Application of Bacillus subtilis BR in organic fertilizer production

Abstract

The present invention provides the application of Bacillus subtilis BR in the production of organic fertilizer. The present invention first finds that Bacillus subtilis BR is mixed with other probiotics (including Candida utilis, Lactobacillus plantarum, and Trichoderma viride), and can ferment pig manure and rice husk powder with high water content into high-quality organic fertilizer that meets national standards, and the organic matter content in the organic fertilizer can reach 55%, the total nutrient (N+P2O5+K2O) content is 6.6%, and the seed germination rate is 89.5%, which is significantly higher than the control sample (organic matter content can reach 45%, total nutrient content is 3.4%, and seed germination rate is 77%) obtained from commercial Bacillus subtilis instead of Bacillus subtilis BR. The use of Bacillus subtilis BR in the production of organic fertilizer has broad application prospects.

Description

Technical Field

[0001] The invention relates to the field of organic fertilizer production, in particular to the application of Bacillus subtilis BR in organic fertilizer production. Background Art

[0002] China is a large livestock breeding country, producing a huge amount of animal manure every year. Animal manure has become one of the largest sources of agricultural pollution, posing a threat to the environment and people's health. Therefore, it is imperative to establish cheap harmless treatment and resource utilization of animal manure. In recent years, a variety of methods for resource treatment of animal manure have been established, but the overall cost is high, and the quality of the farmyard manure or organic fertilizer obtained is poor or unstable. Among them, excessive loss of organic matter and low total nutrient content are the main problems.

[0003] In the early stage, we registered a new high-efficiency Bacillus subtilis strain BR (General Microbiological Center of China Microbiological Culture Collection Administration, strain registration number CGMCC No.23476). The present invention provides a method for quickly obtaining high-quality organic fertilizer by using Bacillus subtilis BR in combination with other probiotics. Summary of the invention

[0004] The invention aims to provide a new use of Bacillus subtilis BR, in particular to use of Bacillus subtilis BR in organic fertilizer production.

[0005] In order to achieve the purpose of the present invention, in a first aspect, the present invention provides an application of Bacillus subtilis BR in the production of organic fertilizer, wherein a mixed bacteria obtained by compounding Bacillus subtilis BR with Candida utilis, Lactobacillus plantarum and Trichoderma viride is inoculated into a mixture containing livestock and poultry manure and agricultural and forestry wastes to ferment and produce organic fertilizer.

[0006] In the present invention, Bacillus subtilis BR can be found in CN114540215A, and its deposit number is CGMCC No.23476.

[0007] The livestock and poultry excrement may be pig excrement, and the agricultural and forestry wastes include but are not limited to rice husk powder and wood chips.

[0008] In a second aspect, the present invention provides a method for producing an organic fertilizer, comprising the following steps:

[0009] (1) mixing 200-300 g (preferably 250 g) of Bacillus subtilis BR, 45-60 g (preferably 50 g) of Candida utilis, 45-60 g (preferably 50 g) of Lactobacillus plantarum and 45-60 g (preferably 50 g) of Trichoderma viride, and the mixture is recorded as A;

[0010] (2) mixing 1300-1800 kg (preferably 1500 kg) of pig manure, 1000-1400 kg (preferably 31200 kg) of rice husk powder and 300-480 kg (preferably 400 kg) of sawdust, and the mixture is recorded as B;

[0011] (3) Mix A and B and let them ferment.

[0012] Furthermore, the bacterial content of the Bacillus subtilis BR agent is 25-35 billion / gram (preferably 30 billion / gram), the bacterial content of the Candida utilis agent is 15-25 billion / gram (preferably 20 billion / gram), the bacterial content of the Lactobacillus plantarum agent is 5-15 billion / gram (preferably 10 billion / gram), and the bacterial content of the Trichoderma viride agent is 15-25 billion / gram (preferably 20 billion / gram).

[0013] Furthermore, the water content of the pig manure is 80%, the water content of the rice husk powder is 10%, and the water content of the sawdust is 11%.

[0014] Further, the mixture is left to ferment for about 90 days.

[0015] In a third aspect, the present invention provides an organic fertilizer prepared according to the above method.

[0016] In a fourth aspect, the present invention provides application of the organic fertilizer in plant cultivation.

[0017] In a fifth aspect, the present invention provides a composite bacterial agent, which is composed of Bacillus subtilis BR, Candida utilis, Lactobacillus plantarum and Trichoderma viride, wherein the viable counts of Bacillus subtilis BR, Candida utilis, Lactobacillus plantarum and Trichoderma viride are 25-35 billion / g (preferably 30 billion / g), 15-25 billion / g (preferably 20 billion / g), 5-15 billion / g (preferably 10 billion / g) and 15-25 billion / g (preferably 20 billion / g), respectively.

