CN108947679B - Microbial organic fertilizer and preparation method thereof - Google Patents
Microbial organic fertilizer and preparation method thereof Download PDFInfo
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- CN108947679B CN108947679B CN201810941544.6A CN201810941544A CN108947679B CN 108947679 B CN108947679 B CN 108947679B CN 201810941544 A CN201810941544 A CN 201810941544A CN 108947679 B CN108947679 B CN 108947679B
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05B—PHOSPHATIC FERTILISERS
- C05B7/00—Fertilisers based essentially on alkali or ammonium orthophosphates
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/60—Biocides or preservatives, e.g. disinfectants, pesticides or herbicides; Pest repellants or attractants
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/80—Soil conditioners
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Abstract
The invention relates to the field of bacterial fertilizers, in particular to a microbial organic fertilizer and a preparation method thereof. The preparation method comprises the following steps: adding bacillus subtilis into the hemp plant degumming waste liquid for aerobic fermentation; stopping supplying oxygen, inoculating EM strain, and performing anaerobic fermentation. According to the invention, the hemp plant degumming waste liquid is used as a raw material, and then bacillus subtilis is added for aerobic fermentation, so that the bacillus subtilis can quickly form a dominant population, thereby inhibiting the growth of pathogenic bacteria or other mixed bacteria; active substances such as subtilin, polymyxin, nystatin, gramicidin and the like are generated in the growth process of the thalli, and the bacillus subtilis has obvious inhibiting effect on pathogenic bacteria or pathogenic bacteria with endogenous infection. Adding EM strain for anaerobic fermentation, wherein the anaerobic EM strain produces organic acid by fermentation, and the pH value is reduced to form subacidity. In addition, the EM is a composite strain, and a plurality of probiotics are fermented and applied to soil to form a symbiotic stable bacterial colony environment, so that the effect is more durable.
Description
Technical Field
The invention relates to the field of bacterial fertilizers, and particularly relates to a microbial organic fertilizer and a preparation method thereof.
Background
Existing microorganisms are commonly used in the preparation of fertilizers or as wastewater treatment.
Common microbial organic fertilizer is prepared by taking straws and/or animal wastes as raw materials and adding strains for fermentation, for example, application number 201410262147.8 discloses a microbial organic fertilizer and a preparation method thereof, and bacillus subtilis is utilized for solid fermentation of agricultureOrganic solid waste, and producing the microbial organic fertilizer for preventing and removing continuous cropping obstacles of the artemisia selengensis. The number of functional microorganisms in the microbial organic fertilizer is more than or equal to 2 multiplied by 108cfu·g-1The water content is less than 30 percent, and the organic matter content is more than 30 percent. The microbial organic fertilizer can accelerate decomposition of artemisia selengensis stubble, inhibit proliferation of soil pathogenic bacteria, accelerate crop growth and reduce harm of artemisia selengensis continuous cropping obstacle. Tests show that after the biological organic fertilizer is applied to continuous cropping obstacle soil, the incidence rate of artemisia selengensis is reduced by 91%, and the yield of artemisia selengensis is increased by 3.3 times; the microbial organic fertilizer can well prevent and remove continuous cropping obstacles of the artemisia selengensis.
The following patents are related to the common microbial treatment of wastewater:
application number 201410659760.3 discloses a production method of a full water-soluble solid bio-organic fertilizer, which comprises the following steps: fermenting by yeast to generate wastewater rich in various nutrient substances, and concentrating the wastewater into a semi-fluid; flash drying the semifluid to a dry powder; and supplementing nitrogen, phosphorus and potassium nutrients in the dry powder, and adding a plant rhizosphere growth promoting microbial agent. According to the invention, the fermentation waste liquid is deeply processed, and the rhizosphere growth promoting microbial inoculum is used, the specially added functional microorganisms can be rapidly propagated by preferentially utilizing amino acid and vitamin, a dominant flora is formed in the soil range reached by the fertilizer, the growth metabolite of the dominant flora can promote the growth of plants, and the propagation of other microorganisms is inhibited or interfered, so that the effects of further improving the fertilizer efficiency and inhibiting the propagation of pathogenic bacteria are achieved.
Application No. 201610710440.5 discloses a method for producing liquid organic fertilizer for improving saline-alkali soil by using starch wastewater, which comprises a series of advanced treatments such as filter screen filtration, anaerobic biofilter fermentation, solar aeration biofilter fermentation, hydrolysis of starch phosphate and poly-phosphorus ferric chloride, microbial fermentation bacteria fermentation and the like, so that the starch wastewater becomes liquid microbial fertilizer rich in beneficial microbial flora and various secondary metabolites, and the problem of microbial population reduction in improving secondary saline-alkali soil after use can be solved, the yield of the secondary saline-alkali soil can be increased by 20-30%, the cost is saved, the soil poor, hardening and salinization are not relieved, and the method accords with the trends of energy conservation, environmental protection and sustainable development.
