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Some of the most pressing health problems of the industrial society are the damage and degeneration of articular cartilage associated with the limited capacity of tissues to regenerate. The review describes the existing and developing technologies for the recovery and replacement of damaged joint cartilage tissue. The results obtained are analyzed covering two major areas: the stimulation of regeneration of damaged cartilage tissue and the growing of cartilage tissue elements in bioreactors.

In this work, we describe an optimized procedure based on gradual hemolysis for the isolation of hemoglobin derived from bovine slaughterhouse erythrocytes in a membrane bioreactor. The membrane bioreactor system provided high yields of hemoglobin (mainly oxyhemoglobin derivate) and its separation from the empty erythrocyte membranes (ghosts). Ten different concentrations of hypotonic media were assessed from the aspect of the extent of hemolysis, hematocrit values of the erythrocyte suspensions, cell swelling and membrane deformations induced by decreased salt concentration. Effective gradual osmotic hemolysis with an extent of hemolysis of 83% was performed using 35 mM Na-phosphate/ NaCl buffer of pH 7.2-7.4. Under these conditions most of the cell membranes presented the appearance of the normal ghosts under phase contrast microscope. The results show that isolation process yielded predominantly to oxyhemoglobin. Kinetic studies showed that maximal concentration of hemoglobin was reached after 40 min, but the process cycle at which recovery of 83% was achieved lasted for 90 min. [ABSTRACT FROM AUTHOR]

The biosynthesis of L-lactate oxidase in the Yarrowia lipolytica yeast during submerged cultivation in laboratory bioreactors ANKUM-2M has been studied. It has been shown under optimal conditions of yeast cultivation with L-lactate that 24.5 U/L enzyme accumulated in the medium and the yield was 2.0 U/(L h). An increase in the biosynthesis of L-lactate oxidase to 75 U/L and the yield to 3.2 U/(L h) was achieved in the medium with L-lactate (1%) and glucose (2%). The enzyme was purified 251 times to homogeneity by hydrophobic and ion exchange chromatography state with a yield of 45% and a specific activity of 55.3 U/mg. Techniques of gel filtration and denaturing electrophoresis showed that L-lactate oxidase from Y. lipolytica is a tetramer with a molecular mass of 200–230 kDa. The enzyme showed a strict specificity to L-lactate and did not oxidize fumarate, pyruvate, succinate, ascorbate, dihydroxyacetone, glycolate, D-lactate, D, L-2-hydroxybutyrate and D, L-alanine or D-serine.

The article concerns converting waste from vegetable processing facilities into methane in anaerobic reactors with a small amount of inoculum (8.4%). Anaerobic digestion of vegetable waste with a high content of organic acids and carbohydrates makes it possible to achieve a methanogenesis productivity of 273–436 L CH4/kg of volatile solidis, which is comparable to or higher than the productivity of such reactors in the world (according to the literature). The contents of ammonia nitrogen and soluble phosphorus in the form of on undiluted substrate basis in the digested vegetable wastes ranged from 3.39 to 5.06 and from 0.78 to 1.03 g/L respectively. Thus, mineralized vegetable waste can be used as an organic fertilizer with a high nutrient content. The results show the feasibility of the technology of conversion of organic waste from vegetable processing facilities into methane and organic fertilizer in anaerobic fermenters (digesters).

The subject of investigation are the peculiarities of yeast metabolism of Saccharomyces cerevisiae M-5 strain in the alcoholic fermentation of molasses wort of increased concentration of dry substances in terms of periodic and continuous fermentation. For periodic alcohol fermentation of wort concentrate having 27 % dry substance content it was established, that 60g /dm³ is the optimal value for inoculate yeast. This results in the maximal accumulation of ethanol in the mature mash (11.5 vol. %), in the deep assimilation of carbohydrates and by the generation of fewer amount of glycerin. By industrial uninterrupted-mode investigations the increase in activity of starting enzymes of glycolysis and strengthening in the capacity of recirculated yeast have been demonstrated. For the first time we show, that this is a consequence of adaptation of yeast to the cultural medium as a result of long-term stay of yeast in it. The increase of the inflow speed of molasses wort into the main fermenter of the battery from 0.43 to 0.57 h⁻¹ improves physiological parameters of recirculated yeast. Also, this reduces the accumulation of secondary products of fermentation (ethers, aldehydes, higher alcohols, glycerin). At the same time, this approach allows to increase the yield of alcohol from the saccharose by 1.84 % and to enhance the productivity with respect to ethanol from 2.3 to 3.8 g/dm³ ×h.

