[FORMATION AND FUNCTIONING OF SYMBIOTIC SYSTEMS AND RHIZOSPHERE MICROBIOCENISIS OF SOYBEAN UNDER VARIOUS FUNGICIDES APPLICATION].

The Relevance. At the recent years in soybean crops the quantity of plant pathogenic fungi has increased. The fungicides of systemic and contact action have been applicated intensively against of them. After introducing into the soil fungicides and/or their deg- radation products can to disrupt the activities of non-target objects - beneficial soil mi- croorganisms, inhibit nodulation process and the nitrogen-fixing activity of diazotrophs. The purpose of the work was to investigate the impact of combined application of fungi- cides with inoculation on the soybean symbiotic system and rhizosphere microorganisms. The Methods. The microbiological and statistical methods, gas chromatography method. The Results. Inoculation of seeds by the highly active Bradyrhizobiumjaponicum UCM B-6035, UCM B-6018 and UCM B-6023 strains the activity of nitrogen-fixing symbiotic systems increased by 1.4-3.4 times in comparison with the variant without of fungicides application and bacterization. Seed treatment by Vitavaks 200 FF fungicide caused a de- crease of'nitrogen-fixing activity of rhizobia industrial strains in symbiosis with soybean by 3-5 times. The seeds inoculation by B. japonicum UCM B-6035 strain promoted to reduce the negative impact of the Maxim Star 025 FS fungicide on the nitrogenase activity of nodulation apparatus. The positive effect of seeds bacterization was observed in the in- crease of the quantity of rhizosphere microorganisms of main ecological trophic groups. In the variant with seed treatment by Maxim Star 025 FS and Kinto duo fungicides was found a decrease in the number of microorganisms of studied groups in comparison with the control variant; the Vitavaks 200 FF fungicide application promoted to improve of these microorganisms development compared with the variant without the fungicides application and bacterization. At the inoculation of rhizobia industrial strains the negative effect of the Maxim Star 025 FS and Kinto duo fungicides to oligoazotrophic and prototrophic micro- organisms was not observed. The Conclusion. The symbiotic system of variant with the combined application of the Kinto duo fungicide with B. japonicum UCM B-6023 strain characterized by the highest nodulation and nitrogen-fixing activity.

[Biological Properties of Nocardia vaccinii IMV B-7405 Surfactants Synthesized on Fried Sunflower Oil].

Aim: To study of antimicrobial and antiadhesive properties of the surface-active substances (surfactants) Nocardia vaccinii IMV B-7405 synthesized on different oil-containing substrates (refined and waste after frying meat and potatoes sunflower oil). Methods: Surfactants were extracted from supernatant of cultural liquid by mixture of chloroform and methanol (2 : 1). Antimicrobial against bacteria and yeast properties of the surfactant was determined by index of the minimum inhibitory concentration (MIC). The number (%) of attached cells (adhesion) was determined as a ratio of the optical density of the suspension obtained from the materials treated with surfactants to the optical density of the control samples (100 %). Results: It was established that surfactants synthesized N. vaccinii IMV B-7405 on fried oil, at a concentration of 0.02 - 0.04 mg/ml decreased adhesion of bacteria (Escherichia coli IEM-1, Bacillus subtilis BT-2) on plastic, dutch tile, glass and linoleum by 25-90 %, and yeast Candida albicans Д-6 - by 15 - 65 %. The minimum inhibitory concentration with respect to studied test cultures surfactants synthesized on waste after frying potato oil was 8 - 67 mg/ml, and against phytopathogenic bacteria Pectobacterium carotovorum UCM B-1095, Pseudomonas syringae pv. atrofaciens UCM B-1015 and Xanthomonas campestris pv. campestris UCM B-1049 - 14 - 52 mg/ml. Conclusions: Replacing traditional substrates on fried oil can not only reduce cost of N. vaccinii IMV B-7405 surfactant biosynthesis, but also to obtain the final product with high antiadhesive and antimicrobial properties. The dependence of antiadhesive and antimicrobial potential of surfactants on the nature of the carbon source indicate the need for studies effect of cultivation conditions of producer on biological properties of synthesized surfactants.

[Antimicrobial Properties Surfactants Synthesized under Different Cultivation Conditions of Acinetobacter calcoaceticus EMV B-7241].

