A composite aquatic ecosystem restoration agent and its preparation method

Abstract

This invention relates to the field of microbial inoculant technology, and proposes a water ecological composite remediation agent and its preparation method. The water ecological composite remediation agent comprises the following components in parts by weight: 60-195 parts of microbial bacteria, 1-10 parts of biological enzymes, and 1-10 parts of nutrients; the microbial bacteria comprise the following components in parts by weight: 5-15 parts of ammonia-oxidizing denitrifying bacteria, 10-35 parts of denitrifying denitrifying bacteria, 10-30 parts of nitrifying bacteria, 10-40 parts of Bacillus subtilis, 10-40 parts of photosynthetic bacteria, 10-20 parts of Bacillus licheniformis, and 5-10 parts of yeast. Through the above technical solution, the problem of poor wastewater treatment effect of water ecological composite remediation agents in related technologies is solved.

Description

Technical Field

[0001] This invention relates to the field of microbial preparation technology, specifically to a water ecological composite restoration agent and its preparation method. Background Technology

[0002] This compound microbial strain possesses both anaerobic and aerobic characteristics, boasting advantages such as high recovery rate, strong environmental adaptability, and a large number of live bacteria. It includes bacterial cells and their metabolites, as well as algae-inhibiting active peptides. It utilizes selected, highly effective organic matter decomposition strains, formulated through biotechnology, to decompose organic matter and sludge in water in a purely biological manner. This reduces the content of ammonia nitrogen, COD, total phosphorus, and total nitrogen, while increasing dissolved oxygen. It plays a role in oxidation, nitrification, denitrification, and phosphorus accumulation in the water, decomposing organic matter into non-toxic substances such as carbon dinitrate, nitrates, and sulfates. This reduces residual organic pollutants, eliminates black water, oily water, and foul-smelling sediment, and prevents water quality deterioration. Simultaneously, while degrading harmful organic matter in the water, it also significantly degrades sediment, enhancing the water body's self-purification capacity.

[0003] In river and lake ecological restoration and wastewater treatment, the use of microorganisms for wastewater treatment is low-cost, produces no secondary pollution, and is environmentally friendly. In practical applications, microbial treatment ultimately converts pollutants in wastewater into carbon dioxide, nitrogen, water, and sludge. Therefore, microorganisms can not only remove organic matter, pathogens, and harmful substances, but also remove odors, improve transparency, and reduce color. However, existing aquatic ecological restoration agents made from composite microorganisms still have the problem of poor wastewater treatment effects. Summary of the Invention

[0004] This invention proposes a water ecological composite remediation agent and its preparation method, which solves the problem that the water ecological composite remediation agent has poor effect on treating sewage in related technologies.

[0005] The technical solution of the present invention is as follows:

[0006] This invention proposes a composite aquatic ecological restoration agent, comprising the following components in parts by weight: 60-190 parts of microbial bacteria, 1-10 parts of biological enzymes, and 1-10 parts of nutrients;

[0007] The microbial strains comprise the following components by weight: 5-15 parts of ammonia-oxidizing denitrifying bacteria, 10-35 parts of denitrifying denitrifying bacteria, 10-30 parts of nitrifying bacteria, 10-40 parts of Bacillus subtilis, 10-40 parts of photosynthetic bacteria, 10-20 parts of Bacillus licheniformis, and 5-10 parts of yeast.

[0008] As a further technical solution, the microorganisms also include Bacillus thuringiensis.

[0009] As a further technical solution, the mass ratio of the ammonia-oxidizing denitrifying bacteria to Bacillus thuringiensis is 1:0.3~3.

[0010] As a further technical solution, the bio-enzyme is obtained by starch-catalyzed hydrolysis; the nutrients include the following components in parts by weight: 10-20 parts ammonium sulfate, 10-15 parts ammonium chloride, 5-10 parts sodium nitrate, 3-5 parts dipotassium hydrogen phosphate, 3-5 parts ammonium dihydrogen phosphate, 10-15 parts food-grade glucose, 2-5 parts potassium carbonate, 5-8 parts ferric chloride, 0.5-1 part potassium peroxide, 3-5 parts potassium chloride, 5-10 parts magnesium oxide, 0.5-1 part copper sulfate pentahydrate, 0.5-1 part manganese oxide, 1-2 parts zinc oxide, 1-1.5 parts sodium molybdate, and 885.5-936 parts water.

[0011] As a further technical solution, the nutrient also includes the following components in parts by weight: 9-11 parts of succinate;

[0012] The succinate component comprises a mixture of disodium succinate and potassium succinate sulfonate in a mass ratio of 1:3 to 4.

[0013] This invention also proposes a method for preparing the aforementioned aquatic ecological composite restoration agent, comprising the following steps:

[0014] S1. The components of the microbial bacteria are activated to obtain activated microbial bacteria;

[0015] S2. The components of the nutrients are cooked, cooled, and then the activated microorganisms are added and mixed to obtain a mixture.

[0016] S3. The mixture is subjected to an oxidative polymerization reaction to obtain a water ecological composite restoration agent.

[0017] As a further technical solution, in step S2, the ripening temperature is 70~80℃, the rotation speed is 140~160r / min, and the time is 3~5h; the cooling is carried out at 30℃; the mixing rotation speed is 140~160r / min, and the time is 20~28h.

[0018] As a further technical solution, in step S3, the temperature of the oxidative polymerization reaction is 30~50℃ and the time is 3~4h; the composite microbial liquid is stored at 4℃.

[0019] As a further technical solution, the following steps are also included:

[0020] The aquatic ecological composite restoration agent is dried to obtain aquatic ecological composite restoration agent powder.

[0021] As a further technical solution, the drying is spray drying, with a temperature of -35~-80℃ and a time of 4~16h.

[0022] As a further technical solution, the aforementioned aquatic ecological composite restoration agent is not used in conjunction with disinfectants, antibacterial drugs, or strong oxidants.

[0023] As a further technical solution, the specific sources and cultivation processes of the components of the microorganisms are as follows:

[0024] (1) Preparation of ammonia-oxidizing denitrifying bacteria:

[0025] Ammonia-oxidizing bacteria oxidize ammonia nitrogen into nitrogen gas and water through the oxidation process of endogenous subgranular cells, and further convert nitrate nitrogen into nitrogen gas for discharge, thereby achieving effective removal of nitrogen from wastewater.