The review considers features of efferent therapy employing extracorporeal systems, the devices known as "artificial liver" and "bioartificial liver" in the treatment of liver insufficiency. Analysis of literature data shows the need for further development of these biomedical studies and the search for optimal solutions in the selection of the source of hepatocytes, the development of bioreactors and biomaterials forming the basis of devices like "bioartificial liver". Taking into consideration certain advantages and disadvantages typical for various methods of extracorporeal support of the functional state of the liver one can evaluate prior experience in the treatment of liver diseases and approaches to the development of new, more effective medical technologies.

Methods of intensifying the anaerobic microbial decomposition of the organic fraction of municipal solid waste (MSW) on an MSW landfill and in anaerobic reactors were studied. It was discovered that it is preferable for the initiation and stabilization of the process of anaerobic digestion of organic waste in laboratory bioreactors at 20 and 50 degrees C to use a mixture of activated suspension of soil from the anaerobic zone of the landfill and digested sewage sludge. Stimulation of methanogenesis was shown in field conditions when digested sewage sludge was added directly into the upper layer of anaerobic zone of the landfill. The investigation of methane production during fermentation of concentrated food waste with a mixture of excessive activated sludge in the laboratory under thermophilic conditions (50 degrees C) has shown that the main problem at the first stage of the process was the acidification of the digested mixture due to the accumulation of volatile fatty acids. It was shown that for stable operation of the bioreactor under thermophilic conditions the amount of inoculum added during the start up should be no less than 30%-50%--based on volatile suspended solids. A sharp decrease in the digestion temperature from 50 to 20 degrees C did not cause methanogenesis termination, since the thermophilically fermented biomass contained both thermophilic and mesophilic methanogens.

It has been established that the presence in the medium Ca2+ and Fe2+ ions in 1.2 - 1.3 times increases the assimilation of hydrocarbon by actinobacteria. The possibility of joint growth of binary actinobacteria cultures on mineral medium with diesel fuel without reducing the assimilation of this substrate compared to monocultures it was shown. In the conditions of co-batch culture Rhodococcus erythropolis IMV B-7277 and Dietzia maris IMV B-7278 strains in the laboratory fermenter "Biotec" (stirring speed 500 r.p.m., degree of aeration 0.3 1/1 per min, sulfite number 1.5 g O2 /1 h,) on the medium with 1.0% diesel oil in the presence of 4 mg/l Ca2+, Fe2+ and 1 g/l yeast extract for 30 hours the substrate assimilation rate was 87.8% and the concentration of biomass - 2.0 g/l.

A method for leaching rare earth elements from coal ash in the presence of elemental sulfur using communities of acidophilic chemolithotrophic microorganisms was proposed. The optimal parameters determined for rare element leaching in reactors were as follows: temperature, 45 degrees C; initial pH, 2.0; pulp density, 10%; and the coal ash to elemental sulfur ratio, 10 : 1. After ten days of leaching, 52.0, 52.6, and 59.5% of scandium, yttrium, and lanthanum, respectively, were recovered.

Macroporous monolithic material containing covalently linked ribonuclease A was used to create high-performance flow heterogeneous biocatalysts (bioreactors). The kinetic parameters of the degradation of polycytidylic acid were identified, and the properties of the obtained systems were compared. A HPLC method has been developed for monitoring products of biocatalytic degradation of RNA, and the possibility of using biocatalytic and HPLC columns in RNA degradation processes in a multicomponent mixture of biological molecules was shown.

