A high-salinity organic wastewater biological treatment method

Abstract

The application discloses a high-salt organic wastewater biological treatment method, characterized in that the method comprises the following steps: step one, screening of halophilic bacteria, the required halophilic bacteria are directly screened from the soil of saline-alkali land, and then the mixed culture of the screened bacteria is subjected to rescreening; the method has the beneficial effects that the halophilic bacteria ZC2403-SYL, ZC2404-SYT and salt-tolerant bacteria ZC2402-MT which can still degrade pollutants are screened out in a high-salt environment by using a microbial method, the organic matters triethylamine, n-butylamine and formamide can be degraded in the high-salt environment, the degradation rates of the organic matters can all reach more than 90% within 72 hours, an efficient salt-tolerant biological treatment system and an efficient bacteria (halophilic bacteria) system are constructed for high-salt and refractory organic pollutants, the three kinds of bacteria screened are mixed and cultured to rapidly form an efficient high-salt organic wastewater degrading bacterial agent ZC-SY, the bacterial agent can be used in actual high-salt organic wastewater and can achieve stable operation, and the treatment problem of the actual organic high-salt wastewater can be effectively solved, and the water treatment cost is reduced.

Description

Technical Field

[0001] This invention relates to the field of high-salinity wastewater treatment technology, and specifically to a biological treatment method for high-salinity organic wastewater. Background Technology

[0002] High-salinity wastewater refers to wastewater containing at least 1% salt by mass. In addition to containing a large amount of inorganic salts, this type of wastewater also contains organic matter and total dissolved solids (TDS) with a mass fraction of ≥3.5%, making it one of the most difficult types of wastewater to treat. Organic high-salt wastewater mainly originates from production wastewater in industries such as petrochemicals, food fermentation, and pharmaceuticals. During the production process, a large amount of salt substances, such as sodium chloride, sodium sulfate, and calcium chloride, need to be added to remove oil, impurities, and adjust pH value. Currently, there are many methods for treating high-salt organic wastewater, such as incineration, electrolysis, membrane separation, and ion exchange. However, incineration produces secondary pollutants such as nitrogen oxides and dioxins; electrolysis has high operating costs; in membrane separation, as the feed solution concentrates, the salt concentration increases, leading to membrane defects, and suspended matter in the wastewater easily clogs the membrane; while ion exchange is mostly used for pretreatment in biological processes to remove metal ions that inhibit microorganisms in wastewater, but its organic matter removal efficiency is poor.

[0003] In existing technologies, biofilm methods produce less sludge, are easy to separate from solids and liquids, and have a stable denitrification effect, and are often used to treat high-nitrogen wastewater. However, high sulfate stress in the wastewater causes an imbalance in the osmotic pressure of microbial cells, which greatly reduces the denitrification effect.

[0004] To address the aforementioned technical issues, CN117185502A discloses a biological denitrification method for high-salt and high-nitrogen wastewater. This method rapidly prepares denitrifying compound bacterial agents, avoiding the tedious process of repeatedly streaking to separate single bacterial species and re-combining them. Furthermore, it optimizes the biofilm start-up conditions suitable for the immobilization of bacterial agents in high-salt wastewater, shortening the start-up cycle of the biofilm method for high-salt wastewater. At the same time, it uses denitrifying bacteria to inhibit sulfate reduction, avoiding secondary sulfide pollution. The aforementioned patent also has the following shortcomings: it cannot degrade pollutants in high-salt environments, and the wastewater treatment efficiency is low. Traditional high-salt organic wastewater treatment processes are complex, costly, and difficult to degrade organic pollutants in high-salt environments. Summary of the Invention

[0005] The purpose of this invention is to provide a biological treatment method for high-salt organic wastewater. This method utilizes microbial methods to screen out halophilic and salt-tolerant bacteria that can degrade pollutants in a high-salt environment. This can effectively solve the problem of treating high-salt organic wastewater in practice, reduce water treatment costs, and address the problems mentioned in the background art.

