Growth and Phytase Activities of Bacillus subtilis IMV B-7023 During Cultivation with Sodium Phytate

Bacteria of the genus Bacillus are known for their ability to mineralize organic phosphorus compounds. Phytates constitute up to 60–80% of the total plant phosphorus and almost 50% of soil organic phosphorus. Phytates phosphorus is unavailable for plants. Bacillus can synthesize phosphatases both wide spectrum of action, and highly specific phytases that catalyze the hydrolysis of phytates. Therefore, the aim of this work was to study the growth and phytase activity of Bacillus subtilis IMV B-7023, which is the component of the ”Azogran” complex bacterial preparation for plant growing. Methods. The growth activity of bacteria was studied by cultivation methods, the phytase activity – by measuring the amount of phosphate released from sodium phytate during the enzymatic reaction. Results. It was shown that B. subtilis IMV B-7023 assimilated phytate as the source of phosphorus nutrition during cultivation in media with 0.5, 1.0 and 2.0 g/L of sodium phytate. The highest growth activity of these bacteria was observed after two days of cultivation in medium with 1.0 g/L of phytate. The number of bacteria was (3.91±0.32)×109 CFU/mL under these conditions. At the same time, B. subtilis IMV B-7023 demonstrated a low level of phytate assimilation as a source of carbon nutrition. Thus, after two days of cultivation the number of bacteria increased from (4.12±0.09)×106 CFU/mL to (1.07±0.07–3.11±0.51)×107 CFU/mL in the presence of 0.5–2.0 g/L phytate in the medium and the absence of another carbon source. It was determined that strain B. subtilis IMV B-7023 had phytase activity, the highest activity (221.85±0.12 U/g) was on the first day of their cultivation in medium with inorganic phosphates. It should be noted that B. subtilis IMV B-7023 phytase activity was lower during cultivating in medium with sodium phytate as a source of phosphorus nutrition, than in medium with inorganic phosphates. The obtained fact may be due to phytate hydrolysis by widespecific phosphatases. Higher rates of phytase activity obtained on the first and third days compared to the second and fourth days of bacterial cultivation may indicate the expression of phosphatases genes only in the period required for maximum bacterial development, in the absence of these proteins in the media. At the same time, the phytase activity of B. subtilis IMV B-7023 after 2 days cultivation in a media with 0.5 and 1.0 g/L of sodium phytate (194.80±0.15 U/g and 160.90±0.13 U/g, respectively) as the source of carbon and phosphorus was higher compared to the activity of bacteria on medium with inorganic phosphates (137.79±0.10 U/g). This may be caused by the synthesis of a larger number of highly specific phosphatases (phytases) in bacterial cells at the presence of only phytate in the medium as a substrate. Conclusions. B. subtilis IMV B-7023 strain is characterized by growth on nutrient medium with sodium phytate and phytase activity. Because they are soil microorganisms used as the component of the ”Azogran” complex bacterial preparation for plant growing, the ability to hydrolyze and assimilate phytate is important for functioning of this strain in the rhizosphere. The obtained results extend the understanding of B. subtilis IMV B-7023 influence on phosphorus nutrition and development of plants.

The use of the strain Enterococcus faecium in the technology of yogurt “Carpathian” as an ancillary culture with probiotic properties