A heterogeneous biocatalyst containing a complex of chitinolytic enzymes isolated from the culture medium of bacteria Clostridium paraputrificum on the surface of macroporous monolithic minidisc was obtained. The complex of chitinolytic enzymes was immobilized on the polymer matrix using a multistep method involving the introduction of an intermediate macromolecular spacer. The endochitinase and N-acetylglucosaminidase activity of the heterogeneous biocatalyst was studied.

The possible adaptation of the association of Rhodococcus ruber and Rhodococcus opacus strains immobilized on modified sawdust to oil hydrocarbons in a column bioreactor was investigated. In the bioreactor, the bacterial population showed higher hydrocarbon and antibiotic resistance accompanied by the changes in cell surface properties (hydrophobicity, electrokinetic potential) and in the content of cellular lipids and biosurfactants. The possibility of using adapted Rhodococcus strains for the purification of oil-polluted water in the bioreactor was demonstrated.

Small diameter tissue engineered vascular grafts could be a potential solution to the shortage of vascular substitutes in reconstructive cardiovascular surgery. Previously, we have developed a decellularization method for human umbilical arteries, which could be used as a scaffold in vascular tissue engineering. Objective of the study was to optimize the recellelularization of decellularized scaffolds with mesenchymal stem cells. In the study, the possibility of cell growth on decellularized vessel has been shown. We also has proved that the use of perfusion-bioreactor improves the results of recellularization.

Addition of perfluorodecalin with gas-transporting function to the liquid medium during submerged cultivation of actinomycetes of the genus Streptomyces resulted in higher intensity and level of the biomass synthesis and increased production of streptomycin and daunorubicin. Addition of perfluorodecalin to the medium provided a 2.0-2.3-fold surpass of the maximum antibiotic production (achieved by the 120th-144th hours of the culture growth) vs. the antibiotic accumulation peaks in the control.

A new genus and species of bacteria capable of ammonium oxidation under anaerobic conditions in the presence of nitrite is described. The enrichment culture was obtained from the Moscow River silt by sequential cultivation in reactors with selective conditions for anaerobic ammonium oxidation. Bacterial cells were coccoid, -0.4 x 0.7 mm, with the intracellular membrane structures typical of bacteria capable of anaerobic ammonium oxidation (anammoxosome and paryphoplasm). The cells formed aggregates 5-25 μm in diameter (10 μm on average). They were readily adhered to solid surfaces. The cells were morphologically labile, they easily lost their content and changed their morphology during fixation for electron microscopy. The organism was capable of ammonium oxidation with nitrite. The semisaturation constants Ks for nitrite and ammonium were 0.38 mg N-NO2/L and 0.41 mg N-NH4/L, respectively. The maximal nitrite concentrations for growth were 90 and 75 mg N-NO2/L for single and continuous application, respectively. The doubling time was 32 days, μ(max) = 0.022 day(-1), the optimal temperature and pH were 20 degrees C and 7.8-8.3, respectively. According to the 16S rRNA gene sequencing, the bacterium was assigned to a new genus and species within the phylum Planctomycetes. The proposed name for the new bacterium is Candidatus Anammoximicrobium moscowii gen. nov., sp. nov. (a microorganisms carrying out anaerobia ammonium oxidation, isolated in the Moscow region).

