CN111979152A - Compound microbial agent for deodorizing livestock and poultry manure and preparation method thereof - Google Patents
Compound microbial agent for deodorizing livestock and poultry manure and preparation method thereof Download PDFInfo
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- CN111979152A CN111979152A CN202010867227.1A CN202010867227A CN111979152A CN 111979152 A CN111979152 A CN 111979152A CN 202010867227 A CN202010867227 A CN 202010867227A CN 111979152 A CN111979152 A CN 111979152A
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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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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F11/00—Treatment of sludge; Devices therefor
- C02F11/02—Biological treatment
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/20—Nature of the water, waste water, sewage or sludge to be treated from animal husbandry
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2303/00—Specific treatment goals
- C02F2303/02—Odour removal or prevention of malodour
Abstract
The invention discloses a compound microbial agent for deodorizing livestock and poultry manure and a preparation method thereof, wherein the compound microbial agent contains 12-20% of pseudomonas proteus, 15-23% of brevibacterium frigostolerant, 30-35% of bacillus belvesii, 18-22% of bacillus japonicus and 5-10% of lactobacillus rhamnosus.
Description
Technical Field
The invention relates to the field of microbial environment treatment, in particular to a compound microbial agent for deodorizing livestock and poultry manure and a preparation method thereof.
Background
Livestock and poultry breeding is an important component of agriculture and rural economy in China and is one of the main causes of pollution in rural areas in China. A large amount of livestock and poultry manure can be fermented during the stacking process to generate various malodorous substances, including NH3 、H2S, indole, methyl mercaptan and the like can cause harm to the health of people and livestock after long-term contact, and has potential for chronic poisoning. Therefore, the compost decomposition is accelerated, the emission of malodorous gases such as ammonia gas, hydrogen sulfide and the like is reduced, and the method is one of the research hotspots for solving the existing compost deodorization problem.
At present, the livestock and poultry manure is deodorized at home and abroad mainly by adopting physical, chemical and biological deodorization technologies, so that the object image and the structure of the malodorous gas are changed, and the deodorization effect is further achieved. The chemical method mainly utilizes strong oxidant potassium permanganate, hydrogen peroxide, etc. to oxidize partial odor components into less-odor and odorless substances. Physical methods include mechanical ventilation to reduce odor concentration, or absorption of odors by absorbents. However, the above methods all have the problems of low removal efficiency, easy generation of secondary pollution and high removal cost.
Disclosure of Invention
The invention provides a compound microbial agent for deodorizing livestock and poultry manure and a preparation method thereof, aiming at overcoming the defects of the prior art.
The invention aims to provide a preparation method of a compound microbial agent for deodorizing livestock and poultry manure, which specifically comprises the following steps:
(1) and (4) preparing a seed solution.
Pseudomonas proteus, brevibacterium frigidum, Bacillus belgii, Bacillus japonicus and Lactobacillus rhamnosus are inoculated in a 500ml triangular flask of liquid under the aseptic condition by a test tube mother strain, the liquid loading amount is 200ml, the culture is carried out in a corresponding culture medium, the temperature is set to be 30-32 ℃, and the culture is carried out in a 200r/min shaking table for 2-3 days to obtain a primary seed liquid.
The beef extract peptone broth culture medium for the pseudomonas proteus, the brevibacterium frigosum, the bacillus belgii and the bamboo joint bacillus comprises the following components in percentage by weight: 1-1.2% of beef extract, 0.5-0.8% of peptone, 0.5-0.8% of yeast extract powder, 0.5-0.8% of glucose, 0.05-0.1% of sodium chloride, 0.005-0.008% of sodium hydroxide and the balance of water.
The MRS culture medium special for the lactobacillus rhamnosus comprises the following components in percentage by weight: 1-1.2% of beef extract, 0.5-0.8% of yeast extract, 1-1.2% of peptone, 1-1.2% of glucose, 0.1-0.2% of dipotassium phosphate, 0.3-0.5% of sodium acetate, 0.01-0.02% of magnesium sulfate heptahydrate, 0.003-0.005% of manganese sulfate tetrahydrate, 800.1% of tween, 0.1-0.2% of triammonium citrate, and the balance of water, wherein the pH value is adjusted to 6.3.
