A composite microbial preparation, a preparation method thereof and application thereof

Abstract

The present application relates to a kind of composite microbial preparation and its preparation method and application.The raw material of the composite microbial preparation of the present application includes composite bacterial species fermentation liquor and carrier, wherein the composite bacterial species fermentation liquor is composed of Bacillus cereus fermentation liquor, Bacillus licheniformis fermentation liquor, Rhodotorula fermentation liquor and Sphingolipid fermentation liquor with mass ratio of (1-3) :(1-5) :(1-3) :(1-5).The present application forms dominant flora of low temperature resistance by Bacillus cereus, Bacillus licheniformis, Rhodotorula and Sphingolipid, which symbiotic coordination with each other, and has good degradation effect on ammonia nitrogen and total phosphorus in low temperature sewage (5℃-10℃).At the same time, the carrier formed by combining mineral carrier and biological carrier is further enhanced biological activity and adaptability, improves water treatment effect, and reduces sewage treatment cost.

Description

Technical Field

[0001] This invention specifically relates to a compound microbial preparation, its preparation method, and its application. Background Technology

[0002] Low temperatures affect the cell activity of microorganisms. Therefore, in cold winters, the overall physiological function and activity of microorganisms in water decrease, and the growth and reproduction of most microorganisms are inhibited. Only a few cold-resistant microorganisms can maintain a certain level of activity, leading to poor performance of water microbial treatment systems. Adding appropriate amounts of dominant cold-resistant microbial strains can significantly improve the water treatment effect of water microbial systems. However, there are currently few microbial agents used for low-temperature water treatment, and their treatment efficiency and stability need further improvement. Developing a cold-resistant microbial agent with high efficiency in removing COD and ammonia nitrogen from water under low-temperature conditions is of great significance for wastewater treatment in cold winters. Summary of the Invention

[0003] The purpose of this invention is to provide a low-temperature resistant compound microbial preparation that can effectively reduce the COD and ammonia nitrogen content in low-temperature water bodies.

[0004] Another object of the present invention is to provide a method for preparing the above-mentioned compound microbial preparation.

[0005] Another object of the present invention is to provide the application of the above-mentioned compound microbial preparation in wastewater treatment.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0007] A compound microbial preparation, comprising a compound bacterial strain fermentation broth and a carrier, wherein the compound bacterial strain fermentation broth is composed of Bacillus cereus fermentation broth, Bacillus sp. Q-12 fermentation broth, Rhodotorula glutinis fermentation broth, and Sphingobium fermentation broth. The Bacillus cereus strain is Bacillus sp. Q-12 with accession number CCTCC NO: M2022745; the Bacillus sp. Q-15 strain is Bacillus sp. Q-15 with accession number CCTCC NO: M2022748; the Rhodotorula glutinis strain is Rhodotorula sp. Q-14 with accession number CCTCC NO: M2022747; and the Sphingobium strain is Sphingobium glutinis with accession number CCTCC NO: M2022746. The mass ratio of the Bacillus cereus fermentation broth, the Bacillus pumilus fermentation broth, the Rhodotorula glutinis fermentation broth, and the Sphingosine monophosphate fermentation broth of strain sp. Q-13 is (1-3):(1-5):(1-3):(1-5).

[0008] Preferably, the particle size of the composite microbial preparation is less than or equal to 100 mesh, and / or the viable count in the composite microbial preparation is 1×10⁻⁶. 9 CFU / g ~ 1×10 10 CFU / g.

[0009] Preferably, the mass ratio of the compound microbial fermentation broth to the carrier is 1:(1-3).

[0010] Preferably, the carrier includes a mineral carrier and a biological carrier. The mineral carrier is one or more of bentonite, kaolin, montmorillonite, and diatomaceous earth, and the biological carrier is one or more of rice husk powder, peanut shell, corn flour, soybean flour, and starch.

[0011] More preferably, the mass ratio of the mineral carrier to the biological carrier is 1:(5-10).

[0012] Preferably, the raw materials of the compound microbial preparation further include adjuvants, which are one or more of sodium lignosulfonate, glucose, and peptone, and the adjuvants account for 1% to 5% of the total mass of the compound microbial preparation. Sodium lignosulfonate is used for uniform dispersion of the strain, while glucose and peptone are used as protectants and nutrients, providing rich nutrition to the strain, promoting its growth and reproduction, and efficiently activating the strain.

