An integrated device for mutagenesis and domestication of anammox bacteria for wastewater treatment

Abstract

The utility model discloses an integrated device for sewage treatment anaerobic ammonia oxidation bacteria mutagenesis and domestication. The device comprises a mutagenesis reaction cabin, a domestication culture cabin, a control system and a connecting pipeline. The mutagenesis reaction cabin is used for physical / chemical mutagenesis of bacterial liquid; the domestication culture cabin is an anaerobic environment, and its innovation lies in that it is provided with a water inlet and a water outlet, can be connected with simulated or actual wastewater, and can continuously or sequentially evaluate the denitrification efficiency domestication of mutagenic strains; the two cabins are seamlessly connected through the pipeline, and the bacterial liquid is transferred without oxygen. The utility model integrates three functions of bacterial strain mutagenesis, efficient enrichment and wastewater domestication, effectively avoids inactivation and pollution in bacterial strain transfer, can directly simulate sewage treatment conditions to select efficient strains, greatly shortens the research and development period from bacterial strain breeding to application verification, and is a special equipment for developing high-performance anaerobic ammonia oxidation bacteria for sewage treatment.

Description

Technical Field

[0001] This utility model relates to the field of biological treatment technology for high ammonia nitrogen wastewater, specifically to an integrated laboratory equipment for the breeding and application of functional microorganisms (anaerobic ammonia oxidizing bacteria) for water treatment. This device directly serves the development of new bacterial agents and process optimization for the treatment of high ammonia nitrogen wastewater. Background Technology

[0002] Anaerobic ammonia oxidation (Anammox) technology is considered one of the most promising biological nitrogen removal technologies for wastewater due to its advantages such as requiring no organic carbon source, producing less sludge, and having lower operating costs. It is also a key technology for achieving energy conservation and self-sufficiency in wastewater treatment plants in the future. However, its core functional bacteria—anaerobic ammonia oxidizing bacteria—have inherent limitations, such as an extremely long doubling time (up to 11 days), extreme sensitivity to the environment (strictly anaerobic and photosensitive), and poor tolerance to organic matter, salts, and toxic substances present in actual wastewater. These limitations severely restrict the widespread application of this technology in the treatment of complex industrial wastewater.

[0003] Genetic modification of anaerobic ammonia-oxidizing bacteria through physical (e.g., ultraviolet light) or chemical mutagenesis breeding is an effective way to obtain highly active and resilient engineered strains. However, traditional research and development models have significant drawbacks: the laboratory breeding environment is severely disconnected from the actual wastewater treatment environment. Mutagenic strains screened under ordinary culture media and standard conditions often experience a sharp decrease in efficacy or even failure when transplanted into real wastewater environments with complex compositions and variable conditions, resulting in lengthy research and development cycles and extremely low success rates. Existing laboratory equipment (such as shakers and ordinary anaerobic reactors) has limited functionality, and key steps such as mutagenesis, enrichment, and wastewater acclimatization must be completed by transferring strains between different devices. During the transfer process, strains not only face the risk of contamination by other bacteria but also suffer severe activity loss due to exposure to oxygen, making the screening work much less efficient.

[0004] Therefore, there is an urgent need in this field for a specialized device that can highly simulate wastewater treatment conditions and seamlessly integrate the processes of strain selection, enrichment and scale-up, and wastewater acclimatization assessment into one unit. This device would enable the targeted screening and acclimatization of strains in a near-realistic application environment, thereby accelerating the development of highly efficient anaerobic ammonia-oxidizing bacteria agents that can be used in practical engineering. Summary of the Invention

[0005] The purpose of this invention is to overcome the shortcomings of existing technologies and provide a highly integrated and fully functional device for the mutagenesis and domestication of anaerobic ammonia-oxidizing bacteria in wastewater treatment. This device can simulate actual wastewater treatment conditions within a closed system, completing the entire process from mutagenesis to performance verification, effectively solving the problem of the disconnect between strain development and application scenarios, and significantly improving R&D efficiency and success rate.

