The granular composition containing microorganisms, the method of its preparation and its use

 

(57) Abstract:

The invention relates to Microbiology and relates to a granular composition containing microorganisms, and a method of its production. The obtained granulated composition comprising fine substrate, the polymer layer containing the microorganisms and water. The polymer is a film-forming, water-soluble and practically seamless polymer or film-forming structurally crosslinked polysaccharide containing carboxyl groups and swelling in water in the presence of potassium ions. As substrate composition contains crushed bran, straw, wood dust, cellulose, clay, attapulgite, diatomaceous earth, vermiculite and other substances. As a film-forming water-soluble polymer, it contains a homopolymer or copolymer of polyvinyl alcohol, polyethylene glycol or polyvinylpyrrolidone, polyacrylamide, starch, carrageenan, alginate, xanthan gum, resin carob, methylcellulose. Of microorganisms Pseudomonas, Penicillium and other Method of obtaining the composition provides for the suspension of the polymer or polysaccharide and microorganism and nabryzgivanii the resulting suspension on fine substrate, and removing water to keep her in the room temperature. 2 C. and 41 C.p. f-crystals, 6 PL.

The invention relates to a granular composition containing (1) a water-insoluble solid and fine substrate, (2) water-soluble or swelling in water, film-forming polymer that is not covalently crosslinked or crosslinked with polyvalent cations, (3) organisms and (4) water. The invention further relates to a method for preparing the specified granular composition and to its use for protection of plants from diseases and insects.

Plant protection using spore or vegetative cells (microorganisms) has recently become of great importance. A prerequisite for the use of such agents for biological control is the ability to obtain useful preparative forms, such as suspension concentrates, dispersible powders, granules or in particular scatter granules. Preparation preparative forms, however, is difficult. For example, for the preparation of preparative forms the basis of most plant cells and some dispute cannot be used, the process is carried out at temperatures above 40oC because the microorganisms when this damage is Mami, because it is impossible to avoid loss of viability in the environment, which leads to cell death, or due to the need of storage of the composition at low temperature to avoid loss of viability.

The most famous structures on the basis of microorganisms represent a polymer gels, crosslinked with polyvalent cations and containing these microorganisms. Such a composition is described inter alia in D. R. Fravel et al. in Phytopathology, T. 75, No. 7, 774-777, 1985, in which the polymer is alginate. In the same source described the use of substrates as a necessary component part of such structure. The preparation of these compounds is usually carried out by mixing solutions of natural or synthetic gel-forming polymers, such as alginates, and aqueous solutions of ions of polyvalent metals with obtaining the individual drops, and microorganisms can be suspended in one or both of the reaction solutions. The gel formation begins when the suspension of the microorganism is added dropwise to a solution of gelling agent. These gel particles can then be dried. This method is called the ionotropic gelation. Depending on the degree of drying at containing the microorganisms and the substrate, which are distributed almost uniformly. The particle size can be up to 5 mm

In EP-A1-0097571 described compositions on the basis of partially cross-linked polysaccharides, which in addition to the microorganism can contain finely dispersed silicic acid as a substrate, and the stitching can be performed with ions of Ca++. The water activity of the compounds can be not more than 0.3. In a review article in W. J. Connick et al., on various compositions, in New Directions in Biological Control: Alternatives for Supressing Agricultural Pests and Diseases, pp. 345-372, Alan R. Liss, Inc. (1990) described granular formulations with vermiculite as substrate and compact alginate granules obtained by ionotropic gelation. Such compositions are also described in D. R. Fravel in Pesticide Formulations and Application Systems: 11thVolume, ASTM STP 1112 American Society for Testing and Materials, Philadelphia, 1992, PP 173-179.

