Compound bacteria for biological enhancement and sludge reduction and preparation method thereof

Abstract

The invention provides compound bacteria for biological enhancement and sludge reduction and a preparation method of the compound bacteria. The compound microbial preparation comprises 25 microbial strains capable of widely degrading organic compounds and sludge zoogloea, original strains with specific degradation capability are subjected to multi-step pure culture fermentation under limited conditions and then are compounded to obtain the compound microbial agent. The composite microbial preparation can be used for degrading various organic matters including aromatic hydrocarbons, alcohols, aldehydes, heterocyclic rings, halogenated hydrocarbons, anilines, phenols and the like and aged activated sludge zoogloea, and can be used for synergism and sludge in-situ reduction of biological systems of various industrial wastewater including municipal wastewater, petrochemical wastewater, synthetic chemistry, printing and dyeing wastewater and the like. Due to the excellent extracellular hydrolase producing function of the compound bacteria, the sludge yield of an existing biochemical system can be remarkably reduced, and in-situ reduction of the sludge is achieved.

Description

Technical Field

[0001] The present invention relates to the technical field of sewage and wastewater treatment, and particularly to a composite bacterium for biological enhancement and sludge reduction and a preparation method thereof. Background Art

[0002] High-concentration industrial sewage has complex components, including halogenated alkanes, olefins, alicyclic hydrocarbons, heterocyclic aromatic hydrocarbons, halogenated benzene alkanes, chlorophenols, nitrobenzenes, furans, pyridines, quinolines, and various synthetic organic pollutants. These enter the environment with the wastewater or solid waste discharged from factories. Due to the complexity of the structures of these substances themselves and their strangeness to organisms, the microorganisms cultured and domesticated with activated sludge in traditional wastewater treatment methods can no longer effectively remove these pollutants. The long-term accumulation of these substances in the environment causes great pollution to the ecological environment on which we depend for survival and poses great harm to the physical and mental health of humans.

[0003] Since various toxic and refractory compounds in industrial wastewater, especially chemical industrial wastewater, cannot be completely degraded by traditional activated sludge, and the use of strong oxidation measures such as physical or chemical methods will significantly increase the treatment cost, resulting in high operation and maintenance costs. Therefore, it is of positive significance to develop a microbial product that can efficiently degrade common refractory organic substances and enhance activated sludge. Summary of the Invention

[0004] The present invention provides a composite bacterium for biological enhancement and sludge reduction, which is characterized by being composed of a fermentation broth or freeze-dried powder of original strains mixed according to the following weight ratio components: 50 - 300 parts of Pseudomonas putida, 50 - 250 parts of Sphingomonas yanoikuyae, 10 - 50 parts of Burkholderia sp., 5 - 50 parts of Rhodococcus erythropolis, 10 - 50 parts of Sphingomonas aromaticivorans, 5 - 50 parts of Cupriavidus basilensis, 5 - 50 parts of Cupriavidus sp., 10 - 50 parts of Xanthobacter autotrophicus, 10 - 50 parts of Rhodococcus rhodochrous, 5 - 50 parts of Acetobacter sp., 5 - 20 parts of Sphingomonas subterranea, 5 - 50 parts of Arthrobacter sp., 10 - 50 parts of Pseudomonas fluorescens, 10 - 50 parts of Delftia acidovorans, 10 - 50 parts of Novosphingobium aromaticivorans, 10 - 50 parts of Rhodococcus ruber, 5 - 50 parts of Achromobacter denitrificans, 10 - 50 parts of Rhodococcus pyridinivorans, 5 - 50 parts of Pseudomonas sp., 10 - 50 parts of Pseudomonas stutzeri, 10 - 50 parts of Pseudomonas aeruginosa, 5 - 50 parts of Pseudomonas chlororaphis, 10 - 50 parts of Bacillus altitudinis, 20 - 100 parts of Lysobacter enzymogenes, 20 - 100 parts of Bacillus pumilus.

[0005] Furthermore, it is characterized by being composed of a fermentation broth of original strains mixed according to the following weight ratio components: 200 parts of Pseudomonas putida, 200 parts of Sphingomonas yanoikuyae, 20 parts of Burkholderia sp., 10 parts of Rhodococcus erythropolis, 20 parts of Sphingomonas aromaticivorans, 20 parts of Cupriavidus basilensis, 10 parts of Cupriavidus sp., 20 parts of Xanthobacter autotrophicus, 20 parts of Rhodococcus rhodochrous, 20 parts of Acetobacterium sp., 20 parts of Sphingomonas subterranea, 50 parts of Arthrobacter sp., 20 parts of Pseudomonas fluorescens, 20 parts of Delftia acidovorans, 20 parts of Novosphingobium aromaticivorans, 20 parts of Rhodococcus ruber, 20 parts of Achromobacter denitrificans, 20 parts of Rhodococcus pyridinivorans, 20 parts of Pseudomonas sp., 20 parts of Pseudomonas stutzeri, 20 parts of Pseudomonas aeruginosa, 20 parts of Pseudomonas chlororaphis, 50 parts of Bacillus altitudinis, 100 parts of Lysobacter enzymogenes, 100 parts of Bacillus pumilus.

[0006] Further, among them, the Pseudomonas putida is Pseudomonas putida ( Pseudomonas putida ): DSM6899 Sphingomonas yanoikuyae is Sphingomonas yanoikuyae ( Sphingobiumyanoikuyae ): DSM6900 Burkholderia sp. is Burkholderia sp. ( Burkholderiasp. ): DSM8530 Rhodococcus erythropolis is Rhodococcus erythropolis ( Rhodococcuserythropolis ): DSM6344 Sphingomonas aromaticivorans is Sphingomonas aromaticivorans ( Sphingomonashaloaromaticamans ): CGMCC1.10206 Cupriavidus basilensis is Cupriavidus basilensis ( Cupriavidusbasilensis ): ACCC 10191 Cupriavidus sp. is Cupriavidus sp. ( Cupriavidussp. ): DSM7346 Xanthobacter autotrophicus is Xanthobacter autotrophicus ( Xanthobacterautotrophicus ): DSM3874 Rhodococcus rhodochrous is Rhodococcus rhodochrous ( Rhodococcusrhodochrous ): DSM11097 Acetobacterium sp. is Acetobacterium sp. DSM9077 Sphingomonas subterranea is Sphingomonas subterranea ( Novosphingobiumsubterraneum ): CGMCCC1.3516 Arthrobacter sp. is Arthrobacter sp. ( Arthrobactersp. ): DSM7325 Pseudomonas fluorescens is Pseudomonas fluorescens ( Pseudomonas fluorescens )DSM7155 Delftia acidovorans is Delftia acidovorans ( Delftiaacidovorans )DSM8370 Novosphingobium aromaticivorans is Novosphingobium aromaticivorans ( Novosphingobiumaromaticivorans) CGMCC1.3746 Rhodococcus ruber is Rhodococcus ruber ( Rhodococcusruber )DSM7511 Achromobacter denitrificans is Achromobacter denitrificans ( Achromobacterdenitrificans )DSM11850 Rhodococcus pyridinivorans is Rhodococcus pyridinivorans ( Rhodococcuspyridinivorans )CCTCC KB20081235 Pseudomonas is Pseudomonas sp. DSM6384 Pseudomonas stutzeri is Pseudomonas stutzeri ( Pseudomonas stutzeri )DSM8219 Pseudomonas aeruginosa is Pseudomonas aeruginosa ( Pseudomonas aeruginosa )DSM6279 Pseudomonas chlororaphis is Pseudomonas chlororaphis ( Pseudomonas chlororaphis )DSM6508 Bacillus altitudinis is Bacillus altitudinis ( Bacillus altitudinis )CICC24401 Lysobacter enzymogenes is Lysobacter enzymogenes ( Lysobacterenzymogenes )ATCC 29487 Bacillus pumilus is Bacillus pumilus ( Bacillus pumilus )CICC 20685 All the strains described in the present invention can be purchased from the China General Microbiological Culture Collection Center (CGMCC), China Center for Type Culture Collection (CCTCC), German Collection of Microorganisms and Cell Cultures (DSM), and China Agricultural Culture Collection Center (ACCC). The obtained slants or lyophilized products are activated according to the activation methods and media recommended by the culture collection centers, and then purified and stored on slants.