[0006] To achieve the above objectives, the present invention provides the following technical solution: a biological treatment method for high-salt organic wastewater, comprising the following steps: Step 1: Screening of halophilic bacteria. The desired halophilic bacteria are directly screened from the soil of saline-alkali land. Then, the mixed culture of the bacterial community obtained from the initial screening is screened again. Step 2: Screening of salt-tolerant bacteria. Salt-tolerant bacteria are directly screened from pre-selected activated sludge, and then the bacterial culture obtained from the initial screening is screened again. Step 3: Isolation and purification of the strains. The bacterial culture obtained above is transferred to a high-salt organic medium at an inoculation rate of 2%. The halophilic and halophilic bacteria are further purified by streak plating until single colonies grow. Step four: identification of strains. Two purified halophilic bacteria and one halophilic bacteria were sent to the assay site for 16S rRNA gene sequencing. The sequencing results were compared with the nucleic acid sequences in the GenBank database.

[0007] For example, the screening method in the screening of halophilic bacteria is as follows: The collected soil samples were prepared into a solution using PBS buffer solution and shaken on a shaker for 30 minutes to make a soil suspension. 100 mL of the mixed soil suspension was taken and added to 1 L of primary screening culture medium, followed by secondary screening, with an addition amount of 1-3%.

[0008] For example, the initial screening culture medium consists of 30-60 g / L sodium chloride, 1 g / L glucose, 0.05 g / L nitrogen source, 0.001 g / L phosphorus source, and a pH between 7.2 and 7.8. Culture temperature: 33-37℃, aerobic culture; During the screening process, the COD concentration is monitored regularly. After 3-5 rounds of degradation, the inorganic salt concentration is gradually increased to 60-90 g / L. After another 3-5 rounds of degradation, a microbial culture that can tolerate high-salt wastewater of no more than 90 g / L is obtained.

[0009] For example, the screening method for the salt-tolerant bacteria is as follows: The activated sludge dosage during screening is 5 g / L; The initial screening medium consisted of 30-60 g / L sodium chloride, 1 g / L glucose, 0.05 g / L nitrogen source, and 0.001 g / L phosphorus source, with a pH between 7.2 and 7.8. The culture temperature was 33-37 ℃, using aerobic culture. Furthermore, during the screening process, the COD concentration was monitored regularly. After 3-5 rounds of degradation, the inorganic salt concentration was gradually increased to 60-90 g / L, and after another 3-5 rounds of degradation, a mixed culture of bacteria capable of tolerating high-salinity wastewater not exceeding 90 g / L was obtained.

[0010] For example, the dosage in the secondary screening process is 1-3%; The secondary screening medium consisted of 60-90 g / L sodium chloride, 1 g / L organic nitrogen, and 0.02 g / L phosphorus source, with a pH between 7.2 and 7.8. The culture temperature was 33-37 ℃, and the culture was aerobic.

[0011] For example, during the screening process, the concentrations of total nitrogen and ammonia nitrogen, as well as the amount of ammonia nitrogen released, need to be monitored regularly. After 3-5 rounds of degradation, a mixed culture of bacteria that can tolerate high-salt organic wastewater of no more than 90g / L is obtained. Among them, the organic nitrogen in the secondary screening culture medium consists of n-butylamine, formamide, and triethylamine, all of which can be degraded.

[0012] For example, the formulation of the high-salt organic culture medium is as follows: 30-60 g / L sodium chloride, 0.5 g / L peptone, 0.25 g / L yeast extract, 1 g / L organic matter, n-butylamine:formamide:triethylamine = 1:1:1, 0.001 g / L phosphorus source, pH between 7.2 and 7.8; The culture was incubated at 33°C in a shaker until the logarithmic phase, and then the culture was diluted and spread onto a high-salt organic solid medium using the dilution plating method.

[0013] For example, the high-salt organic solid culture medium contains 30-60 g / L sodium chloride, 0.5 g / L peptone, 0.25 g / L yeast extract, 1 g / L organic matter, n-butylamine:formamide:triethylamine = 1:1:1, 0.001 g / L phosphorus source, 2% agar, and a pH between 7.2 and 7.8. Incubate in a 33℃ constant temperature incubator for 2-3 days.

[0014] For example, during the purification process, single colonies grown in a solid culture medium are separated using an inoculation needle in a clean bench.

