Bacillus combined fungicide and application thereof in treatment of cow dung litter

Abstract

The invention discloses a bacillus combined fungicide and application thereof in cow dung padding treatment, and belongs to the technical field of microorganisms. The bacillus combined microbial agent is prepared from bacillus pumilus (Bacillus pumilus), bacillus altitudinis (Bacillus altitudinis), geobacillus stearothermophilus (Geobacillus stearothermophilus) and bacillus coagulans (Bacillus coagulans), and the bacillus altitudinis (Geobacillus stearothermophilus), the bacillus altitudinis (Geobacillus stearothermophilus) and the bacillus coagulans (Geobacillus coagulans) are used for preparing the bacillus altitudinis, the bacillus altitudinis, the Geobacillus stearothermophilus and the Geobacillus coagulans. The bacillus combined microbial agent disclosed by the invention can be used for rapidly decomposing organic matters in cow dung, reducing emission of harmful gases such as ammonia gas and hydrogen sulfide, reducing peculiar smell and inhibiting growth of pathogenic microorganisms. When the bacillus combined microbial agent is used for treating the cow dung waste padding, the health of cows and the improvement of the production performance are facilitated.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, in particular to a bacillus combination bacterial agent and application thereof in the treatment of dairy cow dung bedding. Background Art

[0002] With the large-scale development of dairy farming, the treatment of dairy manure has become an important environmental and resource utilization issue. Traditional treatment methods often have many disadvantages, such as high treatment costs, waste of resources, and possible environmental pollution. In recent years, the use of manure litter fermentation technology to treat livestock and poultry manure has been widely studied and applied. This technology uses agricultural waste such as husks, sawdust, straw, etc. as bedding, adds microbial agents, ferments and degrades livestock and poultry manure and urine, and finally forms organic fertilizer. It uses microbial treatment technology to achieve zero emissions, no odor, and low cost. This technology provides a new method for solving the environmental pollution problem of farms. However, the existing manure litter fermentation technology still has many disadvantages, such as low efficiency in decomposing manure, unobvious deodorization effect, and low survival ability of fermentation agents. Manure litter that has not been effectively treated will not only breed a large number of pathogens and harmful microorganisms, but also emit unpleasant odors, which will have adverse effects on the health of dairy cows and the breeding environment.

[0003] As a beneficial microorganism, Bacillus has unique physiological characteristics and biological functions. Therefore, developing an efficient, environmentally friendly and fully functional Bacillus combination agent to effectively treat dairy cow manure is of great significance to improve the treatment efficiency and quality of manure litter in the current dairy farming industry, while reducing treatment costs and environmental risks. Summary of the invention

[0004] The purpose of the present invention is to provide a bacillus combination inoculant and its application in the treatment of dairy cow manure bedding to solve the problems existing in the above-mentioned prior art. The bacillus combination inoculant of the present invention can quickly decompose organic matter in dairy cow manure, and the dairy cow manure bedding treated with the bacillus combination inoculant of the present invention is more conducive to the health of dairy cows and the improvement of production performance.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] The invention provides a bacillus combination bacterial agent, which comprises Bacillus pumilus, Bacillus altitudinis, Geobacillus stearothermophilus and Bacillus subtilis.

[0007] Furthermore, the total number of viable bacteria in the Bacillus combination agent is 1×10 10 ~2×10 10 cfu / g.

[0008] Furthermore, the preparation method comprises the following steps:

[0009] S1. Bacillus pumilus, Bacillus brevis, Bacillus stearothermophilus and Bacillus coagulans were cultured to obtain their respective seed solutions;

[0010] S2. The obtained seed solution is mixed and fermented to obtain a fermentation liquid;

[0011] S3. After mixing the fermentation liquid and the wheat bran, a protective agent is added, and the mixture is dried and crushed to obtain the Bacillus sp. combination bacterial agent.

[0012] Furthermore, in step S2, when the obtained seed solutions are mixed, they are mixed in a volume ratio of Bacillus pumilus seed solution: Bacillus trehalose seed solution: Bacillus stearothermophilus seed solution: Bacillus coagulans seed solution = (1-2): (1-3): 2: (1-3).

[0013] Furthermore, in step S3, the mass ratio of the fermentation liquid to the wheat bran is (1-2):1.

[0014] The present invention also provides application of the bacillus combination bacterial agent in treating dairy cow dung bedding.

[0015] The invention also provides a method for treating cow dung bedding, comprising the steps of mixing cow dung with sawdust, adding the bacillus combination bacterial agent, and performing stacking fermentation.

[0016] Furthermore, the addition amount of the Bacillus combination bacterial agent is 0.5% to 1.5% of the total mass of the mixture of cow dung and sawdust.

[0017] Furthermore, the mixing mass ratio of the cow dung to the sawdust is (1-2):1.

[0018] Furthermore, the temperature of the stacking fermentation is 55°C to 60°C, and the time is 7 to 10 days.

