Microbial agent for decomposing pollutants, and preparation method and application thereof

Abstract

The application provides a microbial agent for decomposing pollutants and a preparation method and application thereof, and the microbial agent for decomposing pollutants comprises the following raw materials in parts by weight: 40-80 parts of compound bacteria, 15-25 parts of enzyme preparation, 10-15 parts of growth substrate, 3-6 parts of nutrient element, 1-3 parts of stress resistance regulator, 0-0.15 part of azido-erythro-sphingosine, and 0-0.05 part of polyvinyl caprolactam; wherein the compound bacteria are one or more of Bacillus megaterium, Bacillus subtilis, Bacillus pumilus, Pseudomonas fluorescens, Bacillus licheniformis and Bifidobacterium pseudolongum; and the stress resistance regulator is a mixture of ethoxylated castor oil and dodecyl hydroxyl poly (oxy-1,2-ethanediyl). The microbial agent for decomposing pollutants has less influence on salt components and toxic substances in wastewater, can still achieve high decomposition efficiency in an environment with high salt content and toxic substances, and has good treatment effect on petrochemical wastewater.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of sewage treatment, in particular to a microbial agent for decomposing pollutants and a preparation method and application thereof. BACKGROUND

[0002] Petroleum chemical industry is a chemical treatment process taking petroleum as the main production raw material, mainly including cracking, fractionation, reforming and synthesis of petroleum, etc. In the above production and processing processes, a large amount of petroleum chemical wastewater is formed. The pollutants in the wastewater are of multiple types, high concentration, complex composition, high toxicity and high salt content, and the wastewater has the characteristics of large water quantity, complex water quality, large variation and serious organic pollution, etc., and is very difficult to treat.

[0003] At present, the treatment methods of petroleum chemical wastewater can be mainly divided into three categories of physical treatment, chemical treatment and biological treatment according to the treatment principle. Physical treatment mainly separates and recovers the insoluble suspended pollutants in wastewater through physical action; chemical treatment separates and recovers the pollutants in wastewater through chemical reaction by adding chemical agents to the wastewater; biological treatment converts the organic pollutants in solution, colloid and fine suspended state in wastewater into stable and harmless substances through the metabolic action of microorganisms.

[0004] Among them, the biological treatment is the most difficult to control. Because the petroleum chemical wastewater has high salt content and contains certain toxic substances, some substances toxic to microorganisms, such as hypochlorous acid, hypochlorite ion and isothiazolinone, are introduced or produced at the front end of biological treatment. These substances can inhibit biodegradation, reduce the decomposition efficiency of pollutants and affect the wastewater treatment effect. SUMMARY

[0005] The purpose of the present application is to provide a microbial agent for decomposing pollutants and a preparation method and application thereof, so as to solve the problem of low decomposition efficiency of microorganisms and poor wastewater treatment effect in the prior art when the petroleum chemical wastewater is biologically treated.

[0006] The present application provides the following technical solutions:

[0007] A microbial agent for decomposing pollutants comprises the following raw materials by weight:

[0008] 40-80 parts of complex bacteria, 15-25 parts of enzyme preparation, 10-15 parts of growth substrate, 3-6 parts of nutrient element, 1-3 parts of stress resistance regulator, 0-0.15 part of azido-erythro-sphingosine and 0-0.05 part of polyvinyl caprolactam;

[0009] The complex bacteria are one or more of Bacillus megaterium, Bacillus subtilis, Bacillus pumilus, Pseudomonas fluorescens, Bacillus licheniformis and Bifidobacterium longum.

[0010] The anti-stress regulator is a mixture of ethoxylated castor oil and dodecyl hydroxyl poly (oxy-1, 2-ethanediy).

[0011] Preferably, the complex bacteria are mixed in a weight ratio of 1: (0.5-1): (0.5-0.8): (1-1.05): (1-2): (0.1-0.3) of Bacillus megaterium, Bacillus subtilis, Bacillus pumilus, Pseudomonas fluorescens, Bacillus licheniformis and Bifidobacterium longum.

[0012] Optionally, the anti-stress regulator is a mixture of ethoxylated castor oil and dodecyl hydroxyl poly (oxy-1, 2-ethanediy) in a weight ratio of 1: (0.2-1).

[0013] Preferably, the pollutant-decomposing microbial agent further comprises 0.5-1.0 parts of vitamins; the vitamins include one or more of vitamin A, vitamin B, vitamin D, vitamin E and vitamin K.

[0014] Optionally, the enzyme preparation includes one or more of peroxidase, reductase, cellulase, amylase and protease.

[0015] Optionally, the growth substrate includes one or more of glucose, molasses, corn powder and soybean powder.

[0016] Optionally, the nutritional elements include one or more of calcium, iron, magnesium, zinc, molybdenum and tungsten.

[0017] Preferably, the molybdenum is ammonium molybdate; the tungsten is one or more of ammonium tungstate and ferrous tungstate.

[0018] Preferably, the pollutant-decomposing microbial agent further comprises 0.05-0.08 parts of polyvinyl pyrrolidone.

[0019] The application further provides a preparation method of the pollutant-decomposing microbial agent, comprising the following steps: taking the complex bacteria, enzyme preparation, growth substrate, nutritional elements, anti-stress regulator, azido-erythro-sphingosine and polyvinyl caprolactam in weight parts, and mixing uniformly to obtain the pollutant-decomposing microbial agent.

[0020] Preferably, the complex bacteria have an effective viable count greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of Bacillus megaterium, an effective viable count greater than or equal to 1.0x10 10Bacillus subtilis fermentation broth with CFU / ml of 1.0x10 10 Bacillus pumilus fermentation broth with CFU / ml of 1.0x10 10 Pseudomonas fluorescens fermentation broth with CFU / ml of 1.0x10 10 Bacillus licheniformis fermentation broth with CFU / ml of 1.0x10 10 Bifidobacterium pseudolongum fermentation broth with CFU / ml of 1.0x10

[0021] The complex bacteria has effective viable count of 1.0x10 10 Bacillus megaterium fermentation broth with CFU / ml of 1.0x10 10 Bacillus subtilis fermentation broth with CFU / ml of 1.0x10 10 Bacillus pumilus fermentation broth with CFU / ml of 1.0x10 10 Pseudomonas fluorescens fermentation broth with CFU / ml of 1.0x10 10 Bacillus licheniformis fermentation broth with CFU / ml of 1.0x10 10 Bifidobacterium pseudolongum fermentation broth with CFU / ml of 1.0x10

[0022] Preferably, the preparation method of the complex bacteria comprises the following steps:

