A composite microbial inoculum and its application in treating heparin sodium wastewater

Abstract

The present invention discloses a composite microbial agent and its application in treating heparin sodium wastewater. The composite microbial agent is prepared from Bacillus, Bacteroides, Lactobacillus, Pseudomonas, Acinetobacter, Auxiliary Bacteria and nutrient agent according to a certain weight ratio, and the total viable count of the microbial agent is not less than 1×10 9 cfu / mL. The present invention also provides a method for treating heparin sodium wastewater by using the composite microbial agent. The method comprises the following steps: resuscitating and culturing the composite microbial agent; and then adding the supernatant of the resuscitated composite microbial agent into the heparin sodium wastewater for treatment. The experimental results show that: after treating the heparin sodium wastewater with the composite microbial agent for 1 week, the COD removal rate is close to 90%, and the ammonia nitrogen removal rate reaches about 60%, effectively solving the problems of high COD and high ammonia nitrogen in the heparin sodium wastewater, providing a feasible method for treating the heparin sodium wastewater, and having good application prospects.

Description

Technical Field

[0001] The invention relates to the field of sewage treatment, and in particular to a composite microbial agent and application thereof in treating sodium heparin wastewater. Background Art

[0002] In today's society, with the rapid development of industry and people's further improvement of water quality requirements, the water pollution problem faced by mankind has gradually attracted everyone's attention. A large amount of organic pollutants in industrial wastewater spread in the water body, causing water quality to deteriorate. These organic pollutants are generally difficult to degrade in water, and eventually enter the food chain through biological accumulation, thus threatening human health.

[0003] Among the numerous industrial wastewaters, pharmaceutical wastewater has become a pollution target that needs to be solved urgently due to its large wastewater discharge volume, complex types of organic pollutants contained in the wastewater, and poor biodegradability. In recent years, in the pharmaceutical field, with the application of sodium heparin drugs, its production and demand have increased year by year. In the process of preparing sodium heparin drugs, a large amount of wastewater is also generated. Statistics show that the domestic casing processing wastewater discharge is about 2.7 million tons / year, and the sodium heparin preparation wastewater is about 2.2 million tons / year.

[0004] The prominent features of sodium heparin wastewater are high COD (chemical oxygen demand), high ammonia nitrogen, high salt and easy to rot. If it is discharged directly into the water without treatment, it will pollute the water environment and aggravate the eutrophication of the water. It will not only harm aquatic organisms such as fish, but also enter the human body through the enrichment of the food chain, causing chronic poisoning. And sodium heparin wastewater also contains a pungent odor. If it is discharged into the atmosphere without treatment, it will pollute the air quality and affect human respiratory health. Therefore, how to efficiently treat sodium heparin production wastewater is an urgent problem to be solved for the healthy development of the industry.

[0005] At present, there are two methods for treating sodium heparin wastewater: physical and chemical methods and biological methods. Among them, the physical and chemical method is limited in the actual treatment and promotion of sodium heparin wastewater due to its high energy consumption, high initial investment, secondary pollution and low treatment efficiency. The biological method is increasingly valued by researchers from various countries due to its advantages such as low treatment cost, simple operation, no secondary pollution and thorough treatment. However, when using biological methods to treat sodium heparin wastewater, the treatment effect is poor due to problems such as microbial activity and organic load impact. Summary of the invention

[0006] In order to solve the problems existing in the prior art, the present invention provides an efficient and safe composite microbial agent, which can effectively reduce the COD and ammonia nitrogen indicators in sodium heparin wastewater, improve the sodium heparin wastewater treatment capacity, and provide an effective solution for my country's water environment management.

[0007] In the first aspect, the present invention claims to protect a compound microbial inoculum.

[0008] The active ingredients of the compound microbial inoculum protected by the present invention include Bacillus species, Bacteroides species, Lactobacillus species, Pseudomonas species, Acinetobacter species, Auxiliary Bacteria genus species and nutrient agent;

[0009] The Auxiliary Bacteria genus includes Sphingomonas, Curtobacterium, Azonobacter and Methylobacterium;

[0010] The nutrient agent includes corn steep liquor dry powder, soybean meal powder, broken wall yeast powder, beef extract, fish meal peptone, glucose, magnesium sulfate, dipotassium hydrogen phosphate, manganese sulfate, sodium chloride and Tween 80;

[0011] The total viable count of the compound microbial inoculum is not less than 1×10 9 cfu / mL.