[0026] a. Source: 100 mL of bottom granular sludge from an upflow anaerobic reactor was collected; 100 mL of sludge was obtained from the sedimentation of the sludge-water mixture in the aeration tank; the two were mixed evenly and allowed to settle. 100 mL of this mixed sample was then added to 50 mL of sterile water and poured into a 200 mL Erlenmeyer flask. The flask was sealed and shaken at room temperature for 4 hours, then allowed to stand for 1 hour. The supernatant was collected for separation, screening (primary screening, secondary screening), cultivation, and purification to obtain the bacterial strain. Sequencing and comparison confirmed it to be an ammonia-oxidizing strain, a Gram-positive bacterium, with a bacterial concentration of 10-1. 8 ~10 9 cells / mL;

[0027] b. Cultivation: The ammonia-oxidizing strain was activated by slant inoculation into the culture medium and cultured at 10-30℃ with shaking at 120 r / min and aeration of 1:0.5-1 for 3-5 days to obtain the ammonia-oxidizing denitrifying bacterial solution. The bacterial solution concentration was (1-2) × (10⁻⁶) / 2 × 10⁻⁶. 7 ~10 9 The obtained bacterial solution was dried and sprayed into powder using a pressure spray dryer at -40℃ for 6 hours to obtain ammonia-oxidizing denitrifying bacteria powder. The viable count of the powder was (1~2)×(10⁻⁶) cells / mL. 7 ~10 9 ( ) per mL, no pulverization or sieving required after drying;

[0028] c. The culture medium ingredients are: ammonium sulfate 0.5 g / L, sodium chloride 0.3 g / L, ferrous sulfate 0.03 g / L, dipotassium hydrogen phosphate 1 g / L, magnesium sulfate 0.03 g / L, calcium chloride 7.5 g / L, peptone 5 g / L, beef extract 5 g / L, trace element molybdenum 0.05 g / L, trace element potassium 0.03 g / L, trace element manganese 0.02 g / L, sterile distilled water 1000 mL, pH=8.

[0029] (2) Preparation of denitrifying bacteria:

[0030] Denitrifying bacteria are facultative anaerobic bacteria that utilize molecular oxygen as an electron acceptor. When dissolved oxygen is below 0.15 mg / L, denitrifying bacteria can use nitrates and nitrogen compounds as electron acceptors to reduce nitrates to NO, N2O, or N2, thereby further removing nitrate nitrogen and total nitrogen.

[0031] Aerobic denitrifying bacteria (Paraccoccus pantotrop Hus) can reduce nitrate and nitrite nitrogen to N2 under aerobic conditions.

[0032] Autotrophic denitrifying bacteria, Thiobacillus denitrificans and Thiomicrospira denitrificans.

[0033] These bacteria are Gram-negative. They have good resistance to low temperatures and can grow in temperatures ranging from 2 to 37°C. They are commonly found in soil, lakes, or oceans, and some strains have been domesticated and have developed some adaptability to temperature, salinity, and heavy metals.

[0034] a. Source: Take 500mL of riverbed sludge; after sedimentation and settling, take 100mL of this mixed sample, add 50mL of sterile water, add to a 200mL Erlenmeyer flask, seal, shake at room temperature for 4 hours, and let stand for 1 hour. Take the supernatant for separation, screening (primary screening, secondary screening), culture, and purification to obtain the bacterial strain. After sequencing and comparison, it was determined to be a denitrifying bacteria strain, a Gram-negative bacterium, with a bacterial concentration of (1~2)×(10) 8 ~10 9 ) per mL;

[0035] b. Cultivation: The denitrifying bacteria strain was activated by slant inoculation into the culture medium and cultured at 10~30℃ with shaking at 40r / min for 48h to obtain the denitrifying bacteria solution. The bacterial solution concentration was (3~4)×(10) 8 ~10 9 The obtained bacterial solution was dried and sprayed into powder using a pressure spray dryer at -80℃ for 12 hours to obtain denitrifying bacteria powder. The viable count of the powder was 2 × 10⁻⁶ cells / mL. 9 Cells / mL. No crushing or screening is required after drying, the process is simple, and the purity is high;

[0036] c. The culture medium ingredients are: manganese sulfate 0.02 g / L, disodium hydrogen phosphate 1.2 g / L, potassium dihydrogen phosphate 1.8 g / L, sodium bicarbonate 1 g / L, magnesium sulfate heptahydrate 0.4 g / L, ammonium chloride 0.5 g / L, calcium chloride 0.05 g / L, ferric chloride 0.02 g / L, potassium nitrate 5 g / L, sodium thiosulfate 10 g / L, sodium citrate 5 g / L, peptone 5 g / L, beef extract 5 g / L, trace element molybdenum 0.05 g / L, trace element potassium 0.03 g / L, trace element manganese 0.02 g / L, sterile distilled water 1000 mL, pH=7.0.

[0037] (3) Preparation of nitrifying bacteria:

[0038] Nitrifying bacteria are aerobic, autotrophic microorganisms that can grow in oxygenated water or gravel, playing a crucial role in the nitrogen cycle and water purification process. They include the genera *Nitrosomonas* and *Nitrobacter*.

