CN110205354B - Preparation method of earthworm bioactive fulvic acid - Google Patents
Preparation method of earthworm bioactive fulvic acid Download PDFInfo
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Abstract
The invention discloses a preparation method of earthworm bioactive fulvic acid, which relates to a preparation method of fulvic acid, and aims to solve the problems in the process of converting livestock manure and crop straws into fulvic acid by a biochemical method and extracting the fulvic acid. The existing biochemical method needs to meet the temperature condition, and the obtained fulvic acid has low purity, poor stability and short shelf life. The invention prepares the bait for earthworms by fermenting and processing the livestock manure and straw mixture through 5 earthworm species synergistic microorganism, feeds the high-efficiency earthworm variety bred by autonomous hybridization, extracts the earthworm manure with specific components, and finally obtains the bioactive fulvic acid of the earthworms. The invention is applied to the field of fulvic acid preparation.
Description
Technical Field
The invention relates to a preparation method of fulvic acid.
Background
Fulvic acid is the mineral source fulvic acid: is extracted from low-grade coal (peat, lignite and weathered coal), has a single composition structure and is mainly aromatic hydroxycarboxylic acid substances, so that the activity and stability are strong, but the purification process is complex, the cost is high, and a large amount of pollutants are produced in the process;
the biochemical fulvic acid is extracted from decomposed organic fertilizer obtained by fermenting plant waste by microorganism, and has 4 problems in the production and purification process:
(1) Under the existing biochemical treatment of livestock manure and crop straws, the conversion rate of fulvic acid is low;
(2) The livestock manure and the crop straw are treated by the existing biochemical method to generate more other substances, such as nucleic acid, sugar, amino acid, protein, water-soluble carbohydrate and the like, and the proportion of the substances is larger;
(3) Because the physical and chemical properties of the substances are similar to those of the fulvic acid, the cost of separately enriching and extracting the fulvic acid after mixing is high;
(4) Under the existing biochemical method treatment, the problem that the biochemical fulvic acid can not be extracted by fermentation treatment under the low-temperature condition can not be realized.
In conclusion, the existing biochemical method needs to meet the temperature condition, and the obtained fulvic acid has low purity, poor stability and short shelf life.
The Chinese patent of the invention discloses a harmless extraction process for preparing agricultural biochemical fulvic acid from livestock manure (CN 102816332B), which is characterized in that the fulvic acid and the fulvic acid are extracted from livestock manure fermentation materials by acid dissolution, and meanwhile, the residual antibiotics, hormone, pesticide, pathogenic bacteria, saprophytic bacteria and parasite eggs in the fulvic acid extraction materials are subjected to oxidative detoxification and disinfection harmless treatment by Fenton reaction. Although agricultural biochemical fulvic acid (comprising biochemical liquid fulvic acid and biochemical fulvic acid fine powder) with higher purity can be obtained, and meanwhile, the antibiotic residues are removed. However, the main purpose of the patent is to remove the antibiotic residues, and secondly to prepare biochemical fulvic acid, so that the technology adopts Fenton reaction technology for removing the antibiotic residues as a technical route; the Fenton reaction condition is high, so that the method is not suitable for being carried out in a low-temperature environment, and the acid solution is adopted to directly extract the fulvic acid in the excrement, so that the problems of low fulvic acid extraction rate and environmental pollution are necessarily caused; the patent does not enrich the fulvic acid in the feces to increase the content of the fulvic acid in the extract, and does not filter the influence of other impurities, thereby improving the extraction rate and purity.
The Chinese patent of the invention, "a method for producing fulvic acid by mixed bacteria fermentation" (CN 106960260A) discloses that bacillus subtilis, candida tropicalis and aspergillus niger are adopted as mixed bacteria, straw is treated in an alkaline environment of straw pulp black liquor, and then a carbon source in mushroom residues is utilized to improve the growth speed of the mixed bacteria, so that organic matters of the straw pulp black liquor and the straw are efficiently converted to prepare fulvic acid. The patent utilizes mixed thalli to grow and reproduce fast, and extracts fulvic acid in straw pulp black liquor and organic matters of straw. However, the method adds a plurality of nutrients to improve the rapid growth and propagation of the thalli, the fermentation process of the thalli is complex, and the thalli directly extracts fulvic acid in straw pulp black liquor and organic matters of straw, so that the extraction rate is low. The method does not solve the problem of low extraction rate of fulvic acid caused by lower fulvic acid content of straw pulp black liquor and straw. In addition, the patent does not extract biochemical fulvic acid. The bacteria used in this patent are not suitable for fermentation treatment in the low temperature environment of north.