[0015] For example, during the purification process, the operation is repeated more than three times until the colonies growing on the plate have a uniform morphology, thus obtaining two purified halophilic bacteria and one halophilic bacteria.

[0016] Compared with the prior art, the beneficial effects of the present invention are: Using microbial methods, halophilic bacteria ZC2403-SYL, ZC2404-SYT, and halophilic bacteria ZC2402-MT, which can still degrade pollutants in high-salt environments, were screened out. They can degrade organic compounds such as triethylamine, n-butylamine, and formamide in high-salt environments, with degradation rates exceeding 90% within 72 hours. A highly efficient salt-tolerant biological treatment system and a highly efficient halophilic bacteria system were constructed for high-salt and recalcitrant organic pollutants. The three screened bacterial strains were mixed and cultured to rapidly form a highly efficient degradative bacterial agent for high-salt organic wastewater, ZC-SY. When applied to actual high-salt organic wastewater, it can achieve stable operation and effectively solve the problem of treating high-salt organic wastewater in practice, reducing water treatment costs.

[0017] Other features and advantages of the invention will be set forth in the following description, and will be apparent in part from the description, or may be learned by practicing the invention. The objects and other advantages of the invention may be realized and obtained by means of the structures pointed out in the description and the drawings. Attached Figure Description

[0018] Figure 1 This is a flowchart of the method of the present invention; Figure 2 This is a graph showing the change in organic nitrogen concentration during the degradation experiment of triethylamine wastewater in this invention. Figure 3 This is a graph showing the change in organic nitrogen concentration during the degradation experiment of n-butylamine wastewater in this invention. Figure 4 This is a graph showing the change in organic nitrogen concentration during the degradation experiment of formamide wastewater in this invention. Figure 5 This is a graph showing the change in the organic nitrogen degradation rate of the present invention. Detailed Implementation

[0019] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0020] The preservation information of *Alkalophilus halophytes* involved in the following examples is as follows: Name of the depository: China General Microbiological Culture Collection Center (CGMCC); Address of the depository: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing; Date of deposit: August 22, 2024; Accession number: CGMCC No. 31700; The preservation information of *Moraxella roselliae* involved in the following examples is as follows: Name of the depository: China General Microbiological Culture Collection Center (CGMCC); Address of the depository: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing; Date of deposit: May 10, 2024; Accession number: CGMCC No. 30575. This invention provides a biological treatment method for high-salt organic wastewater, comprising the following steps: Step 1: Screening of halophilic bacteria. The desired halophilic bacteria are directly screened from the soil of saline-alkali land. Then, the mixed culture of the bacterial community obtained from the initial screening is screened again. Halophilic bacteria were directly screened from saline-alkali soil using the following screening method: The collected soil samples were prepared into a solution using PBS buffer solution and shaken on a shaker for 30 minutes to make a soil suspension. 100 mL of the mixed soil suspension was then added to 1 L of primary screening culture medium. The initial screening culture medium consisted of 30-60 g / L sodium chloride, 1 g / L glucose, 0.05 g / L nitrogen source, and 0.001 g / L phosphorus source, with a pH between 7.2 and 7.8. Incubation temperature: 33-37 ℃, aerobic culture; In addition, during the screening process, the COD concentration is monitored regularly. After 3-5 rounds of degradation, the inorganic salt concentration is gradually increased to 60-90 g / L. After another 3-5 rounds of degradation, a microbial culture that can tolerate high-salt wastewater of no more than 90 g / L is obtained.

[0021] The microbial culture obtained from the initial screening is then subjected to a second screening, with an addition rate of 1-3%. The second screening culture medium consists of 60-90 g / L sodium chloride, 1 g / L organic nitrogen, 0.002 g / L phosphorus source, and a pH between 7.2 and 7.8. Incubation temperature: 33-37 ℃, aerobic culture; In addition, during the screening process, COD concentration and ammonia nitrogen release are monitored regularly. After 3-5 rounds of degradation, a microbial culture that can tolerate high-salt organic wastewater of no more than 90g / L is obtained. The organic matter in the secondary screening culture medium consists of n-butylamine, formamide, and triethylamine, all of which can be degraded.

