CN112574921A - Method for preparing aerobic denitrification composite microbial inoculum by utilizing kitchen waste and application thereof - Google Patents
Method for preparing aerobic denitrification composite microbial inoculum by utilizing kitchen waste and application thereof Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/343—Biological treatment of water, waste water, or sewage characterised by the microorganisms used for digestion of grease, fat, oil
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/348—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the way or the form in which the microorganisms are added or dosed
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
- C02F2101/166—Nitrites
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/38—Organic compounds containing nitrogen
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/20—Nature of the water, waste water, sewage or sludge to be treated from animal husbandry
Abstract
The invention discloses a method for producing an aerobic denitrification composite microbial inoculum with a high-efficiency denitrification function by utilizing kitchen waste and application of the microbial inoculum in a sewage treatment and aquaculture system, belonging to the technical field of environmental protection. Book (I)The aerobic denitrifying bacteria strain adopted by the invention is two high-efficiency denitrifying bacteria with heterotrophic nitrification-aerobic denitrification functions separated and screened from the environment: rhodococcus (A)Rhodococcussp.) YY12 strain and Bacillus subtilis (Bacillus subtilis)Bacillus subtilis) The HA2 bacterial strain is prepared by taking kitchen waste as a main raw material, pretreating the kitchen waste to prepare a fermentation culture medium, and preparing a composite microbial inoculum by means of variable temperature culture and continuous inoculation. The composite microbial inoculum is used for biologically enhanced denitrification of a sewage treatment system and purification of aquaculture water, has high denitrification efficiency, simple operation and remarkable environmental benefit, and shows wide application prospect.
Description
Technical Field
The invention relates to a method for producing an aerobic denitrification composite microbial inoculum with a high-efficiency denitrification function by utilizing kitchen waste in a recycling manner and application of the microbial inoculum in a sewage treatment system, belonging to the technical field of environmental protection.
Background
In recent years, the catering industry in China is rapidly developed, and a large amount of kitchen waste is generated every day. According to research statistics, more than 6000 million kitchen waste is generated in cities in China every year, and the treatment cost is high. On the other hand, the kitchen waste contains rich nutrient substances such as starch, cellulose, protein and the like, and has great utilization value. Therefore, the resource utilization of the kitchen waste becomes a current research hotspot.
The over-high total nitrogen content is one of the main factors of water body pollution and eutrophication. The microbial denitrification method is widely accepted due to various advantages of high efficiency, low cost, no secondary pollution and the like, and becomes a hotspot for attention and research in the fields of environmental science and water treatment, but the existing sewage treatment process has poor effect of removing total nitrogen, a large amount of carbon sources are generally required to be added to improve the denitrification effect, and the treatment cost is greatly increased.
Aerobic denitrification refers to a process of denitrifying microorganisms in sewage with certain dissolved oxygen concentration by using nitrate or nitrite as an electron acceptor. The discovery of aerobic denitrifying bacteria breaks through the traditional cognition that the Denitrification reaction can only be carried out under the anaerobic condition, so that the Synchronous Nitrification and Denitrification (SND) becomes possible.
In recent years, the research and application of the microbial agent for purifying water quality are more and more intensive and extensive. The Chinese patent of invention (application number: 201110006790.0) discloses a Rhodococcus pyridinivorans (CPZ-24) strain with preservation number of CGMCC No.4401, which has heterotrophic nitrification and aerobic denitrification effects, and generates 99% of gas N in the denitrification process2And also a small amount of N2O、NO、NO2(ii) a The culture medium of the strain mainly uses sodium succinate to provide a carbon source, (NH)4)2SO4Providing a nitrogen source, and also K2HPO4、NaCl、MgSO4、MnSO4、FeSO4And the like. The Chinese invention patent (application number: 201110416893.4) discloses a Rhodococcus strain (nitre deep-1) with the preservation number of CGMCC No.5277, which has the capability of removing nitrite nitrogen in sewage under low-temperature aerobic conditions. Especially under the condition of 8-12 ℃, the nitrite nitrogen removal rate is improved by about 10 percent when the rhodococcus of the invention is inoculated to the sewage without the inoculated bacteria; the culture medium of the strain takes sodium acetate as a carbon source, (NH)4)2SO4Providing a nitrogen source, and also K2HPO4、NaCl、MgSO4、MnSO4、FeSO4And the like.