Aim: Selection of high-mucoid morphotype of Streptococcus equi subsp. zooepidemicus (Streptococcus zooepidemicus) and study of its morphological, physiological, biochemical and technological characteristics for providing increased secretion of hyaluronic acid (HA)., Materials and Methods: Submerged cultivation was performed in 100 ml glass flasks without baffles or in 1.5 or 10 1 laboratory bioreactors. LB and MRS media were used for cultivation. Mutagenesis was carried out by UV exposure with consequent selection of mucoid phenotype. HA was determined by carbazole method or after exhaustive acid hydrolysis by reaction of N-acetylglucosamine with Morgan-Elson reagent. Total hyaluronidase activity was evaluated by viscosimeter. Determination of cell and capsule size, ability to ferment carbohydrates and other microbiological, physiological and biochemical tests were performed by standard techniques., Results: Instability of capsule phenotype of S. zooepidemicus B-8014 strain was revealed that is explained most probably by formation under certain conditions of bacterial hyaluronidase. This is confirmed by a reduction of HA concentration in cultural medium at pre- and stationary growth phases. Mucoid strain S. zooepidemicus KB-04 was obtained by mutagenesis with subsequent selection that is characterized by increased capsules. The strain was studied for HA formation. Optimization of growth medium composition, physical-chemical conditions and modes of cultivation allowed to significantly increase HA yield., Conclusion: The studies of morphologic, physiologic, biochemical and technological characteristics of the high-mucoid S. zooepidemicus KB-04 strain obtained by mutagenesis with consequent selection were performed, conditions of its cultivation and composition of growth mediu by carbon source and content of bivalent metal ions were optimized.

Several active microbial communities that form biogas via decomposition of cellulose and domestic food waste were identified among 24 samples isolated from different natural and anthropogenic sources. The methane yield was 90-260 ml CH4/g from microbial communities grown on cellulose substrates, office paper, and cardboard at 37 degrees C without preprocessing. Under mesophilic conditions, bioconversion of paper waste yields biogas with a methane content from 47 to 63%; however, the rate of biogas production was 1.5-2.0 times lower than under thermophilic conditions. When microbial communities were grown on DFW under thermophilic conditions, the most stable and effective of them produced 230-353 ml CH4/g, and the methane content in biogas was 54-58%. These results demonstrated the significance of our studies for the development of a technology for the biotransformation of paper waste into biogas and for the need of selection of microbial communities to improve the efficiency of the process.

9alpha-Hydroxy derivatives were prepared from 11 steroids ofandrostane and pregnane series using Rhodococcus erythropolis VKPM Ac-1740 culture with 0.5-20 g/l substrate concentration in the reaction mixture. 9alpha-Monohydroxylation proceeded regardless of the substituent structure at C17. However, the structure of the steroid molecule influenced the time of complete conversion of the substrate and the yield of the transformation product. 9alpha-Hydroxy-androstenedione was obtained in 35 h in a yield of 85% when the maximum concentration of androstenedione (AD) was 10 g/l. 9alpha-Hydroxy-AD was also formed by the actinobacterium cells entrapped in poly(vinyl alcohol) cryogel beads. Nine successive transformation cycles were carried out using immobilized cells at 4.0 g/l concentration of AD in the medium. The yield of 9alpha-hydroxy-AD formed during six cycles (from two to eight with the duration of each cycle for 22-24 h) was 98%.

Peculiarities of synthesis of surface-active substances (SAS) are studied at periodical cultivation of Rhodococcus erythropolis EK-1 in the AK-210 fermenter on medium containing n-hexadecane. Maximum indicators of SAS synthesis (concentration of extra cellular SAS is 7.2 g/l; factor of emulsification of the cultural liquid 50%; SAS yield from the substrate 50%) have been observed at 60-70% concentration of dissolved oxygen from the saturation level with aerial oxygen (pH 8.0) fractional supply of the substrate by portions each being 0.3-0.4% every 5-6 h to a final volume concentration of 2.4% and with the use of 10% inoculate grown until mid-exponential phase on the medium with 1.0 vol % of n-hexadecane. Implementation of the process of SAS biosynthesis with the fermentation equipment provided the possibility to increase almost two-fold the amount of the synthesized SAS and reduce 3.5-fold the time of cultivation of the producer strain compared with the growth in flasks at shake-flask propagator.