[0022] Step 2: Screening of salt-tolerant bacteria. Salt-tolerant bacteria are directly screened from pre-selected activated sludge, and then the bacterial culture obtained from the initial screening is screened again. Salt-tolerant bacteria were directly screened from activated sludge at a dosage of 5 g / L. The screening method is as follows: The initial screening culture medium consisted of 30-60 g / L sodium chloride, 1 g / L glucose, 0.05 g / L nitrogen source, and 0.001 g / L phosphorus source, with a pH between 7.2 and 7.8. Incubation temperature: 33-37 ℃, aerobic culture; In addition, during the screening process, the COD concentration is monitored regularly. After 3-5 rounds of degradation, the inorganic salt concentration is gradually increased to 60-90 g / L. After another 3-5 rounds of degradation, a microbial culture that can tolerate high-salt wastewater of no more than 90 g / L is obtained.

[0023] The microbial culture obtained from the initial screening is then subjected to a second screening, with an addition rate of 1-3%. The second screening culture medium consists of 60-90 g / L sodium chloride, 1 g / L organic nitrogen, and 0.02 g / L phosphorus source, with a pH between 7.2 and 7.8. Incubation temperature: 33-37 ℃, aerobic culture; In addition, during the screening process, the concentrations of total nitrogen and ammonia nitrogen, as well as the amount of ammonia nitrogen released, are monitored regularly. After 3-5 rounds of degradation, a mixed culture of bacteria that can tolerate high-salt organic wastewater of no more than 90g / L is obtained. The organic nitrogen in the secondary screening culture medium consists of n-butylamine, formamide, and triethylamine, all of which can be degraded.

[0024] Step 3: Isolation and purification of the strains. The bacterial culture obtained above is transferred to a high-salt organic medium at an inoculation rate of 2%. The halophilic and halophilic bacteria are further purified by streak plating until single colonies grow. Isolation: Under aseptic conditions, the above-obtained bacterial cultures were transferred at an inoculation rate of 2% to a high-salt organic medium. The high-salt organic medium was formulated as follows: 30-60 g / L sodium chloride, 0.5 g / L peptone, 0.25 g / L yeast extract, 1 g / L organic matter, n-butylamine:formamide:triethylamine = 1:1:1, 0.001 g / L phosphorus source, pH 7.2-7.8. The culture was incubated at 33℃ in a shaker until the logarithmic growth phase. The culture was then diluted and spread onto a high-salt organic solid medium using the dilution plating method. The high-salt organic solid medium consisted of 30-60 g / L sodium chloride, 0.5 g / L peptone, 0.25 g / L yeast extract, 1 g / L organic matter (n-butylamine:formamide:triethylamine = 1:1:1), 0.001 g / L phosphorus source, 2% agar, and a pH between 7.2 and 7.8. Incubate in a 33℃ constant temperature incubator for 2-3 days.

[0025] Purification: In a clean bench, single colonies grown on solid culture medium were isolated using an inoculation needle. The halophilic and halophilic bacteria were further purified by streak plating until single colonies grew. This process was repeated at least three times until the colonies on the plate exhibited a uniform morphology, resulting in two purified halophilic bacteria and one halophilic bacteria strain.

[0026] Step four: identification of strains. Two purified halophilic bacteria and one halophilic bacteria were sent to the assay site for 16S rRNA gene sequencing. The sequencing results were compared with the nucleic acid sequences in the GenBank database.