[0019] The present invention discloses the following technical effects:

[0020] The bacillus combination bacterial agent of the present invention can quickly decompose organic matter in dairy cow manure, reduce the emission of harmful gases such as ammonia and hydrogen sulfide, and reduce odor; at the same time, using the bacillus combination bacterial agent of the present invention to treat dairy cow manure bedding can effectively inhibit the growth of pathogenic microorganisms, improve the safety of dairy cow manure bedding, and at the same time be more conducive to the health of dairy cows and the improvement of production performance. DETAILED DESCRIPTION

[0021] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0022] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0023] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0024] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0025] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0026] The microbial strains used in the embodiments of the present invention are as follows:

[0027] Bacillus pumilus, strain number CICC 22096; Bacillus altitudinis, strain number CICC 20511; Geobacillus stearothermophilus, strain number CICC 22988, all from China Industrial Microbiological Culture Collection Center; Bacillus subtilis, strain number CGMCC 1.10823, from China General Microbiological Culture Collection Center.

[0028] The above strains can be purchased from the above depository institutions.

[0029] Example 1

[0030] 1. Preparation of Bacillus combination agent:

[0031] 1. Inoculate Bacillus pumilus, Bacillus trehalose, Bacillus stearothermophilus and Bacillus coagulans into LB liquid culture medium (mixed with 10 g / L tryptone, 5 g / L yeast extract and 10 g / L NaCl) respectively, and culture on a shaking table at 35°C and 180 rpm for 24 h to obtain their respective seed solutions.

[0032] 2. The above seed liquid was mixed in a volume ratio of Bacillus pumilus: Bacillus maximus: Bacillus stearothermophilus: Bacillus coagulans = 2:1:2:1, and inoculated with 2% of the inoculum into a fermentation medium (20 g / L glucose, 10 g / L peptone, 5 g / L yeast extract, 5 g / L sodium chloride, 0.5 g / L magnesium sulfate, 1 g / L potassium dihydrogen phosphate, 0.05 g / L manganese sulfate, 0.1 g / L calcium chloride), and cultured at 35°C and 180 rpm for 48 h to obtain a fermentation liquid.

[0033] 3. The fermentation liquid and wheat bran were mixed in a mass ratio of 1:1, and a protective agent (5wt% sucrose and 5wt% skim milk) was added. After low-temperature drying and crushing, the viable bacterial count was 1×10 10 ~2×10 10 Bacillus combination inoculants between cfu / g.

[0034] 2. Treatment of dairy cow dung litter:

[0035] The cow dung and sawdust were mixed in a mass ratio of 2:1, the water content was adjusted to 55%, the prepared Bacillus combination bacterial agent was added (the addition amount was 0.5% of the total mass of the mixture of cow dung and sawdust), stirred evenly, piled and fermented, the temperature was controlled at 55°C during the fermentation process, and fermented for 7 days to obtain bedding that can be used for cow beds.

[0036] Example 2

[0037] 1. Preparation of Bacillus combination agent:

[0038] 1. Inoculate Bacillus pumilus, Bacillus trehalose, Bacillus stearothermophilus and Bacillus coagulans into LB liquid culture medium (mixed with 10 g / L tryptone, 5 g / L yeast extract and 10 g / L NaCl) respectively, and culture on a shaking table at 35°C and 180 rpm for 36 h to obtain their respective seed solutions.

[0039] 2. The above seed liquid was mixed in a volume ratio of Bacillus pumilus: Bacillus maximus: Bacillus stearothermophilus: Bacillus coagulans = 2:3:2:3, and inoculated into a fermentation medium (20 g / L glucose, 10 g / L peptone, 5 g / L yeast extract, 5 g / L sodium chloride, 0.5 g / L magnesium sulfate, 1 g / L potassium dihydrogen phosphate, 0.05 g / L manganese sulfate, 0.1 g / L calcium chloride) at an inoculum amount of 1%, and cultured at 35°C and 180 rpm for 72 h to obtain a fermentation liquid.

[0040] 3. The fermentation liquid and wheat bran were mixed in a mass ratio of 2:1, and a protective agent (5wt% sucrose and 10wt% skim milk) was added. After low-temperature drying and crushing, the viable bacterial count was 1×10 10 ~2×10 10 Bacillus combination inoculants between cfu / g.

[0041] 2. Treatment of dairy cow dung litter:

[0042] The cow dung and sawdust were mixed in a mass ratio of 1:1, the water content was adjusted to 60%, the prepared Bacillus combination bacterial agent was added (the addition amount was 1% of the total mass of the mixture of cow dung and sawdust), stirred evenly, piled and fermented, the temperature was controlled at 60°C during the fermentation process, and fermented for 7 days to obtain bedding that can be used for cow beds.

[0043] Example 3

[0044] 1. Preparation of Bacillus combination agent:

[0045] 1. Inoculate Bacillus pumilus, Bacillus trehalose, Bacillus stearothermophilus and Bacillus coagulans into LB liquid medium (mixed with 10 g / L tryptone, 5 g / L yeast extract and 10 g / L NaCl) respectively, and culture on a shaking table at 35°C and 180 rpm for 48 h to obtain their respective seed solutions.