The Chinese patent of the invention, a compound microbial agent and the application thereof in the production of fulvic acid by fermenting humic acid lignite (CN 107815433A), discloses a method for producing fulvic acid by fermenting humic acid lignite by using the compound microbial agent, and the patent method is more environment-friendly and does not produce pollutants compared with a chemical method. However, this patent is based on lignite extraction of fulvic acid, not biochemical fulvic acid. Moreover, the strain is not suitable for fermentation treatment in low-temperature environment, and can not ferment livestock manure and crop straws to extract fulvic acid
Disclosure of Invention
The invention aims to solve 4 problems in the process of converting livestock manure and crop straws into fulvic acid and extracting the fulvic acid by a biochemical method: 1. under the existing biochemical treatment of livestock manure and crop straws, the conversion rate of fulvic acid is low; 2. the livestock manure and the crop straw are treated by the existing biochemical method to generate more other substances, such as nucleic acid, sugar, amino acid, protein, water-soluble carbohydrate and the like, and the proportion of the substances is larger; 3. because the physical and chemical properties of the substances are similar to those of the fulvic acid, the cost of separately enriching and extracting the fulvic acid after mixing is high; 4. under the existing biochemical method treatment, the problem that the biochemical fulvic acid can not be extracted by fermentation treatment under the low-temperature condition can not be realized.
Therefore, the existing biochemical method needs to meet the temperature condition, and the obtained fulvic acid has lower purity, poorer stability and short shelf life.
In order to solve the problems, the invention creatively adopts the synergistic effect of earthworms and various mixed bacteria to jointly extract the fulvic acid of crop straws and the feces of livestock. Through the combined fermentation of a plurality of bacteria, the fermentation product is fed to earthworms, humic acid with large molecular weight and low functional group number is further decomposed into fulvic acid with small molecular weight and high functional group number by the earthworms, and protein, amino acid and sugar in the bait are absorbed to remove unwanted substances. Moreover, by eating the bait by earthworms, crop straws and livestock manure can be effectively enriched into the earthworms manure, and finally the manure is subjected to fulvic acid extraction, so that the biochemical fulvic acid with extremely high content ratio of aromatic hydroxyl and carboxylic acid substances can be obtained.
The invention relates to a preparation method of earthworm bioactive fulvic acid, which is realized according to the following steps:
1. mixing crop straw with livestock manure, fermenting, and taking the fermented product as earthworm bait; wherein, adding the mixed compound microorganism bacterial liquid of 5 microorganism strains according to the inoculation amount of 0.1% in the fermentation treatment; the microorganism is fusarium oxysporum (Fusarium oxysporum), neurospora (Neurospora sp.), phanerochaete chrysosporium (Phanerochaete chrysosporium), bacillus subtilis (Bacillus subtilis) and Bacillus acidophilus (Bacillus acidiceler);
the initial condition of fermentation treatment is that the carbon-nitrogen ratio is 25-30: 1, the humidity is 45-55%, the pH value is 6.5-7.0, the temperature is 8-20 ℃, the rotary turning of the roller is carried out every 24 hours in the fermentation process, and the fermentation is carried out until the fineness of the product is less than or equal to 5 mm;
2. uniformly putting the earthworm baits obtained in the step one into a bioreactor containing earthworms for feeding and culturing; collecting the excrement produced after feeding earthworms in the second step, adding the excrement into a No. 1 reaction kettle, adding deionized water, adjusting the temperature to 60 ℃, starting the reaction kettle to sufficiently stir and dissolve for 30min, and then carrying out solid-liquid separation to obtain a No. 1 humic acid aqueous solution and a No. 1 residue; wherein, the mass ratio of deionized water to earthworm feces is 3:1, a step of;
3. placing the humic acid aqueous solution No. 1 into a reaction kettle No. 2, adding citric acid or a polybasic organic acid, adjusting the pH value to 2, starting the reaction kettle to stir and react for 15min, and performing solid-liquid separation to obtain the humic acid aqueous solution No. 2 and residue No. 2;
