RU2125549C1 - Method of producing biofertilizer - Google Patents

Abstract

FIELD: fertilizers production. SUBSTANCE: substrate based on agricultural and industrial organic wastes is subjected to vermi-composting on a new industrial line with earth-worms obtained by interbreeding of Red California hybrid with Russian earth-worm population Eisenia foetida at 16-24 C for 4-6 months. Resultant biohumus is freed of worms and is advantageously left under natural conditions to be completely ripened. According to invention, immediately after separation of worms or after final ripening of biohumus, microorganisms exhibiting fungicidal activity are added, in particular, Bacterium subtilis stain IPM-215 in concentrations from 1•10⁹ to 1•10¹² spores per 1 kg or culture of micophilic fungus Trichoderma viride Persex S.F.Cray No 16 in concentrations from 1•10⁴ to 1•10⁸ spores per 1 kg of biohumus. EFFECT: improved agrochemical characteristics of soil, and increased fungicidal and growth-stimulation activity for plants. 5 cl, 6 tbl

Description

 The invention relates to the microbiological industry, to the production of biofertilizers and plant protection products.

 Biohumus is used to improve soil quality. Known methods for producing biohumus from organic waste by vermicomposting [1, 2]. Waste processing is carried out using a special industrial line of earthworms "Red California Hybrid." The resulting product has high fertilizing properties, but does not significantly affect soil phytopathogens.

 To protect plants from pathogens, various preparations based on microorganisms, such as Bacillus subtilis [3, 4], and fungi of the genus Trichoderma [5] are used.

 Closest to the proposed invention is a method for producing compost, into which a suspension of microorganisms antagonists of phytopathogens is added: Trichoderma hamatum fungi and Pseudomonas maltophilia or Flavobacterium bacteria [6]. However, this compost has low fertilizing properties and does not stimulate plant growth.

 The aim of the invention is a method for biofertilizer, improving the agrochemical parameters of the soil, with fungicidal and growth-promoting properties for plants.

 The goal is achieved by the fact that in the vermicompost obtained from a mixture of agricultural and industrial wastes using the new line of earthworms "Obolensky hybrid", microorganisms with fungicidal activity are introduced.

The essence of the invention lies in the fact that the substrate based on organic waste from agriculture and industry (animal manure, bird droppings, plant residues, sewage sludge from sewage treatment plants, waste from food processing plants) is processed using the new industrial line of earthworms Obolensky hybrid obtained by crossing the "Red California Hybrid" with the Russian population of earthworms Eisenia foetida. At the final stage of obtaining biohumus, either a Bac bacterial strain is introduced into the finished product. subtilis IPM-215 [7] in concentrations of 1 • 10 ⁶ - 1 • 10 ¹² spores per 1 kg, or a culture of mycophilic fungus Trichoderma viride Pers ex SF Gray N 16 in concentrations of 1 • 10 ⁴ - 1 • 10 ⁸ colony forming units per 1 kg

 Example 1. Obtaining vermicompost.

To obtain vermicompost, a new industrial line of earthworms Obolensky Hybrid is used, which we obtained by crossing the Red California Hybrid with the Russian population of earthworms Eisenia foetida. The new hybrid is adapted to the climatic conditions of Russia, processes a wide variety of organic waste. Various organic agricultural and industrial wastes are used as a substrate for vermicomposting: animal manure, bird droppings, straw, vegetable residues, substandard vegetables, sewage sludge from sewage treatment plants, etc. Mixed waste is pre-composted in equal amounts at t = 30- 60 ^o C and a humidity of 60-85% for 20 days. After adjusting the pH = 6.5-7.5, earthworms are introduced into the obtained substrate. Vermicomposting is carried out at optimal t = 16-24 ^o C and humidity 40-80% up to 6.0 months (table. 1).

 At the end of composting, the worms are separated from the vermicompost. The resulting substrate is dried under natural conditions (ripening stage) for two weeks and sieved through a sieve.

