Microbial remediation method for synchronously removing copper and ammonia pollution in wastewater and application of microbial remediation method

Abstract

The invention relates to the technical field of environmental engineering, and particularly discloses a microbial remediation method for synchronously removing copper and ammonia pollution in wastewater and application of the microbial remediation method, and the microbial remediation method comprises a bacterial culture process, a bacterial phosphorus dissolving process, an environmental pH increasing process, a biomineralization process and a nitrification-denitrification process. The bacterial culture comprises the steps of (1) bacterial activation, (2) bacterial low-temperature preservation, (3) bacterial propagation and (4) bacterial inoculation; the bacterial phosphate solubilizing process specifically comprises the following steps: bacillus megaterium secretes phosphatase to catalyze sodium glycerophosphate to hydrolyze so as to generate PO4 < 3->; in the environment pH increasing process, bacteria release OH <-> to increase the environment pH value; the biological mineralization process specifically comprises the following steps: Cu < 2 + > in the wastewater reacts with PO4 < 3-> and OH <->, and a blue phosphorus copper ore (Cu3 (PO4) 2 (OH) 3) precipitate is generated by taking bacillus megatherium as a nucleation site; the nitrification-denitrification process specifically comprises the following steps: oxidizing NH4 < + > into NO2 <-> and further oxidizing NO3 <-> by the bacillus megatherium through a nitrification process, then reducing the NO3 <-> into N2 through a denitrification process, and finally achieving the purpose of removing the NH4 < + > in the wastewater. Ammonia can be further oxidized into nitrite to threaten human health. According to previous researches, ammonia and copper form a copper-ammonia complex, and meanwhile, the toxicity of copper to bacteria can also inhibit nitrification and denitrification. Therefore, ammonia and copper in the environment are difficult to remove. The bacillus megaterium adopted by the invention has excellent phosphate solubilizing and nitrification-denitrification capabilities. The copper can be solidified through a microorganism induced phosphate precipitation technology, and ammonia can be removed through a nitrification-denitrification process. The microbial remediation method disclosed by the invention can be used for quickly and efficiently treating copper and ammonia pollution in the wastewater, and secondary pollution to the environment is avoided.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of environmental engineering, and particularly relates to a microbial remediation method for simultaneously removing copper and ammonia pollution in wastewater and application thereof. BACKGROUND

[0002] The coexistence of multiple pollutants is a common feature of wastewater, among which common substances include ammonia (NH4 + ) and heavy metals (Cu, Pb, Zn), etc. Most heavy metal ions are released into the environment through industrial processes such as smelting, mining, chemical industry, and garbage dumping. Heavy metal ions in wastewater pose a threat to human health and ecosystems due to their toxicity, persistence, and bioaccumulation. Copper (Cu) produced by modern agricultural activities exists in the surrounding environment and poses a serious threat to all organisms. It can enter the human body through the food chain and cause damage to the brain, kidneys, bones, lungs, liver, and reproductive system. Over the past few decades, various remediation technologies such as flocculation, sedimentation, electrolysis, and electrochemical treatment have been widely used for the remediation of lead-contaminated wastewater. However, these methods have been criticized for their high cost, time-consuming nature, and limited application to certain Cu 2+ concentrations. The strains used in the microbial-induced carbonate precipitation (MICP) technology have poor urea-lyzing ability in acidic environments, and the precipitates generated are generally less chemically stable and can easily be leached out in acidic environments, causing secondary migration of heavy metals to the environment. The decomposition of urea during MICP treatment of heavy metals produces a large amount of ammonia gas, increasing the risk of secondary environmental pollution. Phosphate minerals obtained through microbial-induced phosphate precipitation (MIPP) technology are considered to be the most effective lead stabilizer. MIPP technology involves the catalysis of inorganic or organic phosphorus sources by phosphate-solubilizing microorganisms to release PO4 3- and promote the immobilization of Cu 2+ . Some phosphate-solubilizing microorganisms can secrete organic acids to increase the solubility of inorganic phosphorus sources (hydroxyapatite and phosphate ore) and release PO4 3- for Cu 2+ immobilization. Some can produce phosphatase or phytase to catalyze the hydrolysis of organic phosphorus sources, releasing PO4 3- and promoting the formation of phosphate minerals, which have a solubility product much lower than that of carbonate minerals.