[0007] A composite microbial agent for wastewater bioaugmentation and sludge reduction, characterized in that the slant strains are first activated in a shaking flask with a low-concentration medium, then cultured and domesticated with a specific sole carbon source, and then subjected to three-stage fermentation and expansion culture with low, medium, and high-concentration media. Then, the bacterial solutions are mixed according to the weight ratio, and after adding a protective agent, they are packaged and stored in the warehouse. The low-concentration medium for the slant strains has the following formula: glucose 1.0 g / L, yeast extract powder 1.0 g / L, soy peptone 0.5 g / L, bovine bone peptone 0.5 g / L. The activation culture temperature of the slant strains is 30-35 °C, the shaking speed of the shaker is 120-150 revolutions per minute, and the culture time is 24-72 h.

[0008] According to the characteristics of the degradation of certain specific pollutants that the activated shaking flask liquid strains may possess as recorded in the culture collection center, a suitable sole carbon source medium is selected for domestication culture. The sole carbon source medium described in the present invention refers to a specific organic medium containing one or more specific pollutants and no nutrients such as yeast extract powder and glucose. According to the degradation characteristics of different microorganisms, different pollutants are selected as the sole carbon source to domesticate, screen, and enhance the function of the bacterial solution in shaking flask fermentation, and strengthen the degradation function of the original strains to the target pollutants. Through the previous low-concentration shaking flask culture and this step of sole carbon source culture, the loss of degradation plasmids in the 22 microorganisms described in the present invention can be prevented, and their degradation ability to the target pollutants can be maintained.

[0009] According to the long-term exploration and research of the inventor on various degrading bacteria, the sole carbon source media corresponding to specific strains are as follows: Pseudomonas putida ( Pseudomonas putida ) DSM6899, is cultured and domesticated with one or more mixtures of m-cresol, o-cresol, toluene, and p-xylene as the sole carbon source; Sphingomonas yanoikuyae ( Sphingobiumyanoikuyae ) DSM6900, is cultured and domesticated with one or more mixtures of anthracene, biphenyl, naphthalene, phenanthrene, cyclohexane, and toluene as the sole carbon source; Burkholderia sp. ( Burkholderiasp. ) DSM8530, is cultured and domesticated with one or more mixtures of ethylbenzene, chlorobenzene, 1,4-dichlorobenzene, and bromobenzene as the sole carbon source; Rhodococcus erythropolis ( Rhodococcuserythropolis ) DSM6344, is cultured and domesticated with 1-chlorohexane and 6-chlorohexanol as the sole carbon source; Sphingomonas aromaticivorans ( Sphingomonashaloaromaticamans ) CGMCC1.10206, is cultured and domesticated with one or more mixtures of chlorobenzene, 1,4-dichlorobenzene, and 1,3-dichlorobenzene as the sole carbon source; Cupriavidus basilensis ( Cupriavidusbasilensis )ACCC 10191 was cultured and domesticated using one or more mixtures of 2,4-dichlorophenol, 2,6-dichlorophenol, benzene, toluene, chlorobenzene, and phenol as the sole carbon source; Cupriavidus sp. Cupriavidussp. )DSM7346 was cultured and domesticated using one or more mixtures of benzoic acid, phenol, aniline, 2-, 3-, or 4-chloroaniline, and 4-bromoaniline as the sole carbon source; Xanthobacter autotrophicus Xanthobacterautotrophicus )DSM3874 was cultured and domesticated using one or more mixtures of dichloromethane, methanol, and cyclohexanol as the sole carbon source; Rhodococcus rhodochrous Rhodococcusrhodochrous )DSM11097 was cultured and domesticated using one or more mixtures of naphthalene, toluene, m-xylene, and p-xylene as the sole carbon source; Acetobacterium sp. DSM9077 was cultured and domesticated using one or more mixtures of ethylene glycol, diethylene glycol, triethylene glycol, polyethylene glycol 200, and methoxyethanol as the sole carbon source; Sphingomonas subterranea Novosphingobiumsubterraneum )CGMCCC1.3516 was cultured and domesticated using one or more mixtures of salicylic acid, toluene, o-xylene, p-xylene, biphenyl, dibenzothiophene, and fluorene as the sole carbon source; Arthrobacter sp. Arthrobactersp. )DSM7325 was cultured and domesticated using one or more mixtures of acetanilide, 2-chloroacetanilide, methylaniline, acetamide, and N-ethylacetamide as the sole carbon source; Pseudomonas fluorescens Pseudomonas fluorescens )DSM7155 was cultured and domesticated using one or more mixtures of acrylamide, lactam, and succinonitrile as the sole carbon source; Delftia acidovorans Delftiaacidovorans )DSM8370 was cultured and domesticated using one or more mixtures of aniline, 2-, 3-, or 4-methylaniline as the sole carbon source; Novosphingobium aromaticivorans Novosphingobiumaromaticivorans) CGMCC1.3746 was cultured and domesticated using one or more mixtures of p-cresol, dibenzothiophene, naphthalene, toluene, m-xylene, o-xylene, p-xylene, and biphenyl as the sole carbon source; Rhodococcus ruber Rhodococcusruber )DSM7511 was cultured and domesticated using one or more mixtures of n-hexadecane, n-octadecane, benzene, toluene, phenol, and naphthalene as the sole carbon source; Achromobacter denitrificans Achromobacterdenitrificans ) DSM11850 was cultured and domesticated using one or more mixtures of 3-chloroaniline and 4-chloroaniline as the sole carbon source; Rhodococcus pyridinivorans ( Rhodococcuspyridinivorans ) CCTCC KB 20081235 was cultured and domesticated using one or more mixtures of sodium benzoate, catechol, resorcinol, hydroquinone, and pyridine as the sole carbon source; The Pseudomonas sp. is Pseudomonas ( Pseudomonas sp. ) DSM6384, which was cultured and domesticated using one or more mixtures of furfural, furfuryl alcohol, and quinoline as the sole carbon source; Pseudomonas stutzeri ( Pseudomonas stutzeri ) DSM8219 was cultured and domesticated using one or more mixtures of chlorobenzene, p-dichlorobenzene, and o-dichlorobenzene as the sole carbon source; Pseudomonas aeruginosa ( Pseudomonas aeruginosa ) DSM6279 was cultured and domesticated using one or more mixtures of -bromobenzoic acid, 5-bromosalicylic acid, 3-chlorobenzoic acid, 5-chlorosalicylic acid, 2,5-dibromobenzoic acid, and 2,3-dichlorobenzoic acid as the sole carbon source; Pseudomonas chlororaphis ( Pseudomonas chlororaphis ) DSM6508 was cultured and domesticated using one or more mixtures of benzoic acid, styrene, and toluene as the sole carbon source.

