Complex microbial inoculant for treating underground water as well as preparation method and application of complex microbial inoculant

Abstract

The invention discloses a complex microbial inoculant for treating underground water as well as a preparation method and application thereof, relates to the field of sewage treatment, and aims to solve the problem that TBBPA and nitrate in underground water are difficult to remove at the same time in an existing biological treatment method. The composite microbial agent is prepared from pseudomonas koreensis Y5-11, acinetobacter pidermatum LYS-5, Pseudomonas sp. Y39-6 and psychrophilic bacteria F5-6, and the pseudomonas sp. Y5-11, the acinetobacter pidermatum LYS-5, the Pseudomonas sp. Y39-6, the psychrophilic bacteria F5-6 and the psychrophilic bacteria F5-6 are added into the microbial agent. The four strains are respectively subjected to fermentation culture, and the four fermentation liquids are mixed to prepare the complex microbial inoculant. The four bacteria in the complex microbial inoculant have a stronger metabolism complementation effect, and the effect on composite pollution remediation is more remarkable. The complex microbial inoculant is used for removing TBBPA and nitrate in underground water at the same time, the complex microbial inoculant has biodiversity, and the treatment result conforms to the water quality characteristics of the underground water.

Description

Technical Field

[0001] The present invention relates to the field of sewage treatment, and in particular to a composite bacterial agent for treating groundwater, a preparation method thereof, and an application thereof. Background Art

[0002] Tetrabromobisphenol A (TBBPA) has the characteristics of high persistence, environmental stability, and difficulty in degradation. It easily accumulates in humans and animals and can be transmitted through the food chain, poisoning organisms at higher trophic levels, ultimately causing harm to human health. TBBPA is a type of endocrine disruptor and a typical representative of brominated flame retardants. It is used in plastics, electronic appliances, furniture, interior decoration, transportation, petroleum and mining. Due to excessive emissions into the environment, it has also been detected in soil and water bodies. As an endocrine disruptor, TBBPA can cause neurobehavioral and immunotoxic effects, hepatotoxicity, oxidative stress and cell apoptosis. With the intensification of soil pollution, contaminated soil has also become a continuous source of groundwater pollution. About 70% of the population in my country relies on groundwater as the main source of drinking water, and more than 95% of the rural population in the country drinks groundwater. Groundwater as a water source is threatened by TBBPA pollution. Affected by overproduction in agriculture, groundwater also has the problem of excessive nitrate content. Moreover, groundwater is located in rock layers and often shows nitrates and Fe 2+ 、Mn 2+ The combined pollution of chlorinated pesticides and TBBPA seriously affects water supply safety.

[0003] Biological treatment methods offer advantages over physical and chemical methods such as ion exchange, electrochemical reduction, and electrodialysis, such as environmental friendliness, cost-effectiveness, and significant effectiveness. However, current biological treatment methods lack an effective solution for simultaneously removing TBBPA and nitrate from groundwater. Summary of the Invention

[0004] The present invention aims to solve the problem that existing biological treatment methods are difficult to simultaneously remove TBBPA and nitrate in groundwater, and provides a composite bacterial agent for treating groundwater, a preparation method and an application thereof.

[0005] The composite bacterial agent for treating groundwater of the present invention includes Pseudomonas koreana ( Pseudomonaskoreensis )Y5-11, Acinetobacter pituitus ( Acinetobacterpittii )LYS-5, Pseudomonasextremaustralis Y39-6 and psychrophilic bacteria ( Psychrobactercryohalolentis )F5-6.

[0006] Furthermore, the Korean Pseudomonas aeruginosa in the composite bacterial agent ( Pseudomonaskoreensis )Y5-11, Acinetobacter pituitus ( Acinetobacterpittii )LYS-5, Pseudomonasextremaustralis Y39-6 and psychrophilic bacteria (Psychrobactercryohalolentis ) The bacterial count ratio of F5-6 is (1~3): (2~4): (0.5~2): (1~3).

