microbial polyesters, as a carrier for pesticides (alpha-hexachlorocyclohexane and lindane) for targeted and controlled delivery of these compounds to soil was investigated. The kinetics of polymer degradation and the dynamics of pesticide release from the extended-release formulations was studied. It is shown that pesticides embedded in a degradable polymer (PHA) carrier are released gradually and slowly, without surges, as the polymer is degraded by the soil microflora. The microbial soil component actively responded to the addition of the polymer as an additional nutrient substrate: the latter was degraded and then utilized. The rate of the pesticide release to the soil can be regulated by varying the polymer-pesticide ratio.

The ability of marine luminescent bacteria to synthesize polyesters of hydroxycarboxylic acids (polyhydroxyalkanoates, PHA) as reserve macromolecules was studied. Twenty strains from the collection of the luminescent bacteria CCIBSO (WDSM839) of the Institute of Biophysics, Siberian Branch, Russian Academy of Sciences, assigned to different taxa (Photobacterium leiognathi, Ph. phosphoreum, Vibrio harveyi, and V. fischeri) were analyzed. The most productive strains were identified, and the conditions ensuring high polymer yields in batch culture (40-70% of the cell dry mass weight) were determined. The capacity of synthesizing two- and three-component polymers containing hydroxybutyric acid as the main monomer and hydroxyvaleric and hydroxyhexanoic acids was revealed in Ph. leiognathi and V. harveyi strains. The results allow luminescent microorganisms to be regarded as new producers of multicomponent polyhydroxyalkanoates.

The effect of carbon supply on polyhydroxyalkanoate (PHA) synthesis by bacteria Wautersia eutropha was studied. Synthesis of multicomponent PHA composed of short- and long-chain monomers (C4-C8) by two natural strains (H16 and B5786) under mixotrophic conditions (CO2 + alkane acids as cosubstrates) was demonstrated for the first time. The PHA composition was shown to be dependent on the cosubstrate type. In the presence of odd fatty acids, four- and five-component polymers were synthesized; hydroxybutyrate, hydroxyvalerate, and hydroxyheptanoate were the major monomers, while hydroxyhexanoate and hydroxyoctanoate were minor and irregular. In the presence of even fatty acids, PHA contained not only the corresponding molecules (hydroxyhexanoate and hydroxyoctanoate), but also hydroxyvalerate; synthesis of four-component PHA which contain mainly hydroxybutyrate and hydroxyhexanoate (up to 18 mol %) is therefore possible. A series of four- and five-component PHA was synthesized and their physicochemical characteristics were determined.

The fatty acid composition of the lipids of a Wautersia eutropha polyhydroxyalkanoate-producing strain was studied by chromato-mass spectrometry. A total of 27 fatty acids were identified; their distribution in the cell fractions was determined. In the cytoplasmic membrane, palmitic, palmitoleic, and cis-vaccenic acids were the major components. Long-chain beta-hydroxy acids and myristic acid (components of the lipopolysaccharides of the cell envelope) predominated in the fraction of strongly bound lipids. When the polymer was actively synthesized, the content of cyclopropane acids in the easily extracted lipids increased and the content of the corresponding monoenoic acids decreased. The strongly bound lipids had a high content of long-chain beta-hydroxy acids (more than 50% of the total fatty acids). These results made it possible to determine the source of polyhydroxyalkanoate (PHA) contamination and to choose the strategy for their purification.

Physiological-biochemical, genetic, and cultural properties of the glucose-utilizing mutant strain Ralstonia eutropha B8562 have been compared with those of its parent strain R. eutropha B5786. It has been shown that growth characteristics of the strain cultured on glucose as the sole carbon and energy source are comparable with those of the parent strain. Strain B8562 is characterized by high polyhydroxyalkanoate (PHA) yields on different carbon sources (CO(2), fructose, and glucose). PHA accumulation in the strain batch cultured on glucose under nitrogen deficiency reaches 90 %. The major monomer in the PHA is beta-hydroxybutyric acid (more than 99 mol %); the identified minor components are beta-hydroxyvaleric acid (0.25-0.72 mol %) and beta-hydroxyhexanoic acid (0.08-1.5 mol %). The strain is a promising PHA producer on available sugar-containing media with glucose.