[0023] (1) take each strain, inoculate on the activated culture medium, culture at 35-37℃ for 10-30h, to obtain the activated bacteria;

[0024] (2) inoculate the activated bacteria in step (1) on the flat plate culture medium, culture at 35-37℃ for 22-26h, to obtain the flat plate cultured colonies;

[0025] (3) inoculate the flat plate cultured colonies in step (2) on the slant culture medium, culture at 35-37℃ for 22-26h, to obtain the slant cultured bacteria;

[0026] (4) inoculate the slant cultured bacteria in step (3) in the shake flask culture medium with inoculation amount of 3-5%, culture at 35-37℃ with the rotation speed of 140-160r / min for 22-26h, to obtain the shake flask fermentation bacteria;

[0027] (5) The shake flask fermentation bacteria in step (4) are inoculated into seed culture medium at an inoculation amount of 6-10%, and are fermented at a temperature of 35-37°C and a stirring speed of 140-160 r / min for 46-50 h to obtain seed culture bacteria;

[0028] (6) The seed culture bacteria in step (5) are inoculated into fermentation tank culture medium, and are fermented at a temperature of 35-37°C and a stirring speed of 140-160 r / min for 46-50 h to obtain fermentation liquor of each bacterial strain.

[0029] (7) The fermentation liquor of each bacterial strain in step (6) is prepared into a composite bacteria according to weight parts.

[0030] Preferably, step (7) comprises the following steps:

[0031] (a) The fermentation liquor of Bacillus megaterium, Bacillus subtilis, Bacillus pumilus, Pseudomonas fluorescens, Bacillus licheniformis and Bifidobacterium pseudolongum is taken according to weight parts, respectively, an adsorption medium is added, and the mixture is uniformly mixed to obtain a mixture;

[0032] The adsorption medium is one of flour, corn flour and soybean meal; and the weight ratio of the adsorption medium to the fermentation liquor is (1-3):1.

[0033] (b) The mixture in step (a) is dried to a water content of less than or equal to 10 wt% to obtain solid bacteria of Bacillus megaterium, Bacillus subtilis, Bacillus pumilus, Pseudomonas fluorescens, Bacillus licheniformis and Bifidobacterium pseudolongum.

[0034] (c) The solid bacteria of Bacillus megaterium, Bacillus subtilis, Bacillus pumilus, Pseudomonas fluorescens, Bacillus licheniformis and Bifidobacterium pseudolongum obtained in step (b) are uniformly mixed to obtain the composite bacteria.

[0035] Preferably, in step (a), the adsorption medium further contains bentonite; and the content of the bentonite in the adsorption medium is 10-25 wt%.

[0036] The bentonite is modified bentonite after intercalation treatment of lactamide propyl trimethyl ammonium chloride.

[0037] Preferably, the bentonite is modified bentonite after intercalation treatment of lactamide propyl trimethyl ammonium chloride.

[0038] The modified bentonite is obtained by the following method: sodium-based bentonite, lactamide propyl trimethyl ammonium chloride and deionized water are uniformly mixed according to a weight ratio of 1:(0.05-0.1):(10-20), stirred at 50-80°C for 2-4 h, statically aged for 12-24 h, and then centrifuged, washed and dried to obtain the modified bentonite.

[0039] The application further provides application of the pollutant-decomposing microbial agent in the field of sewage treatment.

[0040] The above scheme of the application has at least the following beneficial effects:

[0041] (1) The pollutant-decomposing microbial agent comprises the following raw materials in parts by weight: 40-80 parts of compound bacteria, 15-25 parts of enzyme preparation, 10-15 parts of growth substrate, 3-6 parts of nutrient element, 1-3 parts of stress resistance regulator, 0-0.15 part of azido-erythro-sphingosine, and 0-0.05 part of polyvinyl caprolactam; wherein the compound bacteria are one or more of Bacillus megaterium, Bacillus subtilis, Bacillus pumilus, Pseudomonas fluorescens, Bacillus licheniformis, and Bifidobacterium pseudolongum; and the stress resistance regulator is a mixture of ethoxylated castor oil and dodecylhydroxy poly (oxy-1,2-ethanediyl). The pollutant-decomposing microbial agent has less influence on salt components and toxic substances in wastewater, and can still achieve high decomposition efficiency in an environment with high salt content and toxic substances, and has good treatment effect on petrochemical wastewater.

[0042] The growth substrate can provide an energy source for the growth of microorganisms in the compound bacteria. The enzyme preparation can promote metabolic conversion of the growth substrate by the microorganisms in the compound bacteria, and can also metabolically convert target pollutants, and can produce a co-metabolic reaction during biochemical reaction. The nutrient element can promote rapid growth of the microorganisms in the compound bacteria.

[0043] The ethoxylated castor oil has lipophilic castor oil fatty acids, which can interact with lipid components on the cell surface and be adsorbed on the cell surface, and the ethoxylated chain has good flexibility, so that the ethoxylated castor oil forms a protective film with certain fluidity and flexibility on the surface of the microorganisms, and plays a good isolation and protection role. The dodecylhydroxy poly (oxy-1,2-ethanediyl) can improve the permeability of the microbial cell membrane, promote the microorganisms to better regulate the internal and external ion balance and discharge of metabolic products, thereby reducing the damage of high salt to the microorganisms. When used together, the two can significantly improve the stress resistance of the microorganisms in the compound bacteria, promote the microorganisms in the compound bacteria to grow rapidly in harsh environments, form a bacterial sludge, weaken the inhibition of high salt content and toxic substances on biodegradation, improve the decomposition efficiency of pollutants, and achieve good wastewater treatment effect.

[0044] The azido group of the azido-erythro-sphingosine has high reactivity, can react with the unsaturated fatty acid chain in the lipid molecules of the cell membrane of the microorganism, and make the sphingosine filled in the lipid bilayer of the cell membrane, the long chain structure of the sphingosine can play a supporting and connecting role in the lipid bilayer, prevent the cell membrane from being broken or having a loophole due to the change of the fatty acid chain, enhance the structural strength of the cell membrane, enhance the tolerance of the cell membrane to toxic substances, maintain the integrity of the cell membrane, maintain the stability of the intracellular environment, and enable the microorganism to maintain normal metabolic and physiological functions in a harsh environment, so as to play a more long-acting role in degrading pollutants. Especially, when used with the stress resistance regulator, the activity of the microorganism in the complex bacteria in the environment with high salt content and toxic substances can be greatly improved, and a good pollutant decomposition effect can be achieved.

[0045] The chain-connected caprolactam ring of the polyvinyl caprolactam can form a dynamic fracture-recombination hydration layer around the microorganism, which helps the microorganism to maintain the cell osmotic pressure balance in a high-osmotic environment and prevent dehydration inactivation.

