RU2286973C2 - Method of preparing biohumus enriched in nitrogen-fixing bacteria - Google Patents

Abstract

FIELD: organic fertilizers.

SUBSTANCE: invention relates to agricultural biotechnology and provides a method of preparing biohumus comprising vermicomposting of organic wastes using hybrid of red California worm with earth worms of Kuban natural population in amount 10⁴ worms per 1 m², said organic wastes being farm animal manure preliminarily neutralized to pH 7-8. Vermicomposting is carried out over a 2-3 month period at 16-32°C under normal conditions. After separation of worms from biohumus, microorganisms capable of providing associative nitrogen fixation are added in number of 10¹² to 10¹⁴ cells per 1 kg biohumus.

EFFECT: increased assortment of microbiological fertilizers, enhanced their biological activity due to increased vitality of microorganisms, reduced expenses due to simplified technology, and reduced product preparation time.

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Description

The invention relates to agricultural biotechnology, in particular to a technology for producing fertilizers for plant growing.

In recent years, vermicompost has been increasingly used in agriculture, a product of vermicomposting various organic wastes of industrial and agricultural production by a specially bred race of earthworms. 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 agricultural crops and increases soil fertility.

A known method of producing biohumus. 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 shovel. If necessary, add lime additionally, adjust the pH to a value of 6.8-7.2. The substrate prepared in this way is laid in a bed and the worms are launched. After the vermicomposting process, the worms are separated from the biohumus obtained and sold (Pat. RF No. 2094413, class C 05 F 11/00, 10.27.1997).

However, the use of lignin as a raw material requires long preparation of the substrate, which is associated with 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, is toxic to worms in large quantities. In addition, the low content of beneficial microorganisms in biohumus makes it not attractive to consumers.

A known method of disposing of organic waste in vermicompost involves preparing a substrate by mixing components, one of which is animal waste, introducing worms and / or their cocoons into it, laying the substrate in a box or ridges and composting at a humidity of 65-80% and aeration, separation worms from biohumus. As the main component, osprey is introduced into the substrate, which is a waste product of cardboard production, in an amount of 20-80% of the total mass of the substrate. As animal waste, cattle manure and / or pig and bird droppings were used, and worms of the species Eisenia foetida were used in the amount of 15-25 thousand animals per 1 m ² . At a low nitrogen content in the substrate, after mixing the components, nitrogen supplements of mineral fertilizers are introduced in the form of urea in an amount of not more than 5% of the total mass of the substrate with the simultaneous addition of limestone flour in an amount that ensures the pH of the finished substrate is not more than 8 (US Pat. RF No. 2057743, C. C 05 F 3/06. 04/10/1996 - prototype).

However, this method has some disadvantages. The introduction of osprey (cardboard production waste) is not always possible due to the lack of raw materials. In addition, the wide range of its input does not allow standardizing the process of vermicomposting, since the osprey fraction 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 biohumus obtained in this way would include the necessary number of beneficial microorganisms, including those capable of associative nitrogen fixation.

Closest to the claimed method is a method of producing bio-fertilizer, which consists in obtaining vermicompost by vermicomposting agricultural and industrial waste using earthworms, separating vermicompost from the worms and drying it out, and from the earthworms, Obolensky hybrid worms obtained by crossing the Red Californian hybrid "with the Russian population of earthworms Eisenia foetida, while vermicomposting is carried out at a temperature of 16-24 ° C for 4-6 months, Acquiring vermicompost introduce microorganisms having fungicidal properties. In the method, microorganisms having fungicidal activity are introduced after separation of the worms or after ripening of vermicompost, in addition, they use a strain of bacteria Bacillus subtilis IPM-215 in concentrations of 1 · 10 ⁹ -1 · 10 ¹² spores per 1 kg of vermicompost or mycophilous culture Trichoderma viride Pers ex SFGray 16 in concentrations of 1 · 10 ⁴ -1 · 10 ⁸ colony forming units per 1 kg of vermicompost (US Pat. RF No. 2125549, class C 05 F 11/08/1999, bull. No. 3 - prototype).

However, this method has several significant disadvantages. Firstly, the use of this hybrid does not allow to everywhere receive high-quality biohumus, since the climatic conditions of our country are very diverse. Secondly, the possibility of using this technology in the southern regions of Russia is doubtful, since the indicated temperature range does not allow cultivation of this hybrid in the warm months, and the cultivation time (4-6 months) does not allow to obtain a sufficient amount of vermicompost. In addition, the nitrogen deficiency in many agrolandscapes requires additional nitrogen nutrition, which can not provide biohumus obtained by the prototype. In addition, a low titer of the drug requires high doses of application to combat diseases, which makes the use of such biohumus economically inefficient.