Aim: Study in CBA line mice of immunogenicity of cold adapted reassortant influenza virus H5N2 vaccine strain samples produced in rollers in MDCK and Vero cell cultures by using plant derived components., Materials and Methods: Antibody levels in blood sera and nasal swabs, lungs and small intestine of experimental vaccine strain sample immunized mice were evaluated by using HI reaction in accordance with WHO recommendations., Result: Reassortant vaccine strain A/17/duck/Potsdam/86/92 (H5N2) produced in MDCK and Vero cells by using plant derived components (rice and soy flour hydrolyzate and plant protease based nutrient medium) after intranasal immunization of mice induced local and humoral antibodies, and the latter not only against homologous virus, but also against highly pathogenic avian influenza virus strains A/ Chicken/Suzdalka/Nov-11/2005 and A/Chicken/Kurgan/05/2005., Conclusion: Immunogenicity studies of reassortant influenza virus A/17/duck/Potsdam/86/92 (H5N2) vaccine strain samples cultivated in MDCK and Vero cells by using media with plant derived components in mice show high levels of humoral and secretory immunity.

Functional testing of the bioreactor for stirring culture in microgravity was performed during the microgravity episodes aboard research airplane Il-76 following the Kepler parabolic flight path. The attempt to produce the gas vortex stirring effect in a given liquid volume was a failure. Nonetheless, the stirrer ventilator is capable to agitate liquid acting as a mechanical stirrer.

An original method of immobilization of nongrowing microorganism cells on xerogel of silicon dioxide containing insoluble hydroxyl compounds of cobalt(III) has been developed. A recombinant strain producing glucose isomerase has been constructed on the basis of Escherichia coli with the use of a gene of Arthrobacter nicotianae. It was revealed that glucose isomerase activity and stability of biocatalysts prepared on the basis of the recombinant E. coli strain was 3-5 times greater compared with the biocatalysts prepared with the use of the donor strain A. nicotianae. Under conditions of continuous hydrolysis of 3 M fructose at 62-65 degrees C in a fixed bed reactor, time of half-inactivation of the biocatalysts prepared from the recombinant strain and A. nicotianae was -60 and -25 days, respectively.

Perspectives for nature protection and energy-saving, by using blue-green algae, are discussed. Utilization of their phyto biomass for biogas manufacture will lead to the environmental normalization of the Transdniestria and allow one to have about 19,000,000 m3 of methane only from the water area of only one Kremenchug water basin each vegetative period (70 days).

Aim: Assessment of characteristics of antigenic complexes of Staphylococcus aureus vaccine strains in different cultivation conditions., Materials and Methods: S. aureus vaccine strains (No. 5, 9, 1986, 1991) were grown in liquid nutrient media--full value and semi-synthetic--as well as on solid medium. Reactor cultivation was performed in the fermenter ANKUM-2M. Complex of antigens were obtained by water extraction method applied to staphylococcal biomass inactivated with acetone and assessed by common methods on protein and carbohydrate content; specific activity was assessed by minimal inhibitory dose in passive hemagglutination inhibition assay. Study of acute toxicity was performed on outbred mice., Results: Using strain no. 1991, model of reactor cultivation in full value medium with separation of biomass by microfiltration was validated on the basis of biomass and semiproduct of antigenic complex (acetone powder) yield as well as productivity of biomass cumulation. Study of antigenic complexes obtained from biomass of 4 strains during reactor cultivation compared with complexes extracted from cultures grown on solid medium revealed increased protein and decreased carbohydrate content but similar specific activity. It was demonstrated that complex of antigens obtained from cultures grown either by reactor cultivation or on solid medium were non-toxic., Conclusion: New technology for manufacturing staphylococcal complex of antigens with reactor cultivation of vaccine strains in full value medium with subsequent purification of antigenic complex from the biomass by microfiltration was developed. Results of the study demonstrated the usefulness of the developed technology for both further studies on a cellular staphylococcal vaccine and manufacture of staphylococcal component of "Immunovac" vaccine.

