Bacillus bacterial composition and application thereof in compost

Abstract

The invention relates to a bacillus bacterial composition and application thereof in compost, the bacillus bacterial composition comprises Vietnam bacillus ISB-206 and Bacillus cereus ISB-200, the preservation number of the Vietnam bacillus ISB-206 is CCTCC NO: M 2019468, the preservation date is June 19, 2019, and the preservation number of the Bacillus cereus ISB-200 is CCTCC NO: M 2019468. The preservation number of the acid-producing bacterium bacillus cereus ISB-200 is CCTCC (China Center For Type Culture Collection) NO: M 2019097, and the preservation date is February 21, 2019. According to the invention, the promotion effect of a compost product on crop growth is greatly enhanced, the crop yield is increased, the crop quality is improved, remarkable economic benefits and environmental benefits are brought to agricultural production, and perfect fusion of a compost technology and sustainable agricultural development is realized.

Description

Technical Field

[0001] The invention belongs to the field of microbial agents, and in particular relates to a bacillus bacterial composition and application thereof in composting. Background Art

[0002] Bacillus fermentation technology is widely used in food, pharmaceuticals, environmental protection, agriculture and other fields. However, due to the ever-changing application environment and conditions, its effect is often difficult to predict. How to improve the adaptability and stability of Bacillus fermentation technology so that it can play a greater role in different fields is a major challenge.

[0003] Bacillus belongs to the bacterial kingdom, with a wide variety of species and different biological characteristics and metabolic functions. During the fermentation process, different types of Bacillus have different adaptability to substrates and environments. Therefore, choosing a suitable strain is crucial for fermentation. However, in practical applications, there are often problems such as insufficient understanding of the types and characteristics of Bacillus and improper selection, resulting in unsatisfactory fermentation results. Specifically, the conditions for Bacillus fermentation are variable, such as temperature, pH value, oxygen concentration, etc., which have an important influence on the fermentation process and product yield. However, in practical applications, the control of these conditions is often difficult to unify, resulting in unstable fermentation results. In addition, different strains have different adaptability to conditions. How to formulate suitable fermentation conditions according to the characteristics of the strain is a major challenge. At the same time, during the fermentation process of Bacillus, the concentration of substrates and products has an important influence on the fermentation efficiency and product quality. However, in practical applications, there is often a problem of imbalance between substrate and product concentrations. For example, too high a substrate concentration may inhibit strain growth, while too high a product concentration may inhibit enzyme activity. How to achieve a balance between substrate and product concentrations and improve fermentation efficiency is a major challenge.

[0004] There may be side reactions during the fermentation process of Bacillus, such as acidification of the fermentation broth and inactivation of enzymes, which will have an adverse effect on the fermentation results. In practical applications, how to reduce the occurrence of side reactions and improve the quality and yield of the product is a major challenge. In the fermentation process of Bacillus, detection and purification methods are crucial to ensure product quality and yield. However, in practical applications, there are often problems with the detection and purification methods, which makes it difficult to compare and evaluate the results. How to establish standardized detection and purification methods to improve product quality and yield is a major challenge.

[0005] In summary, Bacillus fermentation technology faces some problems and challenges in its application in various fields. In order to promote the further development of this technology, researchers need to strengthen the research on the types and characteristics of Bacillus, optimize the fermentation conditions, reduce side reactions, and improve its adaptability and stability. Summary of the invention

[0006] Technical problem to be solved: The present invention provides a Bacillus bacterial composition and its application in composting. The composition makes up for the limitations of a single bacterial agent in the composting process through the synergistic effect of two functional bacteria. It can not only effectively accelerate the decomposition and transformation of organic nitrogen, but also reduce the pH increase and ammonia volatilization loss caused by nitrogen mineralization, thereby increasing the content of fast-acting nitrogen in the compost product. At the same time, the composition increases the content of small molecular organic acids, significantly improves the effect of compost products on promoting crop yields, and achieves a dual improvement in composting efficiency and crop growth benefits.

[0007] Technical solution: A bacillus bacterial composition, including Bacillus vietnamensis ISB-206 and Bacillus cereus ISB-200, wherein the deposit number of Bacillus vietnamensis ISB-206 is CCTCC NO: M 2019468, and the deposit date is June 19, 2019; the deposit number of Bacillus cereus ISB-200 is CCTCC NO: M 2019097, and the deposit date is February 21, 2019. The deposit units are all China Center for Type Culture Collection, and the deposit addresses are all Wuhan University, Wuhan, China.

[0008] Preferably, the weight ratio of the above-mentioned Bacillus vietnam ISB-206 and Bacillus cereus ISB-200 is 1:1.

[0009] Application of the bacterial composition in chicken manure composting fermentation.

[0010] Preferably, the chicken manure composting raw materials are composed of fresh chicken manure, rice bran, zeolite and superphosphate, wherein the weight ratio of fresh chicken manure, rice bran, zeolite and superphosphate is 15:1:0.3:0.5.

[0011] Preferably, the amount of the above-mentioned Bacillus vietnam ISB-206 and Bacillus cereus ISB-200 is 3wt.% of the chicken manure compost raw material.