[0018] In a sixth aspect, the present invention provides application of the composite bacterial agent in organic fertilizer production.

[0019] By means of the above technical solution, the present invention has at least the following advantages and beneficial effects:

[0020] The present invention first discovered that Bacillus subtilis BR is mixed with other probiotics (including Candida utilis, Lactobacillus plantarum, and Trichoderma viride), and can ferment pig manure and rice husk powder with high water content into high-quality organic fertilizer that meets my country's agricultural industry standards, and the organic matter content in the organic fertilizer can reach 55%, the total nutrient (N+P2O5+K2O) content is 6.6%, and the seed germination rate is 89.5% (National Industry Standard for Completely Compound Organic Fertilizer-NY / T,525-2021), which is significantly higher than the control sample (organic matter content can reach 45%, total nutrient (N+P2O5+K2O) content is 3.4%, and seed germination rate is 77%) obtained from the market instead of Bacillus subtilis BR. Using Bacillus subtilis BR for organic fertilizer production has broad application prospects. DETAILED DESCRIPTION

[0021] The invention provides a method for quickly obtaining high-quality organic fertilizer by utilizing Bacillus subtilis BR in combination with other probiotics.

[0022] The invention also provides the use of Bacillus subtilis BR in maintaining high organic matter content in organic fertilizer.

[0023] The invention also provides application of Bacillus subtilis BR in maintaining high total nutrient content of organic fertilizer.

[0024] The invention also provides application of Bacillus subtilis BR in maintaining low impact of organic fertilizer on seed germination rate.

[0025] The invention also provides application of Bacillus subtilis BR in maintaining low Escherichia coli flora in organic fertilizer.

[0026] The application includes: 1) mixing 250g of Bacillus subtilis BR (25-35 billion / g, preferably 30 billion / g), 50g of Candida utilis (15-25 billion / g, preferably 20 billion / g), 50g of Lactobacillus plantarum (5-15 billion / g, preferably 10 billion / g) and 50g of Trichoderma viride (15-25 billion / g, preferably 20 billion / g), and the mixture is recorded as A; 2) the mixed bacteria are mixed evenly with the following pig manure mixture (mixture is recorded as B). Mixture B includes 1500kg of pig manure (water content 80%), 1000-1400kg (preferably 1200kg) of rice husk powder (water content 10%) and 300-480kg (preferably 400kg) of sawdust (water content 11%). Place the mixed A and B in a 5-cubic-meter pool, cover with plastic sheet to prevent rain, and leave for 90 days. Set up 3 replicates in total.

[0027] In the control experiment, commercially available Bacillus subtilis (25-35 billion / g, preferably 30 billion / g) was purchased to replace Bacillus subtilis BR, and all other materials were the same, with a total of 3 replicates.

[0028] It can be operated all year round, with outdoor temperature ranging from -10℃ to 38℃.

[0029] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are all commercially available products.

[0030] Example 1: Fermentation of pig manure with Bacillus subtilis BR and other probiotics to produce organic fertilizer

[0031] 1. Probiotic raw materials: Bacillus subtilis BR agent (30 billion / g) was purchased from Shandong Dehe Mingxing Biotechnology Co., Ltd., Candida utilis agent (20 billion / g) was purchased from Cangzhou Zhongxin Biotechnology Co., Ltd., Lactobacillus plantarum agent (10 billion / g) was purchased from Shandong Zhongke Jiayi Bioengineering Co., Ltd., and Trichoderma viride agent (20 billion / g) was purchased from Guangzhou Lvhui Biotechnology Co., Ltd.

[0032] Commercially available Bacillus subtilis (referred to as strain SS): 100 billion / g (Shandong Weiduofeng Biotechnology Co., Ltd., product number 201902), diluted to 30 billion / g with light calcium carbonate.

[0033] 2. Pig manure: Pig manure from Bincheng pig farm in Binzhou, Shandong, with a water content of 80%.

[0034] 3. Rice husk powder: Rice husk powder has a mesh size of about 30 and a water content of 10%, and was purchased from Shijiazhuang Jinsui Agricultural Products Trading Co., Ltd.

[0035] 4. Sawdust: Sawdust is about 30 mesh, with a water content of 11%, purchased from Lingshou County Derui Mineral Products Processing Plant

[0036] 5. Dosage and mixing of probiotics: Mix 250 g of Bacillus subtilis BR agent (30 billion / g), 50 g of Candida utilis agent (20 billion / g), 50 g of Lactobacillus plantarum agent (10 billion / g) and 50 g of Trichoderma viride agent (20 billion / g), and the mixture is recorded as A.