A two-stage technology for bacterial and chemical leaching of nonferrous metals in a specifically designed laboratory unit has been proposed. At the first stage of leaching, ferric iron formed during the second stage of oxidation of Fe2+ ions by mesophilic chemolithotrophic microorganisms was used. The optimal parameters of the first stage of the process (flow rate, temperature, and the process duration) were 2 l/h, 75 degrees C, and 2 h, respectively. The results of testing of the two-stage technology for leaching copper-zinc raw materials indicated that the depth of zinc and copper leaching can be increased from 70 to 93% and from 40 to 58.8%, respectively, and the process duration can be reduced from 120 to 24 h as compared to the commonly used one-stage technology.

Solid-phase growth of Bacillus subtilis 8130 on cellulose-rich plant substrates (presscakes or pulp) under hypoxic conditions was accompanied by cellulose depolymerization, protein hydrolysis, and degradation of other plant components, including some processes of mixed-type carbohydrate fermentation. The bacterial fermentation yielded propionic, butyric, and hexanoic acids and butyric acid derivatives. The bacterial metabolism and fermentation degree can be characterized by the proportions of fatty acids in the reaction mixture. The product of sea buckthorn cake fermentation has a good sorption quality.

This review discusses the real prospects of using the potential of microorganisms for bioremediation of PCB-polluted natural environments (soil, sediments, and wastewater sludge) under anaerobic and aerobic conditions. A detailed analysis of the research conditions of original works has shown that the efficiency of bioremediation of PCB-polluted objects strongly depends on the character and degree of pollution. In the case of aerobic bioremediation, the best results were obtained with moderately polluted soils and sediments (20 to 700 PCB/kg), in which the level of pollution decreased by 40-75%. These results could be achieved only by repeated inoculation of a consortium of specific microorganisms (isolated or constructed) with concurrent addition of biphenyl as an inducer and of biosurfactants; their effect increased in a liquid phase bioreactor. PCB concentration decreased mainly due to the degradation of congeners with one to three chlorine atoms. The content of highly chlorinated PCB can be noticeably decreased only under sequential anaerobic-aerobic treatment; the best effect was achieved with anaerobic granules. However, only in individual cases, mainly in laboratory experiments with newly introduced PCB at moderate concentrations, was it possible to reduce their content to a level permissible for technogenic soils. The review begins with the description of the main metabolic pathways and patterns of biodegradation of these pollutants in natural and artificial environments.

A biotechnological pilot plant (National Research Centre of Antibiotics) and its technical potentialities in production of various biosynthetic products are described. Some engineer and technological aspects of the fermentation equipment and particularly sterilization of the media and apparatus, fermentation broth aeration under sterile conditions and control of biosynthesis technological parameters (t degrees, pO2, P, pH, foaming, etc.) are considered. The pilot plant is designed for fermentation processes under aseptic conditions with the use practically of any object, from bacteria to tissue cultures.

Parameters of degradation of p-toluenesulfonate (TS) by free and agar-embedded Comamonas testosteroni BS1310 (pBS1010) cells were determined. The maximum rate of TS degradation was 25% lower in by immobilized than free cells, equaling 11 nmol x min(-1) x mg(-1) cells. Degradation of TS by both free and immobilized cells was associated with molecular oxygen consumption (molar ratio, 1 : 2). In a plug-flow reactor, the degradation rate was 10.4 nmol x min(-1) x mg(-1) cells. The results can be applied to designing reactors for TS degradation in sewage and developing biosensors.

Optimal conditions were developed for cultivating the cold-adapted reassortant live influenza vaccine (CARLIV) in MDCK cells, which were in their turn cultivated in fermenters with serum-free medium and microcarrier. The use of MDCK cells meets all national and WHO requirements to continuous cells used in the production of biological preparations. CARLIV cultivated under such conditions well preserve their ts-mutations and mutation, which entail substitutions of amino acids, in all CARLIV genome segments. Provided the cultivation conditions are optimal, the output of multivalent CARLIV in a 101 fermenter can reach 100000 doses.