In summary, there are many reports of strains with denitrification function, and the culture methods are different, but there is no report of culturing complex bacteria with denitrification function by using kitchen waste. The kitchen waste has high oil content, is difficult to be used for culturing general denitrifying bacteria, and can influence the application of the microbial inoculum.
The invention content is as follows:
in order to overcome the defects in the prior art, the inventor of the application separates and screens two high-efficiency denitrifying bacteria with heterotrophic nitrification-aerobic denitrification functions, namely a Rhodococcus (Rhodococcus sp) YY12 strain and a Bacillus subtilis HA2 strain, and lays a foundation for biologically enhancing sewage denitrification.
The inventor of this application uses kitchen garbage as the raw materials, cultivates aerobic denitrification composite bacteria to prepare high-efficient stable microbial inoculum, be used for strengthening sewage treatment process's denitrogenation efficiency with the microbial inoculum, can improve the quality of water, reduce running cost, the practicality is strong.
In order to achieve the purpose, the invention provides the following technical scheme:
the strain used by the invention is 2 aerobic denitrifying bacteria separated and screened from the environment by an inventor, wherein 1 strain is a Rhodococcus sp YY12 strain which can efficiently remove ammonia nitrogen and total nitrogen in sewage, and the strain is preserved in China general microbiological culture Collection center in 12-3.2012 with the preservation number as follows: CGMCC No. 6924; the other 1 strain is a Bacillus subtilis HA2 strain with the functions of efficiently degrading grease and aerobic denitrification, and the strain is preserved in the China general microbiological culture Collection center in 2019, 05 and 20 months, wherein the preservation number is as follows: CGMCC No. 17825.
The aerobic denitrification composite bacterial agent provided by the invention comprises the following production steps:
s1, kitchen waste pretreatment: fully stirring the collected kitchen waste by a homogenizer, and sieving by a 50-mesh sieve; boiling the filtered serous fluid for 10-30 min, standing for 15-50 min to separate oil phase and water phase, and separating oil phase and water phase with separating funnel; recovering the upper layer of grease for the biodiesel raw material and performing additional treatment; the lower aqueous phase is used for preparing a microbial culture medium;
s2, preparation of a culture medium: diluting the water-phase component of the kitchen waste obtained in the step 1 by using tap water until the COD content is 20-80 g/L; adjusting the pH value of the culture medium to 7.0-8.0 by using a sodium hydroxide aqueous solution, and sterilizing the culture medium for 10-30 minutes at 110-126 ℃ to obtain a microbial inoculum production culture medium; mixing a microbial inoculum production culture medium with an LB culture medium at a ratio of 1:1, and sterilizing at 115-126 ℃ for 15-30 minutes to obtain a liquid seed culture medium;
s3, preparing liquid seeds: filling the liquid seed culture medium obtained in the step 2 into a triangular flask according to the volume ratio of 10-20%, respectively inoculating rhodococcus YY12 strain and bacillus subtilis HA2 strain which are preserved on an inclined plane, and performing shaking culture for 18-36 hours at the temperature of 25-35 ℃ at 120-200 rpm to obtain liquid seeds;
s4, liquid fermentation: inoculating 2-7% by volume of bacillus subtilis HA2 liquid seeds into a fermentation tank filled with a microbial inoculum production culture medium, continuously introducing sterile air, starting from the initial temperature of 25-35 ℃, and within 10-15 hours, carrying out gradient temperature rise to 50-65 ℃, and maintaining for 1-2 hours; cooling to the initial temperature, inoculating 5-9% by volume of rhodococcus rhodochrous YY12 liquid seeds, and continuously culturing for 16-20 hours; the fermentation liquor is counted by plate bacteria, and the number of viable bacteria reaches 2.0 multiplied by 109More than CFU/ml, the obtained fermentation liquor can be directly used as a liquid microbial inoculum;
s5, preparing a solid microbial inoculum: and (2) taking diatomite sieved by a 200-mesh sieve as a carrier, adding the fermentation liquor into the diatomite according to the weight ratio of the fermentation liquor to the diatomite of 2: 1-1: 5, fully and uniformly mixing, freeze-drying or naturally ventilating and drying until the water content of the microbial inoculum is below 20%, so as to obtain a solid microbial inoculum, packaging, and storing in a cool and dry place for later use.