[0010] Bacillus altitudinis, Lysobacter enzymogenes, and Bacillus pumilus were cultured and domesticated using sterilized municipal sludge as the sole carbon source.

[0011] When the various organic substances mentioned above were used as the sole carbon source for strain domestication, their concentration in the medium was 50 - 1000 mg / L, and they were added directly after sterilization or by sterile filtration after medium sterilization according to the properties of the organic substances.

[0012] Further, in the stage of shake-flask sole carbon source domestication culture, granular activated carbon with a particle size of 30 - 200 mesh and a concentration of 20 - 200 g / L was added to the medium as a strain attachment carrier, and it could also adsorb and slowly release organic substances.

[0013] Further, in the stage of shake-flask sole carbon source domestication culture, 0.2 - 2.0 g / L ammonium chloride and 0.2 - 2.0 g / L potassium dihydrogen phosphate were added to the medium as supplementary nitrogen and phosphorus.

[0014] Further, in the step of culturing the shake flask with a single carbon source for domestication, it can be subcultured for 2-3 generations in this medium, and the concentration of the single carbon source can be gradually increased to further domesticate the degradation ability of the target microorganism. The first-generation single-carbon-source culture solution can be inoculated into the sterilized second-generation medium at a concentration ratio of 5-30%, and so on, and finally cultured and domesticated for 2-3 generations. When the OD600 of the 2-3 generation bacterial liquid reaches more than 0.3, the domestication culture is completed.

[0015] Further, the shake flask strain after the single carbon source domestication culture is coated and streaked on a co-culture medium plate for purification until a single colony consistent with the original strain is isolated. The co-culture medium is: 0.5 g / L glucose, 0.5 g / L yeast extract powder, 0.2 g / L soy peptone, 0.2 g / L bovine bone peptone, and 50-1000 mg / L single carbon source. By using a co-culture medium prepared with low-concentration nutrients and single-carbon-source organic matter to coat and streak the shake flask strain after domestication culture on a plate for purification, it is possible to avoid the loss of plasmid degradation by the strain, the non-growth of some strains, or the loss of special degradation functions during the separation of bacteria in traditional rich-nutrient media such as LB medium and beef extract peptone medium.

[0016] After the co-culture medium separation and purification, three-stage fermentation and expansion culture are then carried out using low-, medium-, and high-concentration media. The low-concentration medium is: 1.0 g / L glucose, 1.0 g / L yeast extract powder, 0.5 g / L soy peptone, 0.5 g / L bovine bone peptone; the medium-concentration medium is: 2.0 g / L glucose, 2.0 g / L yeast extract powder, 1.0 g / L soy peptone, 1.0 g / L bovine bone peptone; the high-concentration medium is: 4.0 g / L glucose, 4.0 g / L yeast extract powder, 2.0 g / L soy peptone, 2.0 g / L bovine bone peptone.

[0017] In multiple experiments, the inventors found that if many bacteria are directly inoculated into a high-concentration medium, they usually show inhibition and complete non-growth, or the bacterial concentration after culture is extremely low. The present invention proposes a method of three-stage fermentation with low, medium, and high levels to gradually increase the concentration of nutrients in the medium, which can avoid the problems of inhibition and loss of bacterial degradation function that occur when directly using conventional LB or nutrient broth media. By gradually increasing the nutrients in the medium, the target strain can gradually adapt to high osmotic pressure and nutrient components, which is beneficial to maintaining its degradation function. At the same time, at the initial lower medium concentration, it can also prevent contamination by miscellaneous bacteria.

[0018] For the three-stage fermentation and expansion culture of the low-, medium- and high-concentration culture media, the fermentation temperature is 28-35°C, the culture time is 12-48 h, and the dissolved oxygen concentration is maintained at 2.0-6.0 mg / L during the fermentation process. The seed shake flasks and fermenters used in the fermentation are similar to the currently known conventional fermentation methods.

[0019] The viable cell concentration in the bacterial liquid after the original strain is fermented with the culture media of three-stage gradient concentrations is 3x10 7 -5x10 10 CFU / ml. After a single strain completes pure culture fermentation, the 22 strains described in this scheme are mixed and compounded according to the ratio. The effective viable cell concentration in the mixed compound bacteria is 5x10 7 -3x10 10 CFU / ml.

[0020] Furthermore, for the bacterial liquid mixed according to the ratio, protectants are added in the following ratios, including: 2.0-50.0 g / L glycerol, 0.2-2.0 g / L vitamin C, 0.5-2.5 g / L sodium dehydroacetate, and 1.0-5.0 g / L procyanidins. The purpose of adding glycerol is to provide protection for the bacteria in the liquid preparation and facilitate their resistance to adverse environments; the purpose of adding vitamin C is to remove the dissolved oxygen in the bacterial liquid and reduce the aerobic respiration rate of the bacteria; the purpose of adding sodium dehydroacetate is to prevent the overgrowth of some bacteria and at the same time inhibit the growth of other exogenous miscellaneous bacteria; the purpose of adding procyanidins is to remove the hydroxyl radicals generated during the storage of the bacterial liquid and reduce their destructive effect on the bacteria. The procyanidins can be extracted from grape seeds or peanut skins.