201711421916.4 discloses a method for producing multifunctional bio-organic fertilizer by recycling yeast wastewater, which comprises the following steps: s1: treating raw materials; s2: culturing bacillus; s3: fermentation: inoculating bacillus into the yeast liquid wastewater in the step S1, and stirring for 20-40 hours at 32-42 ℃ to obtain fermentation liquor; s4: organic fertilizer fermentation: adding organic fertilizer into fermentation liquor in S3, wherein the organic fertilizer comprises crushed crop straws, excrement, bran shells and weeds, then uniformly stirring, sealing by plastic cloth, performing anaerobic fermentation, the fertilizer efficiency is 30% higher than that of farmyard manure, the mature days of crops can be improved, the soil environment can be improved, not only industrial and agricultural pollution is solved, but also resource recycling can be realized, harmful substances in air and soil can be degraded, and the plant root system is ensured to be green, organic and environment-friendly.
The ramie degumming waste liquid is sewage with high organic matter content, and has the main characteristics of high water temperature, high pH value and deep chroma of the waste water, taking chemical degumming waste water as an example, the water temperature of the waste water discharged by a boiling pot is 90 ℃, the pH value is 14, and the COD can reach more than 15000mg/L, which is a difficult problem troubling sewage treatment of enterprises for a long time.
The ramie degumming waste liquid is also treated by means of microorganisms, chemical treatment, enzyme treatment and the like to reduce the content of pollutants in the waste water, but the ramie degumming waste liquid is discharged after treatment, thereby wasting energy. Therefore, a new disposal method is urgently needed to make the resources more fully utilized.
In view of the above, the present invention is particularly proposed.
Disclosure of Invention
The invention aims to provide a preparation method of a microbial organic fertilizer, which takes degumming waste liquid of hemp plants as a raw material and finally obtains the microbial organic fertilizer containing effective active ingredients by introducing specific strains.
In order to achieve the above purpose of the present invention, the following technical solutions are adopted:
a preparation method of a microbial organic fertilizer comprises the following steps:
adding bacillus subtilis into the hemp plant degumming waste liquid for aerobic fermentation;
and after stopping supplying oxygen, inoculating EM strain for anaerobic fermentation to obtain the microbial organic fertilizer.
The preparation method of the microbial organic fertilizer provided by the invention takes the hemp plant degumming waste liquid as a raw material, and then bacillus subtilis is added for aerobic fermentation, so that the bacillus subtilis can quickly form an advantageous population, thereby inhibiting the growth of pathogenic bacteria or other mixed bacteria; in addition, active substances such as subtilin, polymyxin, nystatin, gramicidin and the like are generated in the growth process of the thalli, and the active substances have obvious inhibiting effect on pathogenic bacteria or conditional pathogenic bacteria of endogenous infection. And then adding EM (effective microorganisms) for anaerobic fermentation, wherein the anaerobic EM can generate organic acid through fermentation, the pH value is reduced, and the pH value is slightly acidic. In addition, the EM is a composite strain, and a plurality of probiotics are fermented, so that a symbiotic stable bacterial colony environment can be formed when the EM is applied to soil, and the EM is better and more durable than a single microorganism.
The microbial inoculum added in the prior degumming waste liquid treatment is usually activated sludge and is used for treating sewage. The sewage treatment process forms redundant activated sludge, and the brown flocculent activated sludge is observed under a microscope to see a large amount of bacteria, fungi, protozoa and metazoa, which form a special ecological system. Although the activated sludge contains a large amount of organic matters and plant nutrients, the activated sludge can provide nutrients beneficial to the growth and development of plants. However, at the same time it also contains pathogenic bacteria, parasites (eggs) and is putrefactive and smelly.
In the invention, the hemp plant degumming waste liquid can be chemical degumming waste liquid, biological degumming waste liquid or degumming waste liquid of other methods.
Further, adding dipotassium hydrogen phosphate and urea into the hemp plant degumming waste liquid before adding the bacillus subtilis. The dipotassium phosphate provides inorganic elements and plays a certain buffering role. The urea with a certain concentration provides nitrogen element, and provides good nutritional support for the microbial thallus to produce active polypeptide.
Further, the addition amount of the dipotassium phosphate is 0.05-0.2% of the mass of the hemp plant degumming waste liquid, and the addition amount of the urea is 0.05-0.2% of the mass of the hemp plant degumming waste liquid.
The pH values of different degumming waste liquids are greatly different, for example, the pH value of the chemical degumming waste liquid is usually higher than 12, the pH value needs to be adjusted, and the pH value of the waste liquid is adjusted to 7-10 before the bacillus subtilis is added; the pH value of the biological degumming waste liquid is generally 7-10, and the pH value is not required to be adjusted. In addition, the chemical degumming liquid usually contains various degumming aids, the components are complex, the influence on subsequent processing is uncertain, and the chemical degumming mode is rarely applied due to environmental protection factors. Therefore, the degumming waste liquid is preferably a biological degumming waste liquid.