[0003] NH4 + not only has a certain toxic effect on the human body and aquatic organisms, but also is a high-oxygen-consuming substance. High concentrations of NH4 + will directly lead to deterioration of groundwater quality. Therefore, the removal of NH4 + from water is crucial for environmental safety and human health. At present, domestic and foreign researches on NH4 +The removal methods mainly include adsorption, electrochemical method, chemical precipitation, ion exchange, advanced oxidation and nitration-denitrification biological method. Compared with other methods, the nitration-denitrification biological method has the advantages of high removal rate, low cost and no pollution, and thus becomes the most popular NH4 + removal method in water treatment. Compared with the traditional two relatively independent aerobic nitrification and anaerobic / amoxic denitrification processes, the nitration-denitrification process can complete the nitrification and denitrification processes simultaneously. The denitrification mechanism of heterotrophic nitrifying bacteria supplements and breaks through the traditional nitrification theory. Therefore, it has great potential and application blank in denitrification. In the work, the microbial induced phosphate precipitation is combined with the nitration-denitrification process, so that the bacillus megaterium can remove Cu 2+ and NH4 + under the two processes. The study provides an efficient and convenient technology for treating Cu 2+ and NH4 + contaminated groundwater. SUMMARY

[0004] The purpose of the present application is to solve the problems existing in the prior art and provide a microbial remediation method for simultaneously removing copper and ammonia pollution in wastewater.

[0005] In order to achieve the above purpose, the present application adopts the following technical scheme: A preparation method of a microbial agent for simultaneously removing copper and ammonia pollution in wastewater, comprising the following steps: Step one, transfer the bacillus megaterium freeze-dried powder to an activation culture medium, and then culture under constant temperature and humidity conditions for 24h to obtain activated bacteria; Step two, mix the activated bacteria solution with glycerol according to the proportion and transfer to a cryopreservation tube, and then store the cryopreservation tube at-20°C; Step three, inoculate the bacteria solution in the cryopreservation tube into a growth culture medium according to a volume ratio of 0.1%, and culture under constant temperature and humidity conditions for 36h to obtain a bacteria solution.

[0006] Preferably, in step one, the activation culture medium comprises 2-4g / L beef extract, 4-6g / L peptone and 4-6g / L NaCl; the constant temperature and humidity bacteria incubator is set to a culture temperature of 25-35°C, a shaking speed of 170-190rpm and a culture time of 24h; in step one, the density of the activated bacteria solution obtained is OD 600 =1.4-1.6.

[0007] Preferably, in step two, the bacteria solution and glycerol are mixed according to a volume ratio of 7:3.

[0008] Preferably, in step three, the growth medium comprises 4.5-6.5 g / L sodium succinate, 0.25-0.35 g / L ammonium chloride, 0.5-1.5 g / L sodium chloride, 0.1-0.3 g / L potassium dihydrogen phosphate, 0.04-0.06 g / L magnesium sulfate heptahydrate, 0.01-0.03 g / L calcium chloride, 0.005-0.015 g / L ferric sulfate heptahydrate, and 0.005-0.015 g / L manganese sulfate monohydrate; in step three, the constant-temperature and constant-humidity bacterial incubator is set to a culture temperature of 25-35℃, a shaking rate of 150-170 rpm, and a culture time of 36 h; in step three, the prepared growth medium is sterilized in a 121℃ high-temperature and high-pressure steam sterilization pot for 30 min; and in step three, the obtained bacterial solution has a density of OD 600 =1.4-1.6.