4. placing the residue No. 2 in a reaction kettle No. 1, and mixing with the residue No. 1 in the reaction kettle No. 1 to obtain a residue mixture; adding deionized water with the mass of 3 times of the mass of the residue mixture, adding mixed acid with the mass of 1/5 of the mass of the residue mixture, starting a reaction kettle to fully stir and dissolve for 30min, and then carrying out solid-liquid separation to obtain a humic acid water solution No. 3 and residue No. 3;
5. mixing the humic acid aqueous solution No. 3 with the humic acid aqueous solution No. 2, and regulating the pH value to 5.5-6.0 to obtain a humic acid aqueous solution containing fulvic acid;
6. and (3) drying and crushing the humic acid aqueous solution containing the fulvic acid, and obtaining the earthworm bioactive fulvic acid powder after drying and crushing.
The invention has the following beneficial effects:
according to the invention, through years of researches on the extraction of fulvic acid from lignite and the preparation of fulvic acid by a biochemical method, the invention discovers that the fulvic acid is generated by complex physiological and biochemical reactions of lignin and nitrogenous organic matters in plant straws. Therefore, the breeding of microorganisms and the hybridization breeding of earthworm varieties play a decisive role in the degradation and conversion of lignin in straws.
The invention firstly mixes crop straws with livestock manure, adopts mixed bacteria to ferment under a low-temperature environment (10-15 ℃), and improves the content specific gravity of macromolecular humic acid in bait by strictly controlling the mixing proportion of the crop straws with the livestock manure, so that the fermented material is thoroughly decomposed after fermentation, and the fineness of the fermented material can reach a range of 5 mm. 1 ton of fermentation material, and the mass of the earthworm bait after fermentation is about 700 kg. Wherein the mass of humic acid is about 200 kg and the mass of fulvic acid is 20-35 kg.
Secondly, the fermentation product is used as bait to feed earthworms, the earthworms eat the bait according to the health condition of the body of the earthworms, the earthworms chew the bait finely through tiny teeth in the oral cavity of the earthworms, then the earthworms enter the intestinal tract and are subjected to comprehensive digestion through various enzymes in the intestinal tract to absorb soluble carbohydrates, proteins, amino acids and saccharides in the bait, and meanwhile, humic acid with large molecular weight and low functional group number is further decomposed into fulvic acid with small molecular weight and high functional group number. Finally, the high-quality wormcast is obtained, the total content of the purifiable fulvic acid is more than 12% of the mass of the initial material, namely the conversion rate of converting organic solid waste into fulvic acid is more than 12%. 1 ton of fermentation material, the mass of the earthworm bait after fermentation is 700 kg, and 400-450 kg of wormcast with specific components can be produced after feeding earthworms. Wherein the mass of humic acid is about 260-280 kg and the mass of fulvic acid is about 115-125 kg.
Finally, the mixed acid of nitric acid and sulfuric acid is added to assist treatment during the treatment in a reaction kettle, so that the intensity of the reaction is reduced to make the reaction stable, the excessive reaction is prevented from influencing the yield of fulvic acid and damaging workers and equipment in the operation process, the combined humic acid is converted into free humic acid by utilizing the stable oxidation reaction of the mixed acid, and the free humic acid is oxidized and decomposed into fulvic acid with small molecular weight and a large number of functional groups. Obtaining humic acid aqueous solution containing fulvic acid (more than 90%), and finally obtaining the earthworm bioactive fulvic acid (powder) through drying treatment. The comprehensive yield of the final fulvic acid is more than 15% of the initial material, namely the conversion rate of the organic solid waste into fulvic acid reaches more than 15%. 1 ton of fermentation material, the mass of the fermentation material is 700 kg after fermentation, and 400-450 kg of wormcast with specific components can be produced after feeding earthworms; then mixed acid is subjected to oxidative decomposition, purification and drying to obtain 150 kg of fulvic acid powder.