Generally, the temperature is higher when the hemp plant degumming waste liquid is discharged, but the temperature is too high to be beneficial to the multiplication of the bacillus subtilis, and the bacillus subtilis is preferably added at the temperature of below 48 ℃. The Bacillus subtilis is generally cooled to 40-47 ℃.
Further, the bacillus subtilis is alkalophilic bacillus subtilis. Because the degumming waste liquid of the bacillus subtilis is alkaline, the alkalophilic bacillus subtilis can form a dominant flora more quickly after being added.
Further, the bacillus subtilis is activated and domesticated to obtain the alkalophilic bacillus subtilis.
After the domesticated bacillus subtilis strain is inoculated into degumming waste liquid for fermentation for 4 hours, the viable bacteria content reaches 100 hundred million/ml to 170 hundred million/ml, and the concentration of the viable bacteria is improved by 100 times compared with the bacterial liquid obtained by the non-domesticated bacillus subtilis strain.
Further, the domestication is: inoculating the activated bacillus subtilis into a hemp plant gum culture medium for culture, wherein the hemp plant gum culture medium comprises the following components: calculated by the volume of water, the hemp plant is 0.05-0.08g/ml, (NH)4)2SO40.008-0.015g/ml，K2HPO4 0.001-0.002g/ml，MgSO4·7H2O0.001-0.002 g/ml, Na2CO3Adjusting pH to 10-11, and sterilizing.
The domestication process can be divided into multiple times, preferably more than 2 times, that is, after the activated bacillus subtilis is inoculated into the hemp plant gum culture medium for culture, a part of the bacterial liquid is taken as a strain to be continuously inoculated into the hemp plant gum culture medium for continuous culture, and domestication is carried out for 2 times, or can be continuously repeated for multiple times to obtain the domesticated bacillus subtilis.
The hemp plant gum culture medium is sterilized and cooled to below 45 ℃ for use.
In the present invention, the hemp plants include ramie, jute, ramie, flax, kendir, and the like.
If the bast fiber plant in the invention is ramie, the bast fiber plant in the components of the bast fiber plant gum culture medium is generally ramie; if the bast fiber plant in the invention is jute, the bast fiber plant in the component of the bast fiber plant gum culture medium is jute generally; and the like, so as to be beneficial to domestication of the bacillus subtilis.
Further, the volume percentage of the activated bacillus subtilis liquid added into the hemp plant gum culture medium is 1-3%.
Further, the culture temperature of the hemp plant gum inoculated into the culture medium is 37 +/-3 ℃.
And further, inoculating a hemp plant gum culture medium, culturing for 8-10 hours, and subculturing for 6-8 hours by using partial bacterial liquid to obtain the alkalophilic bacillus subtilis.
Preferably, the inoculation amount of the alkalophilic bacillus subtilis added into the hemp plant degumming waste liquid is 1-3%.
Further, the activation step of the bacillus subtilis comprises the following steps:
inoculating bacillus subtilis into a glucose culture medium, and culturing to a logarithmic growth phase, wherein the glucose culture medium comprises the following components: calculated by the volume of water, 0.008-0.015g/ml glucose, 0.005-0.008g/ml peptone, 0.002-0.005g/ml yeast extract and K2HPO4 0.001-0.002g/ml，MgSO4·7H2O0.0002-0.0005 g/ml, and adjusting pH to 9-10.
The bacillus subtilis is subjected to an activation step, so that the activity of the bacillus subtilis is enhanced and the adaptability of the bacillus subtilis is improved while the quantity of the bacillus subtilis is increased.
Further, the bacillus subtilis is added into the hemp plant gum culture medium in a liquid state, and the volume percentage of the added bacillus subtilis is 0.3% -0.8%.
Further, the liquid form of Bacillus subtilis is prepared by adding solid bacteria into sterile water at a concentration of 0.05-0.2 g/ml.
Further, the culture temperature of inoculating the bacillus subtilis into the glucose culture medium is 28 +/-2 ℃, and the culture time is 24-48 hours.
Further, the temperature of the aerobic fermentation is 35-45 ℃. As in the case of the different embodiments,
further, the ventilation of the aerobic fermentation is not less than 1.2m3/100L·h。
According to the difference of the inoculation amount, the concentration of the thalli tends to peak value after aerobic fermentation for a period of time, and the colloid is excessively degraded by prolonging the fermentation time, which is not beneficial to the subsequent anaerobic fermentation.
Further, the time of the aerobic fermentation is 4 to 12 hours, preferably 4 to 6 hours.
Further, the EM strain is inoculated after the oxygen supply is stopped for 1-2 hours.
Further, the inoculated EM species are activated EM species.
The EM strain is subjected to an activation step, so that the number of the strains is increased, the activity of the strains is enhanced, and the adaptability of the strains is improved.