[0009] A microbial remediation method for simultaneously removing copper and ammonia pollution in wastewater, comprising inoculating the bacterial solution obtained by the above method into wastewater containing Cu²⁺ and NH4⁺ at a volume ratio of 5%; adding glycerol sodium phosphate to the wastewater; supplementing the growth medium to the wastewater to maintain the basic nutritional requirements for bacterial growth and reproduction, and then culturing under constant-temperature and constant-humidity conditions for 72 h.

[0010] Preferably, the concentration of the glycerol sodium phosphate is 6 g / L.

[0011] Preferably, the growth medium comprises 4.5-6.5 g / L sodium succinate, 0.25-0.35 g / L ammonium chloride, 0.5-1.5 g / L sodium chloride, 0.1-0.3 g / L potassium dihydrogen phosphate, 0.04-0.06 g / L magnesium sulfate heptahydrate, 0.01-0.03 g / L calcium chloride, 0.005-0.015 g / L ferric sulfate heptahydrate, and 0.005-0.015 g / L manganese sulfate monohydrate; and the constant-temperature and constant-humidity bacterial incubator is set to a culture temperature of 25-35℃ and a shaking rate of 150-170 rpm.

[0012] Preferably, the initial concentration of Cu²⁺ in the wastewater is 50-150 mg / L, and the initial concentration of NH4⁺ is 100-400 mg / L.

[0013] The bacterial solution prepared by the above method is used for removing Cu 2+ and NH4 + in wastewater, and the bacterial solution further comprises glycerol sodium phosphate, and the concentration of the glycerol sodium phosphate is 6 g / L.

[0014] A wastewater remediation reagent comprising the bacterial solution prepared by the above method.

[0015] Compared with the prior art, the present application has the following beneficial effects: 1. This invention provides a microbial remediation method for simultaneously removing copper and ammonia nitrogen pollution from wastewater. In this method, Bacillus megaterium produces phosphatase that hydrolyzes sodium glycerophosphate to produce PO4. 3- and OH - Cu 2+ The reaction with these two materials produces blue phosphorus copper ore (Cu3(PO4)2(OH)3), which has higher chemical stability than Cu2(CO3)(OH)2 produced by microbial carbonate precipitation technology. 2. This invention provides a microbial remediation method for simultaneously removing copper and ammonia nitrogen pollution from wastewater. Traditional MIP (Microbial Microparticle Processing) technology hydrolyzes urea to produce CO3. 2- It will also produce the byproduct NH4. + This not only exacerbates the NH4 in wastewater + The degree of pollution, and the NH4 produced. + It will also react with Cu in an alkaline environment. 2+ Complexation produces copper-ammonia complexes, which degrade Cu 2+ The removal efficiency is high. In this method, Bacillus megaterium hydrolyzes sodium glycerophosphate without producing other environmentally polluting byproducts. Furthermore, this bacterium can perform nitrification-denitrification simultaneously with biomineralization, removing NH4+. + It is converted into N2 and released into the air; 3. This invention provides a microbial remediation method for simultaneously removing copper and ammonia nitrogen pollution from wastewater. The sodium glycerophosphate used in this method has a chelating effect on Cu, compared to urea used in traditional MIP (Microbiological Microencapsulation Process) technology. 2+ This chelating ability will reduce Cu in the environment. 2+ Concentration, protecting bacteria from Cu 2+ The toxic effects enhance the bacteria's phosphorus-solubilizing ability and biomineralize Cu. 2+ Capacity and nitrification-denitrification capacity, improving Cu 2+ and NH4 + Removal efficiency.

[0016] 4. The concept and processing method of this invention are reasonable, simple to operate, and easy to promote and apply.

[0017] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description

[0018] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0019] Figure 1 Cu 2+ The concentration is 50 mg / L, NH4 + Results of the remediation test at a concentration of 100 mg / L.

[0020] Figure 2 Cu 2+ The concentration is 150 mg / L, NH4 + Results of the remediation test at a concentration of 100 mg / L.

[0021] Figure 3 Cu 2+ The concentration is 50 mg / L, NH4 + Results of the remediation test at a concentration of 400 mg / L.