The invention also provides a use method of the aerobic denitrification complex microbial inoculum in the sewage treatment process, which comprises the following steps:
the prepared aerobic denitrification composite microbial inoculum is added into an aeration tank of a sewage treatment system, and the denitrification effect of sewage is enhanced under the condition of dissolved oxygen.
After the liquid microbial inoculum provided by the embodiment of the invention is added in the MBR process, the removal rate of total nitrogen can be improved by more than 55%; in A2After the solid microbial inoculum provided by the embodiment of the invention is added in the O sewage treatment process, the removal rate of ammonia nitrogen can be improved by more than 30%, and the removal rate of total nitrogen can be improved by more than 25%.
The third aspect of the invention provides a method for using the aerobic denitrification complex bacterial agent in aquaculture, which comprises the following steps: the prepared solid microbial inoculum is diluted by 10-100 times by using clear water, is uniformly mixed, is uniformly sprayed into a pond for aquaculture according to the dosage of 10-100 milliliters of diluent per cubic meter of water, is sprayed for 1 time in 5-10 days in summer, and is sprayed for 1 time in 10-20 days in other seasons, so that the content of ammonia nitrogen and nitrite nitrogen in the aquaculture water can be remarkably reduced, and the water quality is kept not to exceed the standard.
The application has at least one of the following beneficial effects:
according to the method for preparing the aerobic denitrification composite microbial inoculum by utilizing the kitchen waste, on one hand, the kitchen waste is changed into valuable, the pollution to the environment is reduced, the treatment cost is saved, and the composite microbial inoculum is prepared in a variable temperature culture and continuous inoculation mode; on the other hand, the prepared aerobic denitrification composite bacteria has strong denitrification capability, can greatly reduce the sewage treatment cost when being applied to sewage denitrification, can remove ammonia nitrogen and nitrite nitrogen in the culture water body when being applied to aquaculture, and can keep good water quality.
The specific implementation mode is as follows:
example 1: preparation of liquid aerobic denitrification composite microbial agent from kitchen waste
1. Pretreating kitchen waste: fully stirring the collected kitchen waste by a homogenizer, and sieving by a 50-mesh sieve; boiling the filtered slurry for 20 minutes, standing for 35 minutes to separate an oil phase from a water phase, and separating the oil phase from the water phase by using a separating funnel; recovering the grease on the upper layer; the lower aqueous phase was used to prepare the microbial culture medium.
2. Preparing a culture medium: diluting the water-phase component of the kitchen waste obtained in the step 1 by using tap water until the COD content is 50 g/L; adjusting the pH value to 7.5 by using an aqueous solution of sodium hydroxide, and sterilizing for 15 minutes at 115 ℃ to obtain a microbial inoculum production culture medium; and (3) mixing the microbial inoculum production culture medium with an LB culture medium in a ratio of 1:1, and sterilizing at 115 ℃ for 15 minutes to obtain a liquid seed culture medium.
3. Preparing liquid seeds: and (3) filling the liquid seed culture medium obtained in the step (2) into a triangular flask according to the volume ratio of 15%, respectively inoculating the rhodococcus YY12 strain and the bacillus subtilis HA2 strain which are preserved on the inclined plane, and performing shaking culture at 30 ℃ and 150 rpm for 24 hours to obtain liquid seeds.