[0013] More preferably, the adjuvant is a composition of sodium lignosulfonate, glucose and peptone in a mass ratio of 1:(2-4):(1-3).

[0014] The present invention also provides a method for preparing the aforementioned composite microbial preparation, wherein the fermentation broth of Bacillus cereus, the fermentation broth of Bacillus oryzae, the fermentation broth of Rhodotorula glutinis, and the fermentation broth of Sphingosine monophosphate are mixed to obtain the composite microbial fermentation broth, and then the composite microbial fermentation broth is mixed evenly with the carrier, and then dried and pulverized to obtain the composite microbial preparation.

[0015] Preferably, the drying temperature is 30℃~40℃, and the moisture content of the dried compound microbial agent is 2%~10%.

[0016] The present invention also provides the application of the compound microbial preparation as described above in wastewater treatment.

[0017] More preferably, the temperature of the wastewater to be treated is 5℃~10℃.

[0018] More preferably, the COD concentration of the wastewater to be treated is 150 mg / L to 1000 mg / L, and the ammonia nitrogen concentration is 30 mg / L to 80 mg / L.

[0019] Due to the application of the above technical solution, the present invention has the following advantages compared with the prior art:

[0020] This invention utilizes a combination of Bacillus cereus, Bacillus oryzae, Rhodotorula glutinis, and Sphingosine monocytogenes to form a dominant, low-temperature-resistant bacterial community. These bacteria coexist and coordinate effectively, exhibiting excellent degradation effects on ammonia nitrogen and total phosphorus in low-temperature wastewater (5℃~10℃). Furthermore, the combination of mineral and biological carriers further enhances biological activity and adaptability, improving water treatment efficiency and reducing wastewater treatment costs. Detailed Implementation

[0021] In the following description, only certain exemplary embodiments are briefly described. As those skilled in the art will recognize, the described embodiments can be modified in various ways without departing from the spirit or scope of the embodiments of the invention. Therefore, the description is considered to be exemplary in nature and not restrictive.

[0022] In this invention, all the raw materials described can be obtained commercially or by known means, and unless otherwise specified, they all meet the requirements of standard chemical products.

[0023] Unless otherwise specified, the experimental methods used in this invention are all conventional methods.

[0024] In this invention, the ammonia nitrogen detection method refers to the national standard HJ535-2009 Determination of ammonia nitrogen in water quality - Nessler's reagent spectrophotometric method; the COD detection method refers to the national standard HJ 828-2017 Determination of chemical oxygen demand in water quality - dichromate method.

[0025] In this invention, the Bacillus cereus Q-12 strain was deposited at the China Center for Type Culture Collection on May 27, 2022, with accession number CCTCC M 2022745, and the address of the depository is Wuhan University, Wuhan, China.

[0026] Bacillus sp. Q-15 strain was deposited at the China Center for Type Culture Collection (CCTCC) on May 27, 2022, with accession number CCTCC M 2022748. The depository address is Wuhan University, Wuhan, China.

[0027] The strain of Sphingobium sp. Q-13 was deposited at the China Center for Type Culture Collection on May 27, 2022, with accession number CCTCC M 2022746. The depository address is Wuhan University, Wuhan, China.

[0028] Rhodotorula sp. Q-14 strain was deposited at the China Center for Type Culture Collection (CCTCC) on May 27, 2022, with accession number CCTCC M 2022747. The depository address is Wuhan University, Wuhan, China.

[0029] The Bacillus megaterium used in this invention was purchased from Beihai Yeshengwang Biotechnology Co., Ltd.