[0021] After the protectants are added to the mixed bacterial liquid, it is sealed and packaged and stored in a plastic bucket, placed in a cool and dry place, and can be stored for about 6 months at room temperature, and the viable cell number is maintained above 50%.

[0022] The described composite microbial agent for wastewater bioaugmentation and sludge reduction can be used for the bioaugmentation or biopotentiation treatment of common chemical wastewaters containing phenols, heterocycles, halogenated hydrocarbons, alcohols, etc. It can be used to strengthen the biological treatment systems in multiple fields such as coking wastewater, pharmaceutical wastewater, pesticide wastewater, synthetic chemistry, and printing and dyeing wastewater.

[0023] Furthermore, when the composite microbial agent is put into anaerobic hydrolysis or aerobic aeration, 3.0-30.0 g / L of powdered activated carbon is added as the attachment carrier of the bacteria in the anaerobic hydrolysis tank and the aerobic aeration tank, and the particle size of the powdered activated carbon is 80-400 mesh. The dosing ratio of the composite microbial agent is dosed according to 0.1-5.0% of the effective tank volume, and when treating higher-concentration wastewater, the dosing ratio can be appropriately increased.

[0024] Furthermore, when the composite microbial agent is used for in-situ sludge reduction in the biochemical system, the composite microbial agent is put into the anoxic tank or the front stage of aerobic zone, and the dosing concentration of the bacterial liquid is one ten-thousandth to one thousandth of the effective tank volume, and the dosing frequency is once a month.

[0025] The beneficial effects of the present invention are as follows: The composite microbial flora for sewage treatment of the present invention can adopt a one-time dosing method, which is simple to operate, conducive to production, and can quickly form a dominant flora; the various strains in the formula are rationally compatible, symbiotic and coordinated, and do not antagonize each other, and the survival rate of the microbial agent is high. The method for treating sewage using the microbial agent of this formula is simple and feasible, and has a unique treatment effect on macromolecules, refractory, toxic and harmful substances such as phenols, aromatic hydrocarbons, ammonia nitrogen, phosphorus, high sulfate, etc. The COD removal rate can be increased by 20 - 50% based on the original activated sludge, and it has high treatment stability and utilization rate, and the sludge production of the biochemical system can be reduced by 30 - 60%. Description of the Drawings

[0026] 图1 is the preparation flow chart of the present invention; 图2 is the table of average influent and effluent water conditions within one month before and after inoculation in Example 2; 图3 is the table of average influent and effluent water values before and after inoculation in Example 3; 图4 is the table of average influent water quality in Example 4; 图5 is the table for analyzing the sludge reduction effect in Example 4. Detailed Embodiments

[0027] The following are the preferred embodiments of the present invention. It should be understood that the preferred embodiments described herein are only used to illustrate and explain the present invention, and are not used to limit the present invention.

[0028] Example 1 A composite microbial agent for wastewater biological enhancement and sludge reduction, characterized in that it is composed of a mixture of original strain fermentation broths with the following weight ratio components: 120 parts of Pseudomonas putida, 60 parts of Sphingomonas yanoikuyae, 40 parts of Burkholderia sp., 10 parts of Rhodococcus erythropolis, 15 parts of Sphingomonas aromaticivorans, 20 parts of Cupriavidus metallidurans, 20 parts of Cupriavidus sp., 20 parts of Xanthobacter autotrophicus, 20 parts of Rhodococcus rhodochrous, 20 parts of Acetobacterium sp., 10 parts of Sphingomonas subterranea, 10 parts of Arthrobacter sp., 20 parts of Pseudomonas fluorescens, 20 parts of Delftia acidovorans, 20 parts of Novosphingobium aromaticivorans, 20 parts of Rhodococcus ruber, 20 parts of Achromobacter denitrificans, 15 parts of Rhodococcus pyridinivorans, 10 parts of Pseudomonas sp., 15 parts of Pseudomonas stutzeri, 15 parts of Pseudomonas aeruginosa, 15 parts of Pseudomonas chlororaphis, 45 parts of Bacillus altitudinis, 40 parts of Lysobacter enzymogenes, 30 parts of Bacillus pumilus.

[0029] Among them, the Pseudomonas putida is Pseudomonas putida ( Pseudomonas putida ): DSM6899 The Sphingomonas yanoikuyae is Sphingomonas yanoikuyae ( Sphingobiumyanoikuyae ): DSM6900 The Burkholderia sp. is Burkholderia sp. ( Burkholderia sp. ): DSM8530 The Rhodococcus erythropolis is Rhodococcus erythropolis ( Rhodococcus erythropolis ): DSM6344 The Sphingomonas aromaticivorans is Sphingomonas aromaticivorans ( Sphingomonas haloaromatica mans ): CGMCC1.10206 The Cupriavidus metallidurans is Cupriavidus metallidurans ( Cupriavidus basilensis ): ACCC 10191 The Cupriavidus sp. is Cupriavidus sp. ( Cupriavidus sp. ): DSM7346 The Xanthobacter autotrophicus is Xanthobacter autotrophicus ( Xanthobacter autotrophicus ): DSM3874 The Rhodococcus rhodochrous is Rhodococcus rhodochrous ( Rhodococcus rhodochrous ): DSM11097 The Acetobacterium sp. is Acetobacterium sp. DSM9077 The Sphingomonas subterranea is Sphingomonas subterranea ( Novosphingobium subterraneum ): CGMCCC1.3516 The Arthrobacter sp. is Arthrobacter sp. ( Arthrobacter sp. ): DSM7325 The Pseudomonas fluorescens is Pseudomonas fluorescens ( Pseudomonas fluorescens ): DSM7155 The Delftia acidovorans is Delftia acidovorans ( Delftia acidovorans ): DSM8370 The Novosphingobium aromaticivorans is Novosphingobium aromaticivorans (Novosphingobium aromaticivorans) CGMCC 1.3746 Rhodococcus ruber is Rhodococcus ruber ( Rhodococcus ruber ) DSM 7511 Achromobacter denitrificans is Achromobacter denitrificans ( Achromobacter denitrificans ) DSM 11850 Rhodococcus pyridinivorans is Rhodococcus pyridinivorans ( Rhodococcus pyridinivorans ) CCTCC KB20081235 Pseudomonas is Pseudomonas sp. DSM 6384 Pseudomonas stutzeri is Pseudomonas stutzeri ( Pseudomonas stutzeri ) DSM 8219 Pseudomonas aeruginosa is Pseudomonas aeruginosa ( Pseudomonas aeruginosa ) DSM 6279 Pseudomonas chlororaphis is Pseudomonas chlororaphis ( Pseudomonas chlororaphis ) DSM 6508 Bacillus altitudinis is Bacillus altitudinis ( Bacillus altitudinis ) CICC 24401 Lysobacter enzymogenes is Lysobacter enzymogenes ( Lysobacter enzymogenes ) ATCC 29487 Bacillus pumilus is Bacillus pumilus ( Bacillus pumilus ) CICC 20685 As Figure 1 shown, the above original strains are first activated in a shake flask with a low-concentration medium, then cultured and domesticated with a specific sole carbon source and separated and purified with a co-culture medium, and then subjected to three-stage fermentation and expansion culture with low, medium, and high-concentration media, and then the bacterial solutions are mixed according to the weight ratio, and after adding a protective agent, they are packaged and stored in the warehouse.