The results of research on the technological features of the production of yogurt "Carpathian" are covered in the article. The bacterial preparation of Chr. Hansen series YoFlex Premium 1.0 (L. bulgaricus, S. thermophilus) and Creamy 1.0 (L. bulgaricus, S. thermophilus, L. rhamnosus) and strain E. faecium SB 18, which is isolated from traditional Carpathian fermented products were used to produce research yogurt samples. It was found that when the strains were used together, the microorganisms were compatible, did not show interspecific antagonism and did not inhibit the enzymatic process. Based on yogurt microorganisms and E. faecium SB 18 strain, seven prototypes of yogurt were created: № 1 (100 %) – control, Premium + Creamy; № 2 (100 %) – control, pure culture of E. faecium SB 18; №3 (100:100 %) – control, (Premium + Creamy) + E. faecium SB 18; №4 (50:50 %) – (Premium + Creamy) + Ent. faecium SB 18; № 5 (70:30 %) – (Premium + Creamy) + Ent. faecium SB 18; № 6 (80:20 %) – (Premium + Creamy) + Ent. faecium SB 18; № 7 (90:10 %) – (Premium + Creamy) + Ent. faecium SB 18. The fastest fermentation took place in sample № 1 (pH 4.78 units – 4 h), the slowest in sample №2 (pH 4.81 units – 6 h), where only pure culture of E. faecium SB 18 was used. The fermentation time in sample №3 was initially slower and then more active (pH 4.77 units – 4 h). The acidity increased more moderately in samples № 4, 5, 6, 7 for 4 h, and at the end of fermentation was 4.84 units, 4.76 units, 4.81 units. and 4.75 units. in accordance. According to organoleptic evaluation, the experimental samples were characterized by slight differences. In general, it is noted that the addition of microbial culture of E. faecium SB 18 improves the taste of yogurt. Samples № 6 and № 7 with the addition of E. faecium SB 18 strain in the amount of 20 and 10 % were noted for the best organoleptic properties. According to the score scale, the above-mentioned samples received the highest number of points – 48, out of a possible 50. The dependence of the acidity of yogurt during storage was established on the dose and composition of the bacterial preparation. It was investigated that the acidity of yogurt, which contained an additional strain of E. faecium SB 18 in the ratios of 100:100 (sample 3) and 50:50 (sample 4), tends to increase rapidly in acidity, which is associated with increased lactic acid bacteria. It was found that partial replacement of the amount of traditional yogurt leaven in the ratio of 70:30 (sample 5), 80:20 (sample 6) and 90:10 (sample 7) provides the optimal course of the enzymatic process during fermentation and storage. It was found that the use of traditional yogurt leaven YoFlex Premium 1.0 and Creamy 1.0. together with strain E. faecium SB 18 in a ratio of 80:20, provides excellent consumer properties of the product.

Aplicarea pesticidelor bioraționale la plantațiile de piersici în zona centrală a Republicii Moldova

In this paper, the results of application of plant extracts, bioelicitors – Reglalg, algae extract – Spi-rogira sp., are shown. A mix of unsaturated fatty acids, aldehydes, ketones and other active components - 0.5 l / ha and Paurin - bacterial preparation (based on Agrobacterium tumefaciens) - 2.0 l / ha, which as growth regulators, contribute to increased peach resistance to different diseases.

Efficiency of the Complex Bacterial Preparation Azogran Application in Protecting Potatoes from the Colorado Potato Beetle Depending on the Stage of Its Development

The Colorado potato beetle (Leptinotarsa decemlineana Say) is a particularly dangerous potato phytophage. The aim of this work was to study the effect of complex granulated bacterial preparation Azogran and its components on the spread of the Colorado potato beetle in the potato phytocenosis. Methods. The study was carried out during potato of the Slavyanka variety growing under the conditions of small-plot and laboratory experiments. The complex granulated preparation Azogran was made on the basis of interaction of nitrogen-fixing bacteria Azotobacter vinelandii IMV B-7076, phosphate-mobilizing bacteria Bacillus subtilis IMV B-7023 and clay mineral bentonite. 1 g of the preparation contains more than 108 viable bacteria of each strain. The mass of 1 granule was 0.25 g. The effect of Azogran, its components and 4-hydroxyphenylacetic acid (4-HPAA), a metabolite of B. subtilis IMV B-7023, on the survival of Colorado potato beetle larvae was estimated in laboratory conditions. Results. It was shown that the use of the complex bacterial preparation Azogran, B. subtilis IMV B-7023 strain and its metabolite 4-HPAA in the potato phytocenosis significantly reduces the spread of the Colorado potato beetle imago, as well as the number of egg-laying on the leaf surface of plants. Treatment of eggs clutches of the phytophage with this preparation and its specified components significantly reduces the number of larvae on the leaves. Treatment of 1–4 stages larvae with solutions of 4-HPAA at a concentration of 0.5–50.0 μg/ml, as well as a suspension of B. subtilis ІМV В-7023 bacteria or their culture medium (without cells), leads to significant death of the larvae. Conclusions. The use of the complex bacterial preparation Azogran, as well as its component bacteria B. subtilis IMV B-7023 in the phytocenosis of potatoes stimulates plant growth, significantly reduces the spread of the Colorado potato beetle and the number of eggs clutches on the leaf surface. Treatment of Colorado potato beetle larvae of 1–4 stages of development with a suspension of B. subtilis IMV B-7023 bacteria, as well as their metabolite 4-HPAA leads to the death of more than 65% of larvae. The results obtained indicate that the use of the complex bacterial preparation Azogran is promising for potatoes growing.