[0012] In the second aspect, the present invention claims to protect a preparation method of the above compound microbial inoculum.

[0013] The preparation method of the compound microbial inoculum protected by the present invention includes the following steps:

[0014] 1) Respectively inoculate Bacillus species, Bacteroides species, Lactobacillus species, Pseudomonas species, Acinetobacter species, Auxiliary Bacteria genus species into a liquid medium for seed culture to obtain Bacillus species seed liquid, Bacteroides species seed liquid, Lactobacillus species seed liquid, Pseudomonas species seed liquid, Acinetobacter species seed liquid and Auxiliary Bacteria genus species seed liquid respectively;

[0015] 2) Respectively inoculate the Bacillus species seed liquid, the Bacteroides species seed liquid, the Lactobacillus species seed liquid, the Pseudomonas species seed liquid, the Acinetobacter species seed liquid and the Auxiliary Bacteria genus species seed liquid into the nutrient agent for fermentation culture to obtain Bacillus species fermentation liquid, Bacteroides species fermentation liquid, Lactobacillus species fermentation liquid, Pseudomonas species fermentation liquid, Acinetobacter species fermentation liquid and Auxiliary Bacteria genus species fermentation liquid respectively;

[0016] 3) Mix the Bacillus species fermentation liquid, the Bacteroides species fermentation liquid, the Lactobacillus species fermentation liquid, the Pseudomonas species fermentation liquid, the Acinetobacter species fermentation liquid and the Auxiliary Bacteria genus species fermentation liquid with the nutrient agent, and culture until the viable count is at least 1×10 9 cfu / mL to obtain a compound microbial liquid;

[0017] 4) Freeze-dry the compound microbial liquid to make a compound microbial inoculum.

[0018] In the above method, in the step 1), the liquid medium comprises tryptone, yeast extract powder and NaCl. Further, the solvent of the liquid medium is water, and the solutes and their concentrations are 10.00 g / L of tryptone, 5.00 g / L of yeast extract powder, and 10.00 g / L of NaCl respectively. Even further, the pH of the liquid medium is 7.00.

[0019] The conditions for seed culture can be 30 - 35 °C, the rotation speed is 150 - 180 rpm, and it is cultured for 24 - 48 h under aerobic conditions. Specifically, it can be 30 °C, the rotation speed is 160 rpm, and it is cultured for 24 h under aerobic conditions.

[0020] In the above method, in the step 2), each strain seed liquid is inoculated into the nutrient agent according to a weight ratio of 3.00 - 5.67% (such as 3% or 4% or 5% or 5.67%).

[0021] The solvent of the nutrient agent is water, and the solutes and their concentrations are 4 g / L of corn steep liquor dry powder, 2 g / L of soybean meal powder, 3 g / L of broken wall yeast powder, 3 g / L of beef extract, 2 g / L of fish meal peptone, 5 g / L of glucose, 0.1 g / L of magnesium sulfate, 0.08 g / L of dipotassium hydrogen phosphate, 0.02 g / L of manganese sulfate, 0.1 g / L of sodium chloride, and 0.01 g / L of Tween 80 respectively.

[0022] The culture conditions can be that the tank pressure is 0.02 - 0.04 MPa, the temperature is 30 - 35 °C, the pH is 6.8 - 7.2, the rotation speed is 150 - 180 rpm, and the dissolved oxygen > 6 mg / L. Specifically, it can be 0.02 MPa, the temperature is 30 °C, the pH is 7.0, the rotation speed is 160 rpm, and the dissolved oxygen > 6 mg / L.