[0046] (2) The pollutant-decomposing microbial agent of the present application, the nutrient elements include one or more of calcium, iron, magnesium, zinc, molybdenum and tungsten. The molybdenum is ammonium molybdate; the tungsten is one or more of ammonium tungstate and ferrous tungstate.

[0047] The ammonium molybdate can participate in the redox reaction in the organism, act as a cofactor of enzyme preparation, and further improve the enzyme activity; and the ammonium molybdate has multiple biological activities such as anti-inflammatory, antioxidant and antitumor, and can help the microorganism to maintain normal physiological functions. The ferrous tungstate can act as a catalytic factor for the growth and reproduction of the microorganism, and promote the growth of the microorganism. DETAILED DESCRIPTION

[0048] In the embodiments of the present application, the specific conditions not specified are carried out according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used are not specified by the manufacturer, and are all conventional products that can be obtained by market purchase. Different manufacturers and models of raw materials do not affect the implementation of the technical solutions and the realization of the technical effects of the present application.

[0049] In the following examples, the Bacillus megatherium, Bacillus subtilis, Bacillus pumilus, Pseudomonas fluorescens, Bacillus licheniformis, Bifidobacterium pseudolongum are all products available in the prior art and can be obtained by purchase. For example, the Bacillus megatherium, with the preservation number ZKCC15770 or ATCC 35985; the Bacillus subtilis, with the preservation number CMCC(B)63501; the Bacillus pumilus, with the preservation number CMCC(B)63202; the Pseudomonas fluorescens Migula, with the preservation number ATCC 13525; the Bacillus licheniformis, with the preservation number ATCC 11946; the Bifidobacterium pseudolongum, with the preservation number ZKCC24853.

[0050] The azido-erythro-sphingosine, also known as azido-erythro-sphingosine, (2S, 3R, 4E)-2-azido-4-octadecen-1, 3-diol, has a CAS number of 103348-49-8. The ethoxylated castor oil has a CAS number of 61791-12-6. The dodecylhydroxy poly (oxy-1, 2-ethanediyl), also known as lauryl polyoxyethylene ether, has a CAS number of 9002-92-0. The lactamidopropyl trimethyl ammonium chloride has a CAS number of 93507-51-8. The polyvinyl caprolactam has a CAS number of 25189-83-7.

[0051] Example 1

[0052] The pollutant-decomposing microbial agent of the present example comprises the following raw materials in parts by weight:

[0053] 40 parts of composite bacteria, 20 parts of enzyme preparation, 10 parts of growth substrate, 5 parts of nutritional elements, 1 part of stress resistance regulator, 0.10 part of azido-erythro-sphingosine, and 0.5 part of vitamin;

[0054] The composite bacteria are a mixture of Bacillus megatherium, Bacillus subtilis, Bacillus pumilus, Pseudomonas fluorescens, Bacillus licheniformis, and Bifidobacterium pseudolongum in a weight ratio of 1:1:0.6:1.05:2:0.2. The stress resistance regulator is a mixture of ethoxylated castor oil and dodecylhydroxy poly (oxy-1, 2-ethanediyl) in a weight ratio of 1:0.6.

[0055] The vitamins include vitamin A, vitamin B, vitamin D, vitamin E and vitamin K mixed in a weight ratio of 1:1:1:1:1; the enzyme preparation is peroxidase; the growth substrate is glucose; and the nutritional elements include calcium, iron, magnesium, zinc, molybdenum and tungsten mixed in a weight ratio of 4:1:1:1:3:3. The molybdenum is ammonium molybdate, the tungsten is ammonium tungstate, the calcium is calcium chloride, the iron is iron chloride, the magnesium is magnesium chloride, and the zinc is zinc chloride.

[0056] The preparation method of the microorganism agent for decomposing pollutants comprises the following steps: taking the compound bacteria, enzyme preparation, growth substrate, nutritional elements, stress resistance regulator and azido-erythro-sphingosine by weight, and mixing uniformly to obtain the product.

[0057] The compound bacteria have an effective viable count of greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of Bacillus megaterium, an effective viable count of greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of Bacillus subtilis, an effective viable count of greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of Bacillus pumilus, an effective viable count of greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of Pseudomonas fluorescens, an effective viable count of greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of Bacillus licheniformis, an effective viable count of greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of Bifidobacterium pseudolongum, and mixing to obtain the product. In the embodiment, the effective viable count of each of the fermentation liquors of the compound bacteria is in the range of 1.0x10 10 CFU / ml to 1.2x10 10 CFU / ml.

[0058] It should be noted that the fermentation liquors of each of the bacteria in the compound bacteria can be prepared by using the prior art, and the culture medium, culture conditions and culture steps can also be achieved by using the prior art, as long as the effective viable count can reach the above-mentioned standard.

[0059] In the embodiment, the preparation method of the compound bacteria comprises the following steps:

[0060] (1) taking each of the bacteria, inoculating on an activated culture medium, and culturing at 35℃ for 20h to obtain the activated bacteria;

[0061] (2) inoculating the activated bacteria in step (1) on a flat plate culture medium, and culturing at 35℃ for 22h to obtain the flat plate cultured colonies;

[0062] (3) Inoculate the colonies cultured on the plate in step (2) onto the slant culture medium and culture at 35°C for 22 h to obtain the bacterial moss cultured on the slant.

[0063] (4) The bacterial culture from the slant culture in step (3) is inoculated into the shake flask culture medium at an inoculation rate of 3%, and cultured at a shaker speed of 160 r / min and 35℃ for 22 h to obtain the shake flask fermentation cells;

[0064] (5) The shake-flask fermentation cells from step (4) were inoculated into the seed culture medium at an inoculation rate of 8%, and fermented for 48 hours at a temperature of 35°C and a stirring speed of 160 r / min to obtain the seed culture cells.

[0065] (6) The seed culture cells from step (5) are inoculated into the culture medium of the fermenter and fermented for 48 hours at a temperature of 35℃ and a stirring speed of 160r / min to obtain the fermentation broth of each strain.

[0066] (7) Mix the fermentation liquid of each strain in step (6) evenly according to the weight parts to obtain the compound strain.