The disadvantage of these formulations based on crosslinked gels is to slow the release of biological control agent because the gel does not dissolve in water and are usually formed by large particles with a diameter greater than a few mm If you want more quick selection, the compositions must be pre-processed, usually buffer solutions. It is difficult for the consumer and safely reduces the at g), necessary to reduce the consumption norms, systems typically do not have sufficient stability and to avoid significant losses to be stored at low temperatures. To prepare the compositions of the gel-forming polymer must be dissolved in water, which in some cases is difficult and only possible at elevated temperatures. The formation of drops of the gel is a necessary stage in the process to obtain the appropriate granular formulations. A device for implementing this process on an industrial scale should be considered difficult and expensive. Thus obtained particles still have a high water content, which must be reduced by drying to provide acceptable stability during storage. This stage drying leads to an even greater appreciation of the process, the microorganisms may be at greater risk of destruction, and their viability is reduced even more. Up to the present time were not known stable when stored granular formulations based on soluble or swelling in water of the polymers, brewed without ionotropic gelation.

Suddenly Birnam layer without ionotropic gelation and partly without complete dissolution of the polymer, (b) significantly reduce the loss caused by the death of living cells during drying, (C) to achieve high stability during storage, in particular in the environment, (g) to achieve very high densities of microbial population while providing excellent stability during storage, (d) to achieve a rapid selection of biological control agent and (e) to provide excellent stabilization of the bacterial cells of plant origin due to the application of microorganisms in soluble or swelling in water, the film-forming polymer that is not covalently crosslinked or crosslinked with polyvalent cations, on the substrate or with a substrate, the composition contains at least 0.5 weight. % of water, calculated on the whole composition.

One of the objects of the invention is a granular composition, representing fine substrate and the polymer layer containing the microorganisms, and this polymer is a

(a) a film-forming, water-soluble and practically seamless polymer, while the granular composition contains at least 0.5 wt.% water in terms of composition, or

(b) plenkoobrazuyushchie structurally cross-linked polysaccharide, which contains carboxyl or is it 0.5 wt.% water in terms of composition.

In the context of the present description, the term "virtually seamless" means that you do not add a Monomeric cross-linking agents, which lead to the formation of covalent bonds, or polyvalent cations, which lead to the ionotropic gelation.

"Structurally-linked" in the context of the present description means the formation of a spatial network of a single polymer or a mixture of two polymers with hydrogen bonds or due to the electrostatic interaction of ions of potassium. This forms a thermally reversible spatial structure (gel), which when heated again dissolved. Typical examples are expressed in the form of the double helix structure of carrageenan in the presence of potassium ions or structural formation of a mixture of carrageenan and resin carob. Thermally irreversible structural formation using polyvalent ions do not fall under the above definition.

On a recurring unit of the polysaccharide may have one or more carboxyl or sulfate groups.

"Water-soluble" in the context of the present description means that you can get at least 0.5% by weight aqueous solution of the polymer at a temperature of 1-10 weight. %, in particular 0.3 to 5 wt.%, most preferably 0.5 to 3.0 weight. % in terms of dry substance per 1 kg of the composition. The sum of all components of the granular composition is always 100%.

Population density per cell concentration may be particularly high. Preferably the density of the population of the microorganism is from 1x105to 1x1011CFU (colony forming units) per gram of granulated composition. During storage at room temperature the concentration of living cells can be maintained in the composition according to the invention during the period up to 10 months with only minor losses of microorganisms that make up less than a single number to 10 KOE.

The residual water content is preferably not less than 1 wt.%, more preferably at least 3 wt.% and most preferably at least 5 wt.%. The upper limit of water content is preferably not more than 40 wt.%, more preferably not more than 30 wt.% and most preferably not more than 20 wt.%. The upper limit of water content is regulated by the media, the solubility of the polymer and method of preparing composition. In the coating process, for example, in the fluidized bed, easily the achievement of the commonly ranges from 0.5 to 40 wt.%.