[0007] The present invention provides a method for preparing a composite bacterial agent for treating groundwater, comprising the following steps: 1. Korean Pseudomonas Pseudomonaskoreensis )Y5-11, Acinetobacter pituitus ( Acinetobacterpittii )LYS-5, Pseudomonasextremaustralis Y39-6 and psychrophilic bacteria ( Psychrobactercryohalolentis ) F5-6 were activated respectively; 2. Activate Korean Pseudomonas Y5-11, Acinetobacter Pitt LYS-5, Pseudomonasextremaustralis Y39-6 and psychrophilic bacteria F5-6 were fermented and cultured separately, and the number of bacteria in the fermentation broth of each bacteria was 10 12 pcs / L; 3. Korean Pseudomonas Y5-11 fermentation broth, Acinetobacter Pitt LYS-5 fermentation broth, Pseudomonasextremaustralis The Y39-6 fermentation broth and the psychrophilic bacteria F5-6 fermentation broth were mixed to prepare a composite bacterial agent.

[0008] Furthermore, the activation method of Korean Pseudomonas Y5-11 in step 1 is: inoculating Korean Pseudomonas Y5-11 into a beef extract peptone solid culture medium and activating it at 6-10°C for 24-72 hours; the beef extract peptone culture medium formula is: beef extract 8.0-14.0 g / L, NaCl 8.0-12.0 g / L, peptone 4.5-5.5 g / L, agar 18-20 g / L.

[0009] Furthermore, the activation method of Acinetobacter pitei LYS-5 in step 1 is: inoculating Acinetobacter pitei LYS-5 into beef extract peptone solid culture medium and activating it at 26-36° C. for 24-72 hours.

[0010] Furthermore, in step one Pseudomonasextremaustralis The activation method of Y39-6 is: Pseudomonasextremaustralis Y39-6 was inoculated into beef extract peptone solid medium and activated at 26-30°C for 24-72h.

[0011] Furthermore, the activation method of psychrophilic bacteria F5-6 in step 1 is: inoculating psychrophilic bacteria F5-6 into beef extract peptone solid culture medium and activating at 6-10° C. for 24-72 hours.

[0012] Furthermore, in step 2, the fermentation temperature of Korean Pseudomonas Y5-11 and psychrophilic bacteria F5-6 is 6-10° C., and the culture medium used for the fermentation is beef extract peptone liquid culture medium.

[0013] Furthermore, in step 2, Acinetobacter pituitus LYS-5 and Pseudomonasextremaustralis The fermentation temperature of Y39-6 is 26~36℃, and the culture medium used for fermentation is beef extract peptone liquid culture medium.

[0014] The beef extract peptone liquid culture medium comprises: beef extract 8.0-14.0 g / L, NaCl 8.0-12.0 g / L, and peptone 4.5-5.5 g / L.

[0015] Furthermore, in step 3, the fermentation broth of Korean Pseudomonas aeruginosa Y5-11, the fermentation broth of Acinetobacter pituitus LYS-5, Pseudomonasextremaustralis The bacterial count ratio of Y39-6 fermentation broth and psychrophilic bacteria F5-6 fermentation broth was (1~3):(2~4):(0.5~2):(1~3).

[0016] The composite bacterial agent of the present invention is used for treating groundwater contaminated by nitrate and TBBPA.

[0017] Beneficial effects of the present invention: The psychrophilic bacteria F5-6 in the composite bacterial agent of the present invention has rich extracellular secretions, which act as an adhesive and can also provide additional carbon sources for other strains. The polysaccharides, proteins and other components in the extracellular secretions can act as adhesives, which help to adhere to the surface of the carrier and also help to stick single bacteria together, thereby forming a structured biofilm community, so that a microenvironment with complementary nutrients is formed, and the small molecule nutrients decomposed by bacteria can be used by other strains. Korean Pseudomonas Y5-11 plays a facultative autotrophic denitrification role, and Pseudomonasextremaustralis Combining with Y39-6 resulted in even better nitrate removal. Acinetobacter pituitus LYS-5 also degrades TBBPA. This interaction synergistically improved nitrate and TBBPA removal.

[0018] The composite bacterial agent of the present invention is used for simultaneously removing TBBPA and nitrate from groundwater. The composite bacterial agent has biodiversity, and the treatment result conforms to the water quality characteristics of groundwater. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 A top view of the circulation fixture. DETAILED DESCRIPTION

[0020] The technical solution of the present invention is not limited to the specific embodiments listed below, but also includes any combination of the specific embodiments.