[0067] Example 2

[0068] The microbial agent for decomposing pollutants in this embodiment includes the following raw materials in parts by weight:

[0069] 80 parts of compound bacteria, 15 parts of enzyme preparation, 15 parts of growth substrate, 6 parts of nutrients, 2 parts of stress resistance regulator, 0.15 parts of azido-erythrosine, and 1.0 part of vitamins;

[0070] The compound bacteria are composed of Bacillus megaterium, Bacillus subtilis, Bacillus pumilus, Pseudomonas fluorescens, Bacillus licheniformis, and Bifidobacterium pseudolongum in a weight ratio of 1:0.5:0.8:1:1.5:0.1. The stress-resistance regulator is composed of ethoxylated castor oil and dodecyl hydroxy poly(oxy-1,2-ethylenedimethyl) in a weight ratio of 1:1.

[0071] The vitamins include vitamins A, B, D, E, and K mixed in a weight ratio of 1:3:2:2:1; the enzyme preparation is a reductase; the growth substrate is corn flour; the nutrients include calcium, iron, magnesium, zinc, molybdenum, and tungsten mixed in a weight ratio of 2:2:3:1:3:2. The molybdenum is ammonium molybdate, the tungsten is ferrous tungstate, the calcium is calcium sulfate, the iron is ferric chloride, the magnesium is magnesium chloride, and the zinc is zinc chloride.

[0072] The preparation method of the microorganism agent for decomposing pollutants comprises the following steps: taking the composite bacteria, enzyme preparation, growth substrate, nutrient element, stress resistance regulator and azido-erythro-sphingosine according to weight parts, and mixing uniformly, and then the microorganism agent is obtained.

[0073] The effective viable cell number of the composite bacteria is greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of the bacillus megaterium, the effective viable cell number of the fermentation liquor of the bacillus subtilis is greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of the bacillus pumilus, the effective viable cell number of the fermentation liquor of the pseudomonas fluorescens is greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of the bacillus licheniformis, and the effective viable cell number of the fermentation liquor of the bifidobacterium pseudolongum is greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of the bacillus licheniformis, and the effective viable cell number of the fermentation liquor of the bifidobacterium pseudolongum is greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of the bacillus licheniformis, and the effective viable cell number of the fermentation liquor of the bifidobacterium pseudolongum is greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of the bacillus licheniformis, and the effective viable cell number of the fermentation liquor of the bifidobacterium pseudolongum is greater than or equal to 1.0x10 10 CFU / ml to 1.2x10 10 CFU / ml.

[0074] It should be noted that the fermentation liquor of each bacterial species in the composite bacteria can be prepared by using the prior art, and the culture medium, culture conditions and culture steps can also be achieved by using the prior art, as long as the effective viable cell number can reach the above standard.

[0075] In this embodiment, the preparation method of the composite bacteria comprises the following steps:

[0076] (1) taking each bacterial species, inoculating on the activated culture medium, and culturing at 36℃ for 30h to obtain the activated bacterial body;

[0077] (2) inoculating the activated bacterial body in step (1) on the flat plate culture medium, and culturing at 36℃ for 24h to obtain the flat plate cultured bacterial colony;

[0078] (3) inoculating the flat plate cultured bacterial colony in step (2) on the slant culture medium, and culturing at 36℃ for 24h to obtain the slant cultured bacterial mat;

[0079] (4) inoculating the slant cultured bacterial mat in step (3) in the shake flask culture medium according to the inoculation amount of 5%, and culturing at 36℃ for 24h under the rotation speed of 150r / min to obtain the shake flask fermentation bacterial body;

[0080] (5) The shake flask fermentation bacteria in step (4) are inoculated into a seed culture medium at an inoculation amount of 6%, and are fermented at a temperature of 36℃ and a stirring speed of 150 r / min for 50 h to obtain seed culture bacteria;

[0081] (6) The seed culture bacteria in step (5) are inoculated into a fermentation tank culture medium, and are fermented at a temperature of 36℃ and a stirring speed of 150 r / min for 50 h to obtain fermentation liquor of each strain;

[0082] (7) The fermentation liquor of each strain in step (6) is prepared into a composite bacteria according to the weight parts, and specifically includes:

[0083] (a) The fermentation liquor of the Bacillus megaterium, Bacillus subtilis, Bacillus pumilus, Pseudomonas fluorescens, Bacillus licheniformis and Bifidobacterium pseudolongum is taken according to the weight parts, respectively, an adsorption medium is added, and the mixture is uniformly mixed to obtain a mixture;

[0084] The adsorption medium is corn powder; and the weight ratio of the adsorption medium to the fermentation liquor is 2:1.

[0085] (b) The mixture in step (a) is dried to a water content of less than or equal to 10 wt% to obtain the solid bacteria of the Bacillus megaterium, Bacillus subtilis, Bacillus pumilus, Pseudomonas fluorescens, Bacillus licheniformis and Bifidobacterium pseudolongum;

[0086] (c) The solid bacteria of the Bacillus megaterium, Bacillus subtilis, Bacillus pumilus, Pseudomonas fluorescens, Bacillus licheniformis and Bifidobacterium pseudolongum obtained in step (b) are uniformly mixed to obtain the composite bacteria.

[0087] Example 3

[0088] The pollutant-decomposing microbial agent of the present example comprises the following raw materials by weight:

[0089] 60 parts of composite bacteria, 25 parts of enzyme preparation, 14 parts of growth substrate, 3 parts of nutritional element, 3 parts of stress resistance regulator, 0.01 part of azido-erythro-sphingosine, and 0.8 part of vitamin;

[0090] The composite bacteria are a mixture of Bacillus megaterium, Bacillus subtilis, Bacillus pumilus, Pseudomonas fluorescens, Bacillus licheniformis and Bifidobacterium pseudolongum in a weight ratio of 1:0.7:0.5:1.02:1:0.3. The stress resistance regulator is a mixture of ethoxylated castor oil and dodecylhydroxy poly (oxy-1, 2-ethanediyl) in a weight ratio of 1:0.2.

[0091] The vitamins include vitamin A, vitamin B, vitamin D, vitamin E and vitamin K mixed in a weight ratio of 2:3:1:1:1; the enzyme preparation is amylase; the growth substrate includes glucose and corn flour mixed in a weight ratio of 1:1; and the nutritional elements include calcium, iron, magnesium, zinc, molybdenum and tungsten mixed in a weight ratio of 2:1:1:3:1:2. The molybdenum is ammonium molybdate, the tungsten is ammonium tungstate, the calcium is calcium chloride, the iron is iron chloride, the magnesium is magnesium chloride, and the zinc is zinc chloride.

[0092] The preparation method of the microorganism agent for decomposing pollutants comprises the following steps: taking the compound bacteria, enzyme preparation, growth substrate, nutritional elements, stress resistance regulator and azido-erythro-sphingosine in weight parts, and mixing them uniformly to obtain the product.