[0022] Figure 4 Cu 2+ The concentration is 150 mg / L, NH4 + Results of the remediation test at a concentration of 400 mg / L.

[0023] Figure 5 The results are the repair test results of Control Example 1, Control Example 2, Control Example 3 and Example 1.

[0024] Figure 6 Cu 2+ The concentration is 50 mg / L, NH4 + UV-Vis test results of the precipitate at a concentration of 100 mg / L. Detailed Implementation

[0025] The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments.

[0026] Example 1 This embodiment presents a microbial remediation method for simultaneously removing copper and ammonia contaminants from wastewater, comprising: Step 1: Inoculate the bacterial solution from the cryopreservation tubes into growth medium at a volume ratio of 0.1%. The growth medium comprises 4.5–6.5 g / L sodium succinate, 0.25–0.35 g / L ammonium chloride, 0.5–1.5 g / L sodium chloride, 0.1–0.3 g / L potassium dihydrogen phosphate, 0.04–0.06 g / L magnesium sulfate heptahydrate, 0.01–0.03 g / L calcium chloride, 0.005–0.015 g / L ferric sulfate heptahydrate, and 0.005–0.015 g / L manganese sulfate monohydrate. Control the culture temperature at 25–35°C, the shaking rate at 150–170 rpm, and the culture time at 36 h. The density of the obtained bacterial solution is OD0.05. 600 =1.4~1.6.

[0027] Step 2: Configure Cu 2+ Concentration of 50 mg / L, NH4 + For a copper-containing wastewater with a concentration of 100 mg / L and ammonia, add 6 g / L sodium glycerophosphate to the copper-containing wastewater and mix thoroughly. Step 3: Inoculate the cultured Bacillus megaterium solution at a volume ratio of 5% into a solution containing Cu. 2+ and NH4 + The sample was placed in wastewater and then incubated under constant temperature and humidity conditions for 48 hours. Samples were taken at 0h, 12h, 24h, 36h, and 48h to determine the remaining Cu. 2+ Concentration, NH4 + Concentration, pH value.

[0028] In this embodiment, Cu 2+ The concentration is 50 mg / L, NH4 + At a concentration of 100 mg / L, the pH ranges from 7.00 to 8.84, and the final Cu... 2+ The removal efficiency is 99.8% for NH4. + The removal efficiency is 96.8%.

[0029] Figure 6 The ultraviolet absorption spectra of the mineralized products were collected at 0, 24, and 48 hours. A distinct ultraviolet absorption peak was observed at approximately 500 nm, corresponding to Cu. 2+ It forms a chelate, copper glycerophosphate, with sodium glycerophosphate. The highest peak is observed at 0 h, with an absorbance of 52.3 Abs. As the reaction progresses, the peak value of this copper glycerophosphate and other metal chelates gradually decreases, reaching its lowest point at 72 h. UV-Vis experiments confirm the view that sodium glycerophosphate and other components adsorb and complex with heavy metal copper.

[0030] Example 2 Step 1: Inoculate the bacterial solution from the cryopreservation tubes into growth medium at a volume ratio of 0.1%. The growth medium comprises 4.5–6.5 g / L sodium succinate, 0.25–0.35 g / L ammonium chloride, 0.5–1.5 g / L sodium chloride, 0.1–0.3 g / L potassium dihydrogen phosphate, 0.04–0.06 g / L magnesium sulfate heptahydrate, 0.01–0.03 g / L calcium chloride, 0.005–0.015 g / L ferric sulfate heptahydrate, and 0.005–0.015 g / L manganese sulfate monohydrate. Control the culture temperature at 25–35°C, the shaking rate at 150–170 rpm, and the culture time at 36 h. The density of the obtained bacterial solution is OD0.05. 600 =1.4~1.6.