[0039] a. Source: Take 500mL of activated sludge from a certain aquaculture wastewater treatment plant; after sedimentation, take 100mL of this mixed sample, add 50mL of sterile water, add to a 200mL Erlenmeyer flask, add culture medium, seal, shake at room temperature for 4 hours, and let stand for 1 hour. Take the supernatant for separation, screening (primary screening, secondary screening), culture, separation, and purification to obtain the bacterial strain. After sequencing, comparison, and identification (referring to the "Bergeminal Handbook of Bacteriological Identification" to determine the physiological and chemical indicators of bacteria), it was determined to be an aerobic nitrifying bacteria strain, a Gram-negative bacterium, with a bacterial concentration of (1~2)×(10) 8 ~10 9 ) per mL;

[0040] b. Cultivation: The diluted nitrifying denitrifying bacteria strain was spread onto beef peptone agar plates and incubated at 25-30℃, pH 7.5, with shaking at 150 rpm for 48 h to obtain an aerobic nitrifying denitrifying bacteria culture. The concentration of the culture was (2-3) × (10⁻⁶)⁻¹. 8 ~10 9 The obtained bacterial solution was dried and sprayed into powder using a pressure spray dryer at -80℃ for 12 hours to obtain denitrifying bacteria powder. The viable count of the powder was 3 × 10⁻⁶ cells / mL. 10 Cells / mL. No crushing or screening is required after drying, the process is simple, and the purity is high;

[0041] c. The culture medium ingredients are: peptone 1g / L, yeast extract 0.5g / L, sodium chloride 1g / L, agar 2g / L, beef extract 0.3g / L, glucose 1g / L, manganese sulfate 0.05g / L, potassium dihydrogen phosphate 0.5g / L, sodium bicarbonate 1g / L, magnesium sulfate heptahydrate 0.5g / L, trace element molybdenum 0.05g / L, sterile distilled water 1000mL, pH=7.0.

[0042] (4) Preparation of Bacillus subtilis:

[0043] Bacillus subtilis is a species of Bacillus. It is a Gram-positive bacterium, oval to columnar in shape, with a rough colony surface. This bacterium is widely distributed in soil and decaying organic matter, and readily multiplies in hay infusion, hence its name. It can synthesize various vitamins and enhance cell activity.

[0044] a. Source: Take 500mL of activated sludge from a certain aquaculture wastewater treatment plant; after sedimentation, take 100mL of this mixed sample, add 50mL of sterile water, add to a 200mL Erlenmeyer flask, add culture medium, seal, shake at room temperature for 4 hours, and let stand for 1 hour. Take the supernatant for separation, screening (primary screening, secondary screening), culture, separation, and purification to obtain the bacterial strain. After sequencing, comparison, and identification (referring to the "Bergeminal Handbook of Bacteriology" to determine the physiological and chemical indicators of bacteria), it was identified as Bacillus subtilis strain, a Gram-positive bacterium, with a bacterial concentration of (1~2)×(10) 8 ~10 9 ) per mL;

[0045] b. Cultivation: The nitrifying denitrifying strain was diluted and spread onto beef peptone solid agar plates and incubated at 25-30℃, pH 7.5, with shaking at 150 rpm for 48 h to obtain Bacillus subtilis bacterial suspension. The bacterial suspension concentration was (4-5) × (10⁻⁶) / 10⁻⁶. 8 ~10 9 The obtained bacterial solution was dried and sprayed into powder using a pressure spray dryer at -40℃ for 10 hours to obtain Bacillus subtilis powder. The viable count of the powder was 2 × 10⁻⁶ cells / mL. 10 Cells / mL; no pulverization or sieving is required after drying;

[0046] c. The culture medium ingredients are: 10 g / L peptone, 0.5 g / L yeast extract, 3 g / L sodium chloride, 10 g / L agar, 5 g / L beef extract, 5 g / L sodium citrate, 7 g / L glucose, 0.5 g / L potassium dihydrogen phosphate, 0.03 g / L potassium, 0.05 g / L manganese, 1000 mL sterile distilled water, pH=6.5~7.5.

[0047] (5) Preparation of photosynthetic bacteria:

[0048] Photosynthetic bacteria are the earliest and most ubiquitous prokaryotes in nature, possessing a primitive photosynthetic system. They are a type of bacteria with multiple functions, including denitrification, nitrogen fixation, carbon fixation, and sulfide oxidation. Photosynthetic bacteria can grow in environments ranging from 10 to 45°C, with an optimal temperature of 30 to 40°C and a pH of 7.0 to 8.0. They are highly efficient at removing BOD and have good nitrogen removal capabilities.

[0049] a. Source: Activated sludge from the bottom of a lake was collected, enriched, screened (primary and secondary screening), cultured, isolated, and purified to obtain the bacterial strain. Sequencing, comparison, and identification (referring to the physiological and chemical indicators of bacteria as determined in the Bergermin Manual of Bacteriology) confirmed it to be a photosynthetic bacterium strain, a Gram-negative bacterium, with a bacterial concentration of (1~2) × (10⁻⁶). 8 ~10 9 ) per mL;

[0050] b. Cultivation: A photosynthetic bacterium was activated by slant inoculation into a culture medium and incubated at 28-36℃ with shaking at 40 rpm under a light intensity of 3000-4000 Rex (LX) for 24-48 hours to obtain a photosynthetic bacterial suspension with a concentration of (1-2) × 10⁻⁶. 9 The obtained bacterial solution was dried and spray-dried in a pressure spray dryer at -40℃ for 4 hours to obtain photosynthetic bacteria powder granules with a viable count of 2×10⁻⁶ cells / mL. 10 pcs / mL; no need for crushing and screening after drying, simple process, high purity;

[0051] c. The culture medium ingredients are: 5 g / L peptone, 1 g / L sodium acetate, 1 g / mL yeast extract, 0.5 g / L sodium chloride, 10 g / L agar, 5 g / L sodium citrate, 7 g / L glucose, 0.05 g / L potassium dihydrogen phosphate, 0.03 g / L magnesium sulfate, 0.05 g / L trace element manganese, 1000 mL sterile distilled water, pH=8~8.5.

[0052] (6) Preparation of Bacillus licheniformis:

[0053] Bacillus licheniformis is a Gram-positive bacterium capable of producing various active enzymes and enzymatic factors. It also works synergistically with other beneficial bacteria such as Bacillus species and photosynthetic bacteria. It can effectively improve water quality, reduce ammonia nitrogen, and promote the reproduction and development of microorganisms.