[0093] The compound bacteria have an effective viable count greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of Bacillus megaterium, an effective viable count greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of Bacillus subtilis, an effective viable count greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of Bacillus pumilus, an effective viable count greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of Pseudomonas fluorescens, an effective viable count greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of Bacillus licheniformis, an effective viable count greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of Bifidobacterium longum, and mixing them to obtain the product. 10 In the embodiment, the effective viable count of each of the fermentation liquors of the compound bacteria is in the range of 1.0x10 10 CFU / ml to 1.2x10

[0094] It should be noted that the fermentation liquors of each of the bacteria in the compound bacteria can be prepared by using the prior art, and the culture medium, culture conditions and culture steps can also be achieved by using the prior art, as long as the effective viable count can reach the above-mentioned standard.

[0095] In the embodiment, the preparation method of the compound bacteria comprises the following steps:

[0096] (1) Each of the bacteria is inoculated on an activated culture medium, and cultured at 37°C for 10h to obtain the activated bacterial bodies;

[0097] (2) The activated bacterial bodies in step (1) are inoculated on a flat plate culture medium, and cultured at 37°C for 26h to obtain the flat plate cultured colonies;

[0098] (3) the colonies of the plate culture in step (2) are inoculated on slant medium, and cultured at 37℃ for 26h to obtain the slant-cultured bacterial lawn;

[0099] (4) the slant-cultured bacterial lawn in step (3) is inoculated in a shake flask culture medium at an inoculation amount of 4%, and cultured at 37℃ at a shaking speed of 140r / min for 26h to obtain the shake flask fermentation bacteria;

[0100] (5) the shake flask fermentation bacteria in step (4) are inoculated in a seed culture medium at an inoculation amount of 10%, and fermented at a temperature of 37℃ and a stirring speed of 140r / min for 46h to obtain the seed culture bacteria;

[0101] (6) the seed culture bacteria in step (5) are inoculated in a fermentation tank culture medium, and fermented at a temperature of 37℃ and a stirring speed of 140r / min for 46h to obtain the fermentation liquor of each strain;

[0102] (7) the fermentation liquor of each strain in step (6) is mixed uniformly according to the weight parts to obtain the composite bacteria.

[0103] Example 4

[0104] The pollutant-decomposing microbial agent of the present example comprises the following raw materials by weight:

[0105] 80 parts of composite bacteria, 20 parts of enzyme preparation, 12 parts of growth substrate, 5 parts of nutritional element, 3 parts of stress resistance regulator, 0.08 parts of azido-erythro-sphingosine, and 0.9 parts of vitamin;

[0106] The composite bacteria are a mixture of Bacillus megaterium, Bacillus subtilis, Bacillus pumilus, Pseudomonas fluorescens, Bacillus licheniformis, and Bifidobacterium pseudolongum in a weight ratio of 1:0.8:0.6:1.02:1.5:0.2. The stress resistance regulator is a mixture of ethoxylated castor oil and dodecylhydroxy poly (oxy-1, 2-ethanediyl) in a weight ratio of 1:0.6.

[0107] The vitamin comprises vitamin A, vitamin B, vitamin D, vitamin E, and vitamin K mixed in a weight ratio of 3:4:1:2:1. The enzyme preparation is protease. The growth substrate comprises glucose and corn flour mixed in a weight ratio of 1:3. The nutritional element comprises calcium, iron, magnesium, zinc, molybdenum, and tungsten mixed in a weight ratio of 1:1:3:1:3:2. The molybdenum is ammonium molybdate, the tungsten is ferrous tungstate, the calcium is calcium sulfate, the iron is ferric chloride, the magnesium is magnesium chloride, and the zinc is zinc chloride.

[0108] The preparation method of the microorganism agent for decomposing pollutants comprises the following steps: taking the composite bacteria, enzyme preparation, growth substrate, nutrient element, stress resistance regulator and azido-erythro-sphingosine according to weight parts, and mixing uniformly, and then the microorganism agent is obtained.

[0109] The effective viable count of the composite bacteria is greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of Bacillus megaterium, the effective viable count of the fermentation liquor of Bacillus subtilis is greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of Bacillus pumilus, the effective viable count of the fermentation liquor of Pseudomonas fluorescens is greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of Bacillus licheniformis, and the effective viable count of the fermentation liquor of Bifidobacterium pseudolongum is greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of Bacillus licheniformis, and the effective viable count of the fermentation liquor of Bifidobacterium pseudolongum is greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of Bacillus licheniformis, and the effective viable count of the fermentation liquor of Bifidobacterium pseudolongum is greater than or equal to 1.0x10 10 CFU / ml of the fermentation liquor of Bacillus licheniformis, and the effective viable count of the fermentation liquor of Bifidobacterium pseudolongum is greater than or equal to 1.0x10 10 CFU / ml to 1.2x10 10 CFU / ml.

[0110] It should be noted that the fermentation liquor of each bacterial species in the composite bacteria can be prepared by using the prior art, and the culture medium, culture conditions and culture steps can also be achieved by using the prior art, as long as the effective viable count can reach the above standard.

[0111] In this embodiment, the preparation method of the composite bacteria comprises the following steps:

[0112] (1) each bacterial species is inoculated on an activated culture medium, and cultured at 37℃ for 20h to obtain the activated bacterial body;

[0113] (2) the activated bacterial body in step (1) is inoculated on a flat plate culture medium, and cultured at 37℃ for 24h to obtain the flat plate cultured bacterial colony;

[0114] (3) the flat plate cultured bacterial colony in step (2) is inoculated on a slant culture medium, and cultured at 37℃ for 24h to obtain the slant cultured bacterial mat;

[0115] (4) the slant cultured bacterial mat in step (3) is inoculated in a shaking flask culture medium at an inoculation amount of 4%, and cultured at 37℃ at a shaking speed of 150r / min for 24h to obtain the shaking flask fermentation bacterial body;

[0116] (5) The shake flask fermentation bacteria in step (4) are inoculated into a seed culture medium at an inoculation amount of 8%, and are fermented at a temperature of 37°C and a stirring speed of 150 r / min for 48 h to obtain seed culture bacteria;

[0117] (6) The seed culture bacteria in step (5) are inoculated into a fermentation tank culture medium, and are fermented at a temperature of 37°C and a stirring speed of 150 r / min for 48 h to obtain fermentation liquor of each strain;

[0118] (7) The fermentation liquor of each strain in step (6) is prepared into a composite bacteria according to the weight parts, and the details are as follows:

[0119] (a) The fermentation liquor of Bacillus megaterium, Bacillus subtilis, Bacillus pumilus, Pseudomonas fluorescens, Bacillus licheniformis and Bifidobacterium pseudolongum is taken according to the weight parts, and an adsorption medium is added to obtain a mixture;

[0120] The adsorption medium is soybean meal and bentonite, the weight ratio of the adsorption medium to the fermentation liquor is 2:1, and the content of the bentonite in the adsorption medium is 20 wt%.