SUBSTANTIATION OF THE ADAPTIVE SORTING TECHNOLOGY OF GROWING LEGUMINOUS CROPS IN THE RIGHT-BANK FOREST STEPPE OF UKRAINE

The relevance of the conducted research is based on the tasks of the applied research of Vinnytsia National Agrarian University on the topic: «Development of methods for improving the technology of growing leguminous crops using biofertilizers, bacterial preparations, foliar fertilizers and physiologically active substances». Analysing the literature sources various technological methods of growing leguminous crops for providing the rational use of natural agricultural potential, which will further increase the sown area of major legumes, which are of strategic importance, have been evaluated. The article analyses the varietal diversity of leguminous crops included in the State Register of Plant Varieties of Ukraine. High-yielding varieties for the Right-Bank Forest-Steppe of Ukraine of soybeans – Azymut, Holubka, sowing peas – Tsarevych and Prystan have been chosen for the study; white lupine – Veresnevyi, Chabanskyi; narrow-leaved lupine – Olimp, Peremozhets; chickpeas for sowing – Skarb, Pegas. According to the group of ripeness, early-, medium- and medium-early-ripening varieties of legumes are selected, which have been distinguished among others for their high grain and protein productivity. It has been determined that the types of legumes in terms of set basic economic-valuable peculiarities have significant advantages such as resistance to adverse environmental factors, pests and manufacturability. The field experiments have shown that the combination of inoculation of seeds with a bacterial preparation and treatment of plants for vegetation with a retardant has had a positive effect on increasing the yield of the crops. For the years of the conducted researches, the maximum seed yield in the leguminous crops has been determined. Thus, in sowing peas the most productive type was Prystan (2.6 t/ha), white lupine – Chabanskyi (3.4 t/ha), narrow-leaved lupine – Peremozhets (2.6 t/ha), chickpea – Skarb. 3.0 t/ha) and in soybeans – Azymut (2.6 t/ha). The largest increases in grain productivity were obtained by treating the seeds with the bacterial preparation Rhizohumin and spraying the crops with chlormequat chloride retardant in the budding phase. Keywords: leguminous crops, variety, quality, yield, growing area.

The technology of soft Camembert cheese with the usage of different bacterial preparations

The aim of the work was to develop a technology of soft Camembert cheese from pasteurized cow's milk using various bacterial preparations and to investigate its basic physicochemical and organoleptic properties. Two experimental samples of cheese were made: sample 1 – culture of direct application of mesothermophilic type RSF-742 + culture of white mold (Penicillium candidum + Geotrichum candidum) + rennet enzyme + calcium chloride; sample 2 – culture of direct application of mesophilic type CHN-19 + culture of white mold (Penicillium candidum + Geotrichum candidum) + rennet enzyme + calcium chloride. The finished product analyzed by the organoleptic and physicochemical parameters. Syneretic properties of rennet clots were studied after the process of fermentation and coagulation at a temperature of 32 ºC. The sample with the use of CNH-19 was characterized by the best consistency and pronounced mushroom and creamy aroma. Sample № 1, made with the bacterial preparation RSF-742, was characterized by a less pronounced aroma and structure, typical of Camembert cheese. It was found that the sample №1 (RSF-742) was characterized by higher syneretic properties. According to syneretic properties, lower moisture content characterized sample 1 (62 %) versus sample 2 (64 %). The volume of serum released in 1 hour was 65 % for sample 1 and 62 % for sample 2. The highest values for the fat content was sample 1 – 43 %, and sample 2 – 42 %. The content of salt did not differ, in sample № 1 – 1.8 % and in sample № 2 – 1.75 %. There were changes in the active acidity of the finished product when using different bacterial preparation. Lower pH values in sample 1 – 6.2, and slightly higher in sample 2 – 6.5 were observed. The highest number of points according to the results of the score was given to sample № 2 – 86 points, sample № 1 – 77 points out of possible 100.