The earthworm variety of the invention can be a conventional earthworm variety, or an earthworm variety which can grow at the temperature of 10-35 ℃, or a high-quality earthworm variety in Ji Si Tan area and a high-quality earthworm variety in Russian Yuan Dong area are bred through distant hybridization, the bred earthworms can efficiently process the mixture of livestock manure and straw, the processing efficiency is more than 2 times of that of the common earthworm variety 'Daping No. two' in China, the optimum working temperature is 10-35 ℃ (about 25 ℃ for Daping No. two earthworms), and the earthworms are special varieties with high efficiency and high stress resistance.
Moreover, the bacteria and earthworms adopted by the invention can be suitable for low-temperature environment (more than 10 ℃), so as to carry out growth and propagation. Therefore, the method of the invention can be widely popularized and applied in the northern low-temperature environment.
Drawings
FIG. 1 is a flow chart of the production process of fulvic acid according to the invention;
FIG. 2 is a photograph of a test paddy field;
FIG. 3 is a comparative photograph showing the appearance of rice treated with a biologically active fulvic acid nutrient solution; wherein A is a control group, and B is earthworm bioactive fulvic acid;
FIG. 4 is a graph showing the evaluation result of the appearance quality of rice; wherein A is the biological active fulvic acid of the earthworm in the number of the amblyseius japonica 4, and B is the control in the number of the amblyseius japonica 4.
Detailed Description
The first embodiment is as follows: the preparation method of the earthworm bioactive fulvic acid in the embodiment is realized according to the following steps:
1. mixing crop straw with livestock manure, fermenting, and taking the fermented product as earthworm bait; wherein, adding the mixed compound microorganism bacterial liquid of 5 microorganism strains according to the inoculation amount of 0.1% in the fermentation treatment; the microorganism is fusarium oxysporum (Fusarium oxysporum), neurospora (Neurospora sp.), phanerochaete chrysosporium (Phanerochaete chrysosporium), bacillus subtilis (Bacillus subtilis) and Bacillus acidophilus (Bacillus acidiceler);
the initial condition of fermentation treatment is that the carbon-nitrogen ratio is 25-30: 1, the humidity is 45-55%, the pH value is 6.5-7.0, the temperature is 8-20 ℃, the rotary turning of the roller is carried out every 24 hours in the fermentation process, and the fermentation is carried out until the fineness of the product is less than or equal to 5 mm;
2. uniformly putting the earthworm baits obtained in the step one into a bioreactor containing earthworms for feeding and culturing; collecting the excrement produced after feeding earthworms in the second step, adding the excrement into a No. 1 reaction kettle, adding deionized water, adjusting the temperature to 60 ℃, starting the reaction kettle to sufficiently stir and dissolve for 30min, and then carrying out solid-liquid separation to obtain a No. 1 humic acid aqueous solution and a No. 1 residue; wherein, the mass ratio of deionized water to earthworm feces is 3:1, a step of;
3. placing the humic acid aqueous solution No. 1 into a reaction kettle No. 2, adding citric acid or a polybasic organic acid, adjusting the pH value to 2, starting the reaction kettle to stir and react for 15min, and performing solid-liquid separation to obtain the humic acid aqueous solution No. 2 and residue No. 2;
4. placing the residue No. 2 in a reaction kettle No. 1, and mixing with the residue No. 1 in the reaction kettle No. 1 to obtain a residue mixture; adding deionized water with the mass of 3 times of the mass of the residue mixture, adding mixed acid with the mass of 1/5 of the mass of the residue mixture, starting a reaction kettle to fully stir and dissolve for 30min, and then carrying out solid-liquid separation to obtain a humic acid water solution No. 3 and residue No. 3;
5. mixing the humic acid aqueous solution No. 3 with the humic acid aqueous solution No. 2, and regulating the pH value to 5.5-6.0 to obtain a humic acid aqueous solution containing fulvic acid;
6. and (3) drying and crushing the humic acid aqueous solution containing the fulvic acid, and obtaining the earthworm bioactive fulvic acid powder after drying and crushing.