[0121] The bentonite is modified bentonite treated by lactamide propyl trimethyl ammonium chloride. The modified bentonite is obtained by the following method: sodium-based bentonite, lactamide propyl trimethyl ammonium chloride and deionized water are uniformly mixed according to a weight ratio of 1:0.08:20, stirred at 60°C for 4 h, statically aged for 24 h, and then centrifuged, washed and dried.

[0122] (b) The mixture in step (a) is dried to a water content of less than or equal to 10 wt% to obtain solid bacteria of Bacillus megaterium, Bacillus subtilis, Bacillus pumilus, Pseudomonas fluorescens, Bacillus licheniformis and Bifidobacterium pseudolongum.

[0123] (c) The solid bacteria of Bacillus megaterium, Bacillus subtilis, Bacillus pumilus, Pseudomonas fluorescens, Bacillus licheniformis and Bifidobacterium pseudolongum obtained in step (b) are uniformly mixed to obtain the composite bacteria.

[0124] Example 5

[0125] The pollutant-decomposing microbial inoculant of the present example and example 4 use the same raw materials, the same amount of each raw material, and the same preparation method, and the only difference is that 0.01 parts of polyvinyl caprolactam is also contained in the raw materials.

[0126] The preparation method of the pollutant-decomposing microbial agent comprises the following steps: taking the composite bacteria, enzyme preparation, growth substrate, nutrient element, stress resistance regulator, azido-erythro-sphingosine and polyvinyl hexanolactam by weight, and mixing uniformly to obtain the pollutant-decomposing microbial agent.

[0127] Example 6

[0128] The pollutant-decomposing microbial agent of the present example is prepared by using the same raw materials, the same amount of each raw material and the same method as those of Example 4, and the only difference is that the raw material further contains 0.03 parts of polyvinyl hexanolactam.

[0129] The preparation method of the pollutant-decomposing microbial agent comprises the following steps: taking the composite bacteria, enzyme preparation, growth substrate, nutrient element, stress resistance regulator, azido-erythro-sphingosine and polyvinyl hexanolactam by weight, and mixing uniformly to obtain the pollutant-decomposing microbial agent.

[0130] Example 7

[0131] The pollutant-decomposing microbial agent of the present example is prepared by using the same raw materials, the same amount of each raw material and the same method as those of Example 4, and the only difference is that the raw material further contains 0.05 parts of polyvinyl hexanolactam.

[0132] The preparation method of the pollutant-decomposing microbial agent comprises the following steps: taking the composite bacteria, enzyme preparation, growth substrate, nutrient element, stress resistance regulator, azido-erythro-sphingosine and polyvinyl hexanolactam by weight, and mixing uniformly to obtain the pollutant-decomposing microbial agent.

[0133] Example 8

[0134] The pollutant-decomposing microbial agent of the present example is prepared by using the same raw materials, the same amount of each raw material and the same method as those of Example 4, and the only difference is that the raw material further contains 0.05 parts of polyvinyl hexanolactam.

[0135] The preparation method of the pollutant-decomposing microbial agent comprises the following steps: taking the composite bacteria, enzyme preparation, growth substrate, nutrient element, stress resistance regulator, azido-erythro-sphingosine and polyvinyl hexanolactam by weight, and mixing uniformly to obtain the pollutant-decomposing microbial agent.

[0136] Example 9

[0137] The pollutant-decomposing microbial agent of the present example is prepared by using the same raw materials, the same amount of each raw material and the same method as those of Example 4, and the only difference is that the raw material further contains 0.06 parts of polyvinyl hexanolactam.

[0138] The preparation method of the pollutant-decomposing microbial agent comprises the following steps: taking the composite bacteria, enzyme preparation, growth substrate, nutrient element, stress resistance regulator, azido-erythro-sphingosine, and polyvinyl pyrrolidone according to the weight parts, and mixing uniformly to obtain the pollutant-decomposing microbial agent.

[0139] Example 10

[0140] The pollutant-decomposing microbial agent of the present example and the pollutant-decomposing microbial agent of Example 4 are prepared by using the same raw materials, the same amount of each raw material, and the same method, and the only difference is that the raw materials further contain 0.08 parts of polyvinyl pyrrolidone.

[0141] The preparation method of the pollutant-decomposing microbial agent comprises the following steps: taking the composite bacteria, enzyme preparation, growth substrate, nutrient element, stress resistance regulator, azido-erythro-sphingosine, and polyvinyl pyrrolidone according to the weight parts, and mixing uniformly to obtain the pollutant-decomposing microbial agent.

[0142] Example 11

[0143] The pollutant-decomposing microbial agent of the present example and the pollutant-decomposing microbial agent of Example 4 are prepared by using the same raw materials, the same amount of each raw material, and the same method, and the only difference is that the raw materials further contain 0.03 parts of polyvinyl hexanolactam and 0.06 parts of polyvinyl pyrrolidone.

[0144] The preparation method of the pollutant-decomposing microbial agent comprises the following steps: taking the composite bacteria, enzyme preparation, growth substrate, nutrient element, stress resistance regulator, azido-erythro-sphingosine, and polyvinyl pyrrolidone according to the weight parts, and mixing uniformly to obtain the pollutant-decomposing microbial agent.

[0145] Comparative Example 1

[0146] The pollutant-decomposing microbial agent of the present example and the pollutant-decomposing microbial agent of Example 4 are prepared by using the same raw materials, the same amount of each raw material, and the same method, and the only difference is that the raw materials further contain 0.03 parts of polyvinyl hexanolactam and 0.06 parts of polyvinyl pyrrolidone.

[0147] Comparative Example 2

[0148] The pollutant-decomposing microbial agent of the present example and the pollutant-decomposing microbial agent of Example 4 are prepared by using the same raw materials, the same amount of each raw material, and the same method, and the only difference is that the raw materials further contain 0.03 parts of polyvinyl hexanolactam and 0.06 parts of polyvinyl pyrrolidone.

[0149] Comparative Example 3

[0150] The pollutant-decomposing microbial agent of the present example and the pollutant-decomposing microbial agent of Example 4 are prepared by using the same raw materials, the same amount of each raw material, and the same method, and the only difference is that the raw materials further contain 0.03 parts of polyvinyl hexanolactam and 0.06 parts of polyvinyl pyrrolidone.