[0054] a. Source: Soil from the vicinity of an artificial wetland was collected, and the strain was obtained through enrichment culture and screening (primary screening, secondary screening), culture, isolation, and purification. Sequencing, comparison, and identification (referring to the physiological and chemical indicators of bacteria as determined in the Bergermin Manual of Bacteriology) confirmed it to be a *Bacillus licheniformis* strain, a Gram-positive bacterium, with a bacterial concentration of (1~2) × (10⁻⁶)⁻¹. 8 ~10 9 ) per mL;

[0055] b. Cultivation: A photosynthetic bacterium was activated by slant inoculation into a culture medium and incubated at 28-36℃ with shaking at 40 rpm under a light intensity of 3000-4000 Rex (LX) for 24-48 hours to obtain a photosynthetic bacterial suspension with a concentration of (3-10) × 10⁻⁶. 10 The obtained bacterial solution was dried and spray-dried in a pressure spray dryer at -80℃ for 16 hours to obtain Bacillus licheniformis powder, with a viable count of 2 × 10⁻⁶ cells / mL. 10 pcs / mL; no need for crushing and screening after drying, simple process, high purity;

[0056] c. The culture medium ingredients are: peptone 3.5 g / L, yeast extract 2 g / mL, agar 10 g / L, sodium citrate 0.5 g / L, glucose 5 g / L, potassium dihydrogen phosphate 0.5 g / L, magnesium sulfate 0.5 g / L, trace element manganese 0.05 g / L, sterile distilled water 1000 mL, pH=6.5~7.2.

[0057] (7) Preparation of Bacillus thuringiensis:

[0058] Bacillus thuringiensis is a Gram-positive bacterium that is widely used as a green and environmentally friendly microbial insecticide.

[0059] a. Source: Bacillus thuringiensis was collected from the soil environment surrounding the constructed wetland and screened, isolated, and purified using a sterile water method. Sequencing, comparison, and identification (referring to the physiological and chemical indicators of bacteria as determined by Bergen's Manual of Bacteriological Identification) confirmed it to be a Bacillus thuringiensis strain, a Gram-positive bacterium.

[0060] b. Cultivation: Incubate Bacillus thuringiensis in a culture medium at 25-35℃ using a shaker at 140 rpm for 12-24 hours. The pH of the culture medium should be 6.5-7.5. This will yield a Bacillus thuringiensis bacterial suspension with a concentration of 10⁻⁶. 7 ~10 10 Inoculation rate: 1.5%~3.0% (v / v); culture temperature: 28~30℃; rotation speed: 150 r / min; fermentation time: 12~24 hours; pH: 6~8.

[0061] The obtained bacterial solution was dried and sprayed into powder using a pressure spray dryer at -40℃ for 8 hours to obtain Bacillus thuringiensis bacterial powder granules. The viable count of the bacterial powder was 2×10⁻⁶. 10 pcs / mL; no need for crushing and screening after drying, simple process, high purity;

[0062] c. The culture medium ingredients are: lysine 3g / L, peptone 8g / L, yeast extract 5g / mL, sodium chloride 0.5g / L, agar 5g / L, beef extract 2g, sodium citrate 0.5g / L, glucose 1g / L, potassium dihydrogen phosphate 0.5g / L, magnesium sulfate 0.05g / L, trace elements 0.05g / L, sterile distilled water 1000mL, pH=6.8~7.5.

[0063] (8) Preparation of yeast:

[0064] Yeast can multiply a large number of live yeast cells in a short period of time and is resistant to acid, alcohol and high temperature.

[0065] a. Source: Yeast strains were collected from different fermentation stages of the beer brewing process at a brewery. They were obtained through aseptic water screening, separation, and purification. Sequencing, comparison, and identification (referencing the Bergerming Manual of Bacteriological Identification to determine the physiological and chemical indicators of the bacteria) confirmed them to be yeast strains, specifically *Saccharomyces cerevisiae*, and Gram-positive bacteria.

[0066] b. Cultivation: Spread a yeast strain onto a corresponding SD plate; invert the plate and inoculate at 30℃, shake the medium at 30℃ and 40r / min, control the DO below 2.0mg / L, and incubate on a shaker for 6h to obtain yeast culture. The concentration of the culture is (2~3)×(10) 8 ~10 9 ) per mL.

[0067] The obtained bacterial solution was separated at -35℃, isolated from air and light, and dried until the moisture content was controlled below 6%, yielding yeast powder for later use. The viable cell count of the powder was 3 × 10⁻⁶. 10 cells / mL;

[0068] c. The culture medium ingredients are: 2% peptone, 1% yeast extract, 3% agar, 1.5% potassium dihydrogen phosphate, 7% glucose, 0.05% manganese sulfate, 0.05% potassium chloride, 1000mL sterile distilled water, pH=7.0.

[0069] (9) Preparation of biological enzymes:

[0070] Biological enzymes include various enzymes (amylase, protease, cellulase, hemicellulase, pectinase, lipase, phytase), which are catalysts produced by living cells. With the participation of enzymes, the body's metabolism can proceed in an orderly manner. They are proteins with catalytic functions. Like other proteins, enzyme molecules are composed of long chains of amino acids. Some of these chains are helical, and some are folded sheet structures, with these two parts linked by unfolded amino acid chains, giving the entire enzyme molecule a specific three-dimensional structure. Biological enzymes are produced in living organisms and possess special catalytic functions. Their characteristics include: High efficiency: When used as catalysts, the catalytic efficiency of enzymes is 10 times that of general inorganic catalysts. 3 ~10 6 times.

[0071] a. Source: An enzyme derived from the catalytic hydrolysis of starch, specifically from maltose or glucose. It is prepared using solid-state fermentation technology, and the resulting enzyme is obtained through screening, separation, and purification.

[0072] b. Cultivation: The bio-enzyme bacteria are activated by slant culture, then cultured in shake flasks, fermented under aeration, expanded in seed tanks, fermented with bran, extracted, filtered, precipitated, centrifuged, air-dried, and pulverized to obtain refined amylase. The production process involves controlling the temperature at 25~38℃, the pH at 5.0~7.5, the cultivation time at 24~48 hours, and shaking at 140 rpm to obtain a yellowish-brown or white bio-enzyme powder with an activity ≥50 U / mg.

[0073] c. The culture medium ingredients are: 2% peptone, 1% beef extract, 3% agar, 1.5% potassium dihydrogen phosphate, 7% glucose, 0.05% potassium chloride, 1% sodium chloride, 1000mL sterile distilled water, pH=7.0.

[0074] The working principle and beneficial effects of this invention are as follows:

[0075] In this invention, the water ecological composite remediation agent has strong adaptability and rapidly attaches to biofilm microorganisms. It is mainly based on local microbial culture and nutrient substrate, combined with bioactive substances and a small amount of biological enzymes, to achieve the result of rapidly reducing the formation of activated sludge.

