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INVENTION
Patent of the Russian Federation RU2286979

METHOD OF OBTAINING MICROBIOLOGICAL FERTILIZER BASED ON BIOGUMUS

The name of the inventor: Koshchaev Andrey Georgievich
The name of the patent holder: Kuban State Agrarian University
Address for correspondence: 350044, Krasnodar, ul. Kalinin, 13, KGAU, PIO
Date of commencement of the patent: 2004.12.06

The invention relates to agricultural biotechnology, in particular to the technology of preparing biological fertilizers. The method includes vermicomposting of organic wastes using a red Californian worm hybrid with earthworms of the Kuban natural population in the amount of 10 ⁴ worms per 1 m ² , with the manure of agricultural animals preliminarily neutralized to pH 7-8 as organic waste. Vermicomposting is carried out for 2-3 months at a temperature of 16-32 ° С in natural conditions, and after separation of worms from vermicompost with a vibrating screen with a pore size of 0.5-1.0 cm, the vermicompost is dried to a humidity of 50-60%, packed in packets Of polypropylene, corn extract is added in an amount of 1.0-5.0% of the mass of vermicompost and is autoclaved for 45-75 minutes at 0.8-1.2 atm, the Bacillus megaterium var strain is introduced. Phosphaticum 319 and maintained until reaching a titer of 8 · 10 ¹⁰ . The invention allows to expand the assortment of microbiological fertilizers, to increase their biological activity by increasing the viability of microorganisms, to reduce material costs in production by simplifying the technology of preparation and to shorten the preparation time of the preparation.

DESCRIPTION OF THE INVENTION

The invention relates to agricultural biotechnology, in particular to the technology of preparing biological fertilizers.

In recent years, biohumus, a product of vermicomposting of various organic wastes of industrial and agricultural production, specially extracted by the race of earthworms, has been increasingly used in agriculture. Biohumus contains humic and fulvic acids, macro- and microelements, amino acids and gibberellins and other biologically active substances in a form accessible to plants. Therefore, the use of biohumus in itself increases the yield of crops and increases the fertility of soils.

A method for producing biohumus is known. Hydrolytic lignin is stored in heaps, sprinkled with lime and watered. The process is carried out for 1.5-2.0 months. Periodically watered and shoveled. If necessary, add lime in addition, adjust the pH to a value of 6.8-7.2. Prepared in this way the substrate is laid in the bed and worms are launched. After the completion of the vermicomposting process, the worms are separated from the biohumus obtained and sold (RF patent No. 20944413, class C 05 F 11/00., 27.10.1997).

However, the use of lignin as a raw material requires a long preparation of the substrate, which is due to the peculiarities of the chemical structure of lignin. In addition, lignin has a very low acidity, which requires large amounts of lime, and it in turn in large quantities is toxic to worms. In addition, a low content of beneficial microorganisms in biohumus makes it unattractive for the consumer.

A known method for disposing of organic waste in biohumus involves the preparation of a substrate by mixing components, one of which is wastes of animal origin, inserting worms and / or their cocoons, placing the substrate in a box or ridge and composting at a humidity of 65-80% and aeration, separation Worms from the obtained biohumus. As the main component in the substrate, an osprey, which is a waste of cardboard production, is introduced in an amount of 20-80% to the total mass of the substrate. As animal waste, cattle manure and / or manure of pigs and dung of birds are used, worms of the species Eisenia foetida are used in the amount of 15-25 thousand individuals per 1 m ² . With a low content of nitrogen in the substrate, after mixing the components, add nitrogen fertilizer in the form of urea in an amount of no more than 5% of the total mass of the substrate with the simultaneous addition of limestone flour in an amount that provides the pH of the medium of the finished substrate no more than 8 (RF Patent No. 2057743, (See C 05 F 3/06, 10.04.1996.).

However, this method has some drawbacks. The introduction of the osprey (cardboard waste) is not always possible due to the lack of raw materials. In addition, a wide range of its input does not allow the standardization of the vermicomposting process, since the share of the osprey affects the activity of worms. It is doubtful that urea can provide the claimed effect. In addition, as in the aforementioned analogue, it is unlikely that the vermicompost obtained in this way included the necessary quantity of useful microorganisms, including those capable of hydrolyzing hardly soluble phosphorus forms for plants.