[0006] To achieve the above objectives, the present invention adopts the following technical solution: An integrated device for mutagenesis and acclimatization of anaerobic ammonia-oxidizing bacteria in wastewater treatment mainly includes a mutagenesis reaction chamber, an acclimatization culture chamber, a control system, and a conveying pipeline connecting the mutagenesis reaction chamber and the acclimatization culture chamber. The mutagenesis reaction chamber has a sealed sample inlet at the top, an ultraviolet mutagenesis lamp in the upper part of the chamber, a magnetic stirrer at the bottom, and a sampling port and an outlet connected to the conveying pipeline on the side wall. The acclimatization culture chamber is a closed anaerobic reactor with a wastewater inlet at the lower part of its side wall, an outlet at the top, an air inlet, an exhaust port, and a sealing cap at the top. The chamber contains suspended biological packing material for biofilm formation and a microporous aerator connected to the air inlet. The side wall also has a bacterial liquid inlet connected to the conveying pipeline and a sampling port for the culture chamber. The conveying pipeline is equipped with a conveying pump and a check valve. The control system is electrically connected to the ultraviolet mutagenesis lamp, magnetic stirrer, delivery pump, water inlet pump located at the water inlet, and temperature sensor, pH sensor, and dissolved oxygen sensor located in each chamber.

[0007] The wastewater inlet of the acclimatization and cultivation chamber is connected to the water distribution tank via an inlet pump. The water distribution tank can store simulated wastewater or pretreated actual wastewater with different C / N ratios or different concentrations of ammonia nitrogen and nitrite nitrogen.

[0008] The outlet of the acclimatization and cultivation chamber is connected to the effluent collection tank and can be equipped with an online water quality monitor for real-time monitoring of changes in ammonia nitrogen, nitrite nitrogen, and nitrate nitrogen levels in the effluent.

[0009] The mutagenesis reaction chamber is used to perform mutagenesis treatment on the bacterial strain. The chamber body is preferably made of ultraviolet-transmitting quartz glass or special plexiglass. A sample inlet with a sealed cap is located at the top for injecting the bacterial solution and chemical mutagen. An ultraviolet mutagenesis lamp is installed in the upper part of the chamber to provide a physical mutagenesis source; a magnetic stirrer is installed at the bottom to ensure that the bacterial solution and mutagen are mixed evenly and receive uniform irradiation. Sampling ports and outlets are located on the side walls.

[0010] The acclimatization and cultivation chamber is the core innovation of this device. Essentially a small, advanced anaerobic bioreactor, it features a wastewater inlet at the bottom and an outlet at the top, allowing connection to an external water distribution system. An air inlet and outlet are located at the top, which can be connected to cylinders for mixed gases such as nitrogen and carbon dioxide to precisely create and maintain a strictly anaerobic environment. The chamber is filled with a large amount of suspended biological packing material (such as porous polyethylene or polyurethane packing), with a preferred filling rate of 30%-50%, providing a large surface area for microbial attachment and forming a highly active biofilm. The microporous aeration heads at the bottom are not for aeration and oxygenation, but rather for introducing inert gases (such as N2) to maintain the anaerobic environment and provide gentle stirring, or for introducing CO2 as an inorganic carbon source. The side walls also have a bacterial liquid inlet and multiple sampling ports.

[0011] The suspended biological packing material is a biological carrier made of porous polymer material, and its filling rate is 30%-50% of the effective volume of the acclimatization and cultivation chamber.

[0012] The connecting pipeline is used to connect the outlet of the mutagenesis reaction chamber and the inlet of the bacterial solution in the acclimatization culture chamber. It is equipped with a sterile transfer pump and a check valve, which can automatically and sterilely pump the mutagenized bacterial solution into the acclimatization culture chamber under the command of the control system, eliminating contamination and activity loss during the transfer process.

[0013] The control system, serving as the brain of the entire device, employs a PLC or microcontroller system. It is electrically connected to all sensors (internal temperature, pH, dissolved oxygen) and actuators (UV lamps, stirrers, transfer pumps, inlet pumps, and gas valves). Users can pre-set mutagenesis parameters (time, intensity) and acclimatization procedures (inlet flow rate, reaction cycle, environmental parameters) via a human-machine interface. The system can then automatically complete the entire "mutation-sterile transfer-wastewater acclimatization" cycle and adjust operating parameters based on online monitoring of effluent water quality data, achieving intelligent screening.