Known methods do not allow to obtain biohumus with a high titer of beneficial microorganisms and having associative nitrogen-fixing activity.

The technical solution to the problem is to expand the range of microbiological fertilizers, increase their biological activity by increasing the viability of microorganisms, reducing material costs during production by simplifying the technology and shortening the production time of biohumus, improving the agrochemical parameters of the soil due to the nutrients that make up the biohumus and the drug has nitrogen-fixing activity.

This object is achieved in that in a method for producing vermicompost enriched with nitrogen-fixing bacteria, including vermicomposting of vermicomposting organic waste using earthworms and introducing microorganisms into biohumus, moreover, animal farm manure, previously neutralized to pH 7-8, is used as organic waste. and as the earthworms use a hybrid of the red California worm with the earthworms of the Kuban natural population, e April ¹⁰ per m ^2, wherein the vermicomposting performed for 2-3 months at 16-32 ° C in vivo, and after separating worms from vermicompost making it microorganisms which are capable of associative nitrogen fixation, the cell concentration ^{12 October} - 10 ¹⁴ in 1 kg of biohumus.

The claimed method for producing biohumus enriched with nitrogen-fixing bacteria differs from the prototype in the production modes of biohumus and the kind of microorganisms used.

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 and have not been identified 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 step".

The method of producing biohumus enriched with nitrogen-fixing bacteria is implemented as follows.

Cattle manure is stored in piles measuring 1 · 5 · 1.5 meters. Manure is shoveled until it reaches ambient temperature. Chalk or slaked lime is used to regulate acidity. Alkalization is carried out in layers until manure reaches a pH of 7-8. If the acidity of manure is less than 7 units, then the worms will not develop in it, which means that biohumus will not form. If the acidity of manure is more than 8 units, then the worms will not develop in it either, since this pH value is also not optimal for them, in addition, there will be an excessive consumption of the alkalizing agent, which will lead to additional costs for the preparation of the substrate for the worms. Thus, the optimum acidity of manure for the development of worms is a pH of 7.5 units.

As ringworms, a hybrid of the red California worm with earthworms of the Kuban natural population, obtained by co-cultivating a hybrid of the red California worm with the Kuban population of ringed dung worms, is used. This is due to the fact that the hybrid of the red California worm 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, annular dung worms that live in the soil, although omnivorous, are low productive and retain an instinct for migration, which makes them an uncomfortable object for vermicomposting. Therefore, the use of adapted breed provides a sufficiently high degree of worm productivity and at the same time low susceptibility to external environmental conditions.

Ready-to-use manure is populated with worms at the rate of 10 thousand worms per 1 m ² , previously checked with the test "50 worms", which allows you to evaluate the quality of the substrate. Settlement is carried out in the evening or on a cloudy day, as the worms do not tolerate direct sunlight. Bulk with a worm-infested substrate is covered with grass on top to protect from the sun and reduce water evaporation. Bulk watered periodically, loosen the upper layer of the substrate. Vermicomposting is carried out for 2-3 months. If the cultivation time is less than 2 months, the hybrid of the red California worm and the ringworm of the Kuban population does not process the substrate, which means that the yield of biohumus will be insignificant. If the cultivation time is more than 3 months, the hybrid of the red California worm and the ringworm of the Kuban population, having processed the entire substrate, will begin to be inhibited and lose productivity, and, in addition, it increases the time of the preparation of the biological product, which increases its cost. Therefore, the optimal time for the cultivation of worms in the substrate is 2.5 months. The optimal cultivation of worms in the substrate is carried out at a temperature of 24 ° C. If the cultivation temperature is less than 16 ° C, then the metabolism and growth of hybrid worms slows down, which means that the efficiency of the conversion of the substrate to vermicompost will be low. If the cultivation temperature is more than 32 ° C, then the growth of vermiculture will also be low.

As a result of the growth and reproduction of worms, manure is processed into vermicompost. The worms are separated on vibrating screens, and the worms remaining on the sieve are used for the subsequent production of biohumus from the next batch of manure. The obtained biohumus is dried in the open air to a moisture content of 60-70% and stored in piles.