[0021] Specific embodiment 1: The composite bacterial agent for treating groundwater in this embodiment includes Korean Pseudomonas ( Pseudomonaskoreensis )Y5-11, Acinetobacter pituitus ( Acinetobacterpittii )LYS-5, PseudomonasextremaustralisY39-6 and psychrophilic bacteria ( Psychrobactercryohalolentis )F5-6.

[0022] The Korean Pseudomonas Pseudomonaskoreensis ) Y5-11 was deposited in the General Microbiology Center of China Culture Collection Administration, the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The deposit date is October 29, 2018, and the deposit number is CGMCC No. 16651.

[0023] described Psychrobactercryohalolentis F5-6 was deposited in the General Microbiology Center of the China Culture Collection Administration, with the deposit address being No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The deposit date was October 29, 2018, and the deposit number is CGMCC No. 16653.

[0024] The Acinetobacter pituitus ( Acinetobacterpittii) LYS-5 is deposited in the General Microbiology Center of the China Culture Collection Administration, the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, the deposit number is CGMCC No. 30399, and the deposit date is April 22, 2024.

[0025] Pseudomonas far-antartica Pseudomonasextremaustralis Y39-6 belongs to the genus Pseudomonas ( Pseudomonas ), has been deposited in the General Microbiology Center of China Culture Collection Administration, with the deposit number CGMCC No. 16652 and the deposit date being October 29, 2018.

[0026] Specific embodiment 2: In this embodiment, the Korean Pseudomonas aeruginosa ( Pseudomonaskoreensis )Y5-11, Acinetobacter pituitus ( Acinetobacterpittii )LYS-5, Pseudomonasextremaustralis Y39-6 and psychrophilic bacteria ( Psychrobactercryohalolentis The bacterial count ratio of F5-6 is (1-3): (2-4): (0.5-2): (1-3). Other steps and parameters are the same as those in the first embodiment.

[0027] Specific embodiment 3: The preparation method of the composite bacterial agent for treating groundwater in this embodiment includes the following steps: 1. Korean Pseudomonas Pseudomonaskoreensis )Y5-11, Acinetobacter pituitus ( Acinetobacterpittii )LYS-5, Pseudomonasextremaustralis Y39-6 and psychrophilic bacteria ( Psychrobactercryohalolentis ) F5-6 were activated respectively; 2. Activate Korean Pseudomonas Y5-11, Acinetobacter Pitt LYS-5, PseudomonasextremaustralisY39-6 and psychrophilic bacteria F5-6 were fermented and cultured separately, and the number of bacteria in the fermentation broth of each bacteria was 10 12 pcs / L; 3. Korean Pseudomonas Y5-11 fermentation broth, Acinetobacter Pitt LYS-5 fermentation broth, Pseudomonasextremaustralis The Y39-6 fermentation broth and the psychrophilic bacteria F5-6 fermentation broth were mixed to prepare a composite bacterial agent.

[0028] Specific embodiment 4: In step 1 of this embodiment, the activation method of Korean Pseudomonas Y5-11 is as follows: Korean Pseudomonas Y5-11 is inoculated into a beef extract peptone solid medium and activated at 6-10°C for 24-72 hours. The beef extract peptone medium formula is: beef extract 8.0-14.0 g / L, NaCl 8.0-12.0 g / L, peptone 4.5-5.5 g / L, agar 18-20 g / L. Other steps and parameters are the same as those in specific embodiment 3.

[0029] Specific embodiment 5: In step 1 of this embodiment, the activation method of Acinetobacter pituitarius LYS-5 is as follows: Acinetobacter pituitarius LYS-5 is inoculated into beef extract peptone solid culture medium and activated at 26-36°C for 24-72 hours. Other steps and parameters are the same as those of specific embodiment 3.

[0030] Specific implementation method six: In step one of this implementation method Pseudomonasextremaustralis The activation method of Y39-6 is: Pseudomonasextremaustralis Y39-6 was inoculated into beef extract peptone solid medium and activated at 26-30° C. for 24-72 hours. Other steps and parameters were the same as those in the third embodiment.

[0031] Specific embodiment seven: In step one of this embodiment, the activation method of psychrophilic bacteria F5-6 is as follows: psychrophilic bacteria F5-6 are inoculated into beef extract peptone solid culture medium and activated at 6-10°C for 24-72 hours. Other steps and parameters are the same as those of specific embodiment three.