Fine substrate can have particles with an average size of from 1 μm to 0.8 cm, more preferably from 10 μm to 0.5 cm, and most preferably from 20 μm to 0.2 see, the Substrate may be inorganic or organic material. For fungi, it is preferable to use organic materials and plant cells (bacteria) - inorganic materials. Typical examples of water-insoluble organic materials are crushed bran, straw, sawdust and cellulose. The most suitable inorganic substrates are water-insoluble metallic oxides and salts of metals (SiO2, Al2O3, BaSO4, CaCO3or silicates and aluminosilicates of alkali metals and alkaline earth metals. Among silicates preferred lamellar silicates. Typical examples of silicates are mineral clay, attapulgite, diatomaceous earth, powdered lime, diatomaceous earth, wollastonite, olivine, montmorillonite and vermiculite. Especially preferred vermiculite.

The number of the substrate is usually from 50 to 99 wt.%, preferably from 65 to 95 wt.% and most preferably from 75 to 90 wt.%.

The average particle size of the granulated composition can comp the Yong an average particle size of from 0.5 to 2 mm.

Film-forming water-soluble and practically seamless polymer can be synthetic or natural. Typical examples of synthetic polymers are Homo - and copolymers of polyvinyl alcohol, polyethylene glycol or polyvinylpyrrolidone, and polyacrylamide. Natural polymers are mainly polysaccharides, which can be derivatization. There are a large number of known preferred natural polymers, and they are usually starch, alginates, carragenan, more preferably k-carrageenan, l-carrageenan-carragenan, xanthan gum, resin carob or methylcellulose. You can also use a mixture of polymers.

The polymers must be compatible with the microorganism. Compatibility may be installed by a specialist in the art in a simple way by connecting microorganism and polymer.

Especially preferred alginates and carragenan. Especially preferred combination of media and water-soluble polymer are vermiculite with k-carrageenan.

Film-forming, structurally crosslinked swellable in water, the polymer is a polysaccharide, preferably k-carrageenan, l-carrageenan, smut thermally reversible gels, characterized by intermolecular hydrogen bonds or ionic bonds.

The amount of soluble or swelling in water of the polymer may be from 0.1 to 20 wt.%, preferably from 0.1 to 10 wt.% and most preferably from 0.5 to 5 wt.%.

The molar ratio of potassium ions and carboxyl or sulfate groups of the polymer is from 0.001:1 to 1:1.

Microorganisms which can be used to control pests or plant diseases in agriculture, known and described inter alia in European patent application EP-A-0472494.

Relevant microorganisms are one or mnogokletochnye fungi or bacteria, usually involving Rhizobium spp., Metharizium, Fusarium, Trichoderma, Stryptomyces, Gliocladium, Penicillium, Talaromyces, Verticillium or Colletotrichum. Preferred Pseudomonas spp., Serratia spp., Bacilus spp., Agrobacter spp., Exserohilum spp., Enterobacter spp. Particularly preferred microorganism Pseudomonas aurantiaca, ATTC N 55169.

Weeds, insects and fungal diseases that can be treated with microorganisms, usually are Rhizoctonia solani, Rhizoctonia oryzae, Phytium ultimum, Fusarium oxysporum spp., Alphanomyces laevis, Phytophtora infestans, Botrytis spp., Sclerotinia sclerotiorum, Bacillus sp. , Microdochium nivale, Thielaviopsis basicola, Gaeumanomyces graminis and basically all other diseases, ntiaca ATTC N 55169 is active against a number of diseases, the above for different cultures. It should be noted a protective effect against Rhizoctonia solani in cotton, pumpkin crops, cabbage, geraniums, Impatiens and poinsettias.

In the preparation of traditional fine-grain compositions (see, for example, Connik W. J.: "Formulation of living biological control agents with alginate" in American Chemical Society, ACS Symposium Series 1988, I. 371, pages 241-250; Fravel, D. R., J. J. Marois, R. D. Lumsden, Connik W. J.: "Encapsulation of potential biocontrol agents in alginate" Phytopathology, 1985, issue 75, pp. 774-777; Stormo K. E., Crawford, R. L.: "Preparation of encapsulated microbial cells for environmental application" in Applied and Environmental Microbiology, 1992, pages 727-730) receive granules, practically insoluble or dissolve only very slowly even in a buffer solution, therefore, the selection of microorganisms occurs very slowly or not at all.