4. Liquid state fermentation: inoculating 5 vol% Bacillus subtilis HA2 liquid seed into a fermenter filled with microbial inoculum production culture medium, continuously introducing sterile air, and starting from the initial temperature of 35 deg.CGradient heating to 60 deg.C for 2 hr, and culturing for 16 hr; cooling to 30 ℃, inoculating 6% by volume of rhodococcus erythropolis YY12 liquid seeds, and continuously culturing for 18 hours; the fermentation liquor is counted by plate bacteria, and the number of viable bacteria reaches 3.0 multiplied by 109Above CFU/ml, the obtained fermentation liquor can be directly used as a liquid microbial inoculum.
Example 2: preparation of solid aerobic denitrification composite microbial agent from kitchen waste
1. Pretreating kitchen waste: fully stirring the collected kitchen waste by a homogenizer, and sieving by a 50-mesh sieve; boiling the filtered slurry for 15 minutes, standing for 50 minutes to separate an oil phase from a water phase, and separating the oil phase from the water phase by using a separating funnel; recovering the upper layer of grease for the biodiesel raw material and performing additional treatment; the lower aqueous phase was used to prepare the microbial culture medium.
2. Preparing a culture medium: diluting the water-phase component of the kitchen waste obtained in the step 1 by using tap water until the COD content is 60 g/L; adjusting the pH value to 7.6 by using a sodium hydroxide aqueous solution, and sterilizing at 110 ℃ for 20 minutes to obtain a microbial inoculum production culture medium; and (3) mixing the microbial inoculum production culture medium with an LB culture medium in a ratio of 1:1, and sterilizing at 115 ℃ for 20 minutes to obtain a liquid seed culture medium.
3. Preparing liquid seeds: and (3) filling the liquid seed culture medium obtained in the step (2) into a triangular flask according to the volume ratio of 20%, respectively inoculating the rhodococcus YY12 strain and the bacillus subtilis HA2 strain which are preserved on the inclined plane, and performing shaking culture at 30 ℃ and 180 r/min for 20 hours to obtain liquid seeds.
4. Liquid state fermentation: inoculating 3% by volume of Bacillus subtilis HA2 liquid seeds into a fermentation tank filled with a microbial inoculum production culture medium, continuously introducing sterile air, starting from an initial temperature of 30 ℃, carrying out gradient temperature rise to 65 ℃ for 1 hour, and carrying out total culture for 15 hours; cooling to 30 ℃, inoculating 5% by volume of rhodococcus erythropolis YY12 liquid seeds, and continuously culturing for 20 hours; the fermentation liquor is counted by plate bacteria, and the number of viable bacteria reaches 5.0 multiplied by 109CFU/ml or above, and the obtained fermentation liquor is used for preparing the solid microbial inoculum.
5. Preparing a solid microbial inoculum: adding the fermentation liquor into diatomite according to the weight ratio of the fermentation liquor to the diatomite of 1:1 by taking the diatomite which is sieved by a 200-mesh sieve as a carrier, fully and uniformly mixing, freeze-drying until the water content of the microbial inoculum is below 20 percent to obtain a solid microbial inoculum, packaging, and storing in a cool and dry place for later use.