Antagonistic Activity of Azotobаcter vinelandii IMV B-7076 against Phytopathogenic Microorganisms

Bacteria of the genus Azotobacter are known for their ability to stimulate plant growth and development. Azotobacter vinelandii IMV B-7076 strain was isolated from Zhytomyr region soil of Ukraine. It is one of the components of the "Azogran" complex bacterial preparation for plant growing. It has been established that A. vinelandii IMV B-7076 synthesizes biologically active substances that promote plant development. At the same time, the antagonistic activity of A. vinelandii IMV B-7076 against phytopathogens has not yet been studied, so this became the aim of this work. Methods. The antagonistic activity of A. vinelandii IMV B-7076 was determined by agar well diffusion and agar blocks methods. Results. It was shown that A. vinelandii IMV B-7076 had antagonistic activity against some phytopathogenic fungi. In particular, the diameter of growth inhibition zones of Alternaria alternata 16861, Fusarium avenaceum 50720, Fusarium verticillioides 50463, Fusarium lactis 50719, Fusarium oxysporum 54201, Fusarium poae 50704 was 14-37 mm, Bipolaris sorokiniana 16868 and Fusarium solani – 11-13 mm. Fusarium culmorum 50716 and Fusarium graminearum 50662 were not sensitive to A. vinelandii IMV B-7076 metabolites. Notably, the antagonistic effect was demonstrated in mycelial growth and spore formation inhibition, in fungal mycelium discoloration. It was also demonstrated that A. vinelandii IMV B-7076 did not show antagonistic activity against phytopathogenic bacteria Agrobacterium tumefaciens 8628, Pectobacterium carotovorum subsp. carotovorum 8982, Pseudomonas fluorescens 8573, Pseudomonas syringae pv. syringae 8511, Clavibacter michiganensis subsp. michiganensis 13a, Xanthomonas campestris pv. campestris 8003b. Conclusions. Studied A. vinelandii IMV B-7076 strain is characterized by antagonistic activity against phytopathogenic fungi and does not have antibacterial properties against phytopathogenic bacteria. The antifungal activity of A. vinelandii IMV B-7076, as a component of "Azogran", will be useful for this bacterial preparation application in plant growing.

Forage productivity of bastard alfalfa as affected by cover crop and seed treatment