[0046] 2. The above seed liquid was mixed in a volume ratio of Bacillus pumilus: Bacillus maximus: Bacillus stearothermophilus: Bacillus coagulans = 1:2:2:1, and inoculated into a fermentation medium (20 g / L glucose, 10 g / L peptone, 5 g / L yeast extract, 5 g / L sodium chloride, 0.5 g / L magnesium sulfate, 1 g / L potassium dihydrogen phosphate, 0.05 g / L manganese sulfate, 0.1 g / L calcium chloride) at an inoculum amount of 1.5%, and cultured at 35°C and 180 rpm for 48 h to obtain a fermentation liquid.

[0047] 3. The fermentation liquid and wheat bran were mixed in a mass ratio of 1.5:1, and a protective agent (10wt% sucrose and 5wt% skim milk) was added. After low-temperature drying and crushing, the viable bacterial count was 1×10 10 ~2×10 10 Bacillus combination inoculants between cfu / g.

[0048] 2. Treatment of dairy cow dung litter:

[0049] The cow dung and sawdust were mixed in a mass ratio of 2:1.5, the water content was adjusted to 65%, the prepared Bacillus combination bacterial agent was added (the addition amount was 1.5% of the total mass of the mixture of cow dung and sawdust), stirred evenly, piled and fermented, the temperature was controlled at 55°C during the fermentation process, and fermented for 10 days to obtain bedding that can be used for cow beds.

[0050] Comparative Example 1

[0051] Compared with Example 1, the difference lies in that Bacillus licheniformis (strain number: CICC 10092) is replaced by Bacillus licheniformis.

[0052] Comparative Example 2

[0053] Compared with Example 1, the difference lies in that Bacillus coagulans is replaced by Bacillus megaterium (strain number: CICC 23033).

[0054] Comparative Example 3

[0055] Compared with Example 1, the difference is that in the treatment of cow dung bedding, the addition amount of the Bacillus combination bacterial agent is 2% of the total mass of the mixture of cow dung and sawdust.

[0056] Test example

[0057] Performance Testing:

[0058] The performance of the cow manure litter treated in Examples 1 to 3 and Comparative Examples 1 to 3 was tested, and the test indicators included organic matter decomposition rate, ammonia and hydrogen sulfide emission concentrations, and the number of pathogenic microorganisms. The test results are shown in Table 1.

[0059] Table 1 Test results of the performance of treated dairy cow dung litter

[0060]

[0061] The data in Table 1 show that the treatment methods of Examples 1 to 3 are superior to Comparative Examples 1 to 3 in terms of organic matter decomposition, ammonia and hydrogen sulfide emission control, and pathogenic microorganism inhibition. This indicates that the Bacillus combination bacterial agent provided by Examples 1 to 3 of the present invention is more conducive to improving the performance of dairy cow manure bedding, and has obvious advantages in the treatment of dairy cow manure bedding.

[0062] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A Bacillus combination agent, characterized in that: These include Bacillus pumilus, Bacillus altitudinis, Geobacillus stearothermophilus, and Bacillus subtilis.

2. The Bacillus combination bacterial agent according to claim 1, characterized in that The total number of viable bacteria in the Bacillus combination agent is 1×10 10 ~2×10 10 cfu / g.

3. The Bacillus combination bacterial agent according to claim 1, characterized in that: The preparation method comprises the following steps: S1. Bacillus pumilus, Bacillus brevis, Bacillus stearothermophilus and Bacillus coagulans were cultured to obtain their respective seed solutions; S2. The obtained seed solution is mixed and fermented to obtain a fermentation liquid; S3. After mixing the fermentation liquid and the wheat bran, a protective agent is added, and the mixture is dried and crushed to obtain the Bacillus sp. combination bacterial agent.

4. The Bacillus combination bacterial agent according to claim 3, characterized in that: In step S2, when the obtained seed solutions are mixed, they are mixed in a volume ratio of Bacillus pumilus seed solution: Bacillus trehalose seed solution: Bacillus stearothermophilus seed solution: Bacillus coagulans seed solution = (1-2): (1-3): 2: (1-3).

5. The Bacillus combination bacterial agent according to claim 3, characterized in that: In step S3, the mass ratio of the fermentation liquid to the wheat bran is (1-2):

1.

6. Use of the Bacillus combination agent according to any one of claims 1 to 5 in the treatment of dairy cow manure and bedding.

7. A method for treating cow dung bedding, characterized in that: The method comprises the steps of mixing cow dung and sawdust, adding the bacillus combination bacterial agent according to any one of claims 1 to 5, and performing stacking fermentation.

8. The method according to claim 7, characterized in that The added amount of the bacillus combination bacterial agent is 0.5% to 1.5% of the total mass of the mixture of dairy cow dung and sawdust.

9. The method according to claim 7, characterized in that: The mixing mass ratio of the cow dung to the sawdust is (1-2):

1.

10. The method according to claim 7, characterized in that The temperature of the stacking fermentation is 55° C. to 60° C., and the time is 7 to 10 days.

Images