[0030] In this example, the low-concentration medium is: glucose 1.0 g / L, yeast extract powder 1.0 g / L, soy peptone 0.5 g / L, bovine bone peptone 0.5 g / L; the medium-concentration medium is: glucose 2.0 g / L, yeast extract powder 2.0 g / L, soy peptone 1.0 g / L, bovine bone peptone 1.0 g / L; the high-concentration medium is: glucose 4.0 g / L, yeast extract powder 4.0 g / L, soy peptone 2.0 g / L, bovine bone peptone 2.0 g / L. The co-culture medium is: glucose 0.5 g / L, yeast extract powder 0.5 g / L, soy peptone 0.2 g / L, bovine bone peptone 0.2 g / L, sole carbon source 50 - 1000 mg / L.

[0031] The low-concentration medium shake flask is activated at a culture temperature of 30-32 °C for 48 h, and the shaker speed is 140 rpm; the three-stage fermentation and expansion culture of low, medium, and high-concentration media is carried out at a culture temperature of 30-32 °C for 48 h, and the dissolved oxygen in the fermenter is controlled at 4.0-6.0 mg / L. The fermentation device is consistent with the currently disclosed conventional fermentation means and control methods.

[0032] In this implementation plan, the unique carbon source media corresponding to specific strains are as follows: Pseudomonas putida ( Pseudomonas putida ) DSM6899 is cultured and domesticated using a mixture of 250 mg / L toluene and 250 mg / L p-xylene as the sole carbon source; Sphingomonas yanoikuyae ( Sphingobium yanoikuyae ) DSM6900 is cultured and domesticated using a mixture of 300 mg / L naphthalene and 300 mg / L cyclohexane as the sole carbon source; Burkholderia sp. ( Burkholderia sp. ) DSM8530 is cultured and domesticated using 400 mg / L chlorobenzene as the sole carbon source; Rhodococcus erythropolis ( Rhodococcus erythropolis ) DSM6344 is cultured and domesticated using 250 mg / L 1-chlorohexane as the sole carbon source; Sphingomonas aromaticivorans ( Sphingomonas haloaromatica mans ) CGMCC1.10206 is cultured and domesticated using 250 mg / L 1,4-dichlorobenzene as the sole carbon source; Cupriavidus basilensis ( Cupriavidus basilensis ) ACCC 10191 is cultured and domesticated using 200 mg / L 2,4-dichlorophenol as the sole carbon source; Cupriavidus sp. ( Cupriavidus sp. ) DSM7346 is cultured and domesticated using 200 mg / L 4-chloroaniline as the sole carbon source; Xanthobacter autotrophicus ( Xanthobacter autotrophicus ) DSM3874 is cultured and domesticated using 200 mg / L dichloromethane as the sole carbon source; Rhodococcus rhodochrous ( Rhodococcus rhodochrous ) DSM11097 is cultured and domesticated using 500 mg / L naphthalene as the sole carbon source; Acetobacterium sp. DSM9077 is cultured and domesticated using 500 mg / L diethylene glycol as the sole carbon source; Sphingomonas subterranea ( Novosphingobium subterraneum ) CGMCCC1.3516 is cultured and domesticated using a mixture of 200 mg / L o-xylene and 200 mg / L p-xylene as the sole carbon source; Arthrobacter ( Arthrobacter sp. ) DSM7325 was cultured and domesticated using 450 mg / L of acetanilide as the sole carbon source; Pseudomonas fluorescens ( Pseudomonas fluorescens ) DSM7155 was cultured and domesticated using a mixture of 300 mg / L of acrylamide and 300 mg / L of succinonitrile as the sole carbon source; Delftia acidovorans ( Delftia acidovorans ) DSM8370 was cultured and domesticated using 800 mg / L of aniline as the sole carbon source; Novosphingobium aromaticivorans ( Novosphingobium aromaticivorans) CGMCC1.3746) was cultured and domesticated using 350 mg / L of p-cresol as the sole carbon source; Rhodococcus ruber ( Rhodococcus ruber ) DSM7511 was cultured and domesticated using 600 mg / L of n-hexadecane as the sole carbon source; Achromobacter denitrificans ( Achromobacter denitrificans ) DSM11850 was cultured and domesticated using 200 mg / L of 3-chloroaniline as the sole carbon source; Rhodococcus pyridinivorans ( Rhodococcus pyridinivorans ) CCTCC KB 20081235 was cultured and domesticated using a mixture of 300 mg / L of catechol and 300 mg / L of pyridine as the sole carbon source; Pseudomonas sp. ( Pseudomonas sp. ) DSM6384 was cultured and domesticated using a mixture of 300 mg / L of furfural and 200 mg / L of quinoline as the sole carbon source; Pseudomonas stutzeri ( Pseudomonas stutzeri ) DSM8219 was cultured and domesticated using one or more mixtures of chlorobenzene, p-dichlorobenzene, and o-dichlorobenzene as the sole carbon source; Pseudomonas aeruginosa ( Pseudomonas aeruginosa ) DSM6279 was cultured and domesticated using 200 mg / L of 3-chlorobenzoic acid as the sole carbon source; Pseudomonas chlororaphis ( Pseudomonas chlororaphis ) DSM6508 was cultured and domesticated using 300 mg / L of benzoic acid as the sole carbon source.

[0033] Bacillus altitudinis, Lysobacter enzymogenes, and Bacillus pumilus were cultured and domesticated using 1000 mg / L of sterilized municipal sludge as the sole carbon source.

[0034] After the above 25 strains of bacteria with special degradation functions were cultured and domesticated with a sole carbon source, a functional bacterial solution with an OD value not lower than 0.3 was obtained. Then, the bacterial solution was spread on a synergistic culture medium by the method of dilution coating. After its growth, it was further purified by multiple streak plate methods to obtain rejuvenated single colony on the plate. The synergistic culture medium was: 0.5 g / L glucose, 0.5 g / L yeast extract powder, 0.2 g / L soy peptone, 0.2 g / L bovine bone peptone, and the organic matter and concentration selected for the sole carbon source were the same as those in the sole carbon source domestication culture medium.

[0035] After fermenting the rejuvenated single colony on the plate with low, medium, and high-level culture media, mixing was carried out according to the weight ratio described above in this example. The effective viable bacteria concentration in the mixed composite bacteria was 3x10 9 CFU / ml.