(2) Seed liquid expanded culture
Respectively inoculating the primary seed liquid of the cultured pseudomonas proteus, brevibacterium frigostolerant, bacillus belgii, bacillus japonicus and lactobacillus rhamnosus into 500ml triangular flasks which are pre-filled with 200ml of corresponding culture medium under the aseptic condition for culturing at the temperature of 30-34 ℃ in a shaking table at 180r/min for 2-3 days to obtain secondary seed liquid.
(3) And (4) compounding and co-fermenting.
Mixing the secondary seed liquid of each strain according to the volume percentage: 15-20% of pseudomonas proteus, 15-30% of brevibacterium frigostolerant, 15-20% of bacillus belgii, 15-20% of bamboo joint bacillus and 5-10% of lactobacillus rhamnosus, and then the bacillus belgii and the lactobacillus rhamnosus are aseptically inoculated into a fermentation tank containing a composite culture medium, wherein the inoculation amount is 0.1-1%, the culture temperature is 28-30 ℃, 0.02% of antifoaming agent is added, the rotation speed is 160-200r/min, and the air flow is 1: 0.8; culturing for 36-48 hours to obtain the liquid composite deodorant bacterial agent.
The weight ratio of the composite culture medium is as follows: 1.0 percent of glucose, 0.5 percent of yeast powder, 0.5 percent of peptone, 0.35 percent of sodium citrate, 0.05 percent of sodium chloride, 0.04 percent of ammonium chloride, 0.1 percent of monopotassium phosphate, 0.02 percent of magnesium chloride, 0.02 percent of calcium chloride, 0.001 percent of ferrous chloride and the balance of water, and the pH value is adjusted to 7-7.2.
The invention also aims to provide the compound microbial agent for deodorizing the livestock and poultry manure, which is prepared by the method. The compound microbial agent is characterized in that: the total colony number in the microbial inoculum is 5-25 multiplied by 109CFU/mL, wherein 12-20% of pseudomonas proteus, 15-23% of brevibacterium frigosum, 30-35% of bacillus belgii, 18-22% of bacillus japonicus and 5-10% of lactobacillus rhamnosus.
The biological deodorization method utilizes the physiological metabolic activity of microorganisms to degrade malodorous substances and convert the malodorous substances into odorless substances, has the characteristics of high treatment efficiency, no secondary pollution, convenient operation, low cost and the like, and becomes a main method for the prevention and treatment research and application of the malodorous at home and abroad. The odor can be metabolized by microorganisms to generate water, carbon dioxide, sulfuric acid, nitric acid and nitrous acid, so that the odor is converted into pollution-free substances, harmlessness is realized, and the environment-friendly concept of modern breeding is met.
The invention provides a compound microbial agent for deodorizing livestock and poultry manure and a preparation method thereof, the compound microbial agent is prepared by compounding multiple strains, wherein pseudomonas proteus and brevibacterium frigostolerant are nitrifying bacteria, bacillus belgii and bamboo joint bacillus are sulfur removing bacteria, lactobacillus rhamnosus is lactic acid bacteria, the multiple strains are fermented together, and the multiple microorganisms are symbiotically combined to realize function complementation, so that odor in the livestock and poultry manure is effectively removed. The composite microbial agent strain for deodorizing the livestock and poultry manure provided by the invention is reasonable in compounding, has high odor removal rate in the livestock and poultry manure, can remove more than 70% of harmful ammonia gas and hydrogen sulfide gas within 36h, and effectively reduces odor substances including NH in the manure3And H2The content of S and the lactic acid bacteria contained in the compound microbial inoculum can effectively improve the immunity of livestock and poultry. The microbial inoculum is subjected to compound fermentation, each strain grows harmoniously, the function complementation is realized, the practicability of the microbial inoculum is high, and the microbial inoculum is suitable for large-scale popularization.
