Analysis of the effect of concentration and magnetic field on bioactivity of humic acids from position of open non-equilibrium systems.

On the example of cultures of bacteria Pseudomonas sp. and Bacillus sp. the effect of humic acids in the concentration range (0-15 mg/L) on the viability of these bacteria was studied. Multidirectional effects of humic acids on microorganisms were found, namely, at concentrations of 1 and 5 mg/L, they reduce, and at 15 mg/L, they do not affect the survival of the studied cultures of bacteria. It has been established that the treatment of aqueous solutions of humic acids with a weak magnetic field affects their physicochemical properties and structural parameters, as well as enhances the biological effect in relation to different groups of microorganisms. It was established that the multidirectionality of the biological effect of humic acids on microorganisms is well explained from the position of considering humic acids as complex open non-equilibrium systems with a developed system for energy dissipation, which leads to the formation on their basis of diverse and complex self-organized structures with different physicochemical properties. The obtained concentration dependences confirm the possibility of the existence of humic acids in the studied concentration range in at least two states. Moreover, it is the state of the system (the level of self-organization), and not the relationship with the source of origin of a humic acids, mainly determines its properties. This fact determines the possibility of using the concentration of humic acids to establish the level of organization of the system, which will allow to manage the structural organization of these objects and predict their properties.

[THE INFLUENCE OF B-IRRADIATION ON MICROMYCETES ISOLATED FROM THE ZONE OF RADIONUCLIDE CONTAMINATION].

The resistance of the micromycetes to ß-irradiation had been studied. The study was conducted on two species: Cladosporium sphaerospermum Penz. and Alternaria alternata (Fr.) Keissl., each of them was represented by six strains. Half of strains were isolated from the inner locations of the "Shelter" object (SO), which were characterized by different levels of radionuclide contamination. For C. sphaerospermum strains maximum resistance to ß-radiation was observed among 20-day-old cultures. The greatest resistance to ß-radiation had been demonstrated by two strains: 5-1 (isolated from SO location with the highest level of contamination) and 852 (control). For A. alternata strains maximum resistance to ß-radiation was observed among 40-day-old cultures. As the most resistant were characterized 56 and 105 isolates from the SO inner location with a high level of the radionuclide contamination. Based on the obtained results we concluded about high resistance to irradiation of both studied species of anamorphic fungi, which achieved through different life strategies: C. sphaerospermum strains survive due to rapid and abundant formation of conidia, whereas conidia of A. alternata strains are formed slowly, but they have greater advantage under irradiation due to their multicellular structure and large sizes.

[Veterinary probiotic preparation influence on the biochemical blood indices of young farm animals].

The veterinary probiotic preparation was developed on the basis of aerobic part of bacterial strains of normal human intestinal microflora: 3 strains of Escherichia coli--G35No1-413, G35No2-412, G35No3-411, and one Enterococcus faecalis G35No4-410 strain. The influence of a pilot lot of the prepapration on the biochemical blood indices and the state of intestinal microflora of different age groups of young farm animal (calves and piglets) was studied during the process of the commission tests. It has been shown, that the developed probiotic promotes the improvement of protein and mineral metabolism, normalization of level of enzyme activities, which show state of the native immunity of the young farm animals' organism. It was the result of normalization of the intestinal microflora composition. The optimal doses and periods of intake have been determined. On the basis of executed researches the veterinary probiotic preparation has been presented for registration to the State Department of Veterinary Medicine of Ukraine.

[Effect of metal microelements on biochemical indices of probiotic bacteria].

The influence of metal-microelements (Zn, Cu, Ag, Au) in colloid and ionic form on the main biochemical parameters (ATP-ase activity, transmembrane potential and respiratory activity) of E. coli G35 N#1-413 and Ent. faecalis G35 N#4-410 probiont-strains has been studied with the goal to create complex metal-bearing probiotic preparations. Monotonous dose-dependent inhibitory influence of all mentioned metals in ionic form on the bacteria functional activity has been established. Metals' colloids had certainly stimulating influence on microorganisms' biochemical parameters in the same concentration limits with the following maximum positive effects: for Zn - 2x10(-7)-2x10(-4) mg/ml, for Cu - 8.4x10(-7)-8.4x10(-6) microg/ml, for Ag - 23x10(-7)-23x10(-4) microg/ml, for Au - 5x10(-6) microg/ml by metals. It has been shown, that cultural characteristics of probiont-bacteria in the presence of both ionic and colloid forms of studied metals remain stable. Certain stimulation of probiont-bacteria' main enzymes and life-support processes by colloidal forms of the studied metals-microelements in determined concentration and particle dimension limits is the evidence of a possibility of metals' colloids application in probiotic preparations' composition in order to increase the resistance and functional activity of probiont-microorganisms.