[0036] To the mixed bacterial solution, a bacterial strain protectant was added. The protectant included: 5.0 g / L glycerol, 0.8 g / L vitamin C, 0.8 g / L sodium dehydroacetate, and 1.0 g / L procyanidins. After adding the protectant, it was mixed and stirred evenly again, sealed and packaged in a plastic bucket, and stored at normal temperature in a cool and dry place.

[0037] Example 2 The composite bacterial solution prepared in Example 1 for biological enhancement of wastewater and sludge reduction was used to enhance the biochemical system of a certain resin production wastewater. This enterprise mainly produces ion exchange resins, and the wastewater contains high concentrations of divinylbenzene, styrene, propylenediamine, dimethylamine, toluene, etc., with high water quality toxicity. The operation effect of the biochemical system was poor before the addition of bacteria. The system adopted a regulating tank, an emergency tank, an iron-carbon tower, a comprehensive sedimentation tank, a decalcification tank, a micro-aeration hydrolysis tank, and an A / O process. The daily water treatment capacity of this sewage treatment station was 1500 tons, the influent COD of the biochemical tank was 3000 - 5000 mg / L, the total nitrogen fluctuated between 300 - 400 mg / L, and the total biochemical residence time was 92 h. The composite bacterial solution prepared according to Example 1 of the present invention was added to the AO tank at a dosing concentration of 1.5% of the effective tank volume, and at the same time, 5.0 g / L of powdered activated carbon was added as a carrier for the starting bacterial strain.

[0038] From Figure 2 It can be seen that after adding the composite bacterial solution prepared in Example 1 of the present invention, the biochemical treatment effect of this resin wastewater was significantly improved. The effluent COD of the biochemical tank decreased from 985 mg / L before adding bacteria to below 400 mg / L. At the same time, due to the detoxification and degradation effect of the synergistic bacterial group on toxic organic amines, the nitrification effect was also significantly improved, and the effluent ammonia nitrogen decreased from 87 mg / L before adding bacteria to 22 mg / L.

[0039] Example 3 The composite bacterial solution prepared in Example 1 for biological enhancement of wastewater and sludge reduction was used for biological enhancement treatment of a certain printing and dyeing wastewater. The dyes used by this enterprise include reactive dyes, disperse dyes, acid dyes, etc., and the auxiliaries include weak acids, weak bases, baking soda, etc. Softeners, silicone oils, olive oils, soap oils, anti-dyeing agents, etc. are added to the washing water. The industrial wastewater discharged during the production process contains pollutants such as SS, COD cr , BOD 5 , NH 3 -N, sulfur and other components. The wastewater has high COD and ammonia nitrogen concentrations and complex components. The biochemical section of this wastewater adopts processes such as a comprehensive regulation tank + hydrolysis acidification tank + anoxic tank + aerobic tank + MBR tank. The daily water treatment capacity of this sewage treatment plant is 8,500 tons. The influent COD of the biochemical tank is 3,000 - 4,000 mg / L, and the total nitrogen fluctuates between 200 - 300 mg / L. The total biochemical residence time is 72 h. The composite bacterial solution prepared according to Example 1 of the present invention was added to the anoxic tank and aerobic tank at a concentration of 1.0% of the effective tank volume, and at the same time, 5 g / L of powdered activated carbon was added as a carrier for starting bacteria.

[0040] It can be seen from Figure 3 that after adding the composite bacterial solution prepared in Example 1 of the present invention, the biochemical treatment effect of this printing and dyeing wastewater is significantly improved. The COD of the effluent from the biochemical tank is reduced from 624 mg / L before adding bacteria to below 300 mg / L. At the same time, due to the detoxification and degradation effect of the synergistic bacteria on toxic substances, the nitrification effect is also significantly improved, and the ammonia nitrogen in the effluent is reduced from 65 mg / L before adding bacteria to about 5 mg / L.

[0041] Example 4 Different from Example 1, the composition of different strains of the composite bacteria was adjusted to: 50 parts of Pseudomonas putida, 50 parts of Sphingomonas yanoikuyae, 10 parts of Burkholderia, 5 parts of Rhodococcus erythropolis, 10 parts of Sphingomonas aromaticivorans, 5 parts of Cupriavidus basilensis, 5 parts of Cupriavidus, 10 parts of Xanthobacter autotrophicus, 10 parts of Rhodococcus rhodochrous, 5 parts of Acetobacter, 5 parts of Sphingomonas subterranea, 5 parts of Arthrobacter, 10 parts of Pseudomonas fluorescens, 10 parts of Delftia acidovorans, 10 parts of Novosphingobium aromaticivorans, 10 parts of Rhodococcus ruber, 5 parts of Achromobacter denitrificans, 10 parts of Rhodococcus pyridinivorans, 5 parts of Pseudomonas, 10 parts of Pseudomonas stutzeri, 10 parts of Pseudomonas aeruginosa, 5 parts of Pseudomonas chlororaphis, 50 parts of Bacillus altitudinis, 100 parts of Lysobacter enzymogenes, 100 parts of Bacillus pumilus.

[0042] Other steps and methods are exactly the same as those in Example 1, and a composite bacterial solution for sludge reduction was prepared for a pilot-scale production experiment on sludge reduction in a certain sewage treatment plant.

[0043] A certain small municipal sewage treatment plant adopts the AO activated sludge process and is designed to treat 20,000 m of sewage per day3 , the actual water volume is 16,000 - 21,000 m 3 / d. The average influent water quality is as Figure 4 : The volume of the anoxic tank in its main process stage is 3,500 m 3 , the volume of the aerobic tank is 8,400 m 3 , and the volume of the secondary sedimentation tank is 2,200 m 3 .

[0044] The excess sludge from the secondary sedimentation tank is conditioned by adding chemicals in the sludge thickening tank and conditioning tank, and then enters the belt filter press for dehydration. From November to December 2022, liquid strains prepared according to the method of this embodiment were added to reduce sludge. In the first month, it was added 5 times, and the dosage per time was one ten-thousandth of the designed water volume. A total of 10 tons of liquid strains were added at the start-up. Subsequently, it was added once a month, and the dosage was one ten-thousandth of the designed water volume, that is, 2 tons / month. After the strains were added, sludge discharge was suspended for half a month, and at the same time, the dissolved oxygen at the end was controlled to be not less than 2.0 mg / L.

[0045] Comparing the data from January to October 2022 and from February to November 2023 after sludge reduction and efficiency improvement, the sludge reduction effect is analyzed as Figure 5 .

[0046] From the above Figure 5 It can be seen that when adding the composite bactericide prepared according to this embodiment for in-situ reduction of municipal sewage, without changing the process and increasing energy consumption, the sludge production can be reduced by about 30%, showing a good reduction effect.