An experimental investigation was carried out, and a mathematical model of interaction between invertebrates (infusoria Paramecium caudatum and rotifera Brachionus plicatilis) and algae (Chlorella vulgaris and Scenedesmus quadricauda) in the "producer-consumer" aquatic biotic cycle with spatially divided links was constructed. The model describes the dynamics of a mixed culture of infusoria and rotifera in the "consumer" link, when they consume a mixed culture of algae coming from the "producer" link. A negative influence of products of algae Scenedesmus metabolism upon the reproduction of infusoria P. caudatum was revealed. Taking this into account, a qualitative coincidence of the results of mathematical modeling with experimental data was obtained. It was shown that the co-existence of mixed algae culture in the "producer" link with invertebrates in the "consumer" link in the "producer-consumer" aquatic biotic cycle is impossible because of the displacement of infusoria P. caudatum by rotifera Brachionus plicatilis.

The whole-culture vaccine preparation of Neisseria meningitidis, serogroup B, obtained by cultivation in a computer-controlled bioreactor, was studied. The preparation was shown to contain antigenically active polysaccharide, outer membrane proteins, as well as lipooligosaccharide, faintly pyrogenic and with low toxicity. After oral immunization of rabbits a multiple increase in the levels of IgG antibodies to these antigens in their blood serum was noted during the period of observation (303 days).

The present review describes the data on electrochemical reduction of cytochrome P450. Three generations of enzyme biosensors have been considered. The concept and potentialities of enzyme electrodes--transducers--as the main element on construction of electrochemical biosensors are described. Different types of electrodes for bioelectrochemistry are presented. New experimental approaches for immobilisation of cytochrome P450 based on nanotechnology are reported. Nanobiotechnology in electrochemistry has potential application for production of biosensors and bioreactors for medicine

We have found the ATP production in the Rhodobacter sphaeroides chromatophores illuminated by single short light flash, that is under conditions when the proton gradient formed as a result of electron transport after the second flash, is absent. The ATP synthesis was accompanied by the H2O2 formation. Simultaneous formation of H2O2 is indicative of the oxidative activation of phosphate during the ATP synthesis, as in the model systems with isolated chlorophyll. These data provide a theoretical background to the fitting of illumination parameters in both laboratory and industrial photobioreactors with photosynthetic bacteria used in biotechnological processes.

A two-phase technology has been developed for biodegradation of plant wastes. On the first phase, wastes are subjected to anaerobic fermentation by specially selected microbial associations processing solid wastes into a liquid mixture of organic acids. On the second phase, methanogenic bacteria are used to produce biogas out of the liquid, which then can be dumped out into open space or utilized as a cold propellant.

A highly potent strain of Bacillus licheniformis 103 that synthesized thermostable alpha-amylase with temperature and pH optima of 90-95 degrees C and 6.0-8.5, respectively, was obtained by mutagenesis and selection. The composition of fermentation media and conditions for submerged cultivation of the producer were optimized. alpha-Amylase whose activity reached 260 U/ml was obtained in laboratory fermenters.

Effects of iron compounds on methanogenic fermentation the water polluted with fatty acids were studied. A natural readily available source of iron applicable to biological treatment of liquid wastes was searched for. A positive effect of iron on the methanogenic fermentation of fats and their degradation products--long-chain fatty acids--in aqueous media was demonstrated. It is recommended to add iron-containing clay, as an inexpensive and easily available iron source, in amounts providing the binding of the long-chain fatty acids present in wastewaters.

The available models of the microorganism growth kinetics are analyzed. The possibility of application of the Monod model is studied. It is shown that these models do not permit describing adequately the experimental data at different initial concentrations of biomass and substrate that vary in a wide range. The calculations of the static modes of operation of biochemical react ors differ greatly.

The dependence of toluene elimination capacity on its load was obtained in five small-scale reactors filled with glass beads carrying biocatalyst cells. With increase in operation time the calculated maximal elimination capacity was shown to increase along with biomass density in the biocatalyst bed. Fivefold increase in trickling intensity did not affect the reactor performance. A simplified mathematical model for evaluation of minimal required biocatalyst bed volume at certain loading was developed based on experimental dependence of elimination capacity vs. loading.