Example 3: aerobic denitrification composite bacterial agent enhanced membrane bioreactor denitrification
In order to overcome the defect of poor capability of removing total nitrogen by a membrane bioreactor, the composite microbial inoculum prepared by the invention is properly added into a membrane pool of an MBR, and the removal effect of the system on total nitrogen in sewage can be improved under the aeration condition. The specific method comprises the following steps: the aerobic denitrification liquid microbial inoculum prepared in any embodiment (taking the composite microbial inoculum prepared in embodiment 1 as an example) is added into a normally-operated domestic sewage treatment membrane bioreactor according to the volume ratio of 0.2-1%, and the aerobic denitrification liquid microbial inoculum is added once every 7-15 days. After the microbial inoculum provided by the embodiment of the invention is added, the average removal rate of the membrane bioreactor on ammonia nitrogen is improved by more than 20%, and the average removal rate of total nitrogen is improved by more than 55%, so that the effluent quality of the membrane bioreactor is improved. Comparative operating data are shown in table 1:
table 1: water inlet and outlet quality of membrane bioreactor with different microbial inoculum addition amounts
| Index of water quality | COD(mg/L) | Ammonia nitrogen (mg/L) | Total nitrogen (mg/L) |
| Inflow water | 309±56 | 27±7 | 39±6 |
| Water yielding without adding microbial inoculum | 32±3 | 0.75±0.13 | 23±4 |
| Adding 0.2% of microbial inoculum to obtain effluent | 28±3 | 0.56±0.16 | 8.9±1.2 |
| Adding 1% of microbial inoculum to obtain effluent | 20±4 | 0.33±0.11 | 6.7±1.1 |
Example 4: aerobic denitrification composite bacterial agent enhanced anoxic-aerobic sewage treatment process denitrification
In normal operation A2The aerobic denitrification solid microbial inoculum prepared in any embodiment is added into the O sewage treatment system according to the weight ratio of 0.1-0.5 per mill, and is added once every 10-20 days. After the microbial inoculum provided by the embodiment of the invention is added, the removal rate of ammonia nitrogen can be improved by more than 30%, and the removal rate of total nitrogen can be improved by more than 25%, so that the effluent quality of a sewage treatment system is improved. Comparative operating data are shown in table 2:
table 2: a of different microbial inoculum addition amounts2Water inlet and outlet quality of O process sewage treatment system
| Index of water quality | COD(mg/L) | Ammonia nitrogen (mg/L) | Total nitrogen (mg/L) |
| Inflow water | 288±63 | 28±6 | 36±7 |
| Water yielding without adding microbial inoculum | 37±9 | 4.3±0.9 | 13±3 |
| Adding 0.2 per mill of microbial inoculum to obtain effluent | 30±5 | 2.7±1.3 | 9.7±1.1 |
| Adding 0.5 per mill of microbial inoculum to obtain effluent | 23±6 | 1.4±0.8 | 7.1±1.3 |
Example 5: application of aerobic denitrification composite bacterial agent in improving water quality of aquaculture water
The experimental base is located in the coastal region of Guangdong, 6 high-level aquaculture ponds (each pond occupies 3 mu of land and is 2.5 meters deep) for penaeus vannamei are respectively provided with 3 control ponds without adding microbial inoculum and 3 treatment ponds with adding microbial inoculum, 36 thousands of shrimp seedlings are put in each pond, and the aquaculture modes are the same. Diluting the solid microbial inoculum by 30 times with clear water, mixing uniformly, uniformly sprinkling the solid microbial inoculum into 3 treatment ponds according to the dosage of 100 ml of diluent per cubic meter of water, sprinkling for 1 time in 7 days in summer, and sprinkling for 1 time in 15 days in other seasons. The ammonia nitrogen content of the culture water body of the treatment pond can be stably controlled below 0.5 mg/L, and the nitrite nitrogen can be controlled below 0.7 mg/L; and the ammonia nitrogen content of the aquaculture water in the control pond fluctuates between 0.4 and 1.3 milligrams per liter, the higher the temperature is, the higher the ammonia nitrogen concentration is, the more the nitrite nitrogen content fluctuates between 0.7 and 1.5 milligrams per liter in most of time, and the nitrite nitrogen content is increased along with the temperature rise. When the size of the penaeus vannamei boone reaches 40-50 tails/jin, the penaeus vannamei boone is harvested, and the yield of the penaeus vannamei boone in the treatment pond is improved by about 20 percent compared with that in the control pond.