[0151] Comparative Example 4

[0152] The pollutant-decomposing microbial agent of the present comparative example is prepared by using the same raw materials as those used in Example 11, in the same amounts, and by using the same method, except that the azido-erythro-sphingosine is not included.

[0153] Comparative Example 5

[0154] The pollutant-decomposing microbial agent of the present comparative example is prepared by using the same raw materials as those used in Example 11, in the same amounts, and by using the same method, except that the bentonite is not included.

[0155] Comparative Example 6

[0156] The pollutant-decomposing microbial agent of the present comparative example is prepared by using the same raw materials as those used in Example 11, in the same amounts, and by using the same method, except that the modified bentonite is replaced by sodium-based bentonite.

[0157] Effect Experimental Example

[0158] To verify the technical effects of the pollutant-decomposing microbial agent and the preparation method and application thereof, the following tests were performed.

[0159] The pollutant-decomposing microbial agents obtained in Examples 1-11 and Comparative Examples 1-6 were added to the same filler at a dosage of 1 L / m3 of the liquid microbial agent and 1 kg / m3 of the solid microbial agent, and the same amount of filler was loaded into the same size reaction tank. First, the microorganisms in the reaction tank were cultivated and domesticated, and when the reaction tank could continuously feed water and the treated effluent reached a COD value of less than 50 mg / L and a total nitrogen of less than 15 mg / L, they were used to treat three kinds of wastewater (referred to as first water sample, second water sample, and third water sample) from a chemical enterprise. After 15 days and 30 days of continuous treatment, the average COD value, the average total nitrogen value, and the bacterial concentration of the effluent were measured.

[0160] Among them, in the first water sample, the COD value is 113 mg / L, the total nitrogen is 42 mg / L, the isothiazolinone is 9.30 mg / L, and the salt content is 2 wt%;

[0161] In the second water sample, the COD value is 107 mg / L, the total nitrogen is 39 mg / L, the hypochlorite is 1.3 mg / L, and the salt content is 2 wt%;

[0162] In the third water sample, the COD value is 117 mg / L, the total nitrogen is 41 mg / L, the isothiazolinone is 0.66 mg / L, the hypochlorite is 0.30 mg / L, and the salt content is 6 wt%;

[0163] The COD value of the sewage cultivated by the microorganism is 115 mg / L, the total nitrogen is 40 mg / L, the isothiazolinone is 0.65 mg / L, the hypochlorite is 0.31 mg / L, and the salt content is 0.8 wt %;

[0164] The results are as follows:

[0165] Table 1 shows the treatment results of the first water sample

[0166]

[0167] Table 2 shows the treatment results of the second water sample

[0168]

[0169] Table 3 shows the treatment results of the third water sample

[0170]

[0171] According to the above results, it can be seen that the pollutant-decomposing microbial agent has less influence on salt components and toxic substances in the wastewater, and can still achieve high decomposition efficiency in an environment with high salt content and toxic substances, and has good treatment effect on petrochemical wastewater.

[0172] According to the results of Examples 1-4 and Examples 5-7, in Examples 5-7, the polyvinyl caprolactam is added, and compared with Example 4, the COD value and the total nitrogen value of the effluent are reduced to a certain extent, the treatment effect is improved, and the bacterial concentration changes little and is slightly higher than that of Example 4. However, in the case of high salt content of the third water sample, Examples 1-4 are relatively greatly affected, while Examples 5-7 are relatively less affected. It can be seen that the addition of the polyvinyl caprolactam can weaken the adverse effect of high salt content on the microorganism.

[0173] According to the results of Examples 1-4 and Examples 8-10, in Examples 8-10, the polyvinylpyrrolidone is added, and compared with Example 4, the COD value and the total nitrogen value of the effluent are reduced to a certain extent, the treatment effect is improved, and the bacterial concentration changes little. However, in the case of high isothiazolinone content of the first water sample, Examples 1-4 are relatively greatly affected, while Examples 8-10 are relatively less affected. It can be seen that the addition of the polyvinylpyrrolidone can improve the problem that the activity of the microorganism is inhibited in the environment of the organic sulfur bactericide isothiazolinone.

[0174] According to the results of examples 1-10 and example 11, when polyvinyl caprolactam and polyvinyl pyrrolidone are used together, the obtained microbial inoculant has better comprehensive performance, especially in treating wastewater with high salt content and wastewater with high isothiazolone content.

[0175] According to the results of example 11 and comparative examples 1-3, compared with comparative example 1 without adding the stress resistance regulator, the addition of the stress resistance regulator can improve the treatment efficiency of the microbial inoculant in treating the first water sample, the second water sample and the third water sample, and also can maintain a better bacterial concentration. Especially, compared with comparative example 2 using only ethoxylated castor oil and comparative example 3 using only dodecylhydroxyl poly(oxy-1,2-ethanediyl), when the two are used together, the treatment effect is better.

[0176] According to the results of example 11 and comparative example 4, the addition of the azido-erythro-sphingosine can effectively prolong the high activity of the microbial inoculant, and in the continuous treatment of wastewater with high salt content, high organic sulfur fungicide content and high inorganic chlorine-containing fungicide content, the microbial inoculant can still maintain a good treatment effect for a long time (30 days).

[0177] According to the results of example 11 and comparative examples 5-6, compared with example 11, the microbial inoculants obtained from comparative example 5 without adding bentonite in the adsorption medium and comparative example 6 using modified bentonite instead of sodium-based bentonite have poor comprehensive performance. This may be because the modified bentonite can effectively form a “adsorption-biodegradation” synergistic system as a survival carrier for microorganisms, and the lactamidopropyltrimethylammonium chloride inserted into the interlayer of the bentonite can form hydrogen bonds with the peptidoglycan on the surface of the microorganisms, thereby enhancing the fixation effect of the microorganisms and providing a rich substrate environment for the microorganisms. Compared with unmodified bentonite, it has higher pollutant removal effect and stronger microbial stability in wastewater treatment, and therefore has better bacterial concentration, COD value and total nitrogen value of effluent.

[0178] From the technical common sense, the present application can be realized by other embodiments without departing from the spirit or essential characteristics thereof. Therefore, the above disclosed embodiments are only examples in all aspects, and are not the only ones. All changes within the scope of the present application or within the scope equivalent to the present application are covered by the present application.