[0076] At the same time, it has a rich microbial community, strong shock resistance, and a complex symbiotic microbial community with a wide variety of species. It has strong adaptability, can adapt to various wastewaters, and differentiate and screen out suitable microbial communities. The biofilm formed is not easy to break and has strong adaptability to physical and chemical impacts.

[0077] It also possesses a rich ecosystem, reduces residual sludge, and fosters a mutually reinforcing relationship among microbial communities. The secretions and excretions of some microbial species also serve as nutrients for protozoa, multicellular organisms, and complex bacteria, accelerating or promoting the formation of ecosystems on activated sludge and biofilms. This enhances the removal rate of pollutants by activated sludge while reducing the formation rate of dead sludge, ensuring the activity and removal rate of organic sludge. It is suitable for various anaerobic and aerobic environments, including comprehensive river and lake ecological management and domestic sewage treatment, and can simultaneously remove COD, ammonia nitrogen, total nitrogen, and phosphorus. Detailed Implementation

[0078] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. 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 of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0079] In the following examples and comparative examples: the viable count of the ammonia-oxidizing denitrifying bacteria powder was 2 × 10⁻⁶. 10 cells / mL;

[0080] The viable count of denitrifying bacteria powder was 2 × 10⁻⁶. 9 cells / mL;

[0081] The viable count of nitrifying bacteria powder was 3 × 10⁻⁶. 10 cells / mL;

[0082] The viable count of Bacillus subtilis powder was 2 × 10⁻⁶. 10 cells / mL;

[0083] The viable count of photosynthetic bacteria in the powder was 2 × 10⁻⁶. 10 cells / mL;

[0084] The viable count of Bacillus licheniformis powder was 2 × 10⁻⁶. 10 cells / mL;

[0085] The viable count of Bacillus thuringiensis powder was 2 × 10⁻⁶. 10 cells / mL;

[0086] The live count of yeast powder is 3 × 10⁻⁶. 10 per mL.

[0087] Example 1

[0088] The water ecological composite restoration agent comprises the following components by weight: 92 parts microbial bacteria, 5 parts biological enzymes, and 5 parts nutrients;

[0089] The microbial strains consist of the following components by weight: 8 parts ammonia-oxidizing denitrifying bacteria, 20 parts denitrifying denitrifying bacteria, 12 parts nitrifying bacteria, 25 parts Bacillus subtilis, 10 parts photosynthetic bacteria, 12 parts Bacillus licheniformis, and 5 parts yeast.

[0090] The nutrients consist of the following components by weight: 20 parts ammonium sulfate, 15 parts ammonium chloride, 10 parts sodium nitrate, 5 parts dipotassium hydrogen phosphate, 5 parts ammonium dihydrogen phosphate, 15 parts food-grade glucose, 5 parts potassium carbonate, 8 parts ferric chloride, 1 part potassium peroxide, 5 parts potassium chloride, 10 parts magnesium oxide, 1 part copper sulfate pentahydrate CuSO4·5H2O, 1 part manganese oxide, 2 parts zinc oxide, 1.5 parts sodium molybdate, and 895.5 parts water.

[0091] The preparation method of the aquatic ecological composite restoration agent includes the following steps:

[0092] S1. The components of the microbial bacteria are activated separately to obtain activated microbial bacteria;

[0093] S2. The nutrient components are matured, cooled, and then mixed with activated microorganisms to obtain a mixture. The maturity temperature is 75℃, the rotation speed is 150r / min, and the time is 4h. The cooling temperature is 30℃, and the mixing speed is 150r / min for 24h.

[0094] S3. The mixture is subjected to an oxidative polymerization reaction, spray-dried to obtain a water ecological composite restoration agent, and stored at 4°C; wherein, the temperature of the oxidative polymerization reaction is 40°C and the time is 3h.

[0095] Example 2

[0096] The water ecological compound restoration agent comprises the following components in parts by weight: 60 parts microbial bacteria, 1 part biological enzyme, and 1 part nutrients;

[0097] The microbial strains consist of the following components by weight: 5 parts ammonia-oxidizing denitrifying bacteria, 10 parts denitrifying denitrifying bacteria, 10 parts nitrifying bacteria, 10 parts Bacillus subtilis, 10 parts photosynthetic bacteria, 10 parts Bacillus licheniformis, and 5 parts yeast.

[0098] The nutrients consist of the following components by weight: 10 parts ammonium sulfate, 10 parts ammonium chloride, 5 parts sodium nitrate, 3 parts dipotassium hydrogen phosphate, 3 parts ammonium dihydrogen phosphate, 10 parts food-grade glucose, 2 parts potassium carbonate, 5 parts ferric chloride, 0.5 parts potassium peroxide, 3 parts potassium chloride, 5 parts magnesium oxide, 0.5 parts copper sulfate pentahydrate CuSO4·5H2O, 0.5 parts manganese oxide, 1 part zinc oxide, 1 part sodium molybdate, and 936 parts water.

[0099] The preparation method of the aquatic ecological composite restoration agent includes the following steps:

[0100] S1. The components of the microbial bacteria are activated separately to obtain activated microbial bacteria;

[0101] S2. The nutrient components are matured, cooled, and then mixed with activated microorganisms to obtain a mixture. The maturity temperature is 70℃, the rotation speed is 160r / min, and the time is 3h. The cooling temperature is 30℃, and the mixing speed is 140r / min for 28h.

[0102] S3. The mixture is subjected to an oxidative polymerization reaction to obtain a water ecological composite restoration agent, which is then refrigerated at 4°C; wherein, the temperature of the oxidative polymerization reaction is 30°C and the time is 4 hours.

[0103] Example 3

[0104] The water ecological compound restoration agent comprises the following components in parts by weight: 190 parts microbial bacteria, 10 parts biological enzymes, and 10 parts nutrients.

[0105] The microbial strains consist of the following components by weight: 15 parts ammonia-oxidizing denitrifying bacteria, 35 parts denitrifying denitrifying bacteria, 30 parts nitrifying bacteria, 40 parts Bacillus subtilis, 40 parts photosynthetic bacteria, 20 parts Bacillus licheniformis, and 10 parts yeast.