[0047] Finally, it should be noted that the above are only the preferred embodiments of the present invention and are not used to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, for those skilled in the art, they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements for some of the technical features. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A composite microbial preparation for wastewater bioaugmentation and sludge reduction, characterized in that: The fermentation liquid of the original strains is mixed with the following components in weight proportion: 50-300 parts of Pseudomonas putida, 50-250 parts of Sphingomonas Yano, 10-50 parts of Burkholderia, 5-50 parts of Rhodococcus erythrophilus, 10-50 parts of Sphingomonas erythrophilus, 5-50 parts of Brasilian Cupric Bacteria, 5-50 parts of Cupric Bacteria, 10-50 parts of Autotrophic Yellow Bacteria, 10-50 parts of Rhodococcus roseus, 5-50 parts of Acetobacter, 5-20 parts of Sphingomonas subterranea, 5-50 parts of Arthrobacter, 10-50 parts of Pseudomonas fluorescens 0-50 parts, 10-50 parts of Delftia acidovorans, 10-50 parts of Neosphingobacterium aromaticum, 10-50 parts of Rhodococcus erythrocytes, 5-50 parts of Achromobacter denitrifying, 10-50 parts of Rhodococcus pyridinophilus, 5-50 parts of Pseudomonas, 10-50 parts of Pseudomonas stutzeri, 10-50 parts of Pseudomonas aeruginosa, 5-50 parts of Pseudomonas chlororaphis, 10-50 parts of Bacillus aeruginosa, 20-100 parts of Bacillus zymolyticus, and 20-100 parts of Bacillus brevis.

2. wherein the Pseudomonas putida is Pseudomonas putida ( Pseudomonas putida ):DSM6899 Sphingomonas Yano is Sphingomonas Yano ( Sphingobium yanoikuyae ):DSM6900 Burkholderia is Burkholderia ( Burkholderia sp. ):DSM8530 Rhodococcus rubervillei is Rhodococcus rubervillei ( Rhodococcus erythropolis ):DSM6344 Sphingomonas aromatica is Sphingomonas aromatica ( Sphingomonas haloaromaticamans ):CGMCC1.10206 Brasiliensis is Brasiliensis ( Cupriavidus basilensis ): ACCC 10191 Cupric bacteria are Cupric bacteria ( Cupriavidus sp. ):DSM7346 Autotrophic yellow bacteria are autotrophic yellow bacteria ( Xanthobacter autotrophicus )DSM3874 Rhodococcus rhodochrous bacteria is Rhodococcus rhodochrous bacteria ( Rhodococcus rhodochrous )DSM11097 Acetobacter is Acetobacterium sp. DSM9077 Sphingomonas subterranea is Sphingomonas subterranea ( Novosphingobium subterraneum ):CGMCCC1.3516 Arthrobacter is a genus of the genus Arthrobacter ( Arthrobacter sp. )DSM7325 Pseudomonas fluorescens is Pseudomonas fluorescens ( Pseudomonas fluorescens )DSM7155 Delftia acidovorans is Delftia acidovorans ( Delftia acidovorans )DSM8370 The aromatic hydrocarbon-eating sphingobacterium is the aromatic hydrocarbon-eating sphingobacterium ( Novosphingobiumaromaticivorans) CGMCC1.3746 Rhodococcus is Rhodococcus ( Rhodococcus ruber )DSM7511 Achromobacter denitrificans is Achromobacter denitrificans ( Achromobacter denitrificans )DSM11850 Rhodococcus pyridinophilus is Rhodococcus pyridinophilus ( Rhodococcus pyridinivorans ) CCTCC KB 20081235 Pseudomonas is Pseudomonas sp. DSM6384 Pseudomonas stutzeri is Pseudomonas stutzeri ( Pseudomonas stutzeri ) DSM8219 Pseudomonas aeruginosa is Pseudomonas aeruginosa ( Pseudomonas aeruginosa )DSM6279 Pseudomonas chlororaphis is Pseudomonas chlororaphis ( Pseudomonas chlororaphis )DSM6508 Bacillus subtilis is a high-lying bacillus ( Bacillus altitudinis )CICC24401 Lysozyme-producing Bacillus Lysobacter enzymogenes )ATCC 29487 Bacillus pumilus is a bacterium Bacillus pumilus )CICC 20685 The method for preparing a composite microbial preparation for wastewater bioaugmentation and sludge reduction according to claim 1 is characterized in that: The original strain is first activated by shaking a low-concentration culture medium, then cultured and domesticated by using a specific sole carbon source and separated and purified by a synergistic culture medium, and then expanded by three-stage fermentation using low, medium and high concentration culture media, and then the bacterial liquid is mixed according to weight proportions, and a protective agent is added before packaging and storage.

3. The method for preparing a composite microbial preparation for wastewater bioaugmentation and sludge reduction according to claim 2, characterized in that: The low-concentration culture medium comprises: 1.0 g / L glucose, 1.0 g / L yeast extract powder, 0.5 g / L soy peptone, and 0.5 g / L bovine bone peptone; the medium-concentration culture medium comprises: 2.0 g / L glucose, 2.0 g / L yeast extract powder, 1.0 g / L soy peptone, and 1.0 g / L bovine bone peptone; the high-concentration culture medium comprises: 4.0 g / L glucose, 4.0 g / L yeast extract powder, 2.0 g / L soy peptone, and 2.0 g / L bovine bone peptone; the synergistic culture medium comprises: 0.5 g / L glucose, 0.5 g / L yeast extract powder, 0.2 g / L soy peptone, 0.2 g / L bovine bone peptone, and 50-1000 mg / L as the sole carbon source.