[0031] Step 2: Configure Cu 2+ Concentration of 150 mg / L, NH4 + For a copper-containing wastewater with a concentration of 100 mg / L and ammonia, add 6 g / L sodium glycerophosphate to the copper-containing wastewater and mix thoroughly. Step 3: Inoculate the cultured Bacillus megaterium solution at a volume ratio of 5% into a solution containing Cu. 2+ and NH4 + The sample was placed in wastewater and then incubated under constant temperature and humidity conditions for 48 hours. Samples were taken at 0h, 12h, 24h, 36h, and 48h to determine the remaining Cu. 2+ Concentration, NH4 + Concentration, pH value.

[0032] In this embodiment, Cu 2+ The concentration is 150 mg / L, NH4 + At a concentration of 100 mg / L, the pH ranges from 7.00 to 8.67, and the final Cu... 2+ The removal efficiency was 88.4% for NH4. + The removal efficiency is 92.3%.

[0033] Example 3 Step 1: Inoculate the bacterial solution from the cryopreservation tubes into growth medium at a volume ratio of 0.1%. The growth medium comprises 4.5–6.5 g / L sodium succinate, 0.25–0.35 g / L ammonium chloride, 0.5–1.5 g / L sodium chloride, 0.1–0.3 g / L potassium dihydrogen phosphate, 0.04–0.06 g / L magnesium sulfate heptahydrate, 0.01–0.03 g / L calcium chloride, 0.005–0.015 g / L ferric sulfate heptahydrate, and 0.005–0.015 g / L manganese sulfate monohydrate. Control the culture temperature at 25–35°C, the shaking rate at 150–170 rpm, and the culture time at 36 h. The density of the obtained bacterial solution is OD0.05. 600 =1.4~1.6.

[0034] Step 2: Configure Cu 2+ Concentration of 50 mg / L, NH4 + For a copper-containing wastewater with a concentration of 400 mg / L and ammonia, 6 g / L sodium glycerophosphate was added to the copper-containing wastewater and mixed thoroughly. Step 3: Inoculate the cultured Bacillus megaterium solution at a volume ratio of 5% into a solution containing Cu. 2+ and NH4 + The sample was placed in wastewater and then incubated under constant temperature and humidity conditions for 48 hours. Samples were taken at 0h, 12h, 24h, 36h, and 48h to determine the remaining Cu. 2+ Concentration, NH4 + Concentration, pH value.

[0035] In this embodiment, Cu 2+ The concentration is 50 mg / L, NH4 + At a concentration of 400 mg / L, the pH ranges from 7.00 to 8.63, and the final Cu... 2+ The removal efficiency was 93.6% for NH4. + The removal efficiency was 87.6%.

[0036] Example 4 Step 1: Inoculate the bacterial solution from the cryopreservation tubes into growth medium at a volume ratio of 0.1%. The growth medium comprises 4.5–6.5 g / L sodium succinate, 0.25–0.35 g / L ammonium chloride, 0.5–1.5 g / L sodium chloride, 0.1–0.3 g / L potassium dihydrogen phosphate, 0.04–0.06 g / L magnesium sulfate heptahydrate, 0.01–0.03 g / L calcium chloride, 0.005–0.015 g / L ferric sulfate heptahydrate, and 0.005–0.015 g / L manganese sulfate monohydrate. Control the culture temperature at 25–35°C, the shaking rate at 150–170 rpm, and the culture time at 36 h. The density of the obtained bacterial solution is OD0.05. 600 =1.4~1.6.

[0037] Step 2: Configure Cu 2+ Concentration of 150 mg / L, NH4 + For a copper-containing wastewater with a concentration of 400 mg / L and ammonia, 6 g / L sodium glycerophosphate was added to the copper-containing wastewater and mixed thoroughly. Step 3: Inoculate the cultured Bacillus megaterium solution at a volume ratio of 5% into a solution containing Cu. 2+ and NH4 + The sample was placed in wastewater and then incubated under constant temperature and humidity conditions for 48 hours. Samples were taken at 0h, 12h, 24h, 36h, and 48h to determine the remaining Cu. 2+ Concentration, NH4 +Concentration, pH value.