Unexpectedly, it was found that the granules obtained by the method according to the invention, provide a very quick selection of microorganisms. The composition decomposes in the buffer or in water, depending on the polymer, for from 0.5 to several hours, i.e., the polymer layer is separated, or swells, making all the "microbial" content is released into the soil within 24 hours

Another object of the invention is a method of obtaining granular who eat the polymer is:

a) a film-forming, water-soluble and practically seamless polymer, while the granular composition contains at least 0.5 wt.% water in terms of composition, or

b) forming a structurally cross-linked polysaccharide, which contains carboxyl or sulfate groups and swells in water in the presence of potassium ions, while the granular composition contains at least 0.5 wt.% water in terms of composition, which includes:

(A) for the preparation of granules (a) the suspension or the temperature is not above 95oC dissolution film-forming and water-soluble polymer and the suspension of the microorganism in the suspension or solution after cooling to room temperature,

(B) for the preparation of granules b) the suspension of the polysaccharide containing carboxyl groups or sulfate groups, in an aqueous buffer solution containing potassium ions, and the subsequent suspension of the microorganism in this solution,

(C) nabryzgivanii received suspensions directly on fine substrate or a mixture of these suspensions with fine substrate and

(G) removing water to a concentration that is not less than 0.5 wt.% in terms of the granular composition.

oC. To obtain a solution film-forming and water-soluble polymer temperature is from 25 to 95oC depending on the type of polymer.

Adding microorganism exercise or suspension of the polymer at a temperature below 40oC or cooled to the polymer solution at a temperature below 40oC, preferably below 30oC.

According to another variant of the method, the granular composition (b) prepared by dissolving the polysaccharide containing carboxyl or sulfate groups, in an aqueous buffer solution containing potassium ions, at an elevated temperature, for example, 70oC, or by dissolving in the same way two polymers, which interact with each other. Thermally reversible gel is formed from these chilled solutions. Adding a microorganism to produce shortly before reaching the point of solidification at a temperature below 40oC.

The buffer can be any potassium salt of polyvalent acid. Preferred commercially available phosphate buffers. Depending on the relationship monopotassium phosphate to the secondary acid phosphate pH is preferably from 0.00001 M/l to 1 M/l, preferably from 0.005 M/l to 0.05 M/L.

Water is removed as possible under more mild conditions, preferably at room or slightly elevated temperature, reaching values up to approximately 35oC.

Devices and methods for removal of water known. The preferred method depends on the viscosity of the mixture. Granular compositions according to the invention can be prepared by the known methods in the conventional system. Method nabryzgivaniya for mixing the components, usually used for coating, usually in the reactor with a fluidized bed. In this method, a solution or suspension of the polymer and the microorganism nabryzgivajut on the substrate in the fluidized bed and at the same time is subjected to drying.

Another variant of the method is the preparation of new granular formulations are known by extrusion. This method involves the mixing of all components in the mixer with the required amount of water and passing the mixture through a perforated plate. Then the pellets may be crushed to the desired size and dried.

You can use single-screw extruders, granulators, subgranular, perforated plates, etc., the Method according to izobratetalny microrganism. The resulting product is usually not a single particle is coated, and the agglomerates consisting of a set of substrate particles of irregular shape.

Depending on the chosen method of mixing and drying receive particles of different shapes. Thus, the extrusion process provides obtain cylindrical pellets, in which the substrate and the microorganism contain a coating of polymer is practically independent from each other, while the method nabryzgivaniya in the fluidized bed results in agglomerates of the substrate, in which the particles are covered with a thin layer of polymer containing microorganisms. The particle shape is preferred because of the thin polymer layer is extremely quick selection of the microorganism.

Granular compositions according to the invention in any case are solid engineering mixtures that can be used directly as a scatter granules. They are simple and safe when working with them, because you can download them directly to mechanical devices for use in the field. Consumption rates range from 1 kg to 20 kg depending on the type of microorganism.

Granular compositions according to the invention can be used DVLA, tubers, roots, soil, and weeds, insect pests and diseases found in plants, can be ingibirovany or destroyed.