[0023] In the above method, in step 3), the weight ratio of the fermentation broth of Bacillus spp., the fermentation broth of Bacteroides spp., the fermentation broth of Lactobacillus spp., the fermentation broth of Pseudomonas spp., the fermentation broth of Acinetobacter spp., the fermentation broth of Auxiliary Bacteria spp., and the nutrient can be (56.15-79.65):(0.19-1.31):(0.01-6.36):(0.33-4.77):(0.02-5.05):(3.72-7.61):(15.00-25.00) or 78.86:0.41:0.01:0.33:0.12:5.27:15.00 or 73.91:0.19:6.36:0.80:0.02:3.72:15.00 or 71.91:0.92:0.02:1.40:0.29:5.46:20.00 or 56.15:1.31:0.05:4.77:5.05:7.61:25.00 or 79.65:0.63:0.02:0.57:0.17:3.66:15.00, preferably 56.15:1.31:0.05:4.77:5.05:7.61:25.00.

[0024] In the fermentation broth of Auxiliary Bacteria spp., the weight ratio of the fermentation broth of Sphingomonas spp., the fermentation broth of Curtobacterium spp., the fermentation broth of Azogomonas spp., and the fermentation broth of Methylobacterium spp. can be (40-50):(15-25):(15-25):(15-25) or 40:25:15:15 (9:5:3:3) or 50:15:20:15 (10:3:4:3) or 45:15:20:20 (8:4:3:3).

[0025] The culture conditions can be a tank pressure of 0.02-0.04 MPa, a temperature of 30-35 °C, a pH of 6.8-7.2, a rotation speed of 150-180 rpm, and a dissolved oxygen > 6 mg / L. Specifically, it can be 0.02 MPa, a temperature of 30 °C, a pH of 7.0, a rotation speed of 160 rpm, and a dissolved oxygen > 6 mg / L.

[0026] The culturing is carried out until the viable cell count reaches (1-9)×10 9 cfu / mL.

[0027] In the above method, in step 4), the freeze-drying conditions can be freeze-drying at (-80--60) °C for 48 hours. Specifically, it can be freeze-drying at -80 °C for 48 hours.

[0028] In the third aspect, the present invention claims the new use of the above composite microbial agent or the composite microbial agent prepared by the above method.

[0029] The present invention claims the application of the above-mentioned composite microbial inoculant or the composite microbial inoculant prepared by the above-mentioned method in any one of the following M1)-M4):

[0030] M1) Treating heparin sodium wastewater;

[0031] M2) Reducing the COD content and ammonia nitrogen content in heparin sodium wastewater;

[0032] M3) Preparing a product for treating heparin sodium wastewater;

[0033] M4) Preparing a product for reducing the COD content and ammonia nitrogen content in heparin sodium wastewater.

[0034] In the fourth aspect, the present invention claims a method for treating heparin sodium wastewater.

[0035] The method for treating heparin sodium wastewater claimed by the present invention includes the following steps:

[0036] X1) Adding the above-mentioned composite microbial inoculant or the composite microbial inoculant prepared by the above-mentioned method to distilled water, and performing resuscitation culture under aerobic oscillation conditions to obtain the resuscitated inoculant;

[0037] X2) Adding the supernatant of the resuscitated inoculant to heparin sodium wastewater, and treating the heparin sodium wastewater under aerobic conditions.

[0038] Furthermore, the ratio of the composite microbial inoculant to the heparin sodium wastewater can be (1-10) g: 250 mL, specifically 5 g: 250 mL (1 g: 50 mL).

[0039] Still further, in the above X1), the conditions for the resuscitation culture can be shaking culture at 20-40 °C and 150-200 rpm for 1-4 h, specifically 30 °C, 160 rpm, and shaking resuscitation for 2 h under aerobic conditions.

[0040] In the above X2), the conditions for the treatment can be 20-40 °C, 150-200 rpm, and treatment for 5-7 days under aerobic conditions, specifically 30 °C, 160 rpm, and treatment for 7 days under aerobic conditions.

[0041] Compared with the prior art, the composite microbial inoculant technology of the present invention is simpler and more convenient, easier to operate, lower in cost, higher in effluent quality, and has been verified to be shock-resistant and stable in operation. Specifically, it is reflected in the following aspects:

[0042] 1. In each composite microbial inoculant combination of the present invention, each strain is cultured using a unified culture medium. On the premise of ensuring the optimal propagation conditions of the strains, the preparation steps of the inoculant combination are simplified, and the cost is saved.