Composition and contents of fatty acids in hunchback, fresh, boiled, fried, roasted and boiled in a small amount of water were determined. It was found, that absolute contents of essential polyunsaturated fatty acids--eicosapentaenoic (EPA) and docosahexaenoic (DHA) decreased significantly only in the process of frying. Sum ratio of omega3/omega6 acids was dietetically more favorable in the roasted and fried fish.

The physiological, biochemical, genetic, and cultural characteristics of the glucose-utilizing mutant strain Ralstonia eutropha B8562 were investigated in comparison with the parent strain R. eutropha B5786. The morphological, cultural, and biochemical characteristics of strain R. eutropha B8562 were similar to those of strain R. eutropha B5786. Genetic analysis revealed differences between the 16S rRNA gene sequences of these strains. The growth characteristics of the mutant using glucose as the sole carbon and energy source were comparable with those of the parent strain grown on fructose. Strain B8562 was characterized by high yields of polyhydroxyalkanoate (PHA) from different carbon sources (CO2, fructose, and glucose). In batch culture with glucose under nitrogen limitation, PHA accumulation reached 90% of dry weight. In PHA, beta-hydroxybutyrate was predominant (over 99 mol %); beta-hydroxyvalerate (0.25-0.72 mol %) and beta-hydroxyhexanoate (0.008-1.5 mol %) were present as minor components. The strain has prospects as a PHA producer on glucose-containing media.

The paper deals with the study of the synthesis of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) copolymers by the bacterium Ralstonia eutropha B-5786 grown under different carbon nutrition conditions (growth on carbon dioxide, fructose, and CO2-valerate and fructose-valerate mixtures). The parameters to be analyzed included the yield of biomass, the yield, synthesis rate, and composition of copolymers, the activity of the key enzymes of polyhydroxyalkanoate (PHA) synthesis (beta-ketothiolase, acetoacetyl-CoA reductase, and PHA synthase), the maximum tolerable concentration of valerate to the bacterium, and the conditions that govern the incorporation of hydroxyvalerate to copolymers. This allowed the relationship between cultivation conditions and the proportion of monomers in the copolymers to be deduced. We were able to synthesize a range of 3HB/3HV copolymers and found that the thermal characteristics and the degree of crystallinity of these copolymers depend on the molar fraction of 3HV.

The dynamics of accumulation of polyhydroxybutyrate (PHB) and the activities of the key enzymes of PHB metabolism (beta-ketothiolase, acetoacetyl-CoA reductase, PHA synthase, D-hydroxybutyrate dehydrogenase, and PHA depolymerase) in the hydrogen bacterium Ralstonia eutropha B5786 were studied under various conditions of carbon nutrition and substrate availability. The highest activities of beta-ketothiolase, acetoacetyl-CoA reductase, and PHA synthase were recorded at the stage of acceleration of PHB synthesis. The activities of enzymes catalyzing PHB depolymerization (PHB depolymerase and D-hydroxybutyrate dehydrogenase) were low, being expressed only at stimulated endogenous PHB degradation. The change of carbon source (CO2 or fructose) did not cause any marked changes in the time course of enzyme activity.

Samples of olyhydroxyalkanoates (PHAs), polyhydroxybutyrate (PHB) and copolymers poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) with 4 and 18 mol% hydroxyvalerate, synthesized by the bacteria Ralstonia eutropha B5786, were investigated. PHA films in contact with blood did not activate the hemostasis system at the level of cell response, but they did activate the coagulation system and the complement reaction. To detect biologically-active components in the PHAs, a detailed analysis of the composition of the polymers was conducted. Gas chromatography-mass spectrometry revealed long-chain fatty acids (FAs) in the tested PHAs. Their total concentration in the polymer ranged from tenths of mol% to 2-3 mol%, depending on the purification method. C16:0 constituted the largest proportion, up to 70%. Of the long-chain hydroxy acids, only beta-OH-C14:0 was detected and it did not exceed 0.06 mol%. The analysis of the hemocompatibility properties of the PHAs purified by a specialized procedure, including the quantitative and morphological estimation of platelets adherent to the surface of polymer films, the plasma recalcification time and complement activation studies, indicated that PHB and PHBV can be used in contact with blood. It has been found out that the lipopolysaccharides of bacteria producing PHAs, which contain mostly long-chain hydroxy acids, can be the factor activating the hemostasis systems. Thus, the technology of PHA purification must satisfy rather stringent specific requirements.