The second embodiment is as follows: the first difference between this embodiment and the specific embodiment is that: the livestock manure is one or more of pig manure, sheep manure and cow manure; the crop straw is one or more of corn, rice, soybean and wheat. The other is the same as in the first embodiment.
And a third specific embodiment: the first difference between this embodiment and the specific embodiment is that: the mass ratio of the pig manure to the corn straw is 1:4, the mass ratio of the pig manure to the rice straw is 1:3.5, the mass ratio of the pig manure to the soybean straw is 1:5, and the mass ratio of the pig manure to the wheat straw is 1:6. The other is the same as in the first embodiment.
The specific embodiment IV is as follows: the first difference between this embodiment and the specific embodiment is that: the mass ratio of the cow dung to the corn straw is 1:3, the mass ratio of the cow dung to the rice straw is 1:2, the mass ratio of the cow dung to the soybean straw is 1:4, and the mass ratio of the cow dung to the wheat straw is 1:4.5. The other is the same as in the first embodiment.
Fifth embodiment: the first difference between this embodiment and the specific embodiment is that: the mass ratio of the sheep manure to the corn straw is 1:3, the mass ratio of the sheep manure to the rice straw is 1:2.5, the mass ratio of the sheep manure to the soybean straw is 1:4.5, and the mass ratio of the sheep manure to the wheat straw is 1:5.5. The other is the same as in the first embodiment.
Specific embodiment six: the first difference between this embodiment and the specific embodiment is that: the earthworm species is obtained by hybridization breeding of Eiseniafoetida (Eiseniafoetida) in the Russian far east and Eisenia reesei (Eisenia rosea rosea) in the Ji Si Tan region. The other is the same as in the first embodiment.
Seventh embodiment: the first difference between this embodiment and the specific embodiment is that: after the earthworms are fed, the environment temperature is ensured to be 10-35 ℃ in the process of biological treatment of the earthworms for feeding baits. The other is the same as in the first embodiment.
Eighth embodiment: the first difference between this embodiment and the specific embodiment is that: after the earthworms are fed, the environment temperature is ensured to be 26-28 ℃ in the process of biological treatment of the earthworms to feed baits. The other is the same as in the first embodiment.
Detailed description nine: the first difference between this embodiment and the specific embodiment is that: the mixed acid is prepared from nitric acid and sulfuric acid according to the volume ratio of 7:3, and mixing the materials in proportion. The other is the same as in the first embodiment.
Detailed description ten: the first difference between this embodiment and the specific embodiment is that: and step two, feeding 5 to 6 ten thousand earthworms with 1 cube of mixed earthworm bait. The other is the same as in the first embodiment.
The present invention is not limited to the above embodiments, and the object of the invention can be achieved by one or a combination of several embodiments.
The beneficial effects of the invention are verified by the following examples:
example 1
The verification test of the earthworm bioactive fulvic acid in the embodiment is as follows:
1. test design
The test was carried out in 2015 at the biological liquid fertilizer development plant of the biological technology development company of the university of Heilongjiang in Heilongjiang black river industry park.
The test selects 3 kinds of livestock manure and crop straws of the main agriculture of China, namely pig, cattle and sheep, and 4 kinds of grain crop straws of corn, rice, soybean and wheat. The test designs that 3 kinds of feces and 4 kinds of straws are processed in a permutation and combination mode, and simultaneously, the proportioning treatment of different materials is designed according to the optimal carbon nitrogen ratio of the biological fermentation of the organic fertilizer, and the total of 12 kinds of treatments are respectively marked as A1, A2, A3, A4, B1, B2, B3, B4, C1, C2, C3 and C4, and the details are shown in the following table 1.