[0038] In this embodiment, Cu 2+ The concentration is 150 mg / L, NH4 + At a concentration of 400 mg / L, with a pH range of 7.00–8.45, the final Cu 2+ The removal efficiency was 86.9% for NH4. + The removal efficiency was 84.7%.

[0039] Compare with Example 1 Set up an identical Cu²⁺+NH₄⁺ wastewater system and treat it using traditional MIP technology.

[0040] Step 1: Inoculate the *Pasteurella multocida* solution from the cryopreservation tubes into growth medium at a volume ratio of 0.1%. The growth medium comprises 4.5–6.5 g / L yeast extract, 18–22 g / L ammonium chloride, 8–12 g / L ammonium chloride, 20–28 mg / L nickel chloride, and 8–12 mg / L manganese sulfate monohydrate. Control the culture temperature at 25–35°C, the shaking rate at 150–170 rpm, and the culture time at 36 h. The density of the obtained bacterial solution is OD0.05. 600 =1.8~2.2.

[0041] Step 2: Configure Cu 2+ Concentration of 50 mg / L, NH4 + For a copper-containing wastewater with a concentration of 100 mg / L and ammonia, add 6 g / L of urea to the copper-containing wastewater and mix thoroughly. Step 3: Inoculate the cultured Bacillus megaterium solution at a volume ratio of 5% into a solution containing Cu. 2+ and NH4 + The sample was placed in wastewater and then incubated under constant temperature and humidity conditions for 48 hours. Samples were taken at 0h, 12h, 24h, 36h, and 48h to determine the remaining Cu. 2+ Concentration, NH4 + Concentration, pH value.

[0042] In this comparative example, Cu 2+ The concentration is 50 mg / L, NH4 + At a concentration of 100 mg / L, the pH ranges from 8.91 to 9.15, and the final Cu... 2+ The removal efficiency is 2.5% for NH4. + The removal efficiency is 0%.

[0043] Compare with Example 2 Set each to contain only Cu 2+ Wastewater containing only NH4 + Wastewater.

[0044] Step 1: Inoculate the bacterial solution from the cryopreservation tubes into growth medium at a volume ratio of 0.1%. The growth medium comprises 4.5–6.5 g / L sodium succinate, 0.25–0.35 g / L ammonium chloride, 0.5–1.5 g / L sodium chloride, 0.1–0.3 g / L potassium dihydrogen phosphate, 0.04–0.06 g / L magnesium sulfate heptahydrate, 0.01–0.03 g / L calcium chloride, 0.005–0.015 g / L ferric sulfate heptahydrate, and 0.005–0.015 g / L manganese sulfate monohydrate. Control the culture temperature at 25–35°C, the shaking rate at 150–170 rpm, and the culture time at 36 h. The density of the obtained bacterial solution is OD0.05. 600 =1.4~1.6.

[0045] Step 2: Configure Cu 2+ For copper-containing wastewater with a concentration of 50 mg / L, add 6 g / L sodium glycerophosphate to the copper-containing wastewater and mix thoroughly. Step 3: Configure NH4 + Wastewater containing ammonia with a concentration of 100 mg / L; Step 3: Inoculate the cultured Bacillus megaterium solution at a volume ratio of 5% into a solution containing Cu. 2+ and NH4 + The sample was placed in wastewater and then incubated under constant temperature and humidity conditions for 48 hours. Samples were taken at 0h, 12h, 24h, 36h, and 48h to determine the remaining Cu. 2+ Concentration, NH4 + Concentration, pH value.

[0046] In this embodiment, the copper-containing wastewater contains Cu 2+ The concentration of NH4 in ammonia-containing wastewater is 50 mg / L. + At a concentration of 100 mg / L, the pH is 8.45–8.57, and the final Cu 2+ The removal efficiency is 93.5% for NH4. + The removal efficiency is 98.2%.

[0047] Compare with Example 3 The step-by-step process of "first MIPP copper fixation, then inoculation with denitrifying bacteria for deammoniation" takes the same total time as in Example 1.