Further, the EM strain is inoculated into the culture medium for activation, wherein the culture medium comprises the following components: 8-15% of brown sugar, 0.08-0.15% of salt and 0.08-0.15% of urea by mass of water.
The culture medium can be sterilized and cooled to below 40 deg.C.
Further, the inoculation amount of the EM strain inoculated into the activation culture medium is 0.8% -1.5%.
Further, the EM strain is inoculated into an activation culture medium and then is subjected to anaerobic fermentation at the temperature of 35-37 ℃ until the mouth of the container is opened and the sour and sweet taste is smelled.
Furthermore, the time for anaerobic fermentation after the EM strain is inoculated into the activation culture medium is 2-3 days.
Furthermore, the inoculation amount of the activated EM strain is 1% -3%, and the time for inoculating the EM strain to carry out anaerobic fermentation after oxygen supply is stopped is 2-7 days.
Further, the anaerobic fermentation also comprises a vacuum drying step to form a soluble solid microbial organic fertilizer.
The invention also provides the microbial organic fertilizer prepared by the preparation method.
The microbial organic fertilizer provided by the invention contains a large amount of active cells, efficient nutrient enzymes, rich organic matters, nitrogen, phosphorus and potassium, and various medium and trace elements such as calcium, magnesium, sulfur, boron, zinc, molybdenum, copper, manganese, iron and the like, and also contains various nutrient components such as amino acid, humic acid, water-soluble protein and the like. After the fertilizer is applied to soil, the microorganisms have continuous action, can produce and increase soil nutrients, convert nutrient substances existing in nature into a state capable of being absorbed by plants, and have the capability of remarkably enhancing the disease and pest resistance, cold resistance and drought resistance of crops.
In addition, the microbial organic fertilizer provided by the invention takes the biological degumming wastewater as a raw material, and the obtained microbial organic fertilizer is green and environment-friendly, contains no hormone, is non-toxic, pollution-free and drug residue, and provides a good foundation for developing high-end certified crop product planting.
Compared with the prior art, the invention has the beneficial effects that:
(1) the invention provides a preparation method of a microbial organic fertilizer by taking degumming wastewater as a raw material, realizes the reutilization of the degumming wastewater, and if the biological degumming wastewater is taken as a raw material, the microbial organic fertilizer obtained by production is green and environment-friendly, contains no hormone, is nontoxic, nuisanceless and has no medicine residue, thereby providing a good foundation for developing high-end certified crop product planting.
(2) The invention firstly uses the bacillus subtilis to inoculate and ferment, the bacillus subtilis has strong adaptability, can quickly form dominant population, and generates active substances such as subtilin, polymyxin, nystatin, gramicidin and the like in the growth process of the bacteria, and the active substances have obvious inhibiting effect on pathogenic bacteria or conditional pathogenic bacteria of endogenous infection.
(3) After the oxygen supply is stopped, the bacillus subtilis quickly consumes free oxygen in the fermentation liquor to cause an anaerobic environment, and at the moment, the EM bacteria are inoculated to promote the growth of beneficial anaerobic bacteria and indirectly inhibit the growth of other pathogenic bacteria.
(4) The bio-organic fertilizer developed by the invention contains a large amount of active cells, efficient nutrient enzymes, rich organic matters, nitrogen, phosphorus and potassium, and various medium and trace elements such as calcium, magnesium, sulfur, boron, zinc, molybdenum, copper, manganese, iron and the like, and also has various nutrient components such as amino acid, humic acid, water-soluble protein and the like, so that the disease and pest resistance, cold resistance and drought resistance of crops can be obviously enhanced.
(5) The microbial organic fertilizer provided by the invention is a quick-acting fertilizer and a long-acting fertilizer in field application, and after long-term application, soil naturally forms a loose and breathable granular structure, so that plowing is reduced.
(6) After the microbial organic fertilizer provided by the invention is applied to soil, microorganisms continuously act to produce and increase soil nutrients, nutrient substances existing in nature are converted into a state capable of being absorbed by plants, the input of nitrogen and phosphate fertilizers can be reduced by 60-80%, the pollution of fertilizers to farmlands and foods is reduced, and the effects of low input and high output are achieved.
(7) The microbial organic fertilizer provided by the invention can generate antioxidant substances, eliminate putrefaction, eliminate oxide substances, inhibit pathogenic bacteria, control diseases and inhibit insects, and relieve fertilizer damage, the harmful pathogenic bacteria can be reduced by about 90%, a good environment suitable for growth of animals and plants is formed, and the root amount of plants is increased by about 70%.
Detailed Description
Embodiments of the present invention will be described in detail below with reference to examples, but it will be understood by those skilled in the art that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
Example 1
A preparation method of a microbial organic fertilizer comprises the following steps:
1) culture medium
Glucose culture medium: glucose 1.5g, peptone 1g, yeast extract 0.5g, K2HPO4 0.2g，MgSO4·7H2O0.05 g and water 150ml, adjusting pH to 9.5, sterilizing and cooling.
