An extracellular drug-resistant gene removal method for poultry manure based on waste plastic microporous carbon / nuclease active bacteria composite material

Abstract

This invention discloses a method for removing extracellular drug resistance genes (eARGs) from livestock and poultry manure based on a composite material of microporous carbon from waste plastics and active nuclease bacteria, belonging to the field of environmental pollution control. This invention uses recycled waste plastics as raw materials to prepare microporous carbon materials, enhances their adsorption performance through physical activation and chemical modification, and co-forms them with strains of high nuclease activity to form a composite material for livestock and poultry manure treatment. The composite material achieves highly efficient adsorption and degradation of eARGs, significantly improving the eARG removal rate. This invention features low energy consumption, widely available materials, good ecological safety, and promising prospects for widespread application, especially suitable for resistance risk management in agricultural non-point source pollution control and resource utilization processes.

Description

Technical Field

[0001] This invention belongs to the field of environmental pollution control, specifically relating to a method for removing extracellular drug resistance genes from poultry and livestock manure based on a composite material of waste plastic microporous carbon / nuclease active bacteria. Background Technology

[0002] With the widespread use of antibiotics in animal husbandry, livestock manure has become a significant environmental source of extracellular antibiotic resistance genes (eARGs), and their spread risk is increasingly concerning. eARGs can spread widely in the environment through horizontal gene transfer, increasing pathogen resistance and posing a potential public health threat. Currently, there is a lack of efficient, safe, and environmentally friendly treatment technologies for eARG pollution in the environment. Traditional chlorine disinfection, ozone, or ultraviolet light technologies are not very efficient at removing eARGs and may cause secondary pollution. Therefore, there is an urgent need to develop an economical and environmentally friendly removal technology that can efficiently degrade eARGs. At the same time, the large-scale accumulation of waste plastics also poses a challenge to the ecological environment, and their resource utilization is of significant practical importance. Summary of the Invention

[0003] The purpose of this invention is to provide a method for removing extracellular drug resistance genes in poultry and livestock manure based on a composite material of waste plastic microporous carbon and nuclease-active bacteria. The aim is to construct a composite material based on the synergistic effect of waste plastic microporous carbon and nuclease-active bacteria for the efficient fixation and degradation of eARGs in poultry and livestock manure, thereby achieving the synergistic goal of pollution control and waste resource utilization.

[0004] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0005] A method for removing extracellular drug resistance genes from poultry and livestock manure based on a composite material of microporous carbon / nuclease-active bacteria from waste plastic includes the following steps:

[0006] (1) Preparation of microporous carbon: Waste plastics were pyrolyzed at 480–520℃ for 1.8–2.2 h under a nitrogen atmosphere to obtain a primary product. The primary product was mixed with activator FeCl3 at a mass ratio of 1:2.8–3.2 and activated at 650–750℃ for 60–70 min to obtain microporous carbon particles with a pore size of 1–3 nm, a particle size of 2–5 mm, and a specific surface area of ​​1100–1300 m². 2 / g, and the surface of the microporous carbon is modified with imine to make it positively charged;

[0007] The waste plastics mentioned are one or a mixture of more than one of polyethylene (PE), polypropylene (PP), and polyethylene terephthalate (PET);

[0008] The method for imine modification of microporous carbon surfaces is as follows:

[0009] 1) Surface grafting: Preparation of 4-formylphenyl diazonium salt and grafting onto carbon

[0010] ① Dissolve 0.80–1.2 g of 4-aminobenzaldehyde in 20 mL of 1 M water / EtOH mixed solvent under ice bath conditions to obtain an acidic solution containing 4-aminobenzaldehyde;

[0011] Add 1.0 g of well-dispersed microporous carbon to 50 mL of water / EtOH (the carbon was first ultrasonically dispersed for 10 min) to form a carbon suspension, and stir in an ice bath (0–5 °C);

[0012] ② In another flask, dissolve NaNO2 (equimolar with 4-aminobenzaldehyde) in a small amount of water (5–10 mL) and add it dropwise to an acidic solution containing 4-aminobenzaldehyde. At 0–5 °C, a diazonium salt is formed (the temperature is controlled at 0–5 °C during the dropwise addition process, and the reaction takes 10–20 min).