Ready biohumus has the following chemical composition,%:
Mass fraction of moisture - 54-60
organic matter - 37-46 per dry product
mineral substances - 54-63
humic substances - 24-28
total nitrogen - 3.0-4.0
total phosphorus (according to P ₂ O ₅ ) - 1.0-3.0
total potassium (according to K ₂ O) - 1.0-1.5
pH - 6.5-7.5
Pathogenic microflora - None
Helminth Eggs - Not Identified
Biohumus contains a large amount of humic substances (up to 28% of dry matter), which provides high fertilizing properties. All plant nutrients are in the form of mineral, easily digestible salts. Their amount is up to 63% by dry matter. Biohumus does not contain pathogenic microorganisms and heavy metals. Therefore, the biohumus obtained is a highly effective, environmentally friendly organic fertilizer that improves the agrochemical parameters of the soil. After separation of the worms or after ripening of the biohumus, microorganisms with fungicidal activity are introduced.

 Example 2. Obtaining fertilizers with different content of Bac. subtilis.

The strain of bacteria Bac. subtilis IPM-215 is used for the production of the microbiological preparation of bactofite against plant diseases [8]. Bactofit is recommended to be used in doses of 6 kg / ha with a titer of 2.5 • 10 ¹⁰ spores / g. At a recommended dose of biohumus of 3 t / ha, 2 g of Bac should be added to 1 kg of biohumus. subtilis with a titer of 2.5 • 10 ¹⁰ spores / g, i.e. the originally calculated bacterial spread rate is 2.5 • 10 ¹⁰ spores / g • 2 g = 5.0 • 10 ¹⁰ spores / kg of vermicompost.

 After separation of the worms or the finished vermicompost, 1 kg is packed in pots and a culture of the Bac strain is introduced. subtilis IPM-215 in 3 different concentrations: initially calculated, 2 times smaller and 2 times larger. The repetition of experiments three times.

Biofertilizer components mix well. After 2 weeks of storage at room temperature (20 ± 5) ^o C, fertilizer samples are taken from which suspensions are prepared (10 g per 90 ml of sterile tap water). The prepared suspensions are shaken on a rocking chair for 40 minutes at (28 ± 1) ^o C and a rotation frequency of 220-240 rpm. Then prepare a series of ten-fold dilutions in saline and make inoculations from these dilutions deep into Petri dishes with sucrose peptone agar. Cultivation is carried out at (28 ± 1) ^o C for 48 hours. At the end of this time, the number of grown colonies is taken into account, then recalculation per 1 kg of biofertilizer is carried out (Table 2).

The experiments performed show the possibility of creating a combined fertilizer with different concentrations of Bac culture. subtilis. The most optimal is the introduction of Bac in vermicompost. subtilis in the amount of 2.5 • 10 ¹⁰ spores / kg. When such a quantity of bacteria is introduced into biohumus, their concentration increases 1.5 times over 2 weeks, reaching the necessary value for suppressing soil phytopathogens. Therefore, in subsequent experiments, we used a combined biofertilizer with an initial titer of 2.5 • 10 ¹⁰ spores / kg. Given the multiplication of bacteria, the dose of microorganisms can be reduced in comparison with the calculated. This significantly reduces the cost of combined biofertilizer and, accordingly, the cost of processing the soil from phytopathogens.

 Example 3. Obtaining biofertilizers with different contents of Trichoderma viride Pers ex S.F. Gray N 16.

 Strain N 16 of the fungus Trichoderma viride Pers ex S.F. Gray received by Colombet L.V. as a result of selection work on the grounds of productivity and hyperparasitic activity with the natural Trichoderma viride clone isolated from soil in the Moscow region (the strain was deposited by the Central collection of microorganisms of the State concern "BIOPREPARAT" CCMK "B" 02/15/94, registration number - CCM F-59M )

After separation of the worms or vermicompost, the vermicompost is packaged 1 kg each in pots and a culture of mycophilic fungus Trichoderma viride Pers ex SF Gray N 16 is introduced in 3 different concentrations: 1 • 10 ⁴ , 1 • 10 ⁶ , 1 • 10 ⁸ colony forming units ( CFU) per 1 kg. The repetition of experiments three times.

Biofertilizer components mix well. After 2 weeks of storage at room temperature (20 ± 5) ^o C, fertilizer samples are taken from which suspensions are prepared (10 g per 90 ml of sterile tap water). The prepared suspensions are shaken on a rocking chair for 40 minutes at (26 ± 1) ^o C and a rotation frequency of 220-240 rpm. Then prepare a series of ten-fold dilutions in saline and make seeding of these dilutions on Petri dishes with synthetic nutrient medium (Chapek's medium) containing X-100 newt to limit the growth of fungal colonies. Cultivation is carried out at (26 ± 1) ^o C. After three days, the number of grown colonies is taken into account, then recalculation per 1 kg of biofertilizer is carried out (Table 3).