[0106] The nutrients consist of the following components by weight: 20 parts ammonium sulfate, 15 parts ammonium chloride, 10 parts sodium nitrate, 5 parts dipotassium hydrogen phosphate, 5 parts ammonium dihydrogen phosphate, 15 parts food-grade glucose, 5 parts potassium carbonate, 8 parts ferric chloride, 1 part potassium peroxide, 5 parts potassium chloride, 10 parts magnesium oxide, 1 part copper sulfate pentahydrate CuSO4·5H2O, 1 part manganese oxide, 2 parts zinc oxide, 1.5 parts sodium molybdate, and 895.5 parts water.

[0107] The preparation method of the aquatic ecological composite restoration agent includes the following steps:

[0108] S1. The components of the microbial bacteria are activated separately to obtain activated microbial bacteria;

[0109] S2. The nutrient components are matured, cooled, and then mixed with activated microorganisms to obtain a mixture. The maturity temperature is 80℃, the rotation speed is 140r / min, and the time is 5h. The cooling temperature is 30℃, and the mixing speed is 160r / min for 20h.

[0110] S3. The mixture is subjected to an oxidative polymerization reaction to obtain a water ecological composite restoration agent, which is then refrigerated at 4°C. The oxidative polymerization reaction is carried out at 50°C for 3 hours.

[0111] Example 4

[0112] The only difference between this embodiment and Embodiment 1 is the aquatic ecological composite restoration agent, which includes the following components by weight: 84.8 parts of microbial bacteria, 5 parts of biological enzymes, and 5 parts of nutrients.

[0113] The microbial composition includes the following components by weight: 20 parts denitrifying bacteria, 12 parts nitrifying bacteria, 25 parts Bacillus subtilis, 10 parts photosynthetic bacteria, 12 parts Bacillus licheniformis, 5 parts yeast, and 0.8 parts Bacillus thuringiensis.

[0114] Example 5

[0115] The only difference between this embodiment and Embodiment 1 is the aquatic ecological composite restoration agent, which includes the following components by weight: 92.8 parts of microbial bacteria, 5 parts of biological enzymes, and 5 parts of nutrients.

[0116] The microbial composition includes the following components by weight: 8 parts ammonia-oxidizing denitrifying bacteria, 20 parts denitrifying denitrifying bacteria, 12 parts nitrifying bacteria, 25 parts Bacillus subtilis, 10 parts photosynthetic bacteria, 12 parts Bacillus licheniformis, 5 parts yeast, and 0.8 parts Bacillus thuringiensis.

[0117] Example 6

[0118] The only difference between this embodiment and Embodiment 1 is the aquatic ecological composite restoration agent, which includes the following components by weight: 117.6 parts of microbial bacteria, 5 parts of biological enzymes, and 5 parts of nutrients.

[0119] The microbial composition includes the following components by weight: 8 parts ammonia-oxidizing denitrifying bacteria, 20 parts denitrifying denitrifying bacteria, 12 parts nitrifying bacteria, 25 parts Bacillus subtilis, 10 parts photosynthetic bacteria, 12 parts Bacillus licheniformis, 5 parts yeast, and 25.6 parts Bacillus thuringiensis.

[0120] Example 7

[0121] The only difference between this embodiment and Embodiment 1 is the aquatic ecological composite restoration agent, which includes the following components by weight: 94.4 parts of microbial bacteria, 5 parts of biological enzymes, and 5 parts of nutrients.

[0122] The microbial composition includes the following components by weight: 8 parts ammonia-oxidizing denitrifying bacteria, 20 parts denitrifying denitrifying bacteria, 12 parts nitrifying bacteria, 25 parts Bacillus subtilis, 10 parts photosynthetic bacteria, 12 parts Bacillus licheniformis, 5 parts yeast, and 2.4 parts Bacillus thuringiensis.

[0123] Example 8

[0124] The only difference between this embodiment and Embodiment 1 is the aquatic ecological composite restoration agent, which includes the following components by weight: 116 parts of microbial bacteria, 5 parts of biological enzymes, and 5 parts of nutrients.

[0125] The microbial composition includes the following components by weight: 8 parts ammonia-oxidizing denitrifying bacteria, 20 parts denitrifying denitrifying bacteria, 12 parts nitrifying bacteria, 25 parts Bacillus subtilis, 10 parts photosynthetic bacteria, 12 parts Bacillus licheniformis, 5 parts yeast, and 24 parts Bacillus thuringiensis.

[0126] Example 9

[0127] The difference between this embodiment and Embodiment 8 lies only in that the nutrients include the following components in parts by weight: 20 parts ammonium sulfate, 15 parts ammonium chloride, 10 parts sodium nitrate, 5 parts dipotassium hydrogen phosphate, 5 parts ammonium dihydrogen phosphate, 15 parts food-grade glucose, 5 parts potassium carbonate, 8 parts ferric chloride, 1 part potassium peroxide, 5 parts potassium chloride, 10 parts magnesium oxide, 1 part copper sulfate pentahydrate CuSO4·5H2O, 1 part manganese oxide, 2 parts zinc oxide, 1.5 parts sodium molybdate, 10 parts disodium succinate, and 885.5 parts water.

[0128] Example 10

[0129] The difference between this embodiment and Embodiment 8 lies only in that the nutrients include the following components in parts by weight: 20 parts ammonium sulfate, 15 parts ammonium chloride, 10 parts sodium nitrate, 5 parts dipotassium hydrogen phosphate, 5 parts ammonium dihydrogen phosphate, 15 parts food-grade glucose, 5 parts potassium carbonate, 8 parts ferric chloride, 1 part potassium peroxide, 5 parts potassium chloride, 10 parts magnesium oxide, 1 part copper sulfate pentahydrate CuSO4·5H2O, 1 part manganese oxide, 2 parts zinc oxide, 1.5 parts sodium molybdate, 10 parts potassium succinate sulfonate, and 885.5 parts water.