[0012] Preferably, the Bacillus cereus ISB-200 is added together with Bacillus vietnamese ISB-206 during mixing before composting.

[0013] Preferably, the Bacillus cereus ISB-200 is added during the high temperature period of composting.

[0014] The above composting process also includes regular turning of the pile to promote oxygen exchange and microbial activity, thereby improving composting efficiency.

[0015] The above compost products are fully matured to ensure their stability and fertilizer efficiency.

[0016] A method for improving chicken manure composting efficiency comprises using the above-mentioned bacterial composition for composting.

[0017] Beneficial effects: The bacterial composition provided by the present invention and its innovative application in composting, relying on the unique synergistic mechanism of two selected functional bacteria, not only greatly accelerates the decomposition and transformation process of organic nitrogen, effectively shortens the composting cycle, but also cleverly solves the problems of abnormal increase in pH value and large-scale volatilization of ammonia caused by excessive mineralization of nitrogen in traditional composting, and significantly reduces the loss of nitrogen nutrients. In addition, the composition also significantly increases the content of fast-acting nitrogen in the compost product, while increasing the proportion of small molecule organic acids that are particularly beneficial to crop growth, so that the overall fertilizer effect of the compost product has achieved a qualitative leap. Under the joint action of this series of optimization measures, the promoting effect of compost products on crop growth is greatly enhanced, which not only increases crop yields, but also improves crop quality, bringing significant economic and environmental benefits to agricultural production, and realizing the perfect integration of composting technology and sustainable agricultural development. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Schematic diagram of temperature changes in different treatments.

[0019] Figure 2 Schematic diagram of pH changes in different treatments.

[0020] Figure 3 Schematic diagram for determining the ammonium nitrogen and nitrate nitrogen content of each sample after 49 days of composting.

[0021] Figure 4 Schematic diagram of lettuce yield under different treatments. DETAILED DESCRIPTION

[0022] The following examples will enable those skilled in the art to more fully understand the present invention, but are not intended to limit the present invention in any way.

[0023] Example 1

[0024] Factory Composting Verification of Fermentation and Maturation Bacteria

[0025] Two strains of highly efficient protein-decomposing bacteria isolated in the early stage: Bacillus vietnamensis ISB-206 CCTCC NO: M 2019468 and acid-producing bacteria: Bacillus cereus ISB-200 CCTCC NO: M 2019097, were used to form an optimized decay-promoting agent, and factory-scale composting fermentation experiments and potted fertilizer efficiency tests of the finished compost products were carried out.

[0026] Experimental Materials

[0027] The experiment was conducted at the Haiyan County Composting Plant in Jiaxing City, Zhejiang Province. The composting materials were 15t of fresh chicken manure, 1t of rice bran, 0.3t of zeolite, and 0.5t of superphosphate. Bacillus cereus ISB-200 and Bacillus vietnamese ISB-206 were propagated to 150L in the fermentation tank and the OD600 was unified to 3.30. The physical and chemical properties of the chicken manure are as follows: total nitrogen 2%, total phosphorus 4.3%, total potassium 2.2%, available phosphorus 17874.2mg / kg, and available potassium 17125mg / kg.

[0028] Composting experiment

[0029] The application of microbial agents and the application period were used as control variables. Three treatment groups were set up in the experiment as shown in Table 3.1. Each pile contained 5 tons of chicken manure; the microbial agents were first poured into rice bran (300 kg) and zeolite powder (100 kg) and mixed evenly before adding to the pile; the microbial agents were added at a ratio of 3% of the weight of the chicken manure. 3% of the weight of the chicken manure was added as superphosphate to adjust the pH value.

[0030] Factory Validation of Composting

[0031] Using the existing bacterial agents in the factory as a control, a factory-scale composting verification of the decay-promoting bacterial agents was carried out. Three treatments were set up (Table 1), and dynamic sampling was performed during the composting process.

[0032] Table 1 Processing settings

[0033]

[0034] Arrange the composting test. Collect background samples on the same day. During the composting process, record the temperature of the pile and the environment once in the morning and afternoon every day. Turn the pile on the 1st, 4th, 7th, 11th, 14th, 18th and 21st days and use a hollow PVC pipe to take samples of about 400g from each pile. All three groups set sampling points at four locations of the pile. A total of 84 samples were collected at 7 time points in the first fermentation stage. Turn the pile and take samples once a week (Thursday) in the ripening period until the 49th day, with a total of 48 samples at 4 time points.

[0035] After the samples were collected, they were immediately placed in a 4 °C refrigerator for determination of physical and chemical properties.

[0036] Determination of physical and chemical properties of compost

[0037] Compost pH determination: Take air-dried samples and grind them through a 20-mesh sieve. 2 The pH value was determined by leaching with a pH meter.