[0014] The beneficial effects of this utility model are as follows: 1. Targeted screening with clear application orientation: The first-ever integration of the wastewater acclimatization system directly into the strain selection device allows the strains to undergo pressure screening under simulated actual wastewater conditions from the beginning of the mutagenesis process. This ensures that the resulting superior strains not only have high activity but also possess inherent strong adaptability and excellent treatment efficiency to the target wastewater, greatly improving the targeting and success rate of the research and development.

[0015] 2. Revolutionary reduction in R&D cycle: The traditionally separate and independent "strain development - small-scale testing" R&D stages are perfectly integrated, avoiding the cumbersome transfer and reactivation adaptation of strains between different devices, greatly shortening the R&D cycle of several months or even several years, and accelerating the process of high-performance bacterial agents from the laboratory to engineering applications.

[0016] 3. Highly integrated functions, avoiding secondary pollution: This multi-functional unit integrates mutagenesis, enrichment, wastewater acclimatization, and online water quality monitoring. The fully enclosed and automated operation completely eliminates contamination by other microorganisms and the damage to strictly anaerobic bacteria caused by oxygen intrusion, ensuring the purity and reliability of experimental results.

[0017] 4. Accurate data and strong guidance: The online water quality monitoring system can provide real-time and accurate feedback on changes in the denitrification performance of the bacterial community. The first-hand data obtained truly reflects the ability of the strains in similar application environments, providing a valuable direct basis for process parameter optimization and bacterial agent performance evaluation. Attached Figure Description

[0018] Figure 1 This is a schematic diagram of the overall structure of this utility model.

[0019] Explanation of reference numerals in the attached figures: 1: Mutagenesis Reaction Chamber 101: Sealed sample dispensing port 102: Ultraviolet Mutagenesis Lamp 103: Magnetic stirrer 104: Sampling port of the reaction chamber 105: Exports 2: Acclimatization and Cultivation Chamber 201: Air Intake 202: Exhaust port 203: Sealing cap 204: Suspended biological packing material 205: Microporous Aeration Head 206: Imported bacterial solution 207: Wastewater inlet 208: Water outlet 209: Sampling port of the incubation chamber 210: Inlet pump 211: Mixed Gas Cylinder 212: Sensor 3: Control System 4: Delivery pipeline 401: Transfer Pump 402: Check valve 5: Water distribution tank 6: Water collection tank 601: Online water quality monitoring instrument Detailed Implementation

[0020] The present invention will be further described in detail below with reference to the accompanying drawings and preferred embodiments.

[0021] like Figure 1 As shown, the main body of this utility model device consists of a mutagenesis reaction chamber (1) and an acclimatization and cultivation chamber (2) arranged side by side, connected by a delivery pipeline (4) and controlled by a control system (3).

[0022] Researchers first prepared high-ammonia nitrogen wastewater (ammonia nitrogen concentration ~1000 mg / L) simulating landfill leachate using a water distribution tank (5). Then, a certain concentration of anaerobic ammonia-oxidizing bacteria enrichment solution was injected into the mutagenesis reaction chamber (1) through a sealed sample inlet (101). After sealing, the ultraviolet mutagenesis program was set via the control system (3) (e.g., wavelength 254 nm, intensity 15 W, distance 20 cm, time 2 min), and the magnetic stirrer (103) was started. After the mutagenesis program ended, the control system (3) automatically started the transfer pump (401), pumping the entire bacterial solution through pipeline (4) into the acclimatization culture chamber (2), which had been pre-purified with high-purity nitrogen.

[0023] Next, the control system (3) starts the inlet pump (210) and begins to operate in the sequential batch reactor (SBR) mode: a certain amount of simulated wastewater is added to the acclimatization and cultivation chamber (2) every day. After reacting for several hours, the mixture is allowed to settle, and then the supernatant is discharged through the outlet (208), and the next cycle begins automatically. The online water quality monitor (601) analyzes the effluent water quality in real time, and the data is transmitted back to the control system (3).