The method closest to the claimed method is the preparation of biofertilizer, which consists in obtaining vermicompost by vermicomposting agricultural and industrial waste using earthworms, separating vermicompost from worms and drying it, and worms "Obolensky hybrid" that we obtained by crossing the "Red California hybrid "with the Russian population of earthworms Eisenia foetida, while vermiculture is carried out at a temperature of 16-24 ° C for 4-6 months, microorganisms with fungicidal properties are introduced into the resulting biohumus. In the method, microorganisms possessing fungicidal activity are added after separation of worms or after ripening of vermicompost, in addition, as a strain of bacteria Bacillus subtilis ИПМ-215 in concentrations of 1 × 10 ⁹ -1 × 10 ¹² spores per 1 kg of vermicompost or culture of mycophilic Fungus Trichoderma viride Pers ex SFGray 16 in concentrations of 1 × 10 ⁴ -1 × 10 ⁸ colony forming units per 1 kg of biohumus (Russian Federation patent No. 21255549, class C 05 F11/08, 1999, bulletin No. 3. - prototype ).

However, this method has a number of significant drawbacks. Firstly, the use of this hybrid does not allow us to obtain a high-quality biohumus everywhere, because the climatic conditions of our country are very diverse. Secondly, the possibility of using this technology in southern regions of Russia is questionable, since this temperature range does not allow the cultivation of this hybrid in warm months, and the cultivation time (4-6 months) does not allow obtaining sufficient amount of biohumus. In addition, the deficiency of phosphorus assimilated for plants in many agrolandscapes requires additional introduction of microorganisms that have the ability to convert intractable phosphorus compounds into accessible forms that the proposed preparation can not provide.

The preparation, obtained by the prototype, requires additional input of the adhesive (when used for seed treatment), which requires additional material and human resources, which is not always economically feasible. In addition, the low titer of the drug requires high doses of application for disease control, which makes the use of this bacterial drug economically ineffective.

The known methods do not allow to obtain efficiently a microbiological fertilizer with a high content of plant nutrients and simultaneously with a high titer of useful microorganisms and having the ability to transfer phosphates to plant-accessible forms without application of an additional scraper before processing the seeds.

The technical solution of the problem is to expand the assortment of bacterial fertilizers, reduce material costs during production and increase their biological activity, and increase the shelf life of the preparation, improve the agrochemical parameters of the soil due to the nutrients that make up the biohumus, the ability of the preparation to convert the intractable phosphorus compounds into Available to plant forms and high titers of beneficial microorganisms.

The task is achieved by the fact that in the method for producing a microbiological fertilizer based on biohumus, including obtaining vermicompost by vermicomposting of organic waste using earthworms and introducing microorganisms into the vermicompost, where the manure of farm animals previously neutralized to pH 7-8 is used as organic waste, And as a earthworm the hybrid of the red Californian worm with earthworms of the Kuban natural population is used, in an amount of 10 ⁴ per 1 m ² , while vermicomposting is carried out for 2-3 months at a temperature of 16-32 ° C under natural conditions, and after separation Worms from vermicompost with a vibrating screen with a pore size of 0.5-1.0 cm, the vermicompost is dried to a moisture content of 50-60%, packed in polypropylene bags, corn extract is added in an amount of 1.0-5.0% by weight of biohumus and autoclaved for 45-75 minutes at 0.8-1.2 atm, the strain Bacillus megaterium var. Phosphaticum 319 and maintained until reaching a titer of 8 · 10 ¹⁰ .

The claimed method of obtaining a microbiological fertilizer based on biohumus differs from the prototype by the modes of manufacturing vermicompost and the physiological group of microorganisms used and additional carbohydrates (corn extract) as a source of nutrition and an adhesive for the preparation during seed treatment.

These differences allow us to conclude that the claimed technical solutions meet the criterion of "novelty".

The features that distinguish the claimed technical solution from the prototype are aimed at achieving the task set and are not revealed in the study of patent and scientific and technical literature in this and related fields of science and technology and, therefore, meet the criterion of "inventive level".

The method of obtaining a microbiological fertilizer based on biohumus is as follows.

EXAMPLE 1 Manure of cattle is stored in beads measuring 1 · 5 · 1.5 meters. The manure is shoveled until it reaches the ambient temperature. To regulate acidity, chalk or hydrated lime is used. The alkalinization is carried out layer by layer until the manure reaches pH 7-8. If the acidity of manure is less than 7 units, then worms will not develop in it, so biohumus will not form. If the acidity of the manure is more than 8 units, then the worms will develop and will not be there, since this pH value is not optimal for them, and there will be an over-consumption of the alkalizing agent, which will lead to additional expenses for preparing the substrate for the worms. Thus, the optimum acidity of manure for the development of worms is a pH equal to 7.5 units.