[0027] Simultaneously, a reasonable phylogenetic tree was constructed using the MEGA 5.0 software method; The final identification determined the taxonomic position of the strain; The identification results and physiological characteristics of the strain are shown in the following tables: halophilic bacteria alkalophilic halomonas Gram-negative bacteria; their morphology and structure, as observed under an electron microscope, are rod-shaped, single, without cross-linking, and the colony surface is smooth. halophilic bacteria Bacillus Horikoshi Gram-positive, bacillus, spore-forming, colony morphology is round, raised in the middle, with neat edges, yellow, smooth, and opaque; strictly aerobic or intermittently anaerobic. Salt-tolerant bacteria Marine Rosellia Moraxella The colonies are round or oval with irregular edges, opaque, moist, and dull. The bacterial cells are rod-shaped, heterotrophic aerobic, and Gram-positive. Based on the above, for organic wastewater with a total salt content of 3-8%, halophilic and salt-tolerant bacteria are pre-acclimated and then introduced into a biological treatment tank for biological treatment. Among them, high-salt organic wastewater with poor biodegradability can be pretreated by advanced oxidation, adsorption and other physicochemical methods to improve its biodegradability, and then degraded by domesticated halophilic and halophilic bacteria. When the salinity of the effluent is limited, advanced treatments such as desalination can be used to ensure that the salinity of the effluent meets the standards. The main technical point of this invention is that it uses microbial methods to screen out halophilic and salt-tolerant bacteria that can degrade pollutants in a high-salt environment, which can effectively solve the problem of treating organic high-salt wastewater in practice, reduce water treatment costs, and improve water treatment efficiency.

[0028] The present invention also includes the following embodiments: The following embodiments are applicable to the present invention, but are not intended to limit the scope of the present invention; Unless otherwise specified, the technical means used in the embodiments are conventional means well known to those skilled in the art.

[0029] In the following examples, the method for determining ammonia nitrogen was Nessler's reagent spectrophotometry; the method for determining total nitrogen was ultraviolet spectrophotometry.

[0030] Example 1: Isolation and Purification of Bacterial Strains Domestication and enrichment: 1. Halophilic bacteria: Add 1% of a mixed culture of bacteria that can tolerate high salinity wastewater up to 90g / L, obtained directly from soil in a saline-alkali area. The culture medium formula is: 60g / L sodium chloride, 1g / L organic nitrogen, 0.2g / L phosphorus source, pH 7.2-7.8. The organic nitrogen in the culture medium was successively replaced by n-butylamine, formamide, and triethylamine, all of which could be degraded. Culture temperature: 33℃, aerobic culture. In addition, during the screening process, the total nitrogen, ammonia nitrogen concentration, and ammonia nitrogen release are monitored regularly. After 3-5 rounds of degradation, a mixed culture of bacteria that can tolerate high-salt organic wastewater not exceeding 90g / L is obtained, from which halophilic bacteria that can degrade organic matter can be isolated and purified.

[0031] 2. Salt-tolerant bacteria: A mixed culture of bacteria capable of tolerating high-salt wastewater not exceeding 90 g / L was obtained by screening and adding it to activated sludge. The addition amount was 1%, and the culture medium consisted of 60 g / L sodium chloride, 1 g / L organic nitrogen, and 0.2 g / L phosphorus source, with a pH between 7.2 and 7.8. Culture temperature: 33℃, aerobic culture; The organic nitrogen in the culture medium was successively replaced by n-butylamine, formamide, and triethylamine, all of which were capable of degradation. Culture temperature: 33℃, aerobic culture. In addition, during the screening process, the concentrations of total nitrogen and ammonia nitrogen, as well as the amount of ammonia nitrogen released, are monitored regularly. After 3-5 rounds of degradation, a mixed culture of bacteria that can tolerate high-salt organic wastewater of no more than 90 g / L is obtained, and salt-tolerant bacteria that can degrade organic matter can be isolated and purified.

[0032] Separation: 1. Halophilic bacteria: Under aseptic conditions, the halophilic bacterial culture obtained through domestication and enrichment was transferred to a high-salt organic medium at an inoculation rate of 2%. The high-salt organic medium formula is as follows: 30-60 g / L sodium chloride, 0.5 g / L peptone, 0.25 g / L yeast extract, 1 g / L organic matter (n-butylamine:formamide:triethylamine = 1:1:1), 0.01 g / L phosphorus source, pH between 7.2 and 7.8. The culture was incubated at 33℃ in a shaker until the logarithmic growth phase. The culture was then diluted and spread onto a high-salt organic solid medium using the dilution plating method. The high-salt organic solid medium consisted of 30-60 g / L sodium chloride, 0.5 g / L peptone, 0.25 g / L yeast extract, 1 g / L organic matter (n-butylamine:formamide:triethylamine = 1:1:1), 0.001 g / L phosphorus source, 2% agar, and a pH between 7.2 and 7.8. Incubate in a 33℃ constant temperature incubator for 2-3 days.