[0130] Example 11

[0131] The difference between this embodiment and Embodiment 8 lies only in that the nutrients include the following components in parts by weight: 20 parts ammonium sulfate, 15 parts ammonium chloride, 10 parts sodium nitrate, 5 parts dipotassium hydrogen phosphate, 5 parts ammonium dihydrogen phosphate, 15 parts food-grade glucose, 5 parts potassium carbonate, 8 parts ferric chloride, 1 part potassium peroxide, 5 parts potassium chloride, 10 parts magnesium oxide, 1 part copper sulfate pentahydrate CuSO4·5H2O, 1 part manganese oxide, 2 parts zinc oxide, 1.5 parts sodium molybdate, 2.5 parts disodium succinate, 7.5 parts potassium succinate sulfonate, and 885.5 parts water.

[0132] Example 12

[0133] The difference between this embodiment and Embodiment 8 lies only in that the nutrients include the following components in parts by weight: 20 parts ammonium sulfate, 15 parts ammonium chloride, 10 parts sodium nitrate, 5 parts dipotassium hydrogen phosphate, 5 parts ammonium dihydrogen phosphate, 15 parts food-grade glucose, 5 parts potassium carbonate, 8 parts ferric chloride, 1 part potassium peroxide, 5 parts potassium chloride, 10 parts magnesium oxide, 1 part copper sulfate pentahydrate CuSO4·5H2O, 1 part manganese oxide, 2 parts zinc oxide, 1.5 parts sodium molybdate, 2 parts disodium succinate, 8 parts potassium succinate sulfonate, and 885.5 parts water.

[0134] Comparative Example 1

[0135] The only difference between this comparative example and Example 1 is the aquatic ecological composite restoration agent, which includes the following components by weight: 84 parts of microbial bacteria, 5 parts of biological enzymes, and 5 parts of nutrients.

[0136] The microbial composition includes the following components by weight: 20 parts denitrifying bacteria, 12 parts nitrifying bacteria, 25 parts Bacillus subtilis, 10 parts photosynthetic bacteria, 12 parts Bacillus licheniformis, and 5 parts yeast.

[0137] Application Example 1

[0138] Domestic sewage from a certain campus

[0139] Treatment method: The aquatic ecological composite remediation agent prepared in Example 1 was mixed with water and added to the biological treatment tank. After three days of aeration, the influent flow rate was gradually increased to the design load. After 25 days of operation, water quality samples were taken before and after treatment to test the chemical oxygen demand (COD), ammonia nitrogen content, total phosphorus content, total nitrogen content, and pH. The results are shown in Table 1, and the removal rates of COD, ammonia nitrogen, total phosphorus, and total nitrogen were calculated. The mass-to-volume ratio of the aquatic ecological composite remediation agent to water was 1 kg: 10 L; 500~1000 m³ / h 3 The dosage of the aquatic ecological composite restoration agent for the pond water body is 1 kg.

[0140] Table 1. Water quality test results from on-site sampling before and after treatment.

[0141]

[0142] As shown in Table 1, compared with the original water quality of the sewage collection tank before treatment, the water quality after treatment with the aquatic ecological composite remediation agent prepared in Example 1 had COD of 41.30 mg / L, ammonia nitrogen of 0.35 mg / L, total phosphorus of 0.33 mg / L, total nitrogen of 11.30 mg / L, and pH of 7.25, meeting the discharge standards. Moreover, the aquatic ecological composite remediation agent prepared in Example 1 achieved removal rates of 87.33%, 99.33%, 95.63%, and 84.78% for COD, ammonia nitrogen, total phosphorus, and total nitrogen, respectively.

[0143] Similarly, the campus domestic sewage samples before treatment were treated with the water ecological composite remediation agents prepared in Examples 2-12 and Comparative Example 1 according to the treatment method of the water ecological composite remediation agent prepared in Example 1. The water quality after treatment was sampled and tested, the chemical oxygen demand (COD) was tested, and the COD removal rate was calculated. The results are shown in Table 2.

[0144] Table 2. Water quality test results from on-site sampling after treatment.

[0145]

[0146] Compared with Comparative Example 1, the COD removal rate of the microbial components containing ammonia-oxidizing denitrifying bacteria in Examples 1-12 was higher, indicating that the presence of ammonia-oxidizing denitrifying bacteria in the microbial components can significantly improve the COD removal rate and enhance the wastewater treatment effect.

[0147] Compared with Examples 3-4, the COD removal rate of the water ecological composite remediation agent prepared in Examples 5-8 was higher, indicating that the addition of ammonia-oxidizing denitrifying bacteria and Bacillus thuringiensis to the microbial components can further improve the COD removal rate and improve the sewage treatment effect.

[0148] Compared with Examples 6-7, the COD removal rate of the water ecological composite remediation agent prepared in Examples 8-9 was higher, indicating that the addition of ammonia-oxidizing denitrifying bacteria and Bacillus thuringiensis in a mass ratio of 1:0.3-3 to the microbial components can further improve the COD removal rate and enhance the sewage treatment effect.

[0149] Compared with Examples 8-10, the COD removal rate of the water ecological composite restoration agent prepared in Examples 11-12 was higher, indicating that the addition of a mixture of disodium succinate and potassium succinate sulfonate in a mass ratio of 1:3-4 to the nutrient components can further improve the COD removal rate and enhance the wastewater treatment effect.

[0150] Application Example 2

[0151] The wastewater treatment project of a certain aquaculture farm uses a UASB anaerobic reactor + A2O process.

[0152] Treatment Method: The aquatic ecological composite remediation agent prepared in Example 1 was mixed with water and added to the UASB anaerobic and aerobic tanks of the biological system, respectively. After aeration for five days, the influent flow rate was gradually increased to the design load. After 45 days of operation, water quality samples were taken and tested before and after treatment. The chemical oxygen demand (COD), ammonia nitrogen content, total phosphorus content, and pH were measured. The results are shown in Table 3, and the removal rates of COD, ammonia nitrogen, and total phosphorus were calculated. The mass-volume ratio of the aquatic ecological composite remediation agent to water was 1 kg: 10 L; the dosage in the UASB anaerobic tank was 1 kg of bacterial powder added to 200-500 m³. 3 Add the bacteria to the water in the tank; for aerobic tanks, add 1 kg of bacterial powder to 300-800 m³ of water. 3 Water is added to the pool.

[0153] Table 3. Water quality test results from on-site sampling before and after treatment.