As a ringworm, a hybrid of the red Californian worm with earthworms of the Kuban natural population is used, which was obtained by the joint cultivation of the hybrid of the red Californian worm with the Kuban population of annelidized manureworms. This is due to the fact that the red Californian worm hybrid is very sensitive to environmental conditions and requires an optimal temperature, certain qualities of the substrate, which makes it not technological in our conditions. At the same time, ringed dung worms that live in the soil, though omnivorous, are low-yielding and retain an instinct for migration, which makes them an inconvenient object for vermicomposting. Therefore, the use of an adapted breed provides a sufficiently high level of worm productivity and, at the same time, low susceptibility to environmental conditions.

Ready-to-use manure is populated with worms at a rate of 10,000 worms per 1 m ² , having previously tested the "50 worms" test, which allows to assess the quality of the substrate. The settlement is made in the evening or on a cloudy day, as worms do not tolerate direct sunlight. Navals with a worm-infested substrate from above are covered with grass to protect from the sun and reduce the evaporation of water. Navals periodically watered, loosened the top layer of the substrate. Vermicomposting is carried out within 2-3 months. If the cultivation time is less than 2 months, the hybrid of the red Californian worm and the annelid worm of the Kuban population will not process the substrate, which means that the biohumus output will be insignificant. If the cultivation time is more than 3 months, the hybrid of the red Californian worm and the annelid worm of the Kuban population, after processing the entire substrate, will begin to oppress and lose productivity, and in addition, it increases the time of the preparation process of the biopreparation, which increases its cost price. Therefore, the optimal time for cultivation of worms in the substrate is 2.5 months. Optimal cultivation of worms in the substrate is carried out at a temperature of 24 ° C. If the culture temperature is less than 16 ° C, the metabolism and growth of hybrid worms slow down, and hence the conversion efficiency of the substrate in vermicompost will be low. If the culture temperature is more than 32 ° C, the growth of vermiculture will be low.

As a result of the growth and reproduction of worms, manure is processed into biohumus. Separation of worms is carried out on mechanical vibrating screens, and the worms remaining on the screen are used for the subsequent production of biohumus from the next batch of manure. At the same time, the 0.75-cm screen size is optimal, which makes it possible to obtain pure biohumus that is free of particles of unrefined substrate and debris. If the pore size of the screen is less than 0.5 cm, the sieving speed will be low, which will increase the time of the technological operation. If the pore size is more than 1.0 cm, then together with the particles of biohumus, particles of the unreconcessed substrate will fall into the container with the finished product, which will negatively affect the quality of the finished product.

The resulting biohumus is dried in a stream of hot air to a humidity of 55% and is stored in kraft bags. If the moisture content of the finished biohumus is less than 50%, then the coolant over-consumption will occur during the drying of the product and this will require additional water injection for the comfortable development of phosphorolytic microorganisms introduced into the vermicompost. If the moisture content of the finished biohumus is more than 60%, the quality of the finished product will deteriorate, as the resulting fertilizer will lose its flowability, and in addition, the aeration of the substrate will be low, which will lead to a slow development of microorganisms.

The biohumus obtained in the above way is used simultaneously as a carrier and a substrate for the development and reproduction of phosphorolytic microorganisms. As necessary, biohumus from kraft bags is packaged in polypropylene bags, depending on the volume of use from 1 kg to 5 kg per package. To ensure the carbohydrate nutrition of phosphorolytic microorganisms, and excluding the use of adhesives in the processing of seeds in pre-packaged vermicompost, maize extract is applied at the rate of 3% of the mass of biohumus (optimal concentration), that is, if the mass of the bag with biohumus is 5 kg, then molasses is added 150 g. If The weight of the introduced maize extract will be less than 1%, then the carbohydrate nutrition for the development of phosphorolytic bacteria will be insufficient, and the required culture of microorganisms will not reach the required level. If the weight of the corn extract is more than 5%, an overabundance of corn extract will lead to additional expenses for the preparation of fertilizer and will reduce the quality of the preparation.

After adding the corn extract, the bags are closed with cotton-gauze stoppers and sterilized in an autoclave at 1 atm for one hour, which is the optimal sterilization regime. If the sterilization time is less than 45 minutes and the pressure in the autoclave is less than 0.8 atm, complete sterilization of the substrate will not occur and microorganisms will remain in it that will interfere with the growth and development of phosphorolytic microorganisms, which means that their desired microorganism titer is not reached, that Will reduce the quality of the drug. If the sterilization time is more than 75 minutes and the pressure in the autoclave is more than 1.2 atm, then due to the rigid autoclaving regime in the substrate (biohumus), substances toxic for phosphorolytic microorganisms are formed, including by caramelization of sugars of maize extract, and the quality The drug will decrease, in addition, the cost of the bacterial drug will be high due to over-expenditure of coolants.