[Obtaining of bacterial bioluminescent strain Protobacterium phosphoreum B7071 (lux+) for the determination of zinc ion concentration].

The transconjugate containing hybrid plasmid (Te(R)Zn(R)lux+) was obtained using the conjugation method on Pseudomonasfragi T2(5) (Te(R)ZnR) strain and bioluminescent strain Protobacterium phosphoreum B7071 (lux+). The expression regulation of lux-genes on the obtained plasmid is carried out by promotor-operational area conjointly with zinc resistance genes. The cells of the obtained genetically modified bacteria have the ability to specific induced luminescence, which is a respond to zinc ions' presence in the measuring medium. It was shown that the cells' bioluminescence intensity of trans-conjugate is linearly dependent on zinc ions' concentration within the range of 1-100 microM, that provides the opportunity of using biosensor as a strain for qualitative and quantitative detection of the metal. The low sensitivity limit of this method is 0.5 microM for the metal. Besides having high sensitivity, the developed lux-biosensor is highly specified.

[Structural and functional aspects of bacterial membranes and liposomes interactions].

Effect of exogenous lipids on the morphology and enzymatic activity of Bacillus cereus B4368 membrane has been studied. Specific character of different lipids action on the membrane enzyme activity was found. This peculiarity had been taken into account by using liposomes in biological systems.

[Effect of colloid gold on physiologic-biochemical processes in Escherichia coli 1257]. / Vliianie kolloidnogo zolota na fiziologo-biokhimicheskie protsessy Escherichia coli 1257.

The influence of colloid gold on the growth processes, ATP-ase activity and extrusion of protons in Escherichia coli 1257 was studied. The particles of colloid gold exert nonmonotonous influence on these processes with different direction is such a way that small concentration of this metal (5 x 10(-7)-5 x 10(-6) mg/ml) exert stimulative effect, while higher concentrations of colloid gold result in the suppression of biological activity of the bacterial cells. The discovered peculiarities of colloid gold influence of E. coli strain may be determined by specificity of contact interaction of metal particles with the surface of bacterial cells.

[Obtaining bioluminescent bacterial strains for detection of thallium ions]. / Poluchenie bioliuminestsentnykh shtammov bakterii dlia detektsii ionov talliia.

The possibility of Alcaligenes eutrophus T1 luxphenotype creation by temperature induced mutagenesis has been shown. These mutants are able to emit light after induction by thallium ions. This biological method of thallium detection possesses high specificity and sensitivity (0.5 microM of metal). Such microbial cells can be used to quantify bioavailability of thallium part in the environment.

[Comparative analysis of radial linear growth velocity of micromycetes isolated from different ecotopes]. / Sravnitel'nyi analiz skorosti radial'nogo rosta mikromitsetov, vydelennykh iz razlichnykh ékotopov.

The work aim was the discovering of peculiarities of microscopic fungi responding to radioactive pollution by growth velocity change. The linear growth velocity of a broad range of micromycetes was analyzed. They were chosen in such a way that every species, as far as possible, was presented by the strains isolated from radioactive polluted and clean regions. Most of the studied species were characterized by the absence of reliable growth velocity differences of strains of the same species growing on rich and starving media. Only two species--Aureobasidium pullulans and Penicillium spinulosum were the exception. Their growth velocities on malt-agar were much more then those on starving medium (1.7-3.6. times). Species Hormonema dematioides did not have reliable differences of growth velocity on optimal and starving media, but A. pullulans had this difference (2.3-3.6 times). It is supposed to use this observation to simplify identification of these species. Only strains within species Cladosporium sphaerospermum and especially P. funiculosum isolated from radioactive polluted ecotopes had growth velocity which was less then that of the strains from clean regions. The delay of growth velocity in such conditions can testify to adaptive change of metabolic processes of these strains. The strains which were characterized by delay of growth velocity existed at the substrates with high radioactive pollution for a long time.