Drawings
FIG. 1 shows the change of the concentration of ammonia in the inlet and outlet gases of the deodorizing tower in example 3, and FIG. 2 shows the change of the concentration of hydrogen sulfide in the inlet and outlet gases of the deodorizing tower in example 3.
Detailed Description
The present invention will be described in detail with reference to specific embodiments. The following examples are presented to assist those skilled in the art in further understanding the invention, but are not intended to limit the invention in any manner.
The strain Pseudomonas proteus used in the examples below (Pseudomonas plecoglossicida) Brevibacterium frigidum (B) ((B))Brevibacterium frigoritolerans) Bacillus belgii (B.Bacillus velezensis strain) Bamboo joint bacillus (B) ((B))Arthrobacter bambusae) Lactobacillus rhamnosus (A), (B), (C)Lactobacillus rhamnosus) Is obtained by separating and storing in China Gansu Ministry of Industrial microbial cultures preservation and management center, Gansu province, Central laboratory, of biological research institute of science and institute of Gansu province, and Ministry of science and technology.
Other reagents used in the following examples, unless otherwise specified, were commercially available.
The media formulations used in the following examples were as follows:
beef extract peptone broth culture medium: 10g of beef extract, 5g of peptone, 5g of yeast extract powder, 5g of glucose, 5g of sodium chloride, 1000mL of water and 7.0-7.2 of pH value.
1g, dipotassium phosphate 0.4g, magnesium sulfate 0.58g, manganese sulfate 0.29g, water 1000mL, pH 6.2-6.6. MRS culture medium: 10g of peptone, 10g of beef extract, 5g of yeast extract powder, 20g of glucose, 5g of sodium acetate, 2g of citric acid diamine and 80 g of tween
Compound culture medium: 10g of glucose, 5g of yeast powder, 5g of peptone, 3.5g of sodium citrate, 0.5g of sodium chloride, 0.4g of ammonium chloride, 1g of monopotassium phosphate, 0.2g of magnesium chloride, 0.2g of calcium chloride, 0.01 g of ferrous chloride, 1000ml of water and 7-7.2 of pH value.
The solid formula of the culture medium is to add 1.6 percent of agar into the culture medium.
Example 1
The preparation method of the compound microbial agent for deodorizing the livestock and poultry manure comprises the following specific steps:
1. preparation of seed liquid
Taking pseudomonas proteus, brevibacterium frigostolerant, bacillus belgii and bamboo joint bacillus strain inclined planes, and selecting partial bacterial moss to be respectively inoculated in a beef extract peptone broth culture liquid culture medium in an aseptic environment. Shaking and culturing for 24h at 30 deg.C and 180 r/min. Taking a slant of the strain of the lactobacillus rhamnosus, selecting a part of lawn in an aseptic environment, inoculating the lawn into an MRS culture medium, and performing static culture in an anaerobic incubator at 37 ℃ for 24 hours. Obtaining a first-order seed liquid
2. Seed liquid expanded culture
Respectively inoculating the cultured primary seed liquid of the pseudomonas proteus, brevibacterium frigostolerant, bacillus belgii, bacillus japonicus and lactobacillus rhamnosus test tubes into 500ml triangular flasks containing corresponding culture media under the aseptic condition for culturing (the liquid loading is 200ml, the culture media are sterilized at 121 ℃ for 25 minutes), setting the temperature at 30-34 ℃, and culturing in a shaking table at 180r/min for 2-3 days to obtain secondary seed liquid.
3. Compounding and co-fermentation
And (3) mixing the secondary seed liquid of each strain according to the volume ratio: inoculating 20% of pseudomonas proteus, 30% of brevibacterium frigostolerant, 20% of bacillus belgii, 20% of bamboo joint bacillus and 10% of lactobacillus rhamnosus into a fermentation tank containing a composite culture medium, wherein the inoculation amount is 0.1-1%, 0.02% of defoaming agent is added, the culture temperature is 28-30 ℃, the rotation speed is 160-200r/min, and the ventilation volume is 1:0.8, culturing for 36-48 hours, and obtaining the co-fermentation bacteria solution which is the composite deodorant bacteria agent.