[0032] Specific embodiment 8: In step 2 of this embodiment, the fermentation temperature of Korean Pseudomonas Y5-11 and psychrophilic bacteria F5-6 is 6-10°C, and the fermentation medium used is beef extract peptone liquid medium. Other steps and parameters are the same as those of specific embodiment 1.

[0033] The beef extract peptone liquid culture medium comprises: beef extract 8.0-14.0 g / L, NaCl 8.0-12.0 g / L, and peptone 4.5-5.5 g / L.

[0034] Specific embodiment nine: In step 2 of this embodiment, Acinetobacter pituitus LYS-5 and PseudomonasextremaustralisThe fermentation temperature of Y39-6 was 26-36°C, and the culture medium used for the fermentation was beef extract peptone liquid culture medium. The other steps and parameters were the same as those in the first embodiment.

[0035] The beef extract peptone liquid culture medium comprises: beef extract 8.0-14.0 g / L, NaCl 8.0-12.0 g / L, and peptone 4.5-5.5 g / L.

[0036] Specific embodiment 10: In step 3 of this embodiment, the fermentation liquid of Korean Pseudomonas Y5-11, the fermentation liquid of Acinetobacter Pitt LYS-5, Pseudomonasextremaustralis The bacterial count ratio of the Y39-6 fermentation broth to the psychrophilic bacteria F5-6 fermentation broth is (1-3): (2-4): (0.5-2): (1-3). Other steps and parameters are the same as those in the third embodiment.

[0037] Specific embodiment 11: The composite bacterial agent of this embodiment is used to treat groundwater contaminated by nitrate and TBBPA.

[0038] The following embodiments of the present invention are described in detail. The following embodiments are implemented on the premise of the technical solution of the present invention, and detailed implementation plans and specific operating processes are given, but the protection scope of the present invention is not limited to the following embodiments.

[0039] Example 1: The preparation method of the composite bacterial agent for treating groundwater in this embodiment includes the following steps: 1. Korean Pseudomonas Pseudomonaskoreensis )Y5-11, Acinetobacter pituitus ( Acinetobacterpittii )LYS-5, Pseudomonasextremaustralis Y39-6 and psychrophilic bacteria ( Psychrobactercryohalolentis ) F5-6 were activated respectively; 2. Activate Korean Pseudomonas Y5-11, Acinetobacter Pitt LYS-5, Pseudomonasextremaustralis Y39-6 and psychrophilic bacteria F5-6 were fermented and cultured separately, and the number of bacteria in the fermentation broth of each bacteria was 10 12 pcs / L; 3. Korean Pseudomonas Y5-11 fermentation broth, Acinetobacter Pitt LYS-5 fermentation broth, Pseudomonasextremaustralis Y39-6 fermentation broth and psychrophilic bacteria F5-6 fermentation broth were mixed to prepare a composite bacterial agent; wherein Korean Pseudomonas Y5-11 fermentation broth, Pitt Acinetobacter LYS-5 fermentation broth, Pseudomonasextremaustralis The bacterial count ratio of Y39-6 fermentation broth and psychrophilic bacteria F5-6 fermentation broth was 1:2:2:1.

[0040] The Korean Pseudomonas PseudomonaskoreensisY5-11 was deposited with the General Microbiology Center of the China Culture Collection Administration of Microorganisms at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, on October 29, 2018, with the deposit number CGMCC No. 16651. This strain has been disclosed in a prior patent application.

[0041] described Psychrobactercryohalolentis F5-6 was deposited with the General Microbiology Center of the China Culture Collection Administration of Microorganisms at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, on October 29, 2018, with the deposit number CGMCC No. 16653. This strain has been disclosed in a prior patent application.

[0042] The Acinetobacter pituitii ( Acinetobacterpittii ) LYS-5 is deposited in the General Microbiology Center of China Culture Collection Administration, the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, the deposit number is CGMCC No. 30399, and the deposit date is April 22, 2024.

[0043] Pseudomonas far-antartica Pseudomonasextremaustralis Y39-6 belongs to the genus Pseudomonas ( Pseudomonas ), deposited with the China General Microbiology Center under the China Culture Collection Administration under the deposit number CGMCC No. 16652 on October 29, 2018. This strain has been disclosed in a prior patent application.