Ramie gum culture medium: ramie 10g, (NH)4)2SO4 1.5g，K2HPO4 0.2g，MgSO4·7H2O0.25g, water 150ml, Na2CO3Adjusting pH to 10.5, sterilizing, cooling to below 45 deg.C, and inoculating.
EM strain activation culture medium: 1kg of brown sugar, 10g of salt, 10g of urea and 10kg of water are mixed, sterilized and cooled to below 40 ℃.
2) Treatment of bacterial species
2.1 basophilic Bacillus subtilis
1g of Bacillus subtilis (commercially available) is suspended in 10ml of sterile water, mixed uniformly, and 1ml is inoculated into a glucose culture medium (150ml) and cultured at 28 ℃ for 36 hours to obtain activated Bacillus subtilis.
Inoculating 2ml of activated bacillus subtilis into a ramie gel culture medium (150ml) cooled to below 45 ℃, culturing for 8 hours at 37 +/-3 ℃, and subculturing for 6 hours by using the ramie gel culture medium to obtain the alkalophilic bacillus subtilis.
2.2EM strains
Sterilizing the EM strain activation culture medium, cooling to below 40 ℃, taking 100ml of EM liquid strain, inoculating into the EM strain activation culture medium prepared by 10kg of water, carrying out closed fermentation at the fermentation temperature of 35-37 ℃ for 2 days, opening the mouth of a container, and smelling acid and sweet taste, namely, successfully activating to obtain the activated EM strain.
Adding dipotassium hydrogen phosphate and urea into the hemp plant biological degumming waste liquid, and mixing, wherein the addition amount of the dipotassium hydrogen phosphate is 0.1% of the mass of the hemp plant biological degumming waste liquid, and the addition amount of the urea is 0.1% of the mass of the hemp plant biological degumming waste liquid;
inoculating alkalophilic bacillus subtilis at the waste liquid temperature of below 48 ℃, inoculating alkalophilic bacillus subtilis liquid according to the proportion of 2%, supplying air between 38 ℃ and 42 ℃ for fermentation for 4 hours, wherein the ventilation volume is not less than 1.2 cubic meters per hour per 100L;
stopping oxygen supply for 2 hours, inoculating activated EM strain, wherein the inoculation amount is 2%, performing anaerobic fermentation for 4 days, and the fermentation temperature is 35 +/-2 ℃ to form the microbial organic fertilizer.
Through multiple tests, the prepared microbial organic fertilizer is detected to contain azotobacter, bacillus, actinomycetes, filamentous fungi, photosynthetic bacteria, lactic acid bacteria and the like, and 300 to 350 hundred million probiotics are contained in 1g of the microbial organic fertilizer.
Example 2
A preparation method of a microbial organic fertilizer comprises the following steps:
1) culture medium
Glucose culture medium: glucose 1.2g, peptone 0.75g, yeast extract 0.3g, K2HPO40.15g，MgSO4·7H2O0.03 g and water 150ml, adjusting pH to 9, sterilizing and cooling.
Ramie gum culture medium: ramie 7.5g, (NH)4)2SO4 1.2g，K2HPO4 0.15g，MgSO4·7H2O0.15g, water 150ml, Na2CO3Adjusting pH to 10, sterilizing, cooling to below 45 deg.C, and inoculating.
EM strain activation culture medium: 0.8kg of brown sugar, 8g of salt, 8g of urea and 10kg of water are mixed, sterilized and cooled to below 40 ℃.
2) Treatment of bacterial species
2.1 basophilic Bacillus subtilis
0.5g of bacillus subtilis is suspended in 10ml of sterile water, evenly mixed, 0.5ml is taken to be inoculated into a glucose culture medium (150ml) and cultured for 48 hours at the temperature of 28 +/-2 ℃, and the activated bacillus subtilis is obtained.
4.5ml of the activated bacillus subtilis is inoculated into a ramie gel culture medium (150ml) cooled to below 45 ℃, cultured for 12 hours at 37 +/-3 ℃, and subcultured for 8 hours by using the ramie gel culture medium to obtain the alkalophilic bacillus subtilis.
2.2EM strains
Sterilizing the EM strain activation culture medium, cooling to below 40 ℃, taking 80ml of EM liquid strain, inoculating into the EM strain activation culture medium prepared by 10kg of water, carrying out closed fermentation at the fermentation temperature of 35-37 ℃ for 3 days, opening a container, and smelling acid and sweet taste to obtain the activated EM strain.