Разработка приёмов повышения кормовой продуктивности люцерны изменчивой для обеспечения стабильной кормовой базы интенсивного молочного скотоводства в Удмуртской Республике является актуальной задачей. Предпосевная обработка семян — простой и малозатратный приём, повышающий продуктивность культур за счёт оптимизации минерального питания растений в начале их роста и развития. Целью работы было определение эффективности предпосевной обработки семян и влияния покровной культуры в технологии возделывания люцерны изменчивой. Исследования проводили в 2017–2019 годах на опытном поле АО «Учхоз Июльское ИжГСХА» на дерново-подзолистой среднесуглинистой почве со следующей агрохимической характеристикой пахотного слоя: среднее и повышенное содержание гумуса (2,3–2,8%); очень высокое содержание подвижного фосфора (265–284 мг/кг почвы), повышенное и очень высокое содержание подвижного калия (171–252 мг/кг почвы), обменная кислотность почвы слабая (рН — 5,3–5,5). Схема опыта включала беспокровный посев и посев под однолетние травы на зелёный корм и яровую пшеницу. Предпосевная подготовка семян заключалась в обработке молибденовокислым аммонием (300 г/т), бактериальным препаратом «Ризоторфин» (0,5 л/т), регулятором роста растений НВ-101 (1 мл/т), комплексным удобрением Agree`s «Форсаж» (1,2 л/т). Наибольший сбор сухого вещества получен при посеве люцерны изменчивой под покров однолетних трав на зелёный корм. Исследованиями установлена эффективность предпосевной обработки семян микроудобрением, бактериальным удобрением и регулятором роста растений. К концу вегетации первого года жизни количество клубеньков на корнях люцерны изменчивой колебалось от 9 до 31 шт. на растении, при этом в контрольном варианте их было 9–16 шт., в изучаемых вариантах — 17–23 шт. на растении. При предпосевной обработке семян в первый год пользования за два укоса сбор сухого вещества увеличился на 1,06–2,71 т/га, или на 14–34%, во второй год пользования — на 1,35–3,45 т/га, или на 13–33%, относительно продуктивности посевов без обработки семян. Optimization of alfalfa production for cattle feeding is of high interest in the Udmurt Republic. Seed treatment prior to planting is a simple and low-cost procedure that improves plant mineral nutrition in the beginning of its life cycle and increases its productivity. The investigation aimed at testing the effectiveness of seed treatment and cover crop when growing bastard alfalfa. Soil — sod-podzolic, medium clay content, medium and high humus content (2.3–2.8%), high concentrations of soluble phosphorus (265–284 mg/kg) and potassium (171–252 mg/kg), рН — 5.3–5.5. Annual grasses for green fodder and spring wheat performed as cover crops. Alfalfa seeds were treated with ammonium molybdate tetrahydrate (300 g/t), “Rizotorfin” bacterial preparation (0.5 l/t), NV-101 growth regulator (1 ml/t), Agree`s “Forsazh” complex fertilizer (1.2 l/t). Seeding alfalfa with annual grasses provided the highest yield of dry matter (DM). Microfertilizer, bacterial preparation and growth regulator had positive effect on alfalfa. By the end of the first life cycle alfalfa root nodule number varied within 9–31 pcs/plant, in the control variant — 9–16 pcs/plant, in the experimental variants — 17–23 pcs/plant. In the first year seed treatment increased DM yield by 1.06–2.71 t ha-1 (14–34%) for two cuts, in the second one — by 1.35–3.45 t ha-1 (13–33%).

EFFICIENCY OF NANOCOMPOSITE COMPLEX OF BACTERIAL PREPARATION AZOHRAN IN MILLET CULTIVATION TECHNOLOGY

Objective. Determine the efficiency of bacterization of millet seeds with a complex nanocomposite bacterial preparation Azohran (Azohran-nano) depending on the growing conditions of the culture; establish the effect of mineral fertilizers in combination with a biopreparation on the productivity of agrocenoses. Methods. Field experiment (to assess the influence of the studied factors onthe growth and development of plants and millet yield), measuring- and weight-based, economiccalculations, statistical. Results. During three-year observations, a positive effect of mineral fertilizers and pre-sowing inoculation of millet seeds with Azohran-nano on plant height, plant weightgain, heads weight and crop productivity was established. The height of plants in the variant withbacterization of seeds when growing millet without fertilizers in the stalking phase was higher by5.6 % than the corresponding parameters of the control variant and with the concomitant use offertilizers and bacterization this figure increased to 8.1 %. Seed inoculation had a positive effect onthe weight of millet heads.The efficiency of bacterization in the cultivation of the crop at the background without fertilizers was 6.3 %, and when the plants were cultivated using nutrition with nitrogen during the growing season, this figure increased by 11.5 %. The highest yields of millet were reported for seedtreatment with Azohran-nano. Yield in the control variant was 3.75 t/ha, while after the bacterization of seeds with the preparation, crop productivity increased by 16.5 % Upon the interaction between mineral fertilizers with nanocomposite preparation, parameters increased by 19.9–28.6 %.Under the influence of pre-sowing inoculation of seeds, the cost of 1 tonne of grain decreased, andthe net operating profit increased. Conclusion. The use of nanocomposite complex of the preparation Azohran-nano in millet cultivation technologies promotes the growth and development ofplants, increases crop productivity and is a highly efficient agricultural measure.