Claims (4)
1. A method for preparing an aerobic denitrification complex microbial inoculum by utilizing kitchen waste is characterized by comprising the following specific operation steps:
s1, kitchen waste pretreatment: fully stirring the collected kitchen waste by a homogenizer, and sieving by a 50-mesh sieve; boiling the filtered serous fluid for 10-30 min, standing for 15-50 min to separate oil phase and water phase, and separating oil phase and water phase with separating funnel; recovering the upper layer of grease for the biodiesel raw material and performing additional treatment; the lower aqueous phase is used for preparing a microbial culture medium;
s2, preparation of a culture medium: diluting the water-phase component of the kitchen waste obtained in the step S1 with tap water until the COD content is 20-80 g/L; adjusting the pH value of the culture medium to 7.0-8.0 by using a sodium hydroxide aqueous solution, and sterilizing the culture medium for 10-30 minutes at 110-126 ℃ to obtain a microbial inoculum production culture medium; mixing a microbial inoculum production culture medium with an LB culture medium at a ratio of 1:1, and sterilizing at 115-126 ℃ for 15-30 minutes to obtain a liquid seed culture medium;
s3, preparing liquid seeds: filling the liquid seed culture medium obtained in the step 2 into a triangular flask according to the volume ratio of 10-20%, respectively inoculating rhodococcus YY12 strain and bacillus subtilis HA2 strain which are preserved on an inclined plane, and performing shaking culture for 18-36 hours at the temperature of 25-35 ℃ at 120-200 rpm to obtain liquid seeds;
s4, liquid fermentation: inoculating into a fermentation tank filled with a microbial inoculum production culture mediumContinuously introducing sterile air into bacillus subtilis HA2 liquid seeds with the volume ratio of 2-7%, starting from the initial temperature of 25-35 ℃, and within 10-15 hours, carrying out gradient temperature rise to 50-65 ℃, and maintaining for 1-2 hours; cooling to the initial temperature, inoculating 5-9% by volume of rhodococcus rhodochrous YY12 liquid seeds, and continuously culturing for 16-20 hours; the fermentation liquor is counted by plate bacteria, and the number of viable bacteria reaches 2.0 multiplied by 109More than CFU/ml, the obtained fermentation liquor can be directly used as a liquid microbial inoculum;
s5, preparing a solid microbial inoculum: adding the fermentation liquor into diatomite according to the weight ratio of the fermentation liquor to the diatomite of 2: 1-1: 5 by taking the diatomite which is sieved by a 200-mesh sieve as a carrier, fully and uniformly mixing, freeze-drying or naturally ventilating and drying until the water content of the microbial inoculum is below 20 percent to obtain a solid microbial inoculum, packaging, and storing in a cool and dry place for later use.
2. The method for preparing the aerobic denitrification complex microbial inoculum by utilizing the kitchen waste as claimed in claim 1, is characterized in that: in the step S3, the Rhodococcus YY12 strain and the Bacillus subtilis HA2 strain both have heterotrophic nitrification-aerobic denitrification functions and are preserved in the China general microbiological culture Collection center with the preservation numbers as follows: CGMCC number 6924 and CGMCC number 17825.
3. The use method of the aerobic denitrification complex microbial inoculum in the sewage treatment process as claimed in claim 1, is characterized in that: and adding the aerobic denitrification composite bacterial agent into an aeration tank of the sewage treatment system to enhance sewage denitrification.
4. The method for using the aerobic denitrification complex bacterial agent in aquaculture as claimed in claim 1, which is characterized in that: diluting the solid microbial inoculum by 10-100 times with clear water, uniformly mixing, uniformly splashing 10-100 ml of diluent per cubic meter of water into a pond for aquaculture, 1 time for 5-10 days in summer, and 1 time for 10-20 days in other seasons.
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| CN113005062A (en) * | 2021-03-24 | 2021-06-22 | 中国科学院成都生物研究所 | Facultative ammonia oxidizing bacteria and application thereof |
| CN114317381A (en) * | 2022-03-04 | 2022-04-12 | 东北农业大学 | T-level microbial composite inoculant and preparation method and application thereof |
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| CN110437017A (en) * | 2019-09-10 | 2019-11-12 | 中国科学院生态环境研究中心 | A kind of method that feces of livestock and poultry produces microbial manure and controls agricultural non -point pollution |
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| CN113005062A (en) * | 2021-03-24 | 2021-06-22 | 中国科学院成都生物研究所 | Facultative ammonia oxidizing bacteria and application thereof |
| CN114317381A (en) * | 2022-03-04 | 2022-04-12 | 东北农业大学 | T-level microbial composite inoculant and preparation method and application thereof |
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