[0048] Step 1: Inoculate the bacterial solution from the cryopreservation tubes into growth medium at a volume ratio of 0.1%. The growth medium comprises 4.5–6.5 g / L sodium succinate, 0.25–0.35 g / L ammonium chloride, 0.5–1.5 g / L sodium chloride, 0.1–0.3 g / L potassium dihydrogen phosphate, 0.04–0.06 g / L magnesium sulfate heptahydrate, 0.01–0.03 g / L calcium chloride, 0.005–0.015 g / L ferric sulfate heptahydrate, and 0.005–0.015 g / L manganese sulfate monohydrate. Control the culture temperature at 25–35°C, the shaking rate at 150–170 rpm, and the culture time at 36 h. The density of the obtained bacterial solution is OD0.05. 600 =1.4~1.6.

[0049] Step 2: Configure Cu 2+ For copper-containing wastewater with a concentration of 50 mg / L, add 6 g / L sodium glycerophosphate to the copper-containing wastewater and mix thoroughly. Step 3: Inoculate the cultured Bacillus megaterium solution at a volume ratio of 5% into a solution containing Cu. 2+ The sample was placed in wastewater and then incubated under constant temperature and humidity conditions for 24 hours. Samples were taken at 0 h, 12 h, and 24 h to determine the remaining Cu. 2+ Concentration and pH value.

[0050] Step 4: Add 100 mg / L NH4 to the above solution. + The samples were then incubated under constant temperature and humidity conditions for 24 hours. Samples were taken at 24 hours, 36 hours, and 48 hours to determine the remaining Cu. 2+ Concentration and pH value.

[0051] In this embodiment, 50 mg / L Cu was added sequentially. 2+ and 100 mg / L NH4 + The concentration was [value missing], the pH was 8.43~8.61, and the final Cu [value missing] 2 + The removal efficiency was 89.8% for NH4. + The removal efficiency was 76.4%.

[0052] The above description is merely a preferred embodiment of the present invention and does not constitute any limitation on the present invention. Any simple modifications, alterations, or equivalent structural changes made to the above embodiments based on the technical essence of the invention shall still fall within the protection scope of the present invention.

[0053] Performance Evaluation Cu in the above embodiments 2+ The concentrations were 50 mg / L and 150 mg / L, respectively; NH4 +The concentrations were 100 mg / L and 400 mg / L, respectively. Compared with the traditional MIP technology, Cu after treatment using the above microbial method... 2+ and NH4 + The removal efficiency was significantly improved, mainly due to the fact that Bacillus megaterium secretes phosphatase that catalyzes the hydrolysis of sodium glycerophosphate to produce PO4. 3- And it increases the pH value of the environment; Cu in the wastewater 2+ With PO4 3- OH - The reaction produces azurite (Cu3PO4(OH)3) precipitate using Bacillus megaterium as the nucleation site; simultaneously, Bacillus megaterium converts NH4+ into nitrate through a nitration process. + Oxidized to NO2 - And further oxidize NO3 - Then, NO3 is removed through a denitrification process. - The NH4 in the wastewater is reduced to N2, ultimately achieving the removal of NH4. + The purpose of this invention is to address the issue that, according to previous research, ammonia forms copper-ammonia complexes with copper, and the toxicity of copper to bacteria inhibits nitrification and denitrification, making it difficult to remove both ammonia and copper from the environment. This invention utilizes *Bacillus megaterium*, which possesses excellent phosphate solubilization and nitrification-denitrification capabilities. It can both solidify copper through microbial-induced phosphate precipitation technology and remove ammonia through the nitrification-denitrification process. Figure 5 It can be seen that the removal efficiency of Example 1 is related to that of Comparative Example 1, Comparative Example 2, and Comparative Example 3.

[0054] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A method for preparing a microbial agent for simultaneously removing copper and ammonia contamination from wastewater, characterized in that, Includes the following steps: Step 1: Transfer the Bacillus megaterium lyophilized powder to the activation medium, and then incubate it under constant temperature and humidity conditions for 24 hours to obtain activated bacteria; Step 2: Mix the activated bacterial solution with glycerol in the specified ratio and transfer it to cryovials. Then store the cryovials at -20°C. Step 3: Inoculate the bacterial solution in the cryopreservation tube into the growth medium at a volume ratio of 0.1%, and incubate under constant temperature and humidity conditions for 36 hours to obtain the bacterial solution.