[0154]

[0155] As can be seen from Table 3, compared with the original water quality of the sewage collection tank before treatment, the water quality after treatment with the aquatic ecological composite remediation agent prepared in Example 1 had COD of 46.45 mg / L, ammonia nitrogen of 3.5 mg / L, total phosphorus of 0.24 mg / L, and pH of 7.3, meeting the discharge standards. Moreover, the aquatic ecological composite remediation agent prepared in Example 1 achieved removal rates of 96.96%, 98.92%, and 98.55% for COD, ammonia nitrogen, and total phosphorus, respectively.

[0156] Application Example 3

[0157] A project to remediate slightly polluted water in an artificial lake in a park covers an area of ​​69,600 square meters, with a depth of 1.5 to 2.5 meters and a water volume of approximately 130,000 cubic meters. The lake was experiencing a blue-green algae bloom and excessive emergent plants, resulting in an overall water quality classified as worse than Class V. Before remediation, the water quality was below Class V (samples taken from June 16th to 18th are shown in Table 4 below). The requirement is that after remediation, the main water quality indicators should meet the Class III water quality limits stipulated in the "Surface Water Environmental Quality Standard" (GB3838-2020).

[0158] The aquatic ecological composite remediation agent prepared in Example 1 was mixed with water and added to the polluted water body for treatment starting from June 19, 2023. After 7 days of operation, the treated water quality was sampled and tested, and the chemical oxygen demand (COD), ammonia nitrogen content, total phosphorus content, total nitrogen content, and pH were measured. The results are shown in Table 5, and the removal rates of COD, ammonia nitrogen, total phosphorus, and total nitrogen were calculated. The mass-to-volume ratio of the aquatic ecological composite remediation agent to water was 1 kg: 10 L; 3000~5000 m³ / h 3 The dosage of the aquatic ecological composite restoration agent for the pond water body is 1 kg.

[0159] Table 4. Water quality test results from on-site sampling before treatment.

[0160]

[0161] Table 5. Water quality test results from on-site sampling after treatment.

[0162]

[0163] Tests showed that before and after adding the aquatic ecological composite restoration agent prepared in Example 1, the dissolved oxygen increased from 0.8~1.2 mg / L before addition to 6.6~8.5 mg / L, representing increases of 87.88% and 85.89%, respectively.

[0164] Tables 4 and 5 show that after adding the aquatic ecological composite remediation agent prepared in Example 1, the COD in the water was 12.34–16.57 mg / L, ammonia nitrogen was 0.29–0.31 mg / L, total phosphorus was 0.11–0.13 mg / L, and total nitrogen was 0.96–1.16 mg / L. The water quality was lower than the Class III water quality limits stipulated in the "Surface Water Environmental Quality Standard" (GB3838–2020). Furthermore, the removal rates of COD, ammonia nitrogen, total phosphorus, and total nitrogen after adding the aquatic ecological composite remediation agent prepared in Example 1 reached 72.20%–78.17%, 87.55%–88.45%, 82.43%–86.21%, and 79.79%–82.48%, respectively.

[0165] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A composite aquatic ecosystem restoration agent, characterized in that, It includes the following components by weight: 60-190 parts microbial bacteria, 1-10 parts biological enzymes, and 1-10 parts nutrients; The microbial strains comprise the following components in parts by weight: 5-15 parts of ammonia-oxidizing denitrifying bacteria, 10-35 parts of denitrifying denitrifying bacteria, 10-30 parts of nitrifying bacteria, 10-40 parts of Bacillus subtilis, 10-40 parts of photosynthetic bacteria, 10-20 parts of Bacillus licheniformis, and 5-10 parts of yeast. The microorganisms also include Bacillus thuringiensis; The mass ratio of the ammonia-oxidizing denitrifying bacteria to Bacillus thuringiensis is 1:0.3~3; The nutrients comprise the following components in parts by weight: 10-20 parts ammonium sulfate, 10-15 parts ammonium chloride, 5-10 parts sodium nitrate, 3-5 parts dipotassium hydrogen phosphate, 3-5 parts ammonium dihydrogen phosphate, 10-15 parts food-grade glucose, 2-5 parts potassium carbonate, 5-8 parts ferric chloride, 0.5-1 part potassium peroxide, 3-5 parts potassium chloride, 5-10 parts magnesium oxide, 0.5-1 part copper sulfate pentahydrate, 0.5-1 part manganese oxide, 1-2 parts zinc oxide, 1-1.5 parts sodium molybdate, and 885.5-936 parts water. The nutrients also include the following components by weight: 9-11 parts of succinate; The succinate composition includes a mixture of disodium succinate and potassium succinate sulfonate.

2. The aquatic ecological composite restoration agent according to claim 1, characterized in that, The bio-enzyme is obtained by starch-catalyzed hydrolysis.

3. The aquatic ecological composite restoration agent according to claim 1, characterized in that, The succinate component comprises a mixture of disodium succinate and potassium succinate sulfonate in a mass ratio of 1:3 to 4.

4. A method for preparing a water ecological composite restoration agent according to any one of claims 1 to 3, characterized in that, Includes the following steps: S1. The components of the microbial bacteria are activated to obtain activated microbial bacteria; S2. The nutrient components are cooked, cooled, and then the activated microorganisms are added and mixed to obtain a mixture. S3. The mixture is subjected to an oxidative polymerization reaction to obtain a water ecological composite restoration agent.

5. The method for preparing a water ecological composite restoration agent according to claim 4, characterized in that, In step S2, the ripening temperature is 70~80℃, the rotation speed is 140~160r / min, and the time is 3~5h; the cooling temperature is 30℃; the mixing speed is 140~160r / min, and the time is 20~28h.

6. The preparation method of the aquatic ecological composite restoration agent according to claim 4, characterized in that, In step S3, the temperature of the oxidative polymerization reaction is 30~50℃ and the time is 3~4h.

7. The preparation method of the aquatic ecological composite restoration agent according to claim 4, characterized in that, It also includes the following steps: The aquatic ecological composite restoration agent is dried to obtain aquatic ecological composite restoration agent powder.

8. The preparation method of the aquatic ecological composite restoration agent according to claim 7, characterized in that, The drying process is spray drying, with a temperature of -35 to -80°C and a drying time of 4 to 16 hours.

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