Example 2
The composite deodorant bacterial agent obtained in example 1 was subjected to viable bacteria number detection. The composite deodorizing bacterial agent is used for viable bacteria quantity detection by adopting a gradient dilution coating selective flat plate method. The total colony count uses the nutrient broth culture medium, the pseudomonas proteus and the brevibacterium frigostabilis belong to the heterotrophic nitrification culture medium of the digestive bacteria using ammonium ions as the only nitrogen source, the Bacillus belgii and the bamboo joint bacillus use the sulfur-containing inorganic culture medium of the sulfur-removing bacteria, and the lactobacillus uses the MRS culture medium. The results are shown in the following table.
| Total number of colonies | Pseudomonas proteus | Cold-resistant bacillus brevis | Bacillus belgii | Bamboo joint bacillus | Lactobacillus rhamnosus | |
| Number of viable bacteria (CFU/mL) | 8.9×109 | 2.04×109 | 1.96×109 | 2.65×109 | 1.5×109 | 0.75×109 |
The results show that the effective viable count of various bacteria basically meets the proportional requirement of the deodorizing microbial inoculum, and the complex fermentation is feasible. The compound co-fermentation concentrates the fermentation of single bacteria into one-time fermentation, thereby not only saving time but also reducing cost.
Example 3
The deodorizing effect of the composite deodorizing bacterial agent obtained in example 1 was examined by subjecting NH to a test with odor generated during composting of cow dung3And H2Reduction of concentration of S harmful gasThe degree of the microorganism is actually measured to evaluate the deodorization effect of the composite microorganism deodorization microbial inoculum.
The test site is arranged in a certain cow farm in Zhangye city in Gansu province, and more than 5000 cows exist in the current hurdle. The deodorization equipment comprises a deodorization pool, an air pump, an automatic control system, an ammonia gas and hydrogen sulfide detector and the like.
Diluting the stock solution of the liquid composite deodorant microbial inoculum by 100 times to be used as a microbial inoculum for experiments, and directly adding the microbial inoculum into a deodorant pool. The deodorant microbial inoculum accounts for 1/3 of the volume of the deodorant pool, and the odor is introduced from the lower part of the deodorant pool, and the flow rate is 10 cubic meters per hour. The odor firstly passes through the deodorizing liquid, then overflows from the liquid level and is discharged through the air outlet.
NH in gas is respectively detected at the gas inlet and the gas outlet of the deodorization tower3And H2The concentration of S varies. NH in gas3With ES10B10-NH3Stationary ammonia gas detector, H2ES10B10-H for S2And S, detecting by using a fixed hydrogen sulfide gas detector.
The test results are shown in figure 1 and figure 2, and the test shows that the composite microbial agent for deodorizing livestock and poultry manure provided by the invention can effectively clear the odor in the livestock and poultry manure, and NH is generated after the composite microbial agent is treated for 36 hours3And H2The concentration of S decreased by 92.54% and 87.42%, respectively.
Example 4
Application of the composite deodorant bacterial agent obtained in example 1. The test site is arranged in Jinma fertilizer industry, Limited liability company, Zhangye, Ganyang, province, and the operation range of the company comprises the manufacture of compound fertilizer and organic fertilizer. Diluting the stock solution of the liquid composite deodorizing microbial inoculum by 100 times to be used as a microbial inoculum for experiments, and directly adding the microbial inoculum into a composting pool. The width of the composting pool is 5 meters, the depth is 4 meters, the length is 10 meters, and 2000L of deodorizing microbial inoculum for experiments is needed. After the deodorizing bactericide is applied for 4 days, the odor of the composting site is obviously improved.
Claims (2)
1. A composite microbial inoculum for deodorizing livestock and poultry feces is characterized in that the total colony number in the microbial inoculum is 5-25 multiplied by 109 CFU/mL, wherein pseudomonad 12-20%, Brevibacterium frigosum 15-23%, Bacillus belgii 30-35%, bamboo joint18-22% of bacillus and 5-10% of lactobacillus rhamnosus.