TABLE 1 Material ratio of different faeces to straw (carbon to nitrogen ratio: 30/1)
2. Microbial fermentation
The length of the fermentation stack is about 800cm, the width is about 300 cm, the upper width is about 180 cm, the height is about 150 cm, and a ventilation groove with the width of 40 cm and the depth of 15 cm is arranged in the middle of the bottom of the crenels. Layering according to the volume of the materials to form a strip stack, dividing into 5 layers, spraying earthworm synergistic microbial fermentation inoculant [ Fusarium oxysporum (Fusarium oxysporum), neurospora (Neurospora sp.), phanerochaete chrysosporium (Phanerochaete chrysosporium), bacillus subtilis (Bacillus subtilis) and Bacillus acidi fast-growing (Bacillus acidiceler) ] between each layer, and uniformly mixing with the materials. The total dry weight of each test treatment material is 10000kg, the water content of the material is regulated to be about 65% during stacking, the stacking is respectively carried out during stacking for 3d, 7d and 10d, the total stacking is carried out for 3 times, and the total fermentation time is 10day.
The 12 treatments adopt long strip stack type aerobic stacking fermentation without repetition. After fermentation, 3kg of fully fermented organic fertilizer is collected by a five-point sampling method, and 1kg of fully and uniformly mixed organic fertilizer is sampled by a four-point method for humic acid and fulvic acid content detection. (first batch of 12 treatments)
3. Earthworm biological treatment
The 12 processed earthworm baits after fermentation are respectively and evenly put into a small-sized bioreactor which is provided with earthworm varieties cultivated independently for feeding and cultivating, and the optimal density (5.5 ten thousand earthworms/m) of the earthworms in the reactor is ensured 3 ) And continued to be treated for 72 hours (the optimal period of earthworm treatment "72 h").
And then carrying out drum-type screening on earthworms and the processed wormcast with specific components to obtain earthworms and wormcast, collecting 3kg of fully fermented organic fertilizer for each treatment by using a five-point sampling method, fully stirring uniformly, and sampling 1kg by using a four-division method for humic acid and fulvic acid content detection. (second batch of 12 treatments)
4. Purification of fulvic acid
(1) Respectively sampling 100g of wormcast subjected to microbial fermentation and earthworm biological treatment, putting the wormcast into a magnetic stirrer, simultaneously adding 500ml of deionized water, uniformly stirring, adding citric acid, adjusting the pH value to 2, starting the stirrer, stirring for 10-15 min, and performing solid-liquid separation;
(2) Introducing the solution into a blank beaker, placing the solution, leaving the precipitated residue in a stirrer, adding 250ml of deionized water, simultaneously adding 5ml of mixed acid (nitric acid: sulfuric acid=7:3), starting the stirrer to sufficiently stir and dissolve for 30min, performing solid-liquid separation, and mixing the newly separated solution with the solution placed in the step (1) to obtain a fulvic acid aqueous solution;
(3) Adding a proper amount of potassium hydroxide into the fulvic acid aqueous solution to adjust the pH value to 5.5-6.0, and drying the solution into powder to obtain fulvic acid powder;
the 12 treatments are operated according to the 3 steps to obtain 12 parts of fulvic acid powder with different masses. (third batch of 12 treatments)
5. Assay of humic acid and fulvic acid content
The detection of humic acid content is carried out according to NYT 1971-2010;
the detection of the fulvic acid content was performed according to HG/T3276-2012.
(1) Assaying humic acid and fulvic acid content in 12 treatments, wherein the first batch of assay objects are 12 treatments after microbial fermentation, and the assay results are shown in the following table 2:
table 2 humic acid content (first lot) units in 12 treatments: g/kg
(2) Assaying the humic acid and fulvic acid contents in 12 treatments, wherein the second batch of assay objects are 12 treatments after earthworm biological treatment, and the assay results are shown in the following table 3:
table 3 humic acid content (second lot) units in 12 treatments: g/kg
(3) And (3) carrying out primary purification and oxidative decomposition of the precipitation residues after 12 treatments and further purifying to obtain a fulvic acid aqueous solution, drying the aqueous solution into powder to obtain fulvic acid powder, and finally determining the quality of the fulvic acid powder to obtain the quality of the fulvic acid powder produced by different treatments, thereby obtaining the fulvic acid yield of different treatments. Specific data are shown in table 4 below:
TABLE 4 mass (third batch) units of purified fulvic acid for 12 treatments g
| Project | A1 | A2 | A3 | A4 | B1 | B2 | B3 | B4 | C1 | C2 | C3 | C4 |
| Fulvic acid | 75.2 | 74.2 | 73.8 | 74.5 | 75.5 | 71.8 | 73.9 | 74.3 | 73.8 | 74.6 | 72.1 | 74.8 |
* And (3) injection: the sample mass taken for each treatment was 500g.