[0044] This example uses a circulating fixture to secure the composite inoculum to an activated carbon carrier. Specifically, the activated carbon carrier is filled into the fixture column of the circulating fixture. Each fixture column has an inner diameter of 5 cm and a height of 70 cm, with the activated carbon carrier occupying 60% of the total volume of the fixture column. The circulating fixture is first disinfected with 3% hydrogen peroxide. The composite inoculum is then introduced through the water inlet pipe. The composite inoculum enters the fixture column, submerging the activated carbon carrier, and occupies 30% of the total volume of the fixture column. The composite inoculum then flows out of the outlet pipe and is circulated back to the inlet pipe by a peristaltic pump. The cycle lasts 48 hours. After one cycle, 50% of the composite inoculum is discharged, and then the composite inoculum is replenished to 100%. After three cycles of this process, the loading is complete.

[0045] The top view of the circulating fixing device is as follows Figure 1As shown, the device includes two fixed columns 1, a water inlet pipe 2, two center pipes 3, and a water outlet pipe 4. One side of each fixed column 1 is provided with a water inlet, and the other side is provided with a water outlet. A drain is also provided on the sidewall of each fixed column 1. One end of the water inlet pipe 2 is connected to two water pipes via a tee, and the other ends of the two water pipes are connected to the water inlets of the two fixed columns 1, respectively. Elbows are provided at the corners of the water pipes. One end of the water outlet pipe 4 is connected to one end of the two water pipes via a tee, and the other ends of the two water pipes are connected to the water outlets of the two fixed columns 1, respectively. Elbows are provided at the corners of the water pipes. The drains of the two fixed columns 1 are both connected to the center pipe 3. Valves are provided on the water inlet pipe 2, the water outlet pipe 4, and the center pipe 3. The outlet of the water outlet pipe 4 is connected to the water inlet of the water inlet pipe 2 via a peristaltic pump.

[0046] Four actual groundwater samples were collected from different groundwater sampling sites, each with a volume of 1000 mL. The water quality is shown in Table 1.

[0047] Table 1 The activated carbon carrier loaded with the composite bacterial agent was placed in a beaker containing the groundwater sample to treat nitrates. After 12 hours of treatment, the results, shown in Table 2, showed that the composite bacterial agent removed 90% of the nitrate and 99.5% of TBBPA from the groundwater.