Processing habitats of plants or the plants themselves granulated compositions can be carried out simultaneously or sequentially in conjunction with other chemical agents. Such chemical agents can be fertilizers, micronutrient donors, as well as other substances that affect the growth of plants. You can use selective herbicides as well as insecticides, fungicides, bactericides, nematicides, molluscicides or mixtures thereof.

The invention relates also to the use of granular formulations according to the invention for protecting plants from disease or insect injury. The struggle is with diseases of agricultural crops or ornamental crops in agriculture or horticulture, especially in cereals, cotton, vegetables, grapes, fruit, oilseeds and colors.

Examples of particularly important vegetables are pumpkin, cabbage and legumes, and flower crops - poinsettia, geranium and balsams.

Below the invention is illustrated in the examples.

2HPO4: KH2PO4= 1:0,78, pH 7) to a concentration of 40 ml 100 ml phosphate buffer heated to 70oC and add 0.7 g k-carrageenan to obtain a 0.7% aqueous solution of k-carrageenan in 0.01 M phosphate buffer. The solution is cooled to a temperature slightly above the solidification point, and mixed with a suspension of microorganisms.

Then this mixture is sprayed into the fluidized bed over 100 g of vermiculite, receiving the granulated composition of the following composition:

16% of residual water;

1.5% of the microorganisms in the form of dry substance;

81,9% vermiculite;

0,6% k-carrageenan.

Initial concentration of approximately 1,110 land only10CFU/g (colony forming units).

To achieve stability when stored at appropriate time intervals to determine the concentration.

Get the data in the table. 1.

Example A2

5 g of k-carrageenan mixed with 40 g of 0.01 M phosphate buffer. Then add 10 g of debris (30% solids) Pseudomonas aurantiaca, ATTS N 55169, received a 50-liter fermenter. The polymer together with microorganisms simultaneously mixed with 120 g of vermiculite powder and then ekstragiruyut. The obtained granules are dried in the fluidized bed full of water;

1.8% of the microorganisms in the form of dry substance;

77% of vermiculite;

3,2% k-carrageenan.

Initial concentration of approximately 3,31010CFU/g (see table. 2).

Example A3

250 ml of the broth Luria inoculated with Pseudomonas aurantiaca, ATTC N 55169, after 16 hours of cultivation the cells centrifuged in a shaker, sediment resuspended in 0.01 M phosphate buffer as in example 1 to a concentration of 40 ml.

Suspension of microorganisms is mixed with 100 ml of 3% aqueous solution of sodium alginate in 0.01 M phosphate buffer as in example 2 and is sprayed into the fluidized bed over 100 g of vermiculite.

Get granular composition of the following composition:

12% of residual water;

0,5% of the microorganisms in the form of dry substance;

85,5% vermiculite;

2.5% of sodium alginate.

The initial concentration is about 7,6108CFU/g (see table. 3).

In examples A4 and A5 use spontaneous mutant of Pseudomonas aurantiaca, ATTC N 55169. The mutant was obtained as follows: Pseudomonas aurantiaca, ATTC N 55169, plated on agar plate was assessed, Luria containing 0,00005% rifampicin, and emit a known method of spontaneously resistant mutants that are cultivated further. Thus obtained reefs is 250 ml broth, Luria, inoculated Pseudomonas aurantiaca, ATTS N 55169, (rifampicin resistant), after 16 hours of cultivation the cells centrifuged in a shaker, sediment resuspended in 0.01 M phosphate buffer content 42G analogously to example 1. Suspension of microorganisms is mixed with the same quantity of a solution of polyvinyl alcohol (Mowiol 40-88, 16%) and sprayed into the fluidized bed over 100 g of vermiculite.

Get granular composition of the following composition:

10% residual water;

0,5% of the microorganisms in the form of dry substance;

84% of vermiculite;

5.5% of polyvinyl alcohol.

The initial concentration is approximately 1,110 land only9CFU/g (see table. 4).