[0030] In this invention, there are no special requirements for the preparation method of the fermentation broth of each strain; existing conventional methods can be used. For example, the method used in this invention is as follows: Different functional strains are aseptically transferred to test tube slant culture (slant culture medium formula: glucose 5g / L, peptone 5g / L, beef extract 0.5g / L, sodium chloride 3g / L, agar 20g / L, pH=7.2), and cultured at 10℃ for 24h to activate the strains; one loop of the activated strain is picked and inoculated into a 250mL Erlenmeyer flask containing 50mL of seed culture medium, and cultured at 8-10℃ for 24h. Microscopic examination shows no contamination, thus obtaining the seed culture of each strain; the seed culture is aseptically transferred to a 1000mL Erlenmeyer flask containing 200mL of fermentation culture medium for fermentation culture (seed culture medium and fermentation culture medium formula: glucose 5g / L, peptone 5g / L, beef extract 0.5g / L, sodium chloride 3g / L, pH=7.2), and cultured at 10℃ for 48h in a constant temperature shaking incubator to obtain the fermentation broth of the low-temperature resistant strain.

[0031] Example 1

[0032] This embodiment provides a low-temperature resistant composite microbial preparation 1, which consists of a composite microbial fermentation broth and a carrier, with a mass ratio of 1:1 between the composite microbial fermentation broth and the carrier.

[0033] The preparation method is as follows:

[0034] (1) Mix the fermentation broth of Bacillus cereus, Bacillus pumilus, Rhodotorula glutinis, and Sphingosine mononitrate in a mass ratio of 2:3:2:3 to obtain a compound bacterial fermentation broth.

[0035] (2) Weigh out kaolin, diatomaceous earth, rice husk powder and corn flour in a mass ratio of 1:1:5:5, mix them and use them as a carrier.

[0036] (3) Mix the compound microbial culture fermentation broth and carrier at a mass ratio of 1:3, add 3% of the total mass of the compound microbial culture fermentation broth as an adjuvant, which is a combination of sodium lignosulfonate, glucose, and peptone (mass ratio of sodium lignosulfonate: glucose: peptone is 1:3:2), mix thoroughly, dry at a temperature of 30℃~40℃, pulverize into powder, and pass through a 100-mesh sieve to obtain a viable count of 1.8×10⁻⁶. 91. Low-temperature resistant compound microbial preparation with CFU / g and a water content of 10%.

[0037] Example 2

[0038] This embodiment provides a low-temperature resistant compound microbial preparation 2, which is basically the same as that in embodiment 1, except that the mass ratio of Bacillus cereus fermentation broth, Bacillus oryzae fermentation broth, Rhodotorula glutinis fermentation broth, and Sphingosine mononitrate fermentation broth is 1:4:2:5.

[0039] Example 3

[0040] This embodiment provides a low-temperature resistant composite microbial preparation 3, which is basically the same as that in embodiment 1, except that the mass ratio of the composite strain fermentation broth to the carrier is 1:3.

[0041] Example 4

[0042] This embodiment provides a low-temperature resistant composite microbial preparation 4, which is basically the same as that in embodiment 1, except that the carrier is composed of montmorillonite powder, diatomaceous earth, starch and rice husk powder in a mass ratio of 1:2:6:10.

[0043] Comparative Example 1

[0044] This embodiment provides a low-temperature resistant compound microbial preparation 5, which is basically the same as that in embodiment 1, except that the Rhodotorula glutinis fermentation broth in embodiment 1 is replaced with Bacillus megaterium fermentation broth.

[0045] Comparative Example 2

[0046] This embodiment provides a low-temperature resistant compound microbial preparation 6, which is basically the same as that in embodiment 1, except that the compound bacterial fermentation broth is made by mixing Bacillus cereus fermentation broth, Bacillus megaterium fermentation broth, Bacillus erythrosporum fermentation broth, Rhodotorula glutinis fermentation broth, and Sphingosine mononitrate fermentation broth in a mass ratio of 2:2:3:2:3.

[0047] Comparative Example 3

[0048] This embodiment provides a low-temperature resistant compound microbial preparation 7, which is basically the same as that in embodiment 1, except that the Bacillus cereus fermentation broth in embodiment 1 is replaced with Bacillus megaterium fermentation broth.

[0049] Comparative Example 4

[0050] This embodiment provides a low-temperature resistant compound microbial preparation 8, which is basically the same as that in embodiment 1, except that the mass ratio of Bacillus cereus fermentation broth, Bacillus oryzae fermentation broth, Rhodotorula glutinis fermentation broth, and Sphingosine monophosphate fermentation broth is 2:6:4:3.