Aim: To study the antimicrobial properties of the surface-active agents (surfactants) Acinetobacter calcoaceticus IMV B-7241 depending on the availability of yeast autolysate and trace elements in the composition of ethanol-, n-hexadecane- and glycerol-containing media. Methods: Antimicrobial against bacteria (Escherichia coli IEM-1, Bacillus subtilis BT-2), and yeast (Candida albicans D-6) properties of the surfactant was determined by index of the minimum inhibitory concentration (MIC). Surfactants were extracted from supernatant of cultural liquid by mixture of chloroform and methanol (2:1). Results: The removal from cultivation medium yeast autolysate and trace element mix and replacing them by copper sulfate and iron sulphate in the medium with ethanol and n-hexadecane, and in the medium with glycerol - by potassium chloride, zinc sulfate and copper sulfate accompanied by decreasing antimicrobial properties of surfactants. The most effective antimicrobial agents were surfactant synthesized on ethanol in the presence of yeast autolysate and trace elements (MIC 9-20 µ/ml), whereas the surfactant obtained under similar cultivation conditions on glycerol and n-hexadecane, inhibited growth of tested bacteria and yeast at higher (9-68 and 27-54 µ/ml, respectively) concentrations. The minimum inhibitory concentration of surfactant, synthesized in a medium with ethanol (glycerol, n-hexadecane), yeast autolysate and trace elements, correlated with the activity of NADP+-dependent glutamate dehydrogenase - a key enzyme of aminolipids biosynthesis (610 ± 30, 395 ± 24, 397 ± 24 nM min-1·mg-1 protein, respectively). Conclusions: The higher activity of NADP+-dependent glutamate dehydrogenase when growing the strain IMV B-7241 in a medium with ethanol (n-hexadecane), yeast autolysates and trace elements compared to that in a medium with copper sulfate and iron sulfate, as well as an increase enzyme activity in the presence of zinc cations suggests the possibility of increasing synthesis aminolipids by introducing Zn2+ in the medium with ethanol and n-hexadecan. The obtained data indicate the need for studies depending on biological properties of surfactants of the cultivation conditions of producer.

[Biological Properties of Nocardia vaccinii IMV B-7405 Surfactants Synthesized on Byproduct of Biodiesel Production].

Aim: Comparison of antimicrobial and anti-adhesive activity of Nocardia vaccinii ІMV B-7405 surfactants synthesized on purified and technical glycerol (byproduct of biodiesel production). Methods: Surfactants were extracted from supernatant of cultural liquid by mixture of chloroform and methanol (2:1). Antimicrobial against bacteria and yeast properties of the surfactant was determined by index of the minimum inhibitory concentration (MIC). The number of attached cells was determined spectrophotometrically. Results: The dependence of surfactant antimicrobial and anti-aghesive activity on the degree of glycerol purification (purified, technical), as well as the duration of N. vaccinii IMV B-7405 cultivation on these substrates was established. MIC against studied test cultures surfactants, synthesized on technical glycerol during 5 days, was 15­121 µg/ml, that lower than MIC of surfactant obtained on purified substrate (22.5−180 µg/ml). Increasing duration of N. vaccinii ІMV B-7405 cultivation accompanied by rise the MIC against some test cultures surfactants synthesized on both technical and purified glycerol. Adhesion of the bacteria Escherichia coli ІEM-1, Bacillus subtilis BT-2 (vegetative cells and spores) and the yeast Candida albicans Д-6 on abiotic surfaces treated with surfactants synthesized on technical glycerol for both 5 and 7 days, was an average of 11−12 % less than after materials treatment with surfactants obtained on the purified substrate. Conclusions: Increasing activity of NADP+-dependent glutamate dehydrogenase − key enzyme of aminolipids biosynthesis (effective antimicrobial and anti-adhesion agents) in the presence of K+ and Na+ may indicate the possibility of biosynthesis intensification of these components of surfacrants complex under N. vaccinii ІMV B-7405 cultivation on technical glycerol, which is characterized by a high content of sodium and potassium cations. Replacing refined glycerol on byproduct of biodiesel production will not only reduce the cost of N. vaccinii ІMV B-7405 surfactant biosynthesis, but also to obtain the final product with high antiadhesive and antimicrobial activity.

[INFLUENCE OF FUNGICIDES COMPLEX INOCULUM EKOVITAL ON RHIZOSPHERE MICROBIOCENOSIS, DISEASES RESISTANCE AND SOYBEEN PRODUCTIVITY].