Adding dipotassium hydrogen phosphate and urea into the hemp plant biological degumming waste liquid, and mixing, wherein the addition amount of the dipotassium hydrogen phosphate is 0.2% of the mass of the hemp plant biological degumming waste liquid, and the addition amount of the urea is 0.2% of the mass of the hemp plant biological degumming waste liquid;
inoculating alkalophilic bacillus subtilis at the waste liquid temperature of below 48 ℃, inoculating alkalophilic bacillus subtilis liquid according to the proportion of 3%, supplying air between 35 ℃ and 42 ℃ for fermentation for 6 hours, wherein the ventilation volume is not less than 1.2 cubic meters per hour per 100L;
stopping supplying oxygen for 1 hour, inoculating activated EM strain with the inoculation amount of 1%, performing anaerobic fermentation for 7 days at the fermentation temperature of 35 +/-2 ℃ to form the microbial organic fertilizer.
In the second test, the prepared microbial organic fertilizer is detected to contain azotobacter, bacillus, actinomycetes, filamentous fungi, photosynthetic bacteria, lactic acid bacteria and the like, and 250-320 hundred million probiotics are contained in 1g of the microbial organic fertilizer.
Example 3
A preparation method of a microbial organic fertilizer comprises the following steps:
1) culture medium
Glucose culture medium: glucose 2.25g, peptone 1.2g, yeast extract 0.75g, K2HPO40.3g，MgSO4·7H2O0.075 g, water 150ml, pH 10, sterilization and cooling.
Ramie gum culture medium: ramie 12g (NH)4)2SO4 2.25g，K2HPO4 0.3g，MgSO4·7H2O0.3g, water 150ml, Na2CO3Adjusting pH to 11, sterilizing, cooling to below 45 deg.C, and inoculating.
EM strain activation culture medium: 1.5kg of brown sugar, 15g of salt, 15g of urea and 10kg of water are mixed, sterilized and cooled to below 40 ℃.
2) Treatment of bacterial species
2.1 basophilic Bacillus subtilis
2g of bacillus subtilis is suspended in 10ml of sterile water, evenly mixed, 1.2ml of the mixture is inoculated into a glucose culture medium (150ml) and cultured for 24 hours at the temperature of 28 +/-2 ℃ to obtain the activated bacillus subtilis.
Inoculating 1.5ml of activated bacillus subtilis into a ramie gel culture medium (150ml) cooled to below 45 ℃, culturing for 10 hours at 37 +/-3 ℃, and subculturing for 6 hours by using the ramie gel culture medium to obtain the alkalophilic bacillus subtilis.
2.2EM strains
Sterilizing the EM strain activation culture medium, cooling to below 40 ℃, taking 150ml of EM liquid strain, inoculating the EM liquid strain into the EM strain activation culture medium prepared by 10kg of water, carrying out closed fermentation at the fermentation temperature of 35-37 ℃ for 2 days, opening a container, and smelling the acid and sweet taste to successfully activate the EM strain to obtain the activated EM strain.
Adding dipotassium hydrogen phosphate and urea into the hemp plant biological degumming waste liquid, and mixing, wherein the addition amount of the dipotassium hydrogen phosphate is 0.05 percent of the mass of the hemp plant biological degumming waste liquid, and the addition amount of the urea is 0.05 percent of the mass of the hemp plant biological degumming waste liquid;
inoculating alkalophilic bacillus subtilis at the waste liquid temperature of below 48 ℃, inoculating alkalophilic bacillus subtilis liquid according to the proportion of 1%, supplying air between 35 ℃ and 42 ℃ for fermentation for 12 hours, wherein the ventilation volume is not less than 1.2 cubic meters per hour per 100L;
stopping oxygen supply for 2 hours, inoculating activated EM strain, wherein the inoculation amount is 3%, performing anaerobic fermentation for 2 days, and the fermentation temperature is 35 +/-2 ℃ to form the microbial organic fertilizer.
And (3) detecting to obtain the microbial organic fertilizer containing azotobacteria, bacillus, actinomycetes, filamentous fungi, photosynthetic bacteria, lactic acid bacteria and the like, wherein 1g of the microbial organic fertilizer contains 260-320 hundred million probiotics.
Comparative example 1
Different from the embodiment 1, the bacillus subtilis does not undergo strain activation and domestication, and the inoculation amount is the same; EM strain is not activated; the others are the same.
The thallus is not acclimated, after bacillus subtilis is inoculated and fermented for 8 hours, the content of viable bacteria is 1 hundred million/ml which is far lower than that of the inoculated thallus after acclimation; and the pH of the waste liquor is still greater than 9. The EM is acidophilic anaerobic bacteria, the EM is inoculated in the environment with the pH value being more than 9, the fermentation is easy to fail, the mixed bacteria are bred, and the obtained liquid has peculiar smell and is easy to expand and not easy to store.
Comparative example 2
Different from the example 1, the same as the above example, except that dipotassium hydrogen phosphate and urea were not added to the hemp plant degumming waste liquid. The microbial organic fertilizer is prepared.