Claims

1. A microbial inoculant for decomposing pollutants, characterized by, The raw materials include the following weight parts: 40-80 parts of complex bacteria, 15-25 parts of enzyme preparation, 10-15 parts of growth substrate, 3-6 parts of nutritional elements, 1-3 parts of stress resistance regulator, 0-0.15 parts of azido-erythro-sphingosine, and 0-0.05 parts of polyvinyl caprolactam; The complex bacteria are mixed in a weight ratio of 1: (0.5-1): (0.5-0.8): (1-1.05): (1-2): (0.1-0.3) from Bacillus megaterium, Bacillus subtilis, Bacillus pumilus, Pseudomonas fluorescens, Bacillus licheniformis, and Bifidobacterium pseudolongum. The preservation number of the Bacillus megaterium is ZKCC 15770 or ATCC 35985; the preservation number of the Bacillus subtilis is CMCC(B)63501; the preservation number of the Bacillus pumilus is CMCC(B)63202; the preservation number of the Pseudomonas fluorescens is ATCC 13525; the preservation number of the Bacillus licheniformis is ATCC 11946; and the preservation number of the Bifidobacterium pseudolongum is ZKCC 24853. The stress resistance regulator is mixed in a weight ratio of 1: (0.2-1) from ethoxylated castor oil and dodecyl hydroxy poly (oxy-1, 2-ethanediyl).

2. The pollutant-decomposing microbial agent of claim 1, characterized by, 0.5-1.0 parts of vitamins are further included; the vitamins include one or more of vitamin A, vitamin B, vitamin D, vitamin E, and vitamin K. Optionally, the enzyme preparation includes one or more of peroxidase, reductase, cellulase, amylase, and protease. Optionally, the growth substrate includes one or more of glucose, molasses, corn powder, and soybean powder. Optionally, the nutritional elements include one or more of calcium, iron, magnesium, zinc, molybdenum, and tungsten.

3. The pollutant decomposing microbial agent according to claim 1, characterized by, 0.05-0.08 parts of polyvinylpyrrolidone are further included.

4. A method for preparing the microorganism agent for decomposing pollutants according to any one of claims 1 to 3, characterized by, The method includes the following steps: taking the complex bacteria, enzyme preparation, growth substrate, nutritional elements, stress resistance regulator, azido-erythro-sphingosine, and polyvinyl caprolactam in weight parts, and mixing them uniformly to obtain the product.

5. The pollutant decomposing microbial agent according to claim 4, characterized by, The compound bacteria have an effective viable count greater than or equal to 1.0 x 10⁻⁶. 10 Fermentation broth of Bacillus megaterium with CFU / ml and an effective viable count greater than or equal to 1.0 x 10⁻⁶ cells / ml. 10 Fermentation broth of Bacillus subtilis with CFU / ml and an effective viable count greater than or equal to 1.0 x 10⁻⁶ 10 Fermentation broth of Bacillus pumilus with CFU / ml and an effective viable count greater than or equal to 1.0 x 10⁻⁶ 10 Fermentation broth of *Pseudomonas fluorescens* with CFU / ml and an effective viable count greater than or equal to 1.0 x 10⁻⁶. 10 Fermentation broth of Bacillus licheniformis with CFU / ml and an effective viable count greater than or equal to 1.0 x 10⁻⁶ 10 It is prepared by mixing the fermentation broth of *Bifidobacterium pseudolongum* at CFU / ml; or, The complex bacteria are fermented liquid of Bacillus megaterium with effective viable cell number greater than or equal to 1.0 x 10 10 CFU / ml, fermented liquid of Bacillus subtilis with effective viable cell number greater than or equal to 1.0 x 10 10 CFU / ml, fermented liquid of Bacillus pumilus with effective viable cell number greater than or equal to 1.0 x 10 10 CFU / ml, fermented liquid of Pseudomonas fluorescens with effective viable cell number greater than or equal to 1.0 x 10 10 CFU / ml, and fermented liquid of Bacillus licheniformis with effective viable cell number greater than or equal to 1.0 x 10 10 CFU / ml, respectively, and mixed after being made into solid bacteria. 10 CFU / ml, respectively, and mixed after being made into solid bacteria.

6. The pollutant decomposing microbial agent according to claim 5, characterized by, The preparation method of the complex bacteria includes the following steps: (1) taking each bacterial strain, inoculating it on an activated culture medium, and culturing it at 35-37°C for 10-30h to obtain the activated bacterial body; (2) inoculating the activated bacterial body in step (1) on a flat plate culture medium, and culturing it at 35-37°C for 22-26h to obtain the flat plate cultured bacterial colony; (3) inoculating the flat plate cultured bacterial colony in step (2) on a slant culture medium, and culturing it at 35-37°C for 22-26h to obtain the slant cultured bacterial mat; (4) inoculating the slant cultured bacterial mat in step (3) into a shake flask culture medium at an inoculation amount of 3-5%, and culturing it at 35-37°C at a shaking speed of 140-160r / min for 22-26h to obtain the shake flask fermented bacterial body. (5) the shake flask fermentation bacteria in step (4) are inoculated into seed culture medium at an inoculation amount of 6-10%, and are fermented at a temperature of 35-37 DEG C and a stirring speed of 140-160 r / min for 46-50 h to obtain seed culture bacteria; (6) the seed culture bacteria in step (5) are inoculated into fermentation tank culture medium, and are fermented at a temperature of 35-37 DEG C and a stirring speed of 140-160 r / min for 46-50 h to obtain fermentation liquor of each strain; (7) the fermentation liquor of each strain in step (6) is prepared into a composite bacteria according to weight parts.

7. The pollutant decomposing microbial agent according to claim 6, characterized by, Step (7) comprises the following steps: (a) the fermentation liquor of the bacillus megaterium, bacillus subtilis, bacillus pumilus, pseudomonas fluorescens, bacillus licheniformis and pseudobifidobacterium longum is taken according to weight parts, an adsorption medium is added, and the mixture is uniformly mixed to obtain a mixture; wherein the adsorption medium is one of flour, corn flour and soybean meal; the weight ratio of the adsorption medium to the fermentation liquor is (1-3): 1; (b) the mixture in step (a) is dried to a water content of less than or equal to 10wt% to obtain solid bacteria of the bacillus megaterium, bacillus subtilis, bacillus pumilus, pseudomonas fluorescens, bacillus licheniformis and pseudobifidobacterium longum; (c) the solid bacteria of the bacillus megaterium, bacillus subtilis, bacillus pumilus, pseudomonas fluorescens, bacillus licheniformis and pseudobifidobacterium longum obtained in step (b) are uniformly mixed to obtain a composite bacteria.

8. The pollutant decomposing microbial agent according to claim 7, characterized by, In step (a), the adsorption medium further contains bentonite; the content of the bentonite in the adsorption medium is 10-25wt%; the bentonite is modified bentonite treated by inserting lactamide propyl trimethyl ammonium chloride.

9. Application of the microorganism microbial agent for decomposing pollutants in any one of claims 1-3 in the field of sewage treatment.

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