[0013] ③ The generated diazonium salt solution is added dropwise to the carbon suspension and stirred at 0–5℃ for 30 min, then heated to room temperature and stirred for 1–2 h. During this process, aryl radicals or diazonium compounds will graft onto the carbon surface;

[0014] ④ After the reaction is complete, centrifuge to separate the components, wash repeatedly with water and ethanol until the washing solution is colorless / free of impurities, and vacuum dry to obtain CHO-functionalized carbon (C-CHO);

[0015] 2) Imine (Schiff base) — Condensation of surface-CHO with DMAPA

[0016] ① Suspend 1.0 g of C-CHO in 40–50 mL of ethanol, add DMAPA (excess, 0.6–1.2 mL) and a small amount of acetic acid as a catalyst (0.1–0.3 mL);

[0017] ② Stirring at room temperature or under reflux for 4–12 h will condense to form a surface -C=N-(CH2)3-N(CH3)2 (i.e., the surface imine is connected to a tertiary amine at one end);

[0018] ③ After the reaction, wash away the free amine with a filter / centrifugation, wash with ethanol and water until there is no amine odor or o-benzaldehyde residue in the washing solution, and dry to obtain C-Schiff-N(CH3)2 (containing surface imine and terminal tertiary amine);

[0019] 3) Alkylation → Quaternization (to obtain a positive charge)

[0020] ① Place the material obtained in the previous step into dry anhydrous acetonitrile (20–40 mL), add excess methyl iodine (MeI), and stir at 40–60 °C for 12–24 h (protected from light). The reaction is: RN(CH3)2+MeI→RN^+(CH3)3I^- (surface quaternary ammonium).

[0021] ② After the reaction is complete, filter and wash with ethanol / water to remove free formioiodine and iodide residues, then wash with water until there is no free iodine in the washing solution (residual iodine is detected by trace amount with AgNO3), and finally vacuum dry to obtain surface modified carbon with quaternary ammonium cations (C-Imine-N^+Me3 I^-).

[0022] (2) Strain screening and fixation: After adaptive culture, strains with high nuclease activity were fixed on the surface of microporous carbon by spraying to form a microporous carbon / nuclease-active bacteria composite material.

[0023] The strain with high nuclease activity is one or a mixture of more than one of the genera *Pseudomonas*, *Bacillus*, *Streptomyces*, and *Xanthomonas*.

[0024] In the preparation of the composite material, each nuclease-active strain was adapted to culture in a modified selective medium, DNAagar, at 30°C. After 48 hours of culture, each strain was prepared into a concentration of 10... 7 ~10 9 A CFU / mL suspension was prepared and taken in equal volumes. Then, the suspension was sprayed evenly onto the surface of the microporous carbon at a ratio of 5 mL suspension to 1 mg microporous carbon under sterile conditions at 25 °C. The culture dish was then moistened at 30 °C and incubated for 48 h. The bacteria formed a biofilm on the surface of the microporous carbon, thus obtaining a microporous carbon / nuclease active bacteria composite material.

[0025] (3) Livestock manure wastewater treatment: Microporous carbon / nuclease active bacteria composite material is added to the livestock manure wastewater treatment reactor, and the operating parameters are set as follows: temperature 30-40℃, pH 6.5-7.5, hydraulic retention time (HRT) 48-72h to promote the adsorption, fixation and enzymatic degradation of eARGs;

[0026] The feed ratio of the microporous carbon / nuclease active bacteria composite material to poultry and livestock manure wastewater is 900-1100 mg: 5 L;

[0027] (4) Material regeneration and recycling: After the treatment of poultry and livestock manure wastewater, the composite material is recovered by solid-liquid separation. After washing with PBS buffer, it is regenerated by incubating at pH 5.0 for 30-40 minutes. The composite material can be recycled 5 times, and the removal rate of eARGs remains above 80%. The by-product liquid generated during the regeneration process can be further treated or utilized as a resource.