Example A5

250 ml of the broth Luria inoculated with Pseudomonas aurantiaca, ATTS N 55169, (rifampicin resistant), after 16 hours of cultivation the cells centrifuged in a shaker, sediment resuspended in 0.01 M phosphate buffer to a concentration of 40 ml analogously to example 1. Suspension of microorganisms is mixed with 100 ml of a 3% suspension of k-carrageenan in 0.01 M phosphate buffer as in example 2 and is sprayed on 100 g of vermiculite.

Get granular composition of the following composition:

12% of residual water;

0.5% microorganisms, Eimeria 1,1 109CFU/g (see table. 5).

Example A6

8 g l-carrageenan mixed with 40 ml of 0.01 M phosphate buffer. Then add 5 g of centrifuged spores of Fusarium nygamai, fermented in 50-ml fermenter with the environment Richard for 120 hours, the Mixture of the polymer with the microorganisms are mixed to homogeneity with 120 g of powdered vermiculite and ekstragiruyut. The obtained granulated composition is dried in the fluidized bed to the desired water content.

Get granular composition of the following composition:

13% of residual water;

0,5% of the microorganisms in the form of dry substance;

81% of vermiculite;

5.5% of l-carrageenan. (See further table. 6).

Example B1: Biological tests

The granular composition prepared according to example 1 are its biological activity after storage for various periods of time at room temperature in greenhouse conditions. Standardized test conditions are as follows:

culture:cotton;

pathogen: Rhizoctonia solani.

The granular composition bring in potted soil in the amount of 16 g/l of potting soil.

When stored for 10 months at room temperature not n the first substrate and the polymer layer, containing microorganisms, and the polymer is a) a film-forming, water-soluble and practically seamless polymer, while the granular composition contains at least 0.5 wt.% water in terms of composition, or (b) film-forming structurally cross-linked polysaccharide, which contains carboxyl or sulfate groups and swells in water in the presence of potassium ions, while the granular composition contains at least 0.5 wt.% water in terms of composition.

2. The granular composition under item 1, which contains the microorganisms in amounts of from 0.1 to 10 wt.% based on 1 kg of composition.

3. The granular composition under item 1, which contains the microorganisms in amounts of from 0.3 to 5 wt.% based on 1 kg of composition.

4. The granular composition under item 1, which contains the microorganisms in amounts of from 0.5 to 3 wt.% based on 1 kg of composition.

5. The granular composition under item 1, which contains the microorganisms with a population density of from 1 to 105to 1 1011CFU (colony forming units) per 1 g of the composition.

6. The granular composition under item 1, in which the residual water is not less than 1 wt.% in terms of composition.

7. The granular composition under item 1, in Coto composition under item 1, in which the residual water is not less than 5 wt.% in terms of composition.

9. The granular composition under item 1, in which the maximum water content is not more than 40 wt.% in terms of composition.

10. The granular composition under item 1, in which the average particle size of the fine substrate is from 1 μm to 0.8 cm

11. The granular composition under item 1, in which the average particle size of the fine substrate is from 10 μm to 0.5 cm

12. The granular composition under item 1, in which the average particle size of the fine substrate is from 20 μm to 0.2 see

13. The granular composition under item 1, in which the water-insoluble substrate is an inorganic or organic material.

14. The granular composition according to p. 13, in which the water-insoluble substrate is a crushed bran, straw, wood dust or cellulose.

15. The granular composition according to p. 13, in which the inorganic substrate is a water-insoluble metal oxide, metal salt (SiO2, Al2O3, BaSO4, CaCO3), silicate or aluminosilicate of alkali and alkaline earth metals.

16. tulgit, diatomaceous earth, powdered lime, diatomaceous earth, wollastonite, olivine, montmorillonite or vermiculite.

17. The granular composition according to p. 15, in which the water-insoluble substrate is a vermiculite.

18. The granular composition under item 1, in which the number of substrates are between 50 and 99 wt.% in terms of composition.

19. The granular composition under item 18, in which the number of the substrate is 65 to 95 wt.% in terms of composition.