The effect of elevated temperatures of 35 and 45 degrees C (at the intensities of photosynthetically active radiation 322, 690 and 1104 micromoles m-2 s-1) on the photosynthesis, respiration, and qualitative and quantitative composition of the volatiles emitted by wheat (Triticum aestuvi L., cultivar 232) crops was investigated in growth chambers. Identification and quantification of more than 20 volatile compounds (terpenoids--alpha-pinene, delta 3 carene, limonene, benzene, alpha- and trans-caryophyllene, alpha- and gamma-terpinene, their derivatives, aromatic hydrocarbons, etc.) were conducted by gas chromatograph/mass spectrometry. Under light intensity of 1104 micromoles m-2 s-1 heat resistance of photosynthesis and respiration increased at 35 degrees C and decreased at 45 degrees C. The action of elevated temperatures brought about variations in the rate and direction of the synthesis of volatile metabolites. The emission of volatile compounds was the greatest under a reduced irradiation of 322 micromoles m-2 s-1 and the smallest under 1104 micromoles m-2 s-1 at 35 degrees C. During the repair period, the contents and proportions of volatile compounds were different from their initial values, too. The degree of disruption and the following recovery of the functional state depended on the light intensity during the exposure to elevated temperatures. The investigation of the atmosphere of the growth chamber without plants has revealed the substances that were definitely technogenic in origin: tetramethylurea, dimethylsulfide, dibutylsulfide, dibutylphthalate, and a number of components of furan and silane nature., (c2003 COSPAR. Published by Elsevier Science Ltd. All rights reserved.)

The lipid composition of the green alga Botryococcus was studied at three different cultivation temperatures: suboptimal, optimal, and supraoptimal. Cultivation at the supraoptimal temperature was found to considerably inhibit the synthesis of nearly all intracellular lipids, except for triacylglycerides, and to influence their fatty acid composition. In particular, the content of trienoic fatty acids was significantly lower at the supraoptimal than at the optimal cultivation temperature. At the same time, the fatty acid composition of the extracellular lipids of the alga virtually did not depend on cultivation temperature.

It has been found that the carbon monoxide (CO)-resistant strain of the hydrogen bacteria Ralstonia eutropha B5786 is able to synthesise polyhydroxyalkanoates (PHAs) in the presence of CO under autotrophic conditions. This strain, grown on model gas mixtures containing 5-25% CO (v/v), accumulates up to 70-75% (of absolutely dry matter) PHA, without significant variation in the yield coefficient on hydrogen. No suppression of the activities of the key enzymes of PHA synthesis ( beta-ketothiolase, acetoacetyl-CoA-reductase, butyrate dehydrogenase and poly-3-hydroxybutyrate synthase) was recorded. The PHA synthesised is a co-polymer containing mostly beta-hydroxybutyrate (more than 99 mol%) with trace amounts of beta-hydroxyvalerate. The investigated properties of the polymer (molecular weight, crystallinity, temperature characteristics) do not differ from those of the polymer synthesised on electrolytic hydrogen.

The effects of elevated temperature (35 and 45 degrees C) on photosynthesis, respiration, and both the qualitative and quantitative compositions of volatile emissions (VE) of wheat (Triticum aestuvum L. cultivar 232) cenoses at light intensities of 70, 150, or 240 W/m2 of photosynthetically available radiation (PAR) were studied. At a PAR of 240 W/m2, the thermal stabilities of photosynthesis and respiration increased at 35 degrees C and decreased at 45 degrees C. Elevated temperatures nonuniformly changed the rates and direction of VE syntheses. In this process, the highest increase in VE evolution was observed at 70 W/m2; the lowest, at 240 W/m2 and 35 degrees C. In addition, the concentrations and composition of VE during the repair period differed from the initial values.