[0028] Compared with existing processing methods, the present invention has the following significant advantages:

[0029] (1) The microporous carbon carrier of the present invention is obtained by pyrolysis and FeCl3 activation of waste plastics. It has a high specific surface area and functional groups, and has excellent adsorption performance.

[0030] (2) The nuclease-active bacterial community originates from the soil system, has high nuclease expression capacity, can be artificially cultured, and utilizes the specific degradation capacity of the nuclease-active bacteria to achieve efficient removal of eARGs.

[0031] (3) By spraying and fixing the microbial community onto the surface of microporous carbon through biofilm formation, a stable microbial-carbon material system is formed. The composite material has strong stability, good regeneration performance and reusability, and is easy to promote in engineering and modular application.

[0032] (4) The reaction system is equipped with inlet and outlet, temperature control module and mechanical stirrer, which can be adapted to anaerobic or facultative anaerobic operation mode. It can intelligently collect key parameters such as pH, temperature and dissolved oxygen in real time, adjust reaction conditions and ensure processing efficiency.

[0033] (5) This invention provides an eARGs removal method based on the synergistic effect of microorganisms and solid waste materials, realizing the high-value resource utilization of waste plastics, preparing high-performance adsorption-bacterial integrated materials, which are suitable for the treatment of manure in large-scale poultry and livestock farms, and help to achieve the dual goals of green agriculture and manure reduction, and have broad application prospects. Attached Figure Description

[0034] Figure 1 This is a schematic diagram of the device;

[0035] 1- Livestock manure pretreatment system; 2- pH adjustment element (for adding dilute acid and alkali); 3- Livestock manure wastewater; 4- Flow regulating valve (for adjusting HRT); 5- Inlet; 6- Facultative / anaerobic reaction system; 7- Reaction system cover; 8- pH adjustment element (for adding dilute acid and alkali); 9- Intelligent monitoring module (pH, water temperature, dissolved oxygen); 10- Mechanical stirrer; 11- Solar temperature regulating layer (for adjusting water temperature); 12- Outlet; 13- Large particle composite material filter module; 14- Monitoring port; 15- Medium particle composite material filter module; 16- Monitoring port; 17- Small particle composite material filter module;

[0036] The pH adjustment element, water flow regulating valve, and solar temperature regulating layer constitute the control module, which respectively regulates pH, HRT, and water temperature; the filter modules in the same layer use snap-fit ​​connections for easy replacement and maintenance. Detailed Implementation

[0037] This invention incorporates a multi-stage composite material filtration module within the reaction apparatus. Each filtration module is filled with the waste plastic microporous carbon / nuclease-active bacteria composite material of this invention. The multi-stage filtration module consists of three stages, each filled with composite materials of different particle sizes to achieve gradient adsorption and degradation. Filter modules within the same stage are connected using a snap-fit ​​design for easy replacement and maintenance. The reaction system is equipped with inlet / outlet ports and a temperature control module to maintain bacterial activity and composite material efficiency. Livestock manure is uniformly pretreated and its pH adjusted before being pumped into the reaction apparatus. After treatment by the multi-stage composite material filtration module, the eARGs adsorption and nuclease degradation process is completed. The treated liquid is collected, and the eARGs content is measured. The nuclease-active bacteria are strains with high nuclease activity selected through domestication. Adjusting the pH to 6.5–7.5 is beneficial for maintaining the nuclease activity of the strains.

[0038] The Bacillus subtilis, P. fluorescens, and S. griseus of the genus Bacillus described in Examples 1-3, as well as the Bacillus licheniformis and B. pumilus, P. putida of the genus Pseudomonas, S. coelicolor of the genus Streptomyces, and X. campestris of the genus Xanthomonas described in Examples 4-6, were all purchased commercially.