[0033] 2. Salt-tolerant bacteria: Under aseptic conditions, the salt-tolerant bacterial culture obtained through domestication and enrichment was transferred to a high-salt organic medium at an inoculation rate of 2%. The high-salt organic medium formula was: 30-60 g / L sodium chloride, 0.5 g / L peptone, 0.25 g / L yeast extract, 1 g / L organic matter (n-butylamine:formamide:triethylamine = 1:1:1), 0.01 g / L phosphorus source, pH between 7.2 and 7.8. The culture was incubated at 33℃ in a shaker until the logarithmic growth phase. The culture was then diluted and spread onto a high-salt organic solid medium using the dilution plating method. The high-salt organic solid medium consisted of 30-60 g / L sodium chloride, 0.5 g / L peptone, 0.25 g / L yeast extract, 1 g / L organic matter (n-butylamine:formamide:triethylamine = 1:1:1), 0.01 g / L phosphorus source, 2% agar, and a pH between 7.2 and 7.8. Incubate in a 33℃ constant temperature incubator for 2-3 days.

[0034] 3. Purification In a clean bench, single colonies grown in solid culture medium are isolated using an inoculation needle. The halophilic and halophilic bacteria are further purified by streak plating until single colonies grow. Repeat the operation more than 3 times until the colonies growing on the plate have a uniform morphology, thus obtaining 2 purified halophilic bacteria and 1 halophilic bacteria.

[0035] Example 2, Identification of the strain Two purified halophilic bacteria and one halophilic bacteria were sent to the sequencing site for 16S rRNA gene sequencing. The sequencing results were compared with the nucleic acid sequences in the GenBank database. At the same time, a reasonable phylogenetic tree was constructed using MEGA 5.0 software. The final identification determined the taxonomic position of the strain; The identification results and physiological characteristics of the strain are shown in the table below: halophilic bacteria alkalophilic halomonas Gram-negative bacteria; their morphology and structure, as observed under an electron microscope, are rod-shaped, single, without cross-linking, and the colony surface is smooth. halophilic bacteria Bacillus Horikoshi Gram-positive, bacillus, spore-forming, colony morphology is round, raised in the middle, with neat edges, yellow, smooth, and opaque; strictly aerobic or intermittently anaerobic. Salt-tolerant bacteria Marine Rosellia Moraxella The colonies are round or oval with irregular edges, opaque, moist, and dull. The bacterial cells are rod-shaped, heterotrophic aerobic, and Gram-positive. The CGMCC (China General Museum of Phytosanitary Collection) entry number for *Alkalophilus halophyte* is 31700. Please deposit and specify the following biological material (strain) used for identification: ZC2403-SYL.

[0036] The CGMCC No. 30575 is the depository registration number for *Roseola molasses*. Please deposit and specify the following biological material (strain) used for identification: ZC2402-MT.

[0037] Example 3, method for mixed culture of strains: The method for mixed culture of strains ZC2403-SYL, ZC2404-SYT, and ZC2402-MT includes the following steps: 1. Activation: Single colonies of strains ZC2403-SYL, ZC2404-SYT, and ZC2402-MT from solid plates were transferred to a high-salt organic medium (20 mL). The medium consisted of 60 g / L sodium chloride, 0.5 g / L peptone, 0.25 g / L yeast extract, 1 g / L organic matter, n-butylamine:formamide:triethylamine = 1:1:1, and 0.1 g / L phosphorus source. The medium was incubated at 33°C for 24-48 h at 100 rpm until the logarithmic phase. The single carbon source medium consisted of 0.5 g / L dimethyl phosphite, 5 g / L nitrogen source, 2-10 g / L inorganic salts, and a pH between 7.2 and 7.8.