[0051] Wastewater samples were collected and divided into 9 groups, each containing 400 mL. Eight groups were experimental groups, with compound microbial agents 1 through 8 added at a concentration of 50 ppm. The remaining group served as the control group. The samples were treated in a low-temperature shaker at 8-10℃ for 3 days. The supernatant was then used to determine the ammonia nitrogen and COD values. The results are shown in Table 1.

[0052] Table 1

[0053]

[0054]

[0055] Table 1 shows the removal effects of compound microbial agents 1-4 on ammonia nitrogen and COD after 3 days of addition. Compound microbial agents 1-4 achieved a COD removal rate of over 85% and an ammonia nitrogen removal rate of over 70% in the experimental wastewater, which were significantly better than the treatment effects of compound microbial agents 5-8. The wastewater treatment effects of compound microbial agents 5-8 were relatively weak.

[0056] The above embodiments are only for illustrating the technical concept and features of the present invention, and are intended to enable those skilled in the art to understand the content of the present invention and implement it accordingly. They should not be construed as limiting the scope of protection of the present invention. All equivalent changes or modifications made in accordance with the spirit and essence of the present invention should be covered within the scope of protection of the present invention.

Claims

1. A complex microbial formulation, characterized in that: The raw material comprises a compound bacterial strain fermentation liquor and a carrier, the compound bacterial strain fermentation liquor is composed of a Bacillus cereus fermentation liquor, a Bacillus licheniformis fermentation liquor, a Rhodotorula glutinis fermentation liquor and a Sphingobium fermentation liquor, the Bacillus cereus is a Q-12 strain with a preservation number of CCTCC NO: M2022745 Bacillus cereus the Bacillus licheniformis is a Q-15 strain with a preservation number of CCTCC NO: M2022748 Bacillus sp. the Rhodotorula glutinis is a Q-14 strain with a preservation number of CCTCC NO: M2022747 Rhodotorula sp. the Sphingobium is a Q-13 strain with a preservation number of CCTCC NO: M2022746 Sphingobium sp. The mass ratio of the Bacillus cereus fermentation liquor, the Bacillus licheniformis fermentation liquor, the Rhodotorula glutinis fermentation liquor and the Sphingobium fermentation liquor is (1-3):(1-5):(1-3):(1-5).

2. The complex microbial preparation according to claim 1, characterized by: The particle size of the composite microbial preparation is less than or equal to 100 mesh, and / or the viable bacterial count in the composite microbial preparation is 1×10 9 CFU / g~1×10 10 CFU / g.

3. The complex microbial preparation according to claim 1, characterized by: The mass ratio of the compound microbial strain fermentation liquor and the carrier is 1: (1-3).

4. The complex microbial preparation according to claim 1, characterized by: The carrier comprises a mineral carrier and a biological carrier, the mineral carrier is one or more of bentonite, kaolin, montmorillonite and diatomite, and the biological carrier is one or more of rice husk powder, peanut shell, corn powder, bean powder and starch.

5. The complex microbial preparation according to claim 4, characterized in that: The mass ratio of the mineral carrier and the biological carrier is 1: (5-10).

6. The complex microbial preparation according to claim 1, characterized in that: The raw material further comprises an additive, the additive is one or more of sodium lignosulfonate, glucose and proteose peptone, and the additive accounts for 1%-5% of the total mass of the compound microbial preparation.

7. The method of claim 1, wherein the preparation of the complex microbial formulation is characterized by: The wax-like Bacillus fermentation liquor, the Bacillus castaneus fermentation liquor, the Rhodotorula fermentation liquor and the Sphingobium fermentation liquor are mixed to obtain the compound microbial strain fermentation liquor, and then the compound microbial strain fermentation liquor is mixed with the carrier to obtain the compound microbial preparation.

8. The method for preparing the compound microbial preparation according to claim 7, characterized in that: The drying temperature is 30-40°C, and the water content of the dried compound microbial preparation is 2%-10%.

9. The compound microbial preparation according to any one of claims 1-6 is applied to sewage treatment.

10. Use according to claim 9, characterized in that: The temperature of the sewage to be treated is 5-10°C, and / or the COD concentration of the sewage to be treated is 150-1000 mg / L, and the ammonia nitrogen concentration is 30-80 mg / L.

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