2. A method for preparing the complex microbial inoculant for deodorizing livestock and poultry manure as claimed in claim 1, comprising the following steps:
(1) preparing a seed solution: inoculating pseudomonas proteus, brevibacterium frigosum, bacillus belgii, bacillus japonicus and lactobacillus rhamnosus into a liquid culture medium, and culturing at 30-32 ℃ and 200r/min for 2-3d to obtain a first-stage seed solution;
(2) seed liquid amplification culture: inoculating the first-stage seed liquid into a liquid culture medium for culturing at 30-34 ℃ at 180r/min for 2-3d to obtain a second-stage seed liquid;
(3) compounding and co-fermenting: inoculating the secondary seed solution into a composite culture medium, wherein the inoculation amount is 0.1-1%, the pseudomonads are 15-20%, the brevibacterium frigosum is 15-30%, the bacillus belgii is 15-20%, the bacillus japonicus is 15-20%, the lactobacillus rhamnosus is 5-10%, 0.02% of defoaming agent is added, the mixture is cultured for 36-48h at the temperature of 28-30 ℃, the speed of 160-200r/min and the ventilation volume of 1:0.8, and the liquid composite deodorant microbial inoculum is obtained;
the weight ratio of the composite culture medium is as follows: 1.0 percent of glucose, 0.5 percent of yeast powder, 0.5 percent of peptone, 0.35 percent of sodium citrate, 0.05 percent of sodium chloride, 0.04 percent of ammonium chloride, 0.1 percent of monopotassium phosphate, 0.02 percent of magnesium chloride, 0.02 percent of calcium chloride, 0.001 percent of ferrous chloride, the balance of water and the pH value of 7-7.2.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110106102A (en) * | 2019-03-08 | 2019-08-09 | 大连理工大学 | The preparation method and application of brevibacterium frigoritolerans ZL-2 and its compost cold-starting microbial inoculum |
| CN114292791A (en) * | 2022-01-06 | 2022-04-08 | 领先生物农业股份有限公司 | Composite microbial deodorant and application thereof |
| CN114410512A (en) * | 2021-12-31 | 2022-04-29 | 农业部沼气科学研究所 | Livestock and poultry manure deodorization microbial agent and preparation method and application thereof |
| CN115960970A (en) * | 2022-11-22 | 2023-04-14 | 哈尔滨工业大学 | Method for producing hydrogen by fermenting human excrement |
| CN116606783A (en) * | 2023-07-18 | 2023-08-18 | 山东健源生物科技有限公司 | Composite microbial agent and application thereof in livestock and poultry cultivation deodorization field |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110106102A (en) * | 2019-03-08 | 2019-08-09 | 大连理工大学 | The preparation method and application of brevibacterium frigoritolerans ZL-2 and its compost cold-starting microbial inoculum |
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| CN114410512A (en) * | 2021-12-31 | 2022-04-29 | 农业部沼气科学研究所 | Livestock and poultry manure deodorization microbial agent and preparation method and application thereof |
| CN114410512B (en) * | 2021-12-31 | 2023-07-04 | 农业部沼气科学研究所 | Livestock and poultry manure deodorizing microbial agent and preparation method and application thereof |
| CN114292791A (en) * | 2022-01-06 | 2022-04-08 | 领先生物农业股份有限公司 | Composite microbial deodorant and application thereof |
| CN115960970A (en) * | 2022-11-22 | 2023-04-14 | 哈尔滨工业大学 | Method for producing hydrogen by fermenting human excrement |
| CN116606783A (en) * | 2023-07-18 | 2023-08-18 | 山东健源生物科技有限公司 | Composite microbial agent and application thereof in livestock and poultry cultivation deodorization field |
| CN116606783B (en) * | 2023-07-18 | 2023-10-03 | 山东健源生物科技有限公司 | Composite microbial agent and application thereof in livestock and poultry cultivation deodorization field |
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Application publication date: 20201124 |