[0038] 2. Transfer: The mixed bacterial culture activated in the logarithmic phase of process 1 was transferred to a 1L seed tank at an inoculation rate of 5% for cultivation. The temperature of the seed tank was maintained at 33℃, the rotation speed was maintained at 100 rpm, and the dissolved oxygen (DO) was controlled at 5 mg / L for 48 hours. Organic matter was added in a timely manner, and the concentration of organic matter was gradually increased to 2 g / L. The organic matter composition was n-butylamine:formamide:triethylamine = 1:1:1, which enabled the bacteria to tolerate higher salinity organic wastewater and significantly improved the organic matter degradation rate. The culture medium for the seed tank consisted of the following components by mass ratio: organic matter, n-butylamine:formamide:triethylamine = 1:1:1, 2 g / L, glucose 0.2 g / L, sodium dihydrogen phosphate 0.2 g / L, MgCl2 0.02 g / L, CaCl2 0.03 g / L, NaHCO3 0.5 g / L, and pH 7.2.

[0039] 3. Expansion culture: The high-salt organic nitrogen degrading bacterial culture from the seed tank in process 2 was transferred to a 10L fermenter for expansion culture at an inoculation rate of 5%. The composition of the culture medium in the fermenter was the same as that in the seed tank. The physicochemical parameters were: temperature 33℃, rotation speed 100rpm, dissolved oxygen 8mg / L, and fermentation time 72h. After fermentation, the effective viable bacteria count in the fermentation solution in the tank can reach 10. 9 When the fermentation broth is discharged from the tank and packaged in plastic buckets, a mixed bacterial solution of high-salt organic high-efficiency degrading strains ZC2403-SYL, ZC2404-SYT and ZC2402-MT can be obtained.

[0040] Example 4: Degradation experiment of triethylamine wastewater by mixed bacterial cultures of ZC2403-SYL, ZC2404-SYT, and ZC2402-MT: Add 1% of the mixed bacterial culture of ZC2403-SYL, ZC2404-SYT, and ZC2402-MT cultivated in this invention to a shake flask containing 0.5 g / L triethylamine as a single carbon source in 100 mL of the system. The total nitrogen concentration is 95 mg / L. The culture medium formula includes: 0.5 g / L triethylamine, 0.01 g / L phosphorus source, 60 g / L NaCl, 0.03 g / L CaCl2, and 0.5 g / L NaHCO3. The shaker was set at 100 rpm and the temperature at 33℃. Ammonia nitrogen and total nitrogen concentrations were measured at 0h, 12h, 24h, 36h, 48h, 60h, and 72h. After 72h of incubation, organic nitrogen decreased to 5 mg / L, achieving complete release. Figure 2 As shown.

[0041] Example 5: Degradation experiment of n-butylamine wastewater by mixed bacterial cultures of ZC2403-SYL, ZC2404-SYT, and ZC2402-MT: Add 1% of the mixed bacterial strains ZC2403-SYL, ZC2404-SYT, and ZC2402-MT cultivated in this invention to a shake flask containing a single carbon source medium of 0.5 g / L n-butylamine. The system is 100 mL, and the total nitrogen concentration is 96 mg / L. The medium formula includes: 0.5 g / L n-butylamine, 0.01 g / L phosphorus source, 60 g / L NaCl, 0.03 g / L CaCl2, and 0.5 g / L NaHCO3. The culture was performed on a shaker at 100 rpm and 33℃. Ammonia nitrogen and total nitrogen concentrations were measured at 0h, 12h, 24h, 36h, and 48h. After 48 hours of incubation, the organic nitrogen concentration decreased to 2 mg / L, achieving complete release. Figure 3 As shown.

[0042] Example 6: Degradation test of formamide wastewater by mixed bacterial solutions of ZC2403-SYL, ZC2404-SYT, and ZC2402-MT: A 1% mixture of ZC2403-SYL, ZC2404-SYT, and ZC2402-MT cultures cultivated in this invention was added to 100 mL of a shake flask containing a single carbon source medium with 0.5 g / L formamide. The total nitrogen concentration was 156 mg / L. The medium formulation included: 0.5 g / L formamide, 0.01 g / L phosphorus source, 60 g / L NaCl, 0.03 g / L CaCl2, and 0.5 g / L NaHCO3. The culture was carried out on a shaker at 100 rpm and 33°C. The concentrations of ammonia nitrogen and total nitrogen were measured at 0h, 12h, 24h, 36h, 48h and 60h, respectively. After 48 hours of incubation, the organic nitrogen concentration decreased to 4 mg / L, achieving complete release. Figure 4 As shown.