Detection shows that the prepared microbial organic fertilizer contains azotobacter, bacillus, actinomycetes, filamentous fungi, photosynthetic bacteria, lactic acid bacteria and the like, and 1g of the microbial organic fertilizer contains less than 220 hundred million beneficial biological bacteria. In addition, the microbial organic fertilizer prepared by the embodiment of the invention is rich in N, P, K elements, the application effect is obviously better than that of the microbial organic fertilizer prepared by the comparative example 2, and the difference between the application effects of the two is obvious.
The microbial organic fertilizer provided by the invention contains azotobacter, bacillus, actinomycetes, filamentous fungi, photosynthetic bacteria, lactic acid bacteria and the like, 200-350 billion probiotics are contained in 1g, the quantity of beneficial bacteria in soil can be increased, the balance of microbes in soil is maintained, root diseases are prevented by utilizing the bacteria inhibiting effect, and the microbial organic fertilizer also contains organic acid, small molecular peptide, oligosaccharide, antibiotic substance, amino acid, vitamin, hormone, trace elements and the like, so that the microbial organic fertilizer provides rich nutrition for plants, can be irrigated for use, and can be quickly absorbed by spraying on leaf surfaces.
The fertilizer is quick-acting fertilizer and long-acting fertilizer, and after long-term application, soil naturally forms loose and breathable granular structures, so that plowing is reduced.
Application of the microbial organic fertilizer prepared in the embodiments 1 to 3 of the invention shows that the microbial organic fertilizer can generate antioxidant substances, eliminate putrefaction, eliminate oxide substances, inhibit pathogenic bacteria, control diseases and inhibit insects, relieve fertilizer damage, reduce harmful bacteria by about 90%, form a good environment suitable for growth of animals and plants, and increase the root amount of plants by about 70%. Stimulating and regulating the balanced absorption of nitrogen, phosphorus, potassium and medium and trace elements by crops, obviously improving the quality of the crops, promoting the precocity of the crops, preventing the premature senility of the crops and the like. Has excellent yield increasing effect on solanaceous fruits, melons and fruits, maintains the natural flavor of crops, improves the disease resistance of plants, promotes healthy growth, and forms pollution-free green products.
In addition, the microbial organic fertilizer provided by the invention has the decomposition effect on soil organic matters, and beneficial bacteria can occupy ecological niches and can improve the soil deterioration phenomenon. After being applied to a field, the microbial organic fertilizer can quickly change the soil property, and is represented as follows: firstly, organic acid, micromolecular peptide, oligosaccharide, antibiotic substance and the like secreted by the compound can kill spoilage organisms and occupy ecological niche. And secondly, the microbial compound cenobium can decompose hydrogen in harmful substances such as hydrogen sulfide, methane and the like decomposed by the putrefactive cenobium to change harmful substances into harmless substances, and is fixed and synthesized with acidolysis nitrogen and carbon dioxide to form substances such as saccharides, amino acids, vitamins, hormones and the like, so that the decomposition bacteria are accelerated to propagate, and rich nutrition is provided for plants. And thirdly, the lactic acid bacteria, the actinomycetes, the yeasts, the bacilli and the like in the beneficial bacterium groups can decompose carbohydrates such as cellulose (lignin), starch and the like into various sugars under the action of enzyme, decompose proteins into soluble organic nutrients such as peptone state, peptide state, amino acid and the like, and directly assemble the peptone bodies onto the new plant bodies to form new plant bodies and fruits which do not need photosynthesis. The organic circulation shortcut in the mycelium form does not waste organic matter energy, and carbon, nitrogen, hydrogen, oxygen and the like are assembled on new plants in a team form, so that the crop growth is particularly balanced and rapid. After the beneficial bacteria are applied to the field, the animal and plant residues can be decomposed and assembled into new plants, and the underground darkening growth effect of the beneficial bacteria is several times larger than the overground photosynthesis growth amount.
According to measurement and calculation, the organic matters in the soil belong to an enlarged cycle, and the nutrient utilization rate can reach 150-200%.
The mechanism of the yield increasing effect of the microbial organic fertilizer on vegetables provided by the invention is as follows:
the microbial organic fertilizer provided by the invention is contacted with pests such as root-knot nematodes, Chinese chive maggots, aphids, whiteflies, liriomyza sativae and the like, so that adults cannot lay eggs because of no production of allergenicity (ecdysone), eggs cannot form pupae, and pupae cannot form adults. The lactobacillus and actinomycetes can inhibit putrefying bacteria and virus, and the peptide, antibiotic and polysaccharide formed by decomposing organic matters can prevent and treat diseases such as leaf mold, gray mold, late blight and the like. The probiotics contained in the product can decompose insoluble nutrients such as zinc, boron, calcium, potassium, carbon and the like into soluble state, thereby achieving the effects of disease resistance and yield increase. After the microbial organic fertilizer provided by the invention is applied, the nitrogen and phosphate fertilizer input can be reduced by 60-80%, because of the nitrogen fixation and phosphorus dissolution functions and the direct conversion of organic matters, the pollution of the fertilizer to farmlands and food is reduced, and the effects of low input and high output are achieved.