[0024] After several weeks of operation, by comparing the removal rates of ammonia nitrogen and nitrite nitrogen in the effluent, the bacterial communities with significantly improved performance after mutagenesis-acclimation can be clearly screened. Researchers can periodically obtain biofilm samples from the sampling port (209) for high-throughput sequencing of the microbial community and analyze changes in dominant bacterial species.

[0025] The entire process is completed automatically under closed, anaerobic conditions, and highly efficient, sterile, and reliable screening of anaerobic ammonia-oxidizing strains for high ammonia nitrogen wastewater has been achieved.

[0026] The above description is only a preferred embodiment of the present utility model, but the protection scope of the present utility model is not limited thereto. Any equivalent substitutions or changes made by those skilled in the art within the technical scope disclosed in the present utility model, based on the technical solution and concept of the present utility model, should be included within the protection scope of the present utility model.

Claims

1. An integrated device for mutagenesis and domestication of anaerobic ammonia-oxidizing bacteria in wastewater treatment, characterized in that: The system includes a mutagenesis reaction chamber (1), an acclimatization and cultivation chamber (2), a control system (3), and a conveying pipeline (4) connecting the mutagenesis reaction chamber (1) and the acclimatization and cultivation chamber (2). The mutagenesis reaction chamber (1) has a sealed sample addition port (101) at the top, an ultraviolet mutagenesis lamp (102) at the upper part of the chamber, a magnetic stirrer (103) at the bottom, a reaction chamber sampling port (104) on the side wall, and an outlet (105) connected to the conveying pipeline (4). The acclimatization and cultivation chamber (2) is a closed anaerobic reactor. Its side wall has a wastewater inlet (207) at the lower part, an outlet (208) at the upper part, an air inlet (201) at the top, and an exhaust outlet. The chamber is equipped with an inlet (202) and a sealing cap (203). The chamber is equipped with a suspended biological packing material (204) for biofilm formation and a microporous aeration head (205) connected to the air inlet. The side wall is also equipped with a bacterial liquid inlet (206) connected to the delivery pipeline (4) and a culture chamber sampling port (209). The delivery pipeline (4) is equipped with a delivery pump (401) and a check valve (402). The control system (3) is electrically connected to the ultraviolet mutagenesis lamp (102), the magnetic stirrer (103), the delivery pump (401), the water pump (210) set at the water inlet (207), and the temperature sensor, pH sensor, and dissolved oxygen sensor set in each chamber.

2. The integrated device for mutagenesis and domestication of anaerobic ammonia-oxidizing bacteria in wastewater treatment according to claim 1, characterized in that: The wastewater inlet (207) of the acclimatization and cultivation chamber (2) is connected to the water distribution tank (5) via the water inlet pump (210). The water distribution tank (5) can store simulated wastewater or pretreated actual wastewater with different C / N ratios or different concentrations of ammonia nitrogen and nitrite nitrogen.

3. The integrated device for mutagenesis and domestication of anaerobic ammonia-oxidizing bacteria in wastewater treatment according to claim 2, characterized in that: The outlet (208) of the acclimatization and cultivation chamber (2) is connected to the effluent collection tank (6), and an online water quality monitor (601) can be installed to monitor the changes in ammonia nitrogen, nitrite nitrogen and nitrate nitrogen in the effluent in real time.

4. The integrated device for mutagenesis and domestication of anaerobic ammonia-oxidizing bacteria in wastewater treatment according to claim 1, characterized in that: The suspended biological filler (204) is a biological carrier made of porous polymer material, and its filling rate is 30%-50% of the effective volume of the acclimatization and cultivation chamber (2).

5. The integrated device for mutagenesis and domestication of anaerobic ammonia-oxidizing bacteria in wastewater treatment according to claim 1, characterized in that: The control system (3) is a PLC or a single-chip microcomputer system, and the built-in program can preset and run the complete process cycle of "mutation-transfer-domestication".

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