6. Analysis of results
Through detection data, as long as the mixture ratio of the straw and the manure is ensured to reach the optimal carbon nitrogen ratio, the standardization of microbial fermentation and the standardization of earthworm biological treatment are ensured, and the influence of the mixture ratio of different manure and different straw on the final yield of fulvic acid is not obvious. The method for preparing the fulvic acid through microbial fermentation and earthworm biological treatment is wider in material selection, has no limitation on specific material selection, and needs to ensure that the carbon-nitrogen ratio of the fermented material is controlled to be 30/1 because the carbon-nitrogen ratio influences the efficiency and quality of microbial fermentation, so that the carbon-nitrogen ratio is more important to the material selection.
Example 2
Farmland experiments were performed using the earthworm bioactive fulvic acid prepared in example 1 to verify the growth promoting effect of the earthworm bioactive fulvic acid prepared in example 1 on rice and the yield improving effect.
The experimental object: baiquan Fengcao (Beijing Hequan agricultural technologies Co., ltd.).
The experimental method comprises the following steps: in month 8 of 2018, 1kg of the earthworm bioactive fulvic acid prepared in example 1 was sprayed on 1 hectare farmland.
The results show that: the rice cultivated by the bioactive fulvic acid nutrient solution has the effective tillering increased by 1.3 in average, thousand grain weight increased by 32g, the same ratio increased by 8%, the yield per mu of rice increased by 1325.7 jin and the yield increased by 13.2%.
In order to test the quality of the rice, the rice sample treated by the bioactive fulvic acid nutrient solution and the rice sample treated by the bioactive fulvic acid nutrient solution are mailed to a cereal detection center of the agricultural rural area together for identifying the appearance quality and the taste quality of the rice. The results are shown in Table 5.
Table 5 sample detection
As shown in Table 5, the rice samples treated with the bioactive fulvic acid nutrient solution each exceeded the standard of high-quality rice specified by the Ministry of agriculture, and some of the indexes reached a very high level. The appearance control is shown in fig. 3. The appearance quality of the rice treated by the bioactive fulvic acid nutrient solution is obviously improved.
The international rice appearance quality evaluation criteria were classified into 5 ranks of "S, A, B, C, D". As shown in FIG. 4, the appearance quality of untreated rice was grade D, while the appearance quality of rice treated with the bioactive fulvic acid nutrient solution was grade S, and the appearance quality of rice was directly increased from the lowest grade D to the highest grade S. Proved that the bioactive selenium nutrient solution has extremely obvious effect of improving the appearance quality of rice.