[0038] Determination of ammonium nitrogen and nitrate nitrogen in compost: Weigh 15 g of compost sample that has passed a 2 mm sieve, mix it with ultrapure water at a water-soil ratio of 2:1 (dry soil weight), shake it at room temperature for 30 min, centrifuge it (4000 r / min) and -1) After 20 min, the supernatant was filtered through a 0.45-μm microporous membrane. Nitrate nitrogen was determined by ultraviolet spectrophotometry (ultraviolet-visible spectrophotometer / 752 automatic), and ammonium nitrogen was determined by distillation and titration method (GB / T 3595-2000).

[0039] The temperature change during the first fermentation period was as Figure 1 shown. It can be seen that (1) for treatment 2# with two kinds of inoculants added at one time, the temperature of the compost pile was significantly higher than that of the control from the 4th day, 2-3 °C higher. Paired-sample T-tests for each treatment found that the temperature of treatment 2# (two inoculants added before fermentation) was significantly higher than that of CK and treatment 3# (protein bacteria first and acid-producing bacteria later). There was no significant difference in temperature change between treatment 1# and 3#.

[0040] The pH changes of different treatments were compared, as Figure 2 shown. It was found that the pH value at the end of composting after inoculating the inoculant was significantly lower than that of the control sample. The inoculated treatments (3#: 7.17, 2#: 7.29) < CK (7.44). It was speculated that this might be due to the production of acid by microorganisms, and more small-molecule organic acids accumulated in the compost.

[0041] The ammonium nitrogen and nitrate nitrogen contents of the raw materials and compost samples after 49 days were measured, as Figure 3 shown. It was found that the soluble nitrogen in the inoculant treatment was significantly higher than that in the control treatment (treatment 3# > treatment 2# > CK).

[0042] Fertilizer efficiency in greenhouse pot experiments

[0043] The fertilizer efficiency of the compost products obtained from the above experiments was verified, and four fertilizer samples taken from the compost pile were used as parallel treatments for the experiments to more clearly understand the fertilizer effect.

[0044] The results were as Figure 4 shown. Under potting conditions, compared with the fertilizer of the control treatment, the organic fertilizers fermented by single and multiple inoculations of the selected inoculant could increase the lettuce yield by 2.64 times and 1.8 times respectively. The treatment with one-time addition of inoculant was better than the treatment with multiple inoculations in terms of lettuce yield, quality, etc. Among them, CK was the control treatment without inoculant, 2# was the treatment with one-time inoculation of inoculant, and 3# was the treatment with multiple inoculations of inoculant; H represents the high-dose fertilization (2%) treatment, L represents the low-dose fertilization (1%) treatment, and 1, 2, 3 represent the CK, one-time inoculation of inoculant treatment group, and multiple inoculations of inoculant treatment group respectively.

[0045] The above embodiments are only specific implementation methods of the present invention, which are used to illustrate the technical solutions of the present invention, rather than to limit them. The protection scope of the present invention is not limited thereto. Although the present invention is described in detail with reference to the above embodiments, ordinary technicians in the field should understand that any technician familiar with the technical field can still modify the technical solutions recorded in the above embodiments within the technical scope disclosed by the present invention, or can easily think of changes, or make equivalent replacements for some of the technical features therein; and these replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present invention, and should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention shall be based on the protection scope of the claims.

Claims

1. A Bacillus bacterial composition, characterized in that Including Vietnamese Bacillus ISB-206 ( Bacillus vietnamese ISB-206) and Bacillus cereus ISB-200 ( Bacillus cereus ISB-200), of which the deposit number of Vietnamese Bacillus ISB-206 is CCTCC NO: M 2019468, and the deposit date is June 19, 2019; the deposit number of the Bacillus cereus ISB-200 is CCTCC NO: M 2019097, and the deposit date is February 21, 2019.

2. The bacterial composition according to claim 1, characterized in that The weight ratio of the Vietnamese Bacillus ISB-206 to the Bacillus cereus ISB-200 is 1:

1.

3. Use of the bacterial composition according to claim 1 or 2 in chicken manure composting fermentation.

4. The use according to claim 3, characterized in that: The chicken manure composting raw materials consist of fresh chicken manure, rice bran, zeolite and superphosphate, wherein the weight ratio of the fresh chicken manure, rice bran, zeolite and superphosphate is 15:1:0.3:0.

5.

5. The use according to claim 3, characterized in that: The amount of the Vietnamese Bacillus ISB-206 and the Bacillus cereus ISB-200 is 3wt.% of the chicken manure compost raw material.

6. The use according to claim 3, characterized in that: The Bacillus cereus ISB-200 is added together with the Vietnamese Bacillus ISB-206 during mixing before composting.

7. The use according to claim 3, characterized in that: The Bacillus cereus ISB-200 is added during the high temperature period of composting.

8. The use according to any one of claims 3 to 7, characterized in that The composting process also includes turning the compost regularly to promote oxygen exchange and microbial activity, thereby improving composting efficiency.

9. The use according to any one of claims 3 to 7, characterized in that The compost product is fully matured to ensure its stability and fertilizer efficiency.

10. A method for improving chicken manure composting efficiency, characterized in that: The method comprises using the bacterial composition according to claim 1 or 2 for composting.

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