For sowing biohumus seed from the collection of pure cultures of the Federal State Institution "Krasnodar Biocenter" is used. A tube with a lyophilized pure culture of Bacillus megaterium var. Phosphaticum 319 is revitalized with a nutrient solution. Sift a lively culture into 750 ml flasks with 200-250 ml of a nutrient medium, which is a solution of corn extract, molasses, ammonium sulfate and chalk. Then, the resulting mother liquor is inoculated in 5-liter bottles containing 2.0-2.5 liters of nutrient medium of the same composition.

The thus obtained working solution of Bacillus megaterium var. Phosphaticum strain 319 is sown sterile in vermicompost from the final titer of microorganisms 4 · 10 ⁷ . Stirring is performed mechanically in such a way as not to damage the polypropylene bags. Then the sown packets are placed in a room thermostatted room temperature.

After 10-14 days I check the titer of the sown microorganisms, which should increase by 1-2 orders of magnitude due to the growth of phosphorolytic bacteria on the biohumus and reach 8 · 10 ¹⁰ .

Further storage of drugs in undamaged packages for 3 months did not reduce the titer of the drug based on Bacillus megaterium var. Phosphaticum strain 319 is below 2 · 10 ¹⁰ , which is related to the optimal content of nutrients necessary for microorganisms in biohumus and additional input of carbohydrate components of nutrition, which is used as corn extract. The results of the experiments are presented in Table 1.

EXAMPLE 2 According to a method for producing a microbiological fertilizer based on the biohumus of Example 1, a chicken manure with the addition of chopped straw is used as the raw material for cultivating worms.

EXAMPLE 3 According to the method for producing a microbiological fertilizer based on the biohumus of Example 1, horse manure is used as the raw material for the cultivation of worms.

EXAMPLE 4 According to a method for producing a microbiological fertilizer based on the biohumus of Example 1, manure is used as a raw material for the cultivation of worms in fattening with the addition of chopped straw.

The results of the determination of the number of bacteria Bacillus megaterium var. Phosphaticum strain 319 in biohumus of different origin are presented in Table 2.

The results of laboratory tests of Examples 1, 2, 3, 4 showed that irrespective of the origin of livestock wastes with biohumus obtained on its basis, a high Bacillus megaterium var titer is provided. Phosphaticum strain 319, necessary to obtain a quality bacterial fertilizer for the transfer of complex organophosphorus compounds and intractable mineral phosphates into a form that is accessible to plants.

In addition, it should be noted that the technology of obtaining vermicompost is a simple process, and raw materials (organic agricultural waste) are generally available. Simultaneously with obtaining biohumus, the problem of protecting the environment is also being solved, which is especially important in agrarian regions with developed livestock breeding.

The results of the analysis of the chemical composition of vermicompost and humus obtained from manure of cattle are presented in Table 3.

As can be seen from the table, the use of biohumus allows not only to provide high concentrations of phosphorolytic bacteria, but also to use biohumus as a highly effective source of nutrients for plant growth (the chemical composition of biohumus is given in comparison with the composition of humus).

We should not forget that the use of biohumus as a filler for the preparation of phosphorolytic bacteria not only provides plants with useful microorganisms, but also in itself is a source of nutrients.

Thus, the method of preparation of bacterial fertilizer based on biohumus not only allows to obtain a high titer of Bacillus megaterium var. Phosphaticum strain 319 using the proposed method due to the presence of nutrients in the biohumus, but also simultaneously activate plants using nutrient and stimulating substances of the biohumus itself. In addition, the implementation of the proposed method will expand the range of biopreparations due to the possibility of placing small-scale production directly in the regions and partially solve the problems of utilization of animal wastes.

CLAIM

The method of obtaining a microbiological fertilizer based on biohumus, including the production of vermicompost by vermicomposting of organic waste with the use of earthworms and introduction of microorganisms into the vermicompost, characterized in that organic manure is used as organic waste, previously neutralized to pH 7-8, and as rain Worms use a hybrid of a red Californian worm with earthworms of the Kuban natural population in the amount of 10 ⁴ per 1 m ² , while vermicomposting is carried out for 2-3 months at a temperature of 16-32 ° C in natural conditions, after separation of worms from vermicompost Pore ​​size 0.5-1.0 cm, the vermicompost is dried to a moisture content of 50-60%, packed in polypropylene bags, corn extract is added in an amount of 1.0-5.0% by weight of biohumus and autoclaved for 45-75 minutes 0, 8-1.2 atm, the strain Bacillus megaterium var. Phosphaticum 319 and maintained until reaching a titer of 8 · 10 ¹⁰ .

print version
Date of publication 04.03.2007gg

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