 Our experiments show that the introduction of a culture of mycophilic fungus Trichoderma viride into biohumus leads to an increase in the biomass of the fungus, which indicates that biohumus is a suitable substrate for the growth of fungal mycelium. In this way, it is possible to create a new form of biofertilizer containing a fungicidal component in the form of a Trichoderma viride culture. This allows you to reduce the cost of the production of biofertilizers with fertilizing and fungicidal properties.

 Example 4. The study of the protective properties of fertilizers.

 To study the protective properties of fertilizer, it is applied to the soil in a ratio of 1: 4 by volume. The experiments are carried out in pots, 1 kg of soil mixture in each. The repetition of experiments three times.

 The study is carried out on an artificial infectious background, for which a mixture of phytopathogens is introduced into the soil: Fusarium culmorum, F. moniliforum, F. heterosporum, F. oxysporum, Verticillium dahliae, Rhizoctonia solani, Colletotrichum atraventa, Alternaria brassicae.

 Pre-sprouted cucumber seeds are sown in prepared soil mixtures.

 The decrease in the infectivity of various soil mixtures is evaluated after 1 month by conducting a microbiological analysis of soil samples on various nutrient media. In samples of soil mixtures with fungicidal components (Bac. Subtilis and Trichoderma viride), a significant decrease in the content of phytopathogens is noted. The presence of fungicidal properties of biofertilizers is reflected in morphometric indicators of plant growth (length and weight of green mass, number of leaves, mustache).

 As can be seen from the table. 4, the introduction of biofertilizer into the soil leads to an increase in morphometric indicators, which indicates the protective effect of biofertilizer.

 Example 5. The study of growth-promoting activity of biofertilizer.

Biofertilizer containing a Bac culture. subtilis at a concentration of 2.5 • 10 ¹⁰ spores / kg, contribute to the soil (1: 4 by volume). As control use: turf soil, soil with the introduced culture of Bac. subtilis and soil with vermicompost. The seeds of cucumbers and eggplant are sown in soil samples. The results of plant growth are presented in table. 5 and 6.

 The data show that the culture of Bac. subtilis and vermicompost separately have a weak growth-stimulating effect. In biofertilizer obtained by the proposed method, a synergy of their action is manifested (see tab. 5, 6).

 Thus, biofertilizer containing a strain of bacteria Bac. subtilis IPM-215, has not only high fertilizing and protective properties, but also has high growth-promoting activity.

List of references
1. US patent N 4108625, CL. C 05 F 11/08, 1978.

 2. USSR patent N 1724658 A1, cl. C 05 F 11/00, 1989.

 3. French patent N 8113118, 1981.

 4. Patent of Russia N 2019966 C1, cl. A 01 63/00, 1991.

 5. French Patent N 2545099, C 12 N 1/14, 1984.

 6. US patent N 4642131, CL. C 05 F 11/08, 1985.

 7. Copyright certificate of the USSR N 1144377, 1983.

 8. The state catalog of pesticides permitted on the territory of the Russian Federation. - 1997.

Claims (5)

1. A method of producing bio-fertilizer, which consists in obtaining vermicompost by vermicomposting agricultural and industrial waste using earthworms and separating vermicompost from worms, characterized in that Obolensky hybrid worms obtained by crossing the “Red California Hybrid” with Russian are used in earthworms Vermicomposting is carried out by a population of earthworms Eisenia foetida at 16-24 ^° C for 4-6 months and microorganisms with fungicidal activity are introduced.  2. The method according to claim 1, characterized in that after vermicomposting vermicompost is kept in vivo for ripening.  3. The method according to claims 1 and 2, characterized in that microorganisms having fungicidal activity are introduced after separation of the worms or after ripening of vermicompost. 4. The method according to claims 1 to 3, characterized in that of the microorganisms having fungicidal activity, the bacterial strain Bacillus subtilis IPM-215 is used in concentrations of 1 • 10 ⁹ - 1 • 10 ¹² spores per 1 kg of vermicompost. 5. The method according to claims 1 to 3, characterized in that microorganisms having fungicidal activity use a culture of the microfilm fungus Trichoderma viride Pers ex SFGray N 16 at concentrations of 1 • 10 ⁴ - 1 • 10 ⁸ colony forming units per 1 kg of vermicompost.

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