4. The method for preparing a composite microbial preparation for wastewater bioaugmentation and sludge reduction according to claim 2, characterized in that: The original strain was cultured and domesticated using a specific sole carbon source. The sole carbon source culture medium corresponding to the specific strain is as follows: Pseudomonas putida ( Pseudomonas putida ) DSM6899, cultured and domesticated using one or a mixture of m-cresol, o-cresol, toluene, and p-xylene as the sole carbon source; Sphingomonas Yano Sphingobium yanoikuyae ) DSM6900, using anthracene, biphenyl, naphthalene, phenanthrene, cyclohexane, toluene or a mixture of two or more as the sole carbon source for cultivation and domestication; Burkholderia Burkholderia sp. ) DSM8530, using ethylbenzene, chlorobenzene, 1,4-dichlorobenzene, bromobenzene or a mixture of two or more as the sole carbon source for cultivation and domestication; Rhodococcus ruber Rhodococcus erythropolis )DSM6344, cultivated and domesticated with 1-chlorohexane and 6-chlorohexanol as the sole carbon source; Sphingomonas aromatica Sphingomonas haloaromaticamans ) CGMCC1.10206, using chlorobenzene, 1,4-dichlorobenzene, 1,3-dichlorobenzene or a mixture of two or more as the sole carbon source for cultivation and domestication; Brasiliensis Cupriavidus basilensis ) ACCC 10191, using 2,4-dichlorophenol, 2,6-dichlorophenol, benzene, toluene, chlorobenzene, phenol or a mixture of two or more as the sole carbon source for cultivation and domestication; Cupric bacteria Cupriavidus sp. ) DSM7346, cultured and domesticated using one or a mixture of two or more of benzoic acid, phenol, aniline, 2-, 3- or 4-chloroaniline, 4-bromoaniline as the sole carbon source; Autotrophic Xanthomonas Xanthobacter autotrophicus ) DSM3874, cultivated and domesticated using one or a mixture of two or more of dichloromethane, methanol, and cyclohexanol as the sole carbon source; Rhodococcus rhodochrous Rhodococcus rhodochrous ) DSM11097, cultivated and domesticated using one or a mixture of two or more of naphthalene, toluene, m-xylene, and p-xylene as the sole carbon source; Acetobacterium sp. DSM9077 was cultivated and domesticated using one or a mixture of two or more of ethylene glycol, diethylene glycol, triethylene glycol, polyethylene glycol 200, and methoxyethanol as the sole carbon source; Sphingomonas subterranea ( Novosphingobium subterraneum ), CGMCCC1.3516, using salicylic acid, toluene, o-xylene, p-xylene, biphenyl, dibenzothiophene, fluorene, one or a mixture of two or more as the sole carbon source for cultivation and domestication; Arthrobacter spp. Arthrobacter sp. ) DSM7325, cultured and domesticated using one or a mixture of two or more of acetanilide, 2-chloroacetanilide, methylaniline, acetamide, and N-ethylacetamide as the sole carbon source; Pseudomonas fluorescens ( Pseudomonas fluorescens ) DSM7155, cultured and domesticated using acrylamide, lactam, succinonitrile or a mixture of two or more as the sole carbon source; Delftia acidovorans Delftia acidovorans ) DSM8370, cultivated and domesticated using aniline, 2-, 3- or 4-methylaniline, or a mixture of two or more thereof as the sole carbon source; Neosphingobacterium aromaticum ( Novosphingobium aromaticivorans) CGMCC1.3746, using p-cresol, dibenzothiophene, naphthalene, toluene, m-xylene, o-xylene, p-xylene, biphenyl, one or a mixture of two or more as the sole carbon source for cultivation and domestication; Rhodococcus ruber Rhodococcus ruber ) DSM7511, cultivated and domesticated with one or a mixture of n-hexadecane, n-octadecane, benzene, toluene, phenol, naphthalene as the sole carbon source; Achromobacter denitrificans ( Achromobacter denitrificans ) DSM11850, cultivated and domesticated using 3-chloroaniline, 4-chloroaniline or a mixture of two or more as the sole carbon source; Rhodococcus pyridinophilus ( Rhodococcus pyridinivorans ) CCTCC KB 20081235, using sodium benzoate, catechol, resorcinol, hydroquinone, pyridine or a mixture of two or more as the sole carbon source for cultivation and domestication; Pseudomonas is Pseudomonas ( Pseudomonas sp. ) DSM6384, cultivated and domesticated using furfural, furfuryl alcohol, quinoline or a mixture of two or more as the sole carbon source; Pseudomonas stutzeri ( Pseudomonas stutzeri ) DSM8219, using chlorobenzene, p-dichlorobenzene, o-dichlorobenzene or a mixture of two or more as the sole carbon source for cultivation and domestication; Pseudomonas aeruginosa ( Pseudomonas aeruginosa ) DSM6279, using 1-bromobenzoic acid, 5-bromosalicylic acid, 3-chlorobenzoic acid, 5-chlorosalicylic acid, 2,5-dibromobenzoic acid, 2,3-dichlorobenzoic acid or a mixture of two or more thereof as the sole carbon source for cultivation and domestication; Pseudomonas chlororaphis ( Pseudomonas chlororaphis ) DSM6508, cultivated and domesticated using benzoic acid, styrene, toluene or a mixture of two or more as the sole carbon source; Bacillus altaica, Bacillus enzymolyticus, and Bacillus brevis were cultivated and domesticated using sterilized municipal sludge as the sole carbon source; When the various organic substances used above are used as the sole carbon source for strain domestication, their concentration in the culture medium is 50-1000 mg / L. Depending on the properties of the organic substances, they are sterilized directly or sterilized and then aseptically filtered before being added to the culture medium.

5. The method for preparing a composite microbial preparation for wastewater bioaugmentation and sludge reduction according to claim 2, characterized in that: The concentration of live bacteria in the bacterial liquid after the secondary fermentation of the original strain is 3x10 7 -5x10 10 CFU / ml, the effective live bacteria concentration in the mixed compound bacteria is 5x10 7 -3x10 10 CFU / ml.

6. The method for preparing a composite microbial preparation for wastewater bioaugmentation and sludge reduction according to claim 2, characterized in that: The protective agent includes: 2.0-50.0 g / L glycerol, 0.2-2.0 g / L vitamin C, 0.5-2.5 g / L sodium dehydroacetate, and 1.0-5.0 g / L proanthocyanidin.

7. The method for preparing a composite microbial preparation for wastewater bioaugmentation and sludge reduction according to claim 2, characterized in that: The fermentation culture is carried out in three stages with low, medium and high concentration culture medium. The fermentation culture temperature is 28-35°C, the culture time is 12-48h, and the dissolved oxygen is controlled at 2.0-6.0mg / L during the culture process.

8. The method for using the composite microbial preparation for wastewater bioaugmentation and sludge reduction according to claim 1, characterized in that: When the composite microbial preparation is put into anaerobic hydrolysis or aerobic aeration, 3.0-30.0 g / L of powdered activated carbon is added into the anaerobic hydrolysis tank and the aerobic aeration tank respectively as an attachment carrier of the bacteria, and the particle size of the powdered activated carbon is 80-400 meshes.

9. The method for using the composite microbial preparation for wastewater bioaugmentation and sludge reduction according to claim 1, characterized in that: When used for bio-enhancement of difficult-to-degrade wastewater, the concentration of the bacterial solution added when the composite microbial preparation is added to anaerobic hydrolysis or aerobic aeration is 0.1-5.0% of the effective tank volume.

10. The method for using the composite microbial preparation for wastewater bioaugmentation and sludge reduction according to claim 1, characterized in that: When used for in-situ sludge reduction in biochemical systems, the composite microbial preparation is added to the anoxic tank or the aerobic front section, the concentration of the bacterial solution is one hundred thousandth to one ten thousandth of the effective tank volume, and the frequency of addition is once a month.

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