[0043] 2. Each composite microbial agent combination of the present invention is composed of multiple strains of bacteria. Each strain of bacteria plays a synergistic role in sewage treatment, quickly forming a dominant flora and rapidly decomposing various eutrophic and degradable harmful substances in the heparin sodium wastewater.

[0044] 3. Using the composite microbial agent of the present invention to treat the pollution problem of heparin sodium wastewater has a short treatment cycle and can effectively solve the problems of high COD and high ammonia nitrogen in the heparin sodium wastewater.

[0045] 4. Using the composite microbial agent of the present invention to treat water bodies can achieve the purpose of treating both the symptoms and the root causes and will not cause secondary pollution.

[0046] 5. Using the composite microbial agent of the present invention to treat water bodies has the characteristics of safety, high efficiency, low cost, low investment, and simple operation. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 It is the COD detection results and COD removal rates at different time points during the treatment of heparin sodium wastewater by the composite microbial agent.

[0048] Figure 2 It is the ammonia nitrogen detection results and ammonia nitrogen removal rates at different time points during the treatment of heparin sodium wastewater by the composite microbial agent. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0049] The present invention will be further described in detail below in conjunction with the specific embodiments. The examples given are only for clarifying the present invention and not for limiting the scope of the present invention. The following examples can be used as a guide for those of ordinary skill in the art to make further improvements and do not constitute any limitation to the present invention in any way.

[0050] The experimental methods in the following examples are all conventional methods unless otherwise specified, and are carried out according to the techniques or conditions described in the literature in this field or according to the product instructions. The materials, reagents, etc. used in the following examples can be obtained from commercial sources unless otherwise specified.

[0051] Example 1. Preparation of the composite microbial agent

[0052] The composite microbial inoculant of the present invention comprises Bacillus, Bacteroides, Lactobacillus, Pseudomonas, Acinetobacter, an auxiliary bacterium genus and a nutrient agent. The auxiliary bacterium genus includes Sphingomonas, Curtobacterium, Azotobacter and Methylobacterium. According to different weight ratios of each component in the composite microbial inoculant, composite microbial inoculants A, B, C, D and E are respectively prepared. The specific preparation method is as follows:

[0053] 1. Respectively inoculate the strains of Bacillus, Bacteroides, Lactobacillus, Pseudomonas, Acinetobacter and the auxiliary bacterium genus into a liquid medium, and culture at 30 °C, 160 rpm under aerobic conditions for 24 - 48 h to respectively obtain the seed solutions of Bacillus strains, Bacteroides strains, Lactobacillus strains, Pseudomonas strains, Acinetobacter strains and the auxiliary bacterium genus strains.

[0054] The solvent of the above liquid medium is water, and the solutes and their concentrations are respectively 10.00 g / L of tryptone, 5.00 g / L of yeast extract powder, 10.00 g / L of NaCl, and pH = 7.00.

[0055] 2. Respectively inoculate the seed solutions of each strain obtained in step 1 into a fermentation medium (nutrient agent) according to a weight ratio of 3.00 - 5.67%, and perform fermentation culture. Culture at a tank pressure of 0.02 MPa, a temperature of 30 - 35 °C, a pH of 6.8 - 7.2, a rotation speed of 150 - 180 rpm, and a dissolved oxygen > 6 mg / L for 24 - 48 h to respectively obtain the fermentation broths of Bacillus strains, Bacteroides strains, Lactobacillus strains, Pseudomonas strains, Acinetobacter strains and the auxiliary bacterium genus strains.

[0056] The solvent of the above fermentation medium is water, and the solutes and their concentrations are respectively 4 g / L of corn steep liquor dry powder, 2 g / L of soybean meal powder, 3 g / L of wall - broken yeast powder (wall - broken yeast - yeast product - Xuzhou Saifu Biotechnology Co., Ltd.), 3 g / L of beef extract, 2 g / L of fish meal peptone, 5 g / L of glucose, 0.1 g / L of magnesium sulfate, 0.08 g / L of dipotassium hydrogen phosphate, 0.02 g / L of manganese sulfate, 0.1 g / L of sodium chloride, and 0.01 g / L of Tween 80.