2. The method for preparing a microbial agent for simultaneously removing copper and ammonia contamination from wastewater according to claim 1, characterized in that, In step one, the activation culture medium comprises 2-4 g / L beef extract, 4-6 g / L peptone, and 4-6 g / L NaCl; the constant temperature and humidity bacterial incubator is set to a culture temperature of 25-35℃, a shaking rate of 170-190 rpm, and a culture time of 24 h; in step one, the density of the activated bacterial solution is OD. 600 =1.4~1.

6.

3. The method for preparing a microbial agent for simultaneously removing copper and ammonia contamination from wastewater according to claim 1, characterized in that, In step two, the bacterial solution and glycerol are mixed in a volume ratio of 7:

3.

4. The method for preparing a microbial agent for simultaneously removing copper and ammonia contamination from wastewater according to claim 1, characterized in that, In step three, the growth medium comprises 4.5–6.5 g / L sodium succinate, 0.25–0.35 g / L ammonium chloride, 0.5–1.5 g / L sodium chloride, 0.1–0.3 g / L potassium dihydrogen phosphate, 0.04–0.06 g / L magnesium sulfate heptahydrate, 0.01–0.03 g / L calcium chloride, 0.005–0.015 g / L ferric sulfate heptahydrate, and 0.005–0.015 g / L manganese sulfate monohydrate. In step three, the constant temperature and humidity bacterial incubator is set to a culture temperature of 25–35°C, a shaking rate of 150–170 rpm, and a culture time of 36 h. In step three, the prepared growth medium is sterilized in a 121°C high-temperature, high-pressure steam sterilizer for 30 min. In step three, the density of the obtained bacterial solution is OD0. 600 =1.4~1.

6.

5. A microbial remediation method for simultaneously removing copper and ammonia contaminants from wastewater, characterized in that, The method includes inoculating the bacterial solution obtained by any one of claims 1-4 into wastewater containing Cu²⁺ and NH₄⁺ at a volume ratio of 5%; adding sodium glycerophosphate to the wastewater; supplementing the wastewater with growth medium to maintain the basic nutritional needs for bacterial growth and reproduction; and then culturing under constant temperature and humidity conditions for 72 hours.

6. The microbial remediation method for simultaneously removing copper and ammonia contamination from wastewater according to claim 5, characterized in that, The concentration of the sodium glycerophosphate is 6 g / L.

7. The microbial remediation method for simultaneously removing copper and ammonia contamination from wastewater according to claim 5, characterized in that, The growth medium comprises 4.5~6.5 g / L sodium succinate, 0.25~0.35 g / L ammonium chloride, 0.5~1.5 g / L sodium chloride, 0.1~0.3 g / L potassium dihydrogen phosphate, 0.04~0.06 g / L magnesium sulfate heptahydrate, 0.01~0.03 g / L calcium chloride, 0.005~0.015 g / L ferric sulfate heptahydrate, and 0.005~0.015 g / L manganese sulfate monohydrate; the constant temperature and humidity bacterial incubator is set to a culture temperature of 25~35℃ and a shaking rate of 150~170 rpm.

8. The microbial remediation method for simultaneously removing copper and ammonia contamination from wastewater according to claim 5, characterized in that, The initial concentration of Cu²⁺ in the wastewater is 50~150 mg / L, and the initial concentration of NH⁺ is 100~400 mg / L.

9. The bacterial solution prepared by the method according to any one of claims 1-4 for removing Cu from wastewater 2+ and NH4 + The application of [the technology] is characterized by, The bacterial solution also includes sodium glycerophosphate at a concentration of 6 g / L.

10. A wastewater remediation reagent, characterized in that, Bacterial solutions prepared by any of the methods described in 1-4.

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