The CO-resistant strain B5786 of the hydrogen-oxidizing bacterium Alcaligenes eutrophus was found to be able to synthesize polyhydroxyalkanoates (PHAs) under the conditions of growth limitation by nitrogen deficiency (the factor that promotes PHA synthesis) and growth inhibition by carbon monoxide. The gas mixtures that contained from 5 to 20 vol% CO did not inhibit the key enzymes of PHA synthesis--beta-ketothiolase, acetoacetyl-CoA reductase, hydroxybutyrate dehydrogenase, and PHA synthase. In the presence of CO, cells accumulated up to 70-75 wt% PHA (with respect to the dry biomass) without any noticeable increase in the consumption of the gas substrate. Chromatographic-mass spectrometric analysis showed that the PHA synthesized by A. eutrophus is a copolymer containing more than 99 mol% beta-hydroxybutyrate and trace amounts of beta-hydroxyvalerate. The PHA synthesized under the conditions described did not differ from that synthesized by A. eutrophus cells from electrolytic hydrogen.

The paper describes the composition of extracellular free fatty acids (FFAs) and intracellular fatty acids (FAs) in the enrichment cultures of the prokaryotic alga Spirulina platensis and the eukaryotic alga Chlorella vulgaris grown at optimal, supraoptimal, and suboptimal growth temperatures. With increasing growth temperature, the degree of unsaturation of the intracellular FAs of both algae decreased, while that of the extracellular FFAs of S. platensis increased. The composition of the extracellular FFAs of C. vulgaris practically did not depend on the growth temperature.

The lipid fraction of the green alga Botryococcus cultured in a batch mode was found to contain polar lipids (more than 50% of the total lipids), di- and triacylglycerides, steroids and their esters, free fatty acids, and hydrocarbons. In senescent culture, the content of polar lipids somewhat decreased and that of triacylglycerides increased by more than four times. The content of hydrocarbons in the algal biomass did not exceed 0.9% and depended little on the culture age. Intracellular lipids contained saturated and unsaturated (mono-, di-, and trienoic) fatty acids. The maximum content of C16:3 and alpha-C18:3 fatty acids (up to 35% of the total fatty acids) was detected in the phase of active growth. The extracellular and intracellular lipids of the alga differed in the proportion of particular lipids and in the fatty acid pattern.

Growth of Seliberia carboxydohydrogena was inhibited by CO at 10 to 40% (v/v), resulting in increased substrate utilization and enhanced synthesis of cytochromes and cyclopropane and saturated fatty acids. The bacteria showed increased formation of new membrane structures, with pronounced folding of their cell walls.

The composition of lipids was studied in three strains of carboxydobacteria, viz. Seliberia carboxydohydrogena Z-1062, Pseudomonas gazotropha Z-1156 and Comamonas compransoris Z-1155. The content of lipids extracted with an alcohol--chloroform mixture varied in these strains from 9 to 13.2%. The main classes were phospholipids (from 62.6 to 77.2% of the overall lipid content). The major phospholipids were phosphatidyl ethanolamine and phosphatidyl choline. Moreover, the strains Z-1062 and Z-1156 contained three unidentified phospholipids which constituted 40 and 20% of the total content respectively. The neutral lipids of the carboxydobacteria comprised diglycerides, triglycerides, free fatty acids, methyl esters of fatty acids and a number of compounds which were not identified. The fatty acids of the strains were represented by saturated, monoene and cyclopropanoic acids. The main acids were palmitic, palmitolenic, C17-cyclopropanoic, stearic and octadecenic acids. Polar lipids differed from neutral lipids in their fatty acid composition. The fatty acid composition of firmly bound lipids was characterized by a high degree of saturation, the absence of cyclopropanoic acids and the presence of two unidentified acids that were not found in free lipids; apparently, they belonged to hydroxy acids.

The authors studied the biochemical composition of the meat of broilers, eggs, liver and muscles of laying hens of 3 generations on a 5, 10, 25, 50 and 100% (broilers), 10 and 20% (laying hens) replacement of the animal protein quota in the diet by protein obtained from the hydrogen bacteria Alcaligenes eutrophus L-1. No deterioration of the quality of the produce was found from the standpoint of the main biochemical parameters.