[0057] 3. Mix the fermentation broth of each strain obtained in step 2 with the fermentation medium (nutrient agent) according to the weight ratio (%) of each strain corresponding to compound microbial inoculant A in Table 1, and culture it for 24 - 48 h under the conditions of a tank pressure of 0.02 MPa, a temperature of 30 - 35 °C, a pH of 6.8 - 7.2, a rotation speed of 150 - 180 rpm, and a dissolved oxygen > 6 mg / L until the viable cell count reaches (1 - 9)×10 9 cfu / mL to obtain compound microbial liquid A.

[0058] Mix the strains obtained in step 2 with the fermentation medium (nutrient agent) according to the weight ratio (%) of each strain corresponding to compound microbial inoculant B in Table 1, and culture it for 24 - 48 h under the conditions of a tank pressure of 0.02 MPa, a temperature of 30 - 35 °C, a pH of 6.8 - 7.2, a rotation speed of 150 - 180 rpm, and a dissolved oxygen > 6 mg / L until the viable cell count reaches (1 - 9)×10 9 cfu / mL to obtain compound microbial liquid B.

[0059] Mix the strains obtained in step 2 with the fermentation medium (nutrient agent) according to the weight ratio (%) of each strain corresponding to compound microbial inoculant C in Table 1, and culture it for 24 - 48 h under the conditions of a tank pressure of 0.02 MPa, a temperature of 30 - 35 °C, a pH of 6.8 - 7.2, a rotation speed of 150 - 180 rpm, and a dissolved oxygen > 6 mg / L until the viable cell count reaches (1 - 9)×10 9 cfu / mL to obtain compound microbial liquid C.

[0060] Mix the strains obtained in step 2 with the fermentation medium (nutrient agent) according to the weight ratio (%) of each strain corresponding to compound microbial inoculant D in Table 1, and culture it for 24 - 48 h under the conditions of a tank pressure of 0.02 MPa, a temperature of 30 - 35 °C, a pH of 6.8 - 7.2, a rotation speed of 150 - 180 rpm, and a dissolved oxygen > 6 mg / L until the viable cell count reaches (1 - 9)×10 9 cfu / mL to obtain compound microbial liquid D.

[0061] Mix the strains obtained in step 2 with the fermentation medium (nutrient agent) according to the weight ratio (%) of each strain corresponding to compound microbial inoculant E in Table 1, and culture it for 24 - 48 h under the conditions of a tank pressure of 0.02 MPa, a temperature of 30 - 35 °C, a pH of 6.8 - 7.2, a rotation speed of 150 - 180 rpm, and a dissolved oxygen > 6 mg / L until the viable cell count reaches (1 - 9)×10 9 cfu / mL to obtain compound microbial liquid E.

[0062] In Table 1, the weight ratios of Sphingomonas, Curtobacterium, Azovibrio, and Methylobacterium in the auxiliary bacteria genus are 8:4:3:3.

[0063] Table 1

[0064]

[0065] 4. Freeze-dry the composite microbial inoculum A-E obtained in step 3 at -80 °C for 48 hours to obtain composite microbial agents A-E respectively.

[0066] Example 2. Application of composite microbial agent in treating heparin sodium wastewater

[0067] I. Method for treating heparin sodium wastewater with composite microbial agent

[0068] Treat heparin sodium wastewater with the composite microbial agents A-E prepared in Example 1 respectively. The specific steps are as follows:

[0069] 1. Take 5 g of the composite microbial agent and add it to 50 mL of distilled water. Place it in a shaker and shake for resuscitation for 2 h under the conditions of 30 °C, 160 rpm, and aerobic to obtain the resuscitated agent.

[0070] 2. After the resuscitated agent is allowed to stand for 5 min, take 50 mL of the supernatant and inoculate it all into 250 mL of heparin sodium wastewater (production wastewater of Yinuorui Company). Treat the heparin sodium wastewater under the conditions of 30 °C, 160 rpm, and aerobic for 7 consecutive days. Sample and detect the water quality indexes of COD value (GB11914-89 Determination of Chemical Oxygen Demand in Water - Dichromate Method) and ammonia nitrogen value (GB7479-87 Determination of Ammonium in Water - Nessler's Reagent Colorimetric Method) every 1 day, and calculate the COD removal rate and ammonia nitrogen removal rate.