While particular embodiments of the present invention have been illustrated and described, it would be obvious that various other changes and modifications can be made without departing from the spirit and scope of the invention. It is therefore intended to cover in the appended claims all such changes and modifications that are within the scope of this invention.
Claims (9)
1. The preparation method of the microbial organic fertilizer is characterized by comprising the following steps of:
adding bacillus subtilis into the hemp plant degumming waste liquid for aerobic fermentation;
stopping supplying oxygen for 1-2 hours, inoculating EM strain for anaerobic fermentation to obtain the microbial organic fertilizer;
the degumming waste liquid is biological degumming waste liquid;
adding dipotassium hydrogen phosphate and urea into the hemp plant degumming waste liquid before adding the bacillus subtilis;
the addition amount of the dipotassium phosphate is 0.05-0.2% of the mass of the hemp plant degumming waste liquid, and the addition amount of the urea is 0.05-0.2% of the mass of the hemp plant degumming waste liquid;
the temperature of the aerobic fermentation is 35-45 ℃;
the ventilation of the aerobic fermentation is not less than 1.2m3/100L·h；
The time of aerobic fermentation is 4-12 hours;
the bacillus subtilis is alkalophilic bacillus subtilis obtained after activation and domestication;
the activation step of the bacillus subtilis comprises the following steps: inoculating bacillus subtilis into a glucose culture medium, and culturing to a logarithmic growth phase, wherein the glucose culture medium comprises the following components: calculated by the volume of water, 0.008-0.015g/ml glucose, 0.005-0.008g/ml peptone, 0.002-0.005g/ml yeast extract and K2HPO4 0.001-0.002 g/ml，MgSO4·7H2O0.0002-0.0005 g/ml, and the pH is adjusted to 9-10;
the bacillus subtilis is added into a glucose culture medium in a liquid state, and the volume percentage of the added bacillus subtilis is 0.3-0.8%;
the liquid bacillus subtilis is prepared by adding solid bacteria into sterile water, and the concentration is 0.05-0.2 g/ml;
the culture temperature of the bacillus subtilis inoculated into the glucose culture medium is 28 +/-2 ℃, and the culture time is 24-48 hours;
the domestication step is as follows: inoculating the activated bacillus subtilis into a hemp plant gum culture medium for culture, wherein the hemp plant gum culture medium comprises the following components: calculated by the volume of water, the hemp plant is 0.05-0.08g/ml, (NH)4)2SO4 0.008-0.015g/ml，K2HPO4 0.001-0.002 g/ml，MgSO4·7H2O0.001-0.002 g/ml, Na2CO3Adjusting pH to 10-11, and sterilizing;
the inoculated EM strain is activated EM strain;
the EM strain is inoculated into an activation culture medium for activation, and the activation culture medium contains the following components: 8-15% of brown sugar, 0.08-0.15% of salt and 0.08-0.15% of urea by mass of water;
the inoculation amount of the EM strain inoculated into the activation culture medium is 0.8-1.5%;
inoculating the EM strain into an activation culture medium, and then performing anaerobic fermentation at the temperature of 35-37 ℃ until a container is opened and sour and sweet taste is smelled;
the time for anaerobic fermentation is 2-3 days after the EM strain is inoculated into the activated culture medium;
the inoculation amount of the activated EM strain is 1% -3%, and the time for inoculating the EM strain to carry out anaerobic fermentation after oxygen supply is stopped is 2-7 days.
2. The method for preparing the microbial organic fertilizer as claimed in claim 1, wherein the bacillus subtilis is added at a temperature of below 48 ℃.
3. The method for preparing the microbial organic fertilizer as claimed in claim 1, wherein the volume percentage of the activated bacillus subtilis liquid added into the hemp plant gum culture medium is 1-3%.
4. The preparation method of the microbial organic fertilizer as claimed in claim 1, wherein the temperature for culturing the hemp plant gum inoculated into the culture medium is 37 ℃ ± 3 ℃.
5. The preparation method of the microbial organic fertilizer as claimed in claim 1, wherein the alkaline-philic bacillus subtilis is obtained by inoculating the hemp plant gum culture medium, culturing for 8-10 hours, and subculturing for 6-8 hours with partial bacterial liquid.
6. The method for preparing the microbial organic fertilizer as claimed in claim 1, wherein the time of the aerobic fermentation is 4-6 hours.
7. The method for preparing microbial organic fertilizer according to claim 1, wherein the bast fiber plants comprise ramie, jute, ramie, flax, apocynum venetum and hibiscus hemp.
8. The method for preparing the microbial organic fertilizer as claimed in claim 1, wherein the anaerobic fermentation is followed by a vacuum drying step to form a soluble solid microbial organic fertilizer.
9. The microbial organic fertilizer prepared by the preparation method of the microbial organic fertilizer as claimed in any one of claims 1 to 8.
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