Investigation of the stability of an association of active hydrocarbon-oxidizing microorganisms grown on diesel fuel showed that not all members of the association were competitive in an open flow system. The abundance of some strains considerably decreased during cultivation. One of the strain groups had consistently high cell titers, both in the medium and on a support. This prompts the authors to use these strains as degraders of diesel fuel.

Growing phototrophic microorganisms consume the light energy. These microorganisms most often suffer from light deficiency because of exponential decrease in the energy of light passing through an absorbing medium. Therefore, effective distribution of light within the cultures is needed for their intensive cultivation. This is possible in special devices called photobioreactors. The photobioreactors described in the literature are classified into several types according to their geometric features. Their advantages and drawbacks are analyzed. Criteria currently used for comparing various photobioreactors are specified.

Colonies produced by a consortium of nitrifying bacteria were studied using light and electron microscopy. The colonies were obtained by direct plating of inoculum from a two-stage nonsterile chemostat fermentor and by repeatedly passing the microbial community of the fermentor through selective media containing ammonium or nitrite. The colonies studied can be characterized by a specific combination of six types of cells differing in their ultrastructure and spatial location within the colony. The types of cells occurring within a given colony were found to depend on the nitrogen compound present in the medium. As a result of our study, morphological features of colonial bacterial communities were preliminarily identified. The proposed approach can be viewed as a method to describe microbial associations and communities.

The parameters of the cultivation process and the characteristics of the culture growth have been analyzed. The phases of the population growth have been determined. The existence of short stages differing in the physiological state of the cells, their resistance to damaging factors and metabolic activity within the deceleration phase of the growth of bifidobacteria has been experimentally substantiated. To harvest the biomass, the period of the completion of the cultivation process has been chosen. The lyophilized seed culture in which a high percentage of viable cells is preserved for one year has been obtained.

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.

Our study of the techniques of long-term storage of the biomass of various strains of microorganisms, which cause breakdown or transformation of synthetic organic compounds, demonstrates that desiccated agar beads with immobilized microbial cells can be used for this purpose. In addition, the cells can be stored in desiccated matrices of agar or polyvinyl alcohol, coating synthetic cords. Such dry biocatalysts may be used for quick starting of bioreactors and in other biotechnological processes. The technique is applicable to storage of various strains of Pseudomonas, Corynebacterium, Rhodococcus, and, to a lesser extent, Enterobacteriaceae.

The processes of the cultivation of Bordetella pertussis, immobilized on polyurethane carrier in a fermenter, were carried out and studied. Acellular pertussis preparations were produced from the culture fluid obtained in the batch and multi-cycle cultivation processes with immobilized cells, as well as in the process with interrupted fermentation (for confirming the possibility of the preservation of cell viability). The content of protein and B. pertussis toxin in these preparations, as well as their leukocytes-stimulating and hemagglutinating activity, did not differ from similar characteristics of preparations obtained from culture fluid in homogeneous cultivation.

The authors examined the antigenic complexes obtained from serogroup B Neisseria meningitidis strain 125 cells grown in the bioreactor to different phases of growth in the reaction of gel precipitation and counter immunophoresis in a periodical fashion. A set of serotypic antigens was shown to increase from 3 to 9 with cultural growth. Their largest quantities were seen in the cells grown till the end of the stationary growth phase, the concentration of high-molecular weight peptides rising from 19% (in the exponential growth phase cells) to 54% (in the stationary growth phase ones). The revealed regularities were evidenced by gel precipitation examinations of the sera from rabbits immunized with meningococci grown till different growth phases. At the same time, there were the largest quantities of serotypic antigenic precipitation bands when the sera obtained in the culture grown till the end of the stationary growth phase were used. It cannot be excluded that the results depend on the combined effects of several stressors on Neisseria meningitidis in the transition from the exponential to stationary growth phase, as resulted from the increased synthesis of high-molecular weight proteins that seem to act as protectors.