[0037] 6. Mixing of fermentation raw materials: Evenly mix 1500 kg of pig manure, 1200 kg of rice husk powder and 400 kg of sawdust. The mixture is recorded as B.

[0038] 7. Mix A and B thoroughly, place the mixed A and B in a 5 cubic meter pool, cover with plastic sheet to prevent rain, ferment at an outdoor temperature of not less than -10℃ and not more than 38℃, no need to turn the pile, and leave for 90 days. Set up 3 replicates in total.

[0039] 8. In the control experiment, commercially available Bacillus subtilis SS (30 billion / g) was purchased to replace Bacillus subtilis BR, and all other materials were the same and left for 90 days. A total of 3 replicates were set.

[0040] Example 2 Quality testing of organic fertilizer produced by fermenting pig manure with Bacillus subtilis BR and other probiotics

[0041] The organic fertilizer prepared in Example 1 was tested using national standard or industry standard testing methods. The test results are shown in Table 1.

[0042] Table 1 Test results of various indicators of organic fertilizer

[0043]

[0044]

[0045] **: P<0.01, the difference is extremely significant.

[0046] As can be seen from Table 1, compared with the control group (No. 1 organic fertilizer), the organic matter content, total nutrient content, and seed germination index of the experimental group (No. 2 organic fertilizer) are significantly higher than the control, reaching a very significant level, indicating that bacillus subtilis BR and probiotics and Candida utilis, Lactobacillus plantarum, Trichoderma viride can be used for pig manure fermentation (need to add a certain proportion of wood chips, rice husk powder) with a reasonable ratio, and high-quality organic fertilizer is obtained. Compared with the control, the strain difference used in the experimental group only has bacillus subtilis, and the experimental result proves that bacillus subtilis BR plays an important role in the above-mentioned fermentation process. Bacillus subtilis is a generally recognized probiotic, but different strains have very different functions, and bacillus subtilis BR has a stronger effect with the SS withered grass purchased on the market, which can make organic fertilizer keep high-content organic matter, total nutrients and higher seed germination rate (existing literature reports that some probiotic compositions are used for the processing of different agricultural wastes, but most of them are obtained as farmyard manure, and its application value is lower than organic fertilizer). Its mechanism may be related to the different temperatures, types and contents of volatile gases produced during the fermentation process, which requires further study.

[0047] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements can be made based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. The application of Bacillus subtilis BR in organic fertilizer production is characterized in that: Inoculate 250 grams of Bacillus subtilis BR with a deposit number of CGMCC No. 23476, 50 grams of Candida utilis, 50 grams of Lactobacillus plantarum and 50 grams of Trichoderma viride into a mixture containing livestock and poultry manure and agricultural and forestry wastes to ferment and produce organic fertilizer; The bacterial content of the Bacillus subtilis BR agent is 25-35 billion / gram, the bacterial content of the Candida utilis agent is 15-25 billion / gram, the bacterial content of the Lactobacillus plantarum agent is 5-15 billion / gram, and the bacterial content of the Trichoderma viride agent is 15-25 billion / gram.

2. The use according to claim 1, characterized in that: The livestock and poultry excrement is pig excrement, and the agricultural and forestry waste includes rice husk powder and wood chips.

3. The production method of organic fertilizer is characterized in that, The following steps are involved: (1) 250 g of Bacillus subtilis BR with a deposit number of CGMCC No. 23476 was mixed with 50 g of Candida utilis, 50 g of Lactobacillus plantarum and 50 g of Trichoderma viride, and the mixture was recorded as A; (2) Mix 1300-1800 kg of pig manure, 1000-1400 kg of rice husk powder and 300-480 kg of sawdust, and record the mixture as B; (3) Mix A and B and let them ferment; The bacterial content of the Bacillus subtilis BR agent is 25-35 billion / gram, the bacterial content of the Candida utilis agent is 15-25 billion / gram, the bacterial content of the Lactobacillus plantarum agent is 5-15 billion / gram, and the bacterial content of the Trichoderma viride agent is 15-25 billion / gram.

4. The method according to claim 3, characterized in that The water content of the pig manure is 80%, the water content of the rice husk powder is 10%, and the water content of the sawdust is 11%.

5. The method according to claim 3 or 4, characterized in that: Let it ferment for 70-120 days.

6. The method according to claim 5, characterized in that Let it ferment for 90 days.

7. according to the fertilizer prepared by any one of the described methods of claim 3-6.

8. Use of the organic fertilizer according to claim 7 in plant cultivation.

Images