[0071] COD removal rate = (Initial COD content - COD content on the seventh day) / Initial COD content * 100%.

[0072] Ammonia nitrogen removal rate = (Initial ammonia nitrogen content - Ammonia nitrogen content on the seventh day) / Initial ammonia nitrogen content * 100%.

[0073] II. Effect detection of treating heparin sodium wastewater with composite microbial agent

[0074] 1. COD

[0075] The detection results of COD value and the calculation results of COD removal rate are as Figure 1 shown.

[0076] The results show that after adding five different composite microbial agents to the heparin sodium wastewater respectively, the COD value in the wastewater gradually decreases. After one week, the COD removal effects of the five composite microbial agents on the wastewater all reach more than 85%, among which the composite microbial agents A and D have the best treatment effects, and the removal rate is as high as 92%.

[0077] 2. Ammonia nitrogen

[0078] The detection results of ammonia nitrogen values and the calculation results of ammonia nitrogen removal rates are as Figure 2 shown.

[0079] The results show that after adding five different composite microbial agents to the heparin sodium wastewater respectively, the ammonia nitrogen value in the wastewater basically shows a downward trend. After one week, the ammonia nitrogen removal effects of the three composite microbial agents A, B, and E on the wastewater are not good, only about 10%, but the ammonia nitrogen removal rates of the composite microbial agents C and D reach about 60%.

[0080] Therefore, generally speaking, the composite microbial agent D prepared by the present invention can effectively solve the problems of high COD and high ammonia nitrogen in the heparin sodium wastewater.

[0081] The above has detailed the present invention. For those skilled in the art, without departing from the purpose and scope of the present invention and without unnecessary experiments, the present invention can be implemented within a relatively wide range under equivalent parameters, concentrations, and conditions. Although specific embodiments of the present invention are given, it should be understood that the present invention can be further improved. In short, according to the principle of the present invention, this application intends to include any changes, uses, or improvements to the present invention, including those that depart from the scope disclosed in this application and are made with conventional techniques known in the art. Some basic features can be applied according to the scope of the following appended claims.

Claims

1. A preparation method of a compound microbial inoculum, comprising the following steps: 1) Respectively inoculate Bacillus species, Bacteroides species, Lactobacillus species, Pseudomonas species, Acinetobacter species, and Auxiliary Bacteria species into a liquid medium for seed culture to obtain Bacillus species seed liquid, Bacteroides species seed liquid, Lactobacillus species seed liquid, Pseudomonas species seed liquid, Acinetobacter species seed liquid, and Auxiliary Bacteria species seed liquid respectively; 2) Respectively inoculate the Bacillus species seed liquid, the Bacteroides species seed liquid, the Lactobacillus species seed liquid, the Pseudomonas species seed liquid, the Acinetobacter species seed liquid, and the Auxiliary Bacteria species seed liquid into a nutrient agent for fermentation culture to obtain Bacillus species fermentation liquid, Bacteroides species fermentation liquid, Lactobacillus species fermentation liquid, Pseudomonas species fermentation liquid, Acinetobacter species fermentation liquid, and Auxiliary Bacteria species fermentation liquid respectively; 3) Mix the fermentation broth of the Bacillus species, the fermentation broth of the Bacteroides species, the fermentation broth of the Lactobacillus species, the fermentation broth of the Pseudomonas species, the fermentation broth of the Acinetobacter species and the fermentation broth of the auxiliary bacteria species with the nutrient agent, and culture until the viable count is at least 1×10 9 cfu / mL to obtain a composite microbial liquid; 4) Freeze-dry the compound microbial liquid to make a compound microbial inoculum; The Auxiliary Bacteria species consists of Sphingomonas species, Curtobacterium species, Azogloea species, and Methylobacterium species; The nutrient agent consists of corn steep liquor dry powder, soybean meal powder, broken wall yeast powder, beef extract, fish meal peptone, glucose, magnesium sulfate, dipotassium hydrogen phosphate, manganese sulfate, sodium chloride, and Tween 80; In step 3), the weight ratio of the Bacillus species fermentation liquid, the Bacteroides species fermentation liquid, the Lactobacillus species fermentation liquid, the Pseudomonas species fermentation liquid, the Acinetobacter species fermentation liquid, the Auxiliary Bacteria species fermentation liquid, and the nutrient agent is 78.86 : 0.41 : 0.01 : 0.33 : 0.12 : 5.27 : 15.00 or 73.91 : 0.19 : 6.36 : 0.80 : 0.02 : 3.72 : 15.00 or 71.91 : 0.92 : 0.02 : 1.40 : 0.29 : 5.46 : 20.00 or 56.15 : 1.31 : 0.05 : 4.77 : 5.05 : 7.61 : 25.00 or 79.65 : 0.63 : 0.02 : 0.57 : 0.17 : 3.66 : 15.00; In the Auxiliary Bacteria species fermentation liquid, the weight ratio of the Sphingomonas species fermentation liquid, the Curtobacterium species fermentation liquid, the Azogloea species fermentation liquid, and the Methylobacterium species fermentation liquid is 9:5:3:3 or 10:3:4:3 or 8:4:3:

3.

2. The method according to claim 1, wherein: In step 1), the liquid medium includes tryptone, yeast extract powder, and NaCl.

3. The method according to claim 1, characterized in that: In step 1), the conditions for the seed culture are culturing at 30 - 35 °C, 150 - 180 rpm, under aerobic conditions for 24 - 48 h.

4. The method according to claim 1, characterized in that: In the above (2), each strain seed solution is inoculated into the nutrient agent according to a weight ratio of 3.00 - 5.67%.

5. The method according to claim 1, wherein: In the above (2), the solvent of the nutrient agent is water, and the solutes and their concentrations are respectively 4 g / L of corn steep liquor powder, 2 g / L of soybean meal powder, 3 g / L of broken wall yeast powder, 3 g / L of beef extract, 2 g / L of fish meal peptone, 5 g / L of glucose, 0.1 g / L of magnesium sulfate, 0.08 g / L of dipotassium hydrogen phosphate, 0.02 g / L of manganese sulfate, 0.1 g / L of sodium chloride, and 0.01 g / L of Tween 80.

6. The method according to claim 1, wherein: In the above (2), the culture conditions are a tank pressure of 0.02 - 0.04 MPa, a temperature of 30 - 35 °C, a pH of 6.8 - 7.2, a rotation speed of 150 - 180 rpm, and a dissolved oxygen > 6 mg / L.

7. The method according to claim 1, characterized in that: In the above (3), the culture conditions are a tank pressure of 0.02 - 0.04 MPa, a temperature of 30 - 35 °C, a pH of 6.8 - 7.2, a rotation speed of 150 - 180 rpm, and a dissolved oxygen > 6 mg / L.

8. The method according to claim 1, characterized in that: In the above (3), the cultivation is carried out until the viable cell count reaches (1-9)×10 9 cfu / mL.

9. The method according to claim 1, wherein: In the above (4), the temperature of the freeze-drying is (-80 - -60) °C for 48 hours of freeze-drying.

10. Application of the composite microbial inoculant prepared by the method according to any one of claims 1 - 9 in treating heparin sodium wastewater.

11. The application according to claim 10, wherein: The treatment of heparin sodium wastewater is manifested as reducing the COD content and ammonia nitrogen content in the heparin sodium wastewater.

12. A method for treating heparin sodium wastewater, comprising the following steps: X1) Adding the composite microbial inoculant prepared by the method according to any one of claims 1 - 9 into distilled water, and performing resuscitation culture under aerobic oscillation conditions to obtain the resuscitated inoculant; X2) Adding the supernatant of the resuscitated inoculant into the heparin sodium wastewater, and treating the heparin sodium wastewater under aerobic conditions.

13. The method according to claim 12, wherein: In the above X1), the ratio of the composite microbial inoculant to the heparin sodium wastewater is (1 - 10) g : 250 mL.

14. The method according to claim 12, characterized in that: In the above X1), the conditions for the resuscitation culture are shaking culture for 1 - 4 h under aerobic conditions at 20 - 40 °C and 150 - 200 rpm.

15. The method according to claim 12, characterized in that: In the above X2), the conditions for the treatment are 20 - 40 °C, 150 - 200 rpm, and treatment for 5 - 7 days under aerobic conditions.

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