It has been investigated the effect of fungicide with systemic action of Vitavaks 200 FF, Maxim Star 025 FS, Kinto duo and the complex inoculum Ekovital on rhizospheric microbial communities, diseases resistance and soybean productivity of Annushka biovar. The combined use of fungicides and inoculation has contributed to better formation of nodulation apparatus (the number of nodules on the roots was increased in 1.3-2.8 times), resistance increase to soybean pathogens septoriosis and ascochitosis, plant productivity increase on 17.4-32.1% relatively to the variant with Ekovital. The efficiency of the combined treatment of seeds against septoriosis and ascochitosis has become 45.8-64.1% and 82.0-95.1% respectively, in the flowering-early fruiting stage and 38.1-60.6% and 70.3-82.1% respectively, in the loading beans phase.

[BIOCONVERSION OF CRUDE GLYCEROL AND MOLASSES MIXTURE IN BIOSURFACTANTS OF NOCARDIA VACCINII IMB B-7405].

The possibility of replacing glucose and pure glycerol in mixed substrates for surtace-active substances (SAS, biosurfactants) biosynthesis of Nocardia vaccinii IMB B-7405 on molasses (sugar production waste) and crude glycerol (by-product of biodiesel production) was established. It was established that the increasing concentration of crude glycerol to 6% in mixture with 1.0% molasses was accompanied by increase of amount of SAS synthesized more than twice, and the increasing content of molasses to 3.0% in mixture with 1.0% crude glycerol--by some decrease in the level of surfactant as compared to that in a medium containing 1.0% monosubstrates. It was shown that the increasing concentration of sodium nitrate to 2-fold in medium cultivation of N. vaccinii IMB B-7405 allowed to increase to 7.0% content of grude glycerol in mixture with 1.0% molasses. Under such conditions of cultivation concentration of exocellular SAS synthesized was 7,5 g/l, that to 1,3 fold higher than in basic medium with a lower content of nitrogen source.

[ANTIMICROBIAL ACTION OF NOCARDIA VACCINII IMV B-7405 SURFACTANTS].

AIM: To study the effect of Nocardia vaccinii IMV B-7405 surfactants on some bacteria (including pathogens of genera Proteus, Staphylococcus, Enterobacter), yeast of Candida species and fungi (Aspergillus niger R-3, Fusarium culmorum T-7). METHODS: The antimi- crobial properties of surfactant were determined in suspension culture by Koch method and also by index of the minimum inhibitory concentration. Surfactants were extracted from supernatant of cultural liquid by mixture of chloroform and methanol (2:1). RESULTS: It is shown that the antimicrobial properties of N. vaccinii IMV B-7405 surfactant depended on the degree of purification (supernatant, solution of surfactant), concentration and exposure. Survival of Escherichia coli IEM-1 and Bacillus subtilis BT-2 (both vegetative cells and spores) after treatment for 1-2 hours with surfactants solution and the supernatant (the surfactant concentration 21 µg/ml) was 3-28%. Minimum inhibitory concentrations of N. vaccinii IMV B-7405 surfactants on studied bacteria, yeast and micromycetes were 11.5-85.0; 11.5-22.5 and 165.0-325.0 µ/ml respectively. CONCLUSIONS: Minimum inhibitory concentrations of N. vaccinii IMV B-7405 surfactants are comparable to those of the known microbial surfactants. The possibility of using the supernatant of culture liquid as an effective antimicrobial agent noticeably simplifies and reduces the cost of the technology of its obtaining.

Elaboration of natural polyfunctional preparations with antiparasitic and biostimulating properties for plant growing.

Producer of macrolide antibiotic avermectin Streptomyces avermitilis UCM Ac-2179 has been isolated from Ukrainian chernozem soil, its biosynthetic activity has been increased by the traditional selection and chemical mutagenesis methods. Streptomyces avermitilis UCM Ac-2179 synthesizes avermectin with the content of anti-parasitic B-components more than 40%. Addition of exogenous Na-pyruvate (1.5 mg/L) in cultural medium promotes a 2.5-fold augmentation of the avermectin synthesis. The preparation Avercom has been obtained by the method of ethanol extraction from the producer biomass. This preparation includes antibiotic avermectin and other biologically active substances: free amino acids, lipids, phytohormones. Avercom has high nematicidic activity and raises plant resistance to fungal and viral diseases. On the base of Avercom and plant growth regulators the complex preparations Actinolan and Ascoldia have been elaborated. The effectiveness of the biopreparations as nematicidic and plantstimulating means under experimental and industrial conditions was confirmed.