Claims (9)
1. The preparation method of the earthworm bioactive fulvic acid is characterized by comprising the following steps of:
1. mixing crop straw with livestock manure, fermenting, and taking the fermented product as earthworm bait; wherein, adding the mixed compound microorganism bacterial liquid of 5 microorganism strains according to the inoculation amount of 0.1% in the fermentation treatment; the microorganism is fusarium oxysporum (Fusarium oxysporum), neurospora (Neurospora sp.), phanerochaete chrysosporium (Phanerochaete chrysosporium), bacillus subtilis (Bacillus subtilis) and Bacillus acidophilus (Bacillus acidiceler); the initial condition of fermentation treatment is that the carbon-nitrogen ratio is 25-30: 1, the humidity is 45-55%, the pH value is 6.5-7.0, the temperature is 8-20 ℃, the rotary turning of the roller is carried out every 24 hours in the fermentation process, and the fermentation is carried out until the fineness of the product is less than or equal to 5 mm;
2. uniformly putting the earthworm baits obtained in the step one into a bioreactor containing earthworms for feeding and culturing;
collecting the excrement produced after feeding earthworms in the second step, adding the excrement into a No. 1 reaction kettle, adding deionized water, adjusting the temperature to 60 ℃, starting the reaction kettle to sufficiently stir and dissolve for 30min, and then carrying out solid-liquid separation to obtain a No. 1 humic acid aqueous solution and a No. 1 residue; wherein, the mass ratio of deionized water to earthworm feces is 3:1, a step of;
3. placing the humic acid aqueous solution No. 1 into a reaction kettle No. 2, adding citric acid, adjusting the pH value to 2, starting the reaction kettle to stir and react for 15min, and performing solid-liquid separation to obtain the humic acid aqueous solution No. 2 and residue No. 2;
4. placing the residue No. 2 in a reaction kettle No. 1, and mixing with the residue No. 1 in the reaction kettle No. 1 to obtain a residue mixture; adding deionized water with the mass of 3 times of the mass of the residue mixture, adding mixed acid with the mass of 1/5 of the mass of the residue mixture, starting a reaction kettle to fully stir and dissolve for 30min, and then carrying out solid-liquid separation to obtain a humic acid water solution No. 3 and residue No. 3; the mixed acid is prepared from nitric acid and sulfuric acid according to the volume ratio of 7:3, mixing the materials in proportion;
5. mixing the humic acid aqueous solution No. 3 with the humic acid aqueous solution No. 2, and regulating the pH value to 5.5-6.0 to obtain a humic acid aqueous solution containing fulvic acid;
6. and (3) drying and crushing the humic acid aqueous solution containing the fulvic acid, and obtaining the earthworm bioactive fulvic acid powder after drying and crushing.
2. The method for preparing the earthworm bioactive fulvic acid, according to claim 1, wherein the livestock manure is one or more of pig manure, sheep manure and cow manure; the crop straw is one or more of corn, rice, soybean and wheat.
3. The method for preparing the earthworm bioactive fulvic acid, according to claim 2, is characterized in that the mass ratio of pig manure to corn straw is 1:4, the mass ratio of pig manure to rice straw is 1:3.5, the mass ratio of pig manure to soybean straw is 1:5, and the mass ratio of pig manure to wheat straw is 1:6.
4. The method for preparing the earthworm bioactive fulvic acid, according to claim 2, which is characterized in that the mass ratio of cow dung to corn straw is 1:3, the mass ratio of cow dung to rice straw is 1:2, the mass ratio of cow dung to soybean straw is 1:4, and the mass ratio of cow dung to wheat straw is 1:4.5.
5. The method for preparing the earthworm bioactive fulvic acid, according to claim 2, is characterized in that the mass ratio of sheep manure to corn straw is 1:3, the mass ratio of sheep manure to rice straw is 1:2.5, the mass ratio of sheep manure to soybean straw is 1:4.5, and the mass ratio of sheep manure to wheat straw is 1:5.5.
6. The method for preparing the earthworm bioactive fulvic acid according to claim 1, wherein the earthworm species is obtained by hybridization breeding of a red Eisenia (Eisenia rosea rosea) in Jier Jikistan region and a red Eisenia foetida (Eisenia foetida) earthworm species in Russian far east region.
7. The method for preparing the earthworm bioactive fulvic acid according to claim 1 or 6, wherein after feeding earthworms, the environment temperature is ensured to be 10-35 ℃ in the process of earthworm biological treatment feeding baits.
8. The method for preparing the earthworm bioactive fulvic acid according to claim 1 or 6, wherein after feeding earthworms, the environment temperature is ensured to be 26-28 ℃ in the process of earthworm biological treatment feeding baits.
9. The method for preparing the earthworm bioactive fulvic acid, according to claim 1, wherein in the second step, 1 cube of the mixed earthworm bait is fed to 5-6 ten thousand earthworms.
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