[0043] Example 7, Method for rapid formation of bacterial agent from mixed bacterial solutions of ZC2403-SYL, ZC2404-SYT and ZC2402-MT: The steps for rapid sludge formation using mixed bacteria are as follows; 1. Add inoculum sludge to the reactor, and transport wastewater from the bottom device. Add 1 g / L of mixed bacterial solution of ZC2403-SYL, ZC2404-SYT and ZC2402-MT at one time. The temperature inside the device is 33℃, the rotation speed is maintained at 150 rpm, the dissolved oxygen (DO) value is 4 mg / L, and the pH is 7.5.

[0044] 2. Set the retention time to 24 hours and add nutrients and trace elements; The nutrient dosage is as follows: NaCl 6%, NaHCO3 0.2%, MgSO4 0.05%, CaCl2 0.02%, yeast extract 0.01%, and the pH is maintained between 7.2 and 7.8. The dosage of trace elements is as follows: Fe 10 mg / L, Cu 0.3 mg / L, Mo 0.1 mg / L, Zn 0.1 mg / L, Co 0.1 mg / L, Mn 0.1 mg / L.

[0045] 3. Add the mixed bacterial solution of ZC2403-SYL, ZC2404-SYT and ZC2402-MT to the reactor in a single addition method. The temperature inside the device is 33℃ and the pH is controlled at 7.2. The bacterial strain was inoculated into the sludge at a dosage of 10% for rapid propagation, and the effective viable bacteria count in the reactor solution could reach 10. 9 When the number of bacteria per ml is above a certain level, a mixed bacterial solution of ZC2403-SYL, ZC2404-SYT and ZC2402-MT, which are highly efficient degrading bacteria for high-salt organic wastewater, is obtained and named as ZC-SY, which can be used in subsequent wastewater treatment processes. The reactor height-to-diameter ratio H / D is 5.

[0046] Example 8: Application of ZC-SY, a highly efficient degrading microbial agent for high-salt organic wastewater, in actual high-salt organic wastewater: To further verify the application effect of the high-salt organic wastewater efficient degradation agent ZC-SY used in this invention in actual high-salt organic wastewater, high-salt organic nitrogen wastewater was obtained from the production workshop of the pharmaceutical company in question. The concentration of organic nitrogen in the wastewater was measured to be approximately 200 mg / L. The ZC-SY bacterial solution, a highly efficient degrading agent for high-salt organic wastewater, obtained in Example 7, was directly added to the wastewater at an inoculation rate of 10%, and the pH of the system was adjusted to 7.2 to meet the needs of normal microbial growth. Under room temperature conditions, the changes in organic nitrogen in the system were monitored by taking samples at regular intervals of 0h, 16h, 24h, 48h, and 72h. Figure 5 As shown; After 24 hours, the organic nitrogen concentration had dropped to 2 mg / L, and the rate of organic nitrogen released as ammonia nitrogen was as high as 99%. The above experimental data indicate that the highly efficient degrading bacterial agent ZC-SY for high-salt organic wastewater also has good treatment effects in actual high-salt organic wastewater and has broad application prospects.

[0047] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A biological treatment method for high-salt organic wastewater, characterized in that, The high-salt organic wastewater is triethylamine, n-butylamine, or formamide wastewater. It is treated using a mixed bacterial solution composed of *Halomonas alkaliphila*, *Bacillus hormonei*, and *Moraxella rosenbergii*. The *Halomonas alkaliphila* strain is named ZC2403-SYL, deposited at the China General Microbiological Culture Collection Center (CGMCC) on August 22, 2024, at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. The deposit number is CGMCC No. 31700. The Bacillus horikoshii strain was named ZC2404-SYT; The species *Rossellomorea aquimaris* is named ZC2402-MT. The depositary institution is the China General Microbiological Culture Collection Center (CGMCC); address: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing; deposit date: May 10, 2024; accession number: CGMCC No. 30575.

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