20. The granular composition under item 1, in which the number of the substrate is 75 to 90 wt.% in terms of composition.

21. The granular composition under item 1, in which the average particle size is from 0.01 to 8 mm

22. The granular composition according to p. 21, in which the average particle size is from 0.2 to 4 mm

23. The granular composition according to p. 21, in which the average particle size is from 0.5 to 2 mm.

24. The granular composition under item 1, in which the film-forming, water-soluble and practically seamless polymer is a synthetic or natural polymer.

25. The granular composition under item 1, in which the film-forming water-soluble and practically seamless polymer is a homopolymer or copoly. The granular composition under item 1, in which the film-forming water-soluble and practically seamless polymer is a polysaccharide or its derivative.

27. The granular composition according to p. 26, in which the film-forming water-soluble and practically seamless polymer include starch, alginate, carrageenan, k-carrageenan, l-carrageenan, xanthan gum, resin carob or methyl cellulose or a mixture thereof.

28. The granular composition according to p. 27, in which the film-forming water-soluble and practically seamless polymer are k-carrageenan, l-carrageenan or alginate.

29. The granular composition under item 1, in which the film-forming structurally crosslinked swellable in water polymer containing a carboxyl or sulfate groups, is k-carrageenan, l-carrageenan, xanthan gum or resin mix carob and xanthan gum.

30. The granular composition under item 1, in which the film-forming structurally stitched swelling in the form of a polymer containing a carboxyl or sulfate groups, is k-carrageenan, l-carrageenan.

31. The granular composition under item 1, which contains water-soluble or swelling in water of the polymer in an amount of from 0.1 to 20 wt.% in terms of composition.

33. The granular composition under item 1, in which the microorganism is chosen from the group consisting of Rhizobium spp., Metharizium, Fusarium, Trichoderma, Streptomyces, Gliocladium, Penicillum, Talaromyces, Verticillium or Colletotrichum, Pseudomonas spp., Serratia spp., Exserohilum spp., Bacillus spp., Agrobacter spp., Enterobacter spp. and Pseudomonas aurantiaca ATTC N 55169.

34. The granular composition under item 1, in which the microorganism is Pseudomonas aurantiaca ATTC N 55169.

35. The method of preparation of the granulated composition comprising fine substrate and the polymer layer containing the microorganisms, and the polymer is a) a film-forming, water-soluble and practically seamless polymer, while the granular composition contains at least 0.5 weight. % of water, calculated on the composition, or (b) film-forming structurally cross-linked polysaccharide, which contains carboxyl or sulfate groups and swells in water in the presence of potassium ions, while the granular composition contains at least 0.5 wt.% water in terms of composition, which includes (A) for the preparation of granules (a) the suspension or the temperature is not above 95oWith the dissolution of the film-forming and water-soluble polymer and the suspension of the microorganism in the suspension or solution after cooling to room temperature, (B) the group's in aqueous buffer solution containing potassium ions, and the subsequent suspension of the microorganism in the solution; (C) nabryzgivanii received suspensions directly on fine substrate or a mixture of these suspensions with fine substrate and (G) removing water to a concentration not less than 0.5 wt.% in terms of the granular composition.

36. The method according to p. 35, which in the case of use for preparing a granular composition (a) suspension of film-forming and water-soluble polymer of this suspension get preferably at a temperature of from 10 to 30oC.

37. The method according to p. 35, which in the case of use for preparing a granular composition (a) solution of film-forming and water-soluble polymer of this solution get preferably at a temperature of from 25 to 95oC.

38. The method according to p. 35, in which the microorganism is added at a temperature of less than 40oWith the solution or suspension of the polymer.

39. The method according to p. 35, in which the microorganism is added at a temperature of less than 30oC.

40 the Method according to p. 35, in which the buffer is a mixture of monopotassium phosphate and potassium secondary hydrogen phosphate potassium.

43. The granular composition according to PP.1 to 34, characterized in that it possesses insecticidal and fungicidal activity.

 

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