[Peculiarities of linear growth of the melanin-containing fungi Cladosporium sphaerospermum Penz. and Alternaria alternata (Fr.) Keissler]. / Osobennosti lineinogo rosta melaninsoderzhashchikh gribov Cladosporium sphaerospermum Penz. i Alternaria alternata (Fr.) Keissler.

Results on determination of radial (linear) growth rate of melanin-containing fungi Cladosporium sphaerospermum Penz. and Alternaria alternata (Fr.) Keissler concerning place of their isolation and ecologic peculiarities are presented in the paper. Essential differences, as to all studied indices of the strains of C. sphaerospermum strains, isolated from premices of the ChNPP 4th unit from the control strains of this species have been established. Only strain 34 of C. sphaerospermum, isolated from the cable driving region of the "Shelter" object, which did not differ from the control strains as to all the studied indices, made the exception. The radial growth rate was 2-4 times as low and variability amplitude 10-30 times as high in strains 60 and 5-1 of C. sphaerospermum, (which suffered high radiation load), as in the control strains of this species. The radial growth rate of C. sphaerospermum studied strains was an order lower as a whole than that of A. alternata strains. All the above said evidence for implementation of K-type vital strategy by C. sphaerospermum species and in this connection one can suppose its active growth on the surface of walls and ferroconcrete structures of the ChNPP 4th unit. Absence of differences, as to the studied radial growth indices, between A. alternata strains isolated from the ChNPP 4th unit premices and control strains of the species can evidence for the existence of A. alternata species under the conditions of high radioactivity in the surviving state.

[The effect of gamma irradiation on the physiological-biochemical properties of strains of Cladosporium cladosporioides (Fres.) de Vries differing by the trait of radiotropism]. / Vliianie gamma-oblucheniia na fiziologo-biokhimicheskie svoistva shtammov Cladosporium cladosporioides (Fres.) de Vries, razlichaiushchikhsia po priznaku radiotropizma.

The influence of the low-intensity gamma-irradiation on the process of lipid peroxidation and the activities of the antioxidant glutathione-dependent system (catalase, glutathione-transferase) has been investigated in a number of Cladosporium cladosporioides (Fres.) de Vries strains. The dark-pigmented strains isolated from the habitats with different degree of radionuclide contamination, and the nonpigmented alb-mutant of the same species have been used in our work. The studied properties have been analyzed with the respect of the radiotropism property and of the presence of melanin pigment in the cell wall of these strains. The lipid perioxidation level under the effect of the low-intensity gamma-irradiation was greatly increased in Cladosporium cladosporioides 396 strain only. This strain was isolated from the radiation-pure soil. Catalase activity in a number of the studies strains correlated neither with their pigmentation, nor with the property of positive radiotropism. The correlation between the strain pigmentation and activity of glutathione transferase has been found.

[The protein-synthesizing activity of a number of strains of Cladosporium cladosporioides (Fres.) de Vries grown under conditions of continuous low-intensity gamma irradiation]. / Beloksinteziruiushchaia aktivnost' riada shtammov Cladosporium cladosporioides (Fres.) de Vries, vyrashchennykh v usloviiakh nepreryvnogo gamma-oblucheniia maloi intensivnosti.

Phenomena of melanization of radioactively polluted soils, due to prevalence of melanin-containing species and positive radiotropism of some micromycetes have been found during monitoring of mycobiota of the 30-km alienation zone of the Chernobyl NPP for 10 years. To elucidate the contribution of the melanin system to the cell protection against irradiation, the influence of gamma-irradiation on the activity of protein synthesis in four Cladosporium cladosporioides (Fres.) de Vries strains has been investigated. Two strains isolated from radioactively polluted substrates were characterised by the presence of positive radiotropism. Laboratory strain 396 and its alb-mutant (with blocked melanin synthesis), did not possess this feature. The protein synthesizing activity was assayed by incorporation of 14C-leucine in the protein fraction of mycelium, grown during 7 days under continuous gamma-irradiation of low intensity and without it. The protein synthesis was activated in the radioactively treated mycelium of dark-pigmented C. cladosporioides strains and it was suppressed in similarly treated mycelium of alb-mutant of C. cladosporioides. The rate of 14C-leucine incorporation into biomass of investigated strains correlated with positive radiotropism. The dependence between protein synthesis intensity and the availability of melanin protection system in micromycetes is assumed.