For sowing vermicompost, seed is used from the collection of pure crops of the Federal State Institution "Krasnodar Biocenter". A tube of lyophilized pure culture of Agrobacterium radiobacter strain 204 is revitalized using a nutrient solution. The lively culture is inoculated into 850 ml flasks of 250-300 ml of culture medium, which is a solution of sucrose and mineral salts. Then, the resulting mother liquor is inoculated in 5 liter bottles containing 2.0-2.5 liters of nutrient medium, including corn extract, molasses and mineral salts.

Thus obtained working solution of the culture of Agrobacterium radiobacter strain 204 is inoculated with vermicompost located in the piles. Moreover, in such a way that the concentration of inoculated bacteria is at least 10 ¹⁰ cells per 1 kg of biohumus. Mixing is performed mechanically until uniform mixing and cover with an insulating material. After 7-10 days, I check the titer of the seeded microorganisms, which should increase by 1-2 orders of magnitude due to the growth of bacteria on vermicompost. The optimal culture titer of Agrobactenum radiobacter strain 204 should be 10 ¹³ cells in 1 kg of vermicompost. If the number of cells is less than 10 ¹² per 1 kg of vermicompost, then the quality of the drug will be low and it is necessary to increase the time of cultivation of bacteria in vermicompost. If the number of cells of Agrobacterium radiobacter strain 204 is more than 10 ¹⁴ per 1 kg of biohumus, then the quality of the drug will be reduced due to the high consumption of nutrients of biohumus by bacterial cells.

Example 2. According to the technology for producing microbiological fertilizer according to example 1, Agrobacterium chroococcum strain 10 is used as microorganism.

Example 3. According to the technology for producing microbiological fertilizer according to example 1, Azospirillum Upoferum strain Az-12 is used as a microorganism.

The results of the determination of the titer of associative nitrogen-fixing microorganisms in the proposed preparation based on biohumus in table 1.

The results of laboratory tests in examples 1, 2, 3 showed that regardless of the nature of the strain of associative nitrogen-fixing microorganisms, the proposed vermicompost provides a high titer of the desired bacteria.

Table 1
The titer of associative nitrogen-fixing microorganisms in the proposed preparation based on vermicompost Example Microorganism culture Microorganism titer one Agrobacterium radiobacter strain 204 2.7 × 10 ¹² 2 Agrobacterium chroococcum strain 10 9.4 × 10 ¹² 3 Azospirillum lipoferum strain Az-12 7.6 × 10 ¹²

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

The results of the analysis of the chemical composition of vermicompost and humus obtained from cattle manure are presented in table 2.

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

It should not be forgotten that the use of biohumus as a filler for the preparation of associative nitrogen-fixing bacteria not only provides plants with beneficial microorganisms, but also is itself a source of nutrients.

table 2
Chemical characteristics of vermicompost and humus (%) Name Vermicompost Humus Medium acidity 6.7 7.8 Organic matter 44.9 23.6 Humic acids 3.4 2,3 Fulvic Acids 2.2 0.6 Organic carbon 3.31 1.7 Nitrogen 3.22 1,54 Phosphorus 0.49 0.35 C: N 1,04 1.10 Electrical conductivity - a measure of the relative salinity of the soil or the amount of soluble salts 12.1 3.60

Thus, the method for producing biohumus enriched with nitrogen-fixing bacteria not only allows one to obtain a high titer of associative nitrogen-fixing bacteria due to the presence of nutrients in it, but also to activate the treated plants due to the presence of nutrients and stimulants of the biohumus itself, but also to expand the range of fertilizers , due to the possibility of placing small-tonnage production directly in the regions and partially solve the problems of recycling livestock.

Claims (1)

A method of producing biohumus enriched with nitrogen-fixing bacteria, including the production of biohumus by vermicomposting organic waste using earthworms and introducing microorganisms into biohumus, characterized in that farm animal manure previously neutralized to pH 7-8 is used as organic waste, and rain animal manure of worms use a hybrid of a red California worm with earthworms of the Kuban natural population in the amount of 10 ⁴ per 1 m ² , while vermicom Posting is carried out for 2-3 months at a temperature of 16-32 ° C under natural conditions, and after separation of the worms from vermicompost, microorganisms capable of associative nitrogen fixation are introduced into it, and the cell concentration is 10 ¹² -10 ¹⁴ in 1 kg of vermicompost.