[0048] Table 2 Example 2: Removal effect of different combinations of bacterial agents on nitrate nitrogen and tetrabromobisphenol A 1. Preparation of single strain inoculant: Korean Pseudomonas Y5-11, Pitt's Acinetobacter LYS-5, Pseudomonasextremaustralis Y39-6 and psychrophilic bacteria F5-6 were activated respectively; the activated Korean Pseudomonas Y5-11, Pitt Acinetobacter LYS-5, Pseudomonasextremaustralis Y39-6 and psychrophilic bacteria F5-6 were fermented and cultured respectively to obtain four single strain inoculants, namely, the concentration of each single strain inoculant was 10 12 pcs / L; 2. Preparation of compound bacterial agent of two strains: Korean Pseudomonas Y5-11, Pitt's Acinetobacter LYS-5, Pseudomonasextremaustralis Y39-6 and psychrophilic bacteria F5-6 were activated respectively; the activated Korean Pseudomonas Y5-11, Pitt Acinetobacter LYS-5, Pseudomonasextremaustralis Y39-6 and psychrophilic bacteria F5-6 were fermented and cultured separately, and the number of bacteria in the fermentation broth of each bacteria was 10 12 pcs / L; The fermentation broth of Acinetobacter pietro LYS-5 and the fermentation broth of Pseudomonas aeruginosa Y5-11 were mixed at a bacterial ratio of 2:1 to prepare a Y5-11+LYS-5 composite bacterial agent; the fermentation broth of Acinetobacter pietro LYS-5 and Pseudomonasextremaustralis The Y39-6 fermentation broth was mixed at a bacterial count ratio of 2:2 to prepare a Y39-6+LYS-5 composite bacterial agent; the Acinetobacter pitei LYS-5 fermentation broth and the psychrophilic bacteria F5-6 fermentation broth were mixed at a bacterial count ratio of 2:1 to prepare an F5-6+LYS-5 composite bacterial agent; 3. Preparation of three bacterial strain compound agents: Korean Pseudomonas Y5-11, Pitt's Acinetobacter LYS-5, Pseudomonasextremaustralis Y39-6 and psychrophilic bacteria F5-6 were activated respectively; the activated Korean Pseudomonas Y5-11, Pitt Acinetobacter LYS-5, Pseudomonasextremaustralis Y39-6 and psychrophilic bacteria F5-6 were fermented and cultured separately, and the number of bacteria in the fermentation broth of each bacteria was 10 12 pcs / L; Acinetobacter pitei LYS-5 fermentation broth, Pseudomonasextremaustralis The fermentation liquid of Y39-6 and the fermentation liquid of Korean Pseudomonas Y5-11 were mixed, and the bacterial count ratio of the fermentation liquid was 2:2:1 to prepare the LYS-5+Y5-11+Y39-6 composite bacterial agent; the fermentation liquid of Acinetobacter Pitt LYS-5, the fermentation liquid of Korean Pseudomonas Y5-11 and the fermentation liquid of psychrophilic bacteria F5-6 were mixed, and the bacterial count ratio of the fermentation liquid was 2:1:1 to prepare the LYS-5+Y5-11+F5-6 composite bacterial agent; the fermentation liquid of Acinetobacter Pitt LYS-5, Pseudomonasextremaustralis The fermentation broth of Y39-6 and the fermentation broth of psychrophilic bacteria F5-6 were mixed at a bacterial count ratio of 2:2:1 to prepare a LYS-5+Y39-6+F5-6 composite bacterial agent; 4. Preparation of compound bacterial agent with four strains: Korean Pseudomonas Y5-11, Pitt's Acinetobacter LYS-5, Pseudomonasextremaustralis Y39-6 and psychrophilic bacteria F5-6 were activated respectively; the activated Korean Pseudomonas Y5-11, Pitt Acinetobacter LYS-5, Pseudomonasextremaustralis Y39-6 and psychrophilic bacteria F5-6 were fermented and cultured separately, and the number of bacteria in the fermentation broth of each bacteria was 10 12 / L; Korean Pseudomonas Y5-11 fermentation broth, Pitt Acinetobacter LYS-5 fermentation broth, Pseudomonasextremaustralis The fermentation broth of Y39-6 and the fermentation broth of psychrophilic bacteria F5-6 were mixed, and the bacterial count ratio of the fermentation broth was 1:2:2:1 to prepare a composite bacterial agent of four strains.

[0049] In order to explore the removal effects of four different bacteria in different combinations on nitrate nitrogen and tetrabromobisphenol A, the bacterial agents of different combinations prepared in this example were fixed on an activated carbon carrier.

[0050] Wastewater to be treated with initial nitrate nitrogen and tetrabromobisphenol A concentrations of 50 mg / L and 1 mg / L was selected. The wastewater was treated using activated carbon carriers immobilized with different bacterial agents for 12 hours. The residual concentrations of nitrate nitrogen and tetrabromobisphenol A were tested. The test results are shown in Tables 3 to 5.

[0051] Table 3 Sewage treatment effect of single strain Table 4 Sewage treatment effect of two bacterial strain compound agents Table 5 Sewage treatment effects of three-strain compound bacterial agents and four-strain compound bacterial agents After the four strains were compounded, NO3 - The removal effect of nitrate and TBBPA was significantly better than that of other combinations. The strains promoted each other and synergistically improved the removal effect of nitrate and TBBPA.

[0052] Korean Pseudomonas ( Pseudomonaskoreensis ) Y5-11 has the function of aerobic facultative autotrophic denitrification; Acinetobacter pituitii ( Acinetobacterpittii LYS-5 has the ability to degrade TBBPA; Pseudomonasextremaustralis Y39-6 can be used to remove nitrates; psychrophilic bacteria ( Psychrobactercryohalolentis ) F5-6 has psychrophilic characteristics and the ability to process nitrate under low temperature conditions. Pseudomonasextremaustralis Y39-6 and psychrophilic bacteria ( Psychrobactercryohalolentis ) F5-6 can bind the other strains and protect the environment. At the same time, the intermediate products produced by LYS-5 degrading TBBPA and the extracellular secretions produced by F5-6 can provide carbon sources for Y39-6 and Y5-11, removing nitrates in the water. Y39-6 can use Fe 2+ and Mn 2+ , acting as an electron donor to enhance nitrate degradation. Microbial agents have stronger metabolic complementarity and are more effective in remediating complex pollution.