[0039] Example 1

[0040] (1) Preparation of microporous carbon: Waste PE was pyrolyzed at 500℃ for 2 hours under a nitrogen atmosphere to obtain a primary product. The primary product was mixed with activator FeCl3 at a mass ratio of 1:3 and activated at 700℃ for 60 minutes to obtain microporous carbon particles. The surface of the microporous carbon was then modified with imine to make it positively charged. The specific method is as follows:

[0041] 1) Surface grafting: Preparation of 4-formylphenyl diazonium salt and grafting onto carbon

[0042] ① Under ice bath conditions, 1 g of 4-aminobenzaldehyde was dissolved in 20 mL of 1 M water / EtOH mixed solvent to obtain an acidic solution containing 4-aminobenzaldehyde;

[0043] Add 1.0 g of well-dispersed microporous carbon to 50 mL of water / EtOH (the carbon was first ultrasonically dispersed for 10 min) to form a carbon suspension, and stir in an ice bath (0 °C);

[0044] ② In another flask, dissolve NaNO2 (equimolar with 4-aminobenzaldehyde) in a small amount of water (5 mL), and add it dropwise to an acidic solution containing 4-aminobenzaldehyde. At 0°C, a diazonium salt is formed (the temperature is controlled at 0°C during the dropwise addition process, and the reaction takes 10 min).

[0045] ③ The generated diazonium salt solution is added dropwise to the carbon suspension, stirred at 0°C for 30 min, then heated to room temperature and stirred for 1.5 h. During this process, aryl radicals or diazonium compounds will graft onto the carbon surface;

[0046] ④ After the reaction is complete, centrifuge to separate the components, wash repeatedly with water and ethanol until the washing solution is colorless / free of impurities, and vacuum dry to obtain CHO-functionalized carbon (C-CHO);

[0047] 2) Imine (Schiff base) — Condensation of surface-CHO with DMAPA

[0048] ① Suspend 1.0g of C-CHO in 40mL of ethanol, add DMAPA (excess, 1.2mL) and a small amount of acetic acid as a catalyst (0.2mL);

[0049] ② Stirring at room temperature or under reflux for 8 hours will condense to form a surface -C=N-(CH2)3-N(CH3)2 (i.e., the surface imine is connected to a tertiary amine at one end);

[0050] ③ After the reaction, wash away the free amine with a filter / centrifugation, wash with ethanol and water until there is no amine odor or o-benzaldehyde residue in the washing solution, and dry to obtain C-Schiff-N(CH3)2 (containing surface imine and terminal tertiary amine);

[0051] 3) Alkylation → Quaternization (to obtain a positive charge)

[0052] ① Place the material obtained in the previous step into a dry anhydrous acetonitrile (20 mL), add excess methyl iodine (MeI), and stir at 50 °C for 18 h (protected from light). The reaction is: RN(CH3)2 + MeI → RN^+(CH3)3I^- (surface quaternary ammonium).

[0053] ② After the reaction is complete, filter and wash with ethanol / water to remove free formioiodine and iodide residues, then wash with water until there is no free iodine in the washing solution (residual iodine is detected by trace amount with AgNO3), and finally vacuum dry to obtain surface modified carbon with quaternary ammonium cations (C-Imine-N^+Me3 I^-).

[0054] (2) Strain screening and fixation: Nuclease-high expression strains (Bacillus subtilis, Pseudomonas fluorescens, and Streptomyces S. griseus) from the soil system were loaded onto the surface of a microporous carbon carrier to form a microporous carbon / nuclease-active bacteria composite material.

[0055] In the preparation of the composite material, each nuclease-active strain was adapted to culture in a modified selective medium, DNAagar, at 30°C. After 48 hours of culture, each strain was prepared into a concentration of 10... 8 A CFU / mL suspension was prepared and an equal volume (5 mL per strain) was taken. Subsequently, the suspension was sprayed evenly onto the surface of the microporous carbon at a ratio of 5 mL suspension to 1 mg microporous carbon under aseptic conditions at 25°C. Then, the culture dish was moistened at 30°C and incubated for 48 h. The bacteria formed a biofilm on the surface of the microporous carbon, thus obtaining a microporous carbon / nuclease-active bacteria composite material.

[0056] (3) Treatment of livestock manure wastewater: 400 mg of microporous carbon / nuclease active bacteria composite material was added to a treatment reactor containing 2 L of pig manure wastewater. The operating parameters were set as follows: temperature 37℃