Claims

1. A composite bacterial agent for treating groundwater, characterized in that: The composite bacterial agent includes Korean Pseudomonas ( Pseudomonaskoreensis )Y5-11, Acinetobacter pituitus ( Acinetobacterpittii )LYS-5, Pseudomonas extremaustralis Y39-6 and psychrophilic bacteria ( Psychrobactercryohalolentis )F5-6; The Acinetobacter pituitus ( Acinetobacter pittii) LYS-5 is deposited in the General Microbiology Center of the China Culture Collection Administration, the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, the deposit number is CGMCC No. 30399, and the deposit date is April 22, 2024.

2. The composite bacterial agent for treating groundwater according to claim 1, characterized in that: The Korean Pseudomonas ( Pseudomonaskoreensis )Y5-11, Acinetobacter pituitus ( Acinetobacterpittii )LYS-5, Pseudomonas extremaustralis Y39-6 and psychrophilic bacteria ( Psychrobactercryohalolentis ) The bacterial count ratio of F5-6 is (1~3): (2~4): (0.5~2): (1~3).

3. The method for preparing the composite bacterial agent for treating groundwater according to claim 1, wherein: The method comprises the following steps:

1. Korean Pseudomonas Pseudomonaskoreensis )Y5-11, Acinetobacter pituitus ( Acinetobacterpittii )LYS-5, Pseudomonas extremaustralis Y39-6 and psychrophilic bacteria ( Psychrobactercryohalolentis ) F5-6 were activated respectively; 2. Activate Korean Pseudomonas Y5-11, Acinetobacter Pitt LYS-5, Pseudomonas extremaustralis Y39-6 and psychrophilic bacteria F5-6 were fermented and cultured separately, and the number of bacteria in the fermentation broth of each bacteria was 10 12 pcs / L; 3. Korean Pseudomonas Y5-11 fermentation broth, Acinetobacter Pitt LYS-5 fermentation broth, Pseudomonas extremaustralis The Y39-6 fermentation broth and the psychrophilic bacteria F5-6 fermentation broth were mixed to prepare a composite bacterial agent.

4. The method for preparing the composite bacterial agent for treating groundwater according to claim 3, characterized in that: The activation method of Korean Pseudomonas Y5-11 in step 1 is as follows: Korean Pseudomonas Y5-11 is inoculated into beef extract peptone solid culture medium and activated at 6-10° C. for 24-72 hours.

5. The method for preparing the composite bacterial agent for treating groundwater according to claim 3, characterized in that: The activation method of Acinetobacter pitei LYS-5 in step 1 is: inoculating Acinetobacter pitei LYS-5 into beef extract peptone solid culture medium and activating it at 26-36° C. for 24-72 hours.

6. The method for preparing the composite bacterial agent for treating groundwater according to claim 3, characterized in that: Step 1 Pseudomonas extremaustralis The activation method of Y39-6 is: Pseudomonas extremaustralis Y39-6 was inoculated into beef extract peptone solid medium and activated at 26-30°C for 24-72h.

7. The method for preparing the composite bacterial agent for treating groundwater according to claim 3, characterized in that: The activation method of psychrophilic bacteria F5-6 in step 1 is as follows: inoculating psychrophilic bacteria F5-6 into beef extract peptone solid culture medium and activating at 6-10° C. for 24-72 hours.

8. The method for preparing the composite bacterial agent for treating groundwater according to claim 3, characterized in that: In step 3, the fermentation broth of Korean Pseudomonas Y5-11 and Acinetobacter Pitt LYS-5, Pseudomonas extremaustralis The bacterial count ratio of Y39-6 fermentation broth and psychrophilic bacteria F5-6 fermentation broth was (1~3):(2~4):(0.5~2):(1~3).

9. The method for preparing the composite bacterial agent for treating groundwater according to claim 3, characterized in that: In step 2, the fermentation temperature of Korean Pseudomonas Y5-11 and psychrophilic bacteria F5-6 is 6~10℃, and the fermentation temperature of Acinetobacter Pitt LYS-5 and Pseudomonas extremaustralis The fermentation temperature of Y39-6 is 26~36℃, and the culture medium used for fermentation is beef extract peptone liquid culture medium.

10. The composite bacterial agent according to claim 1 is used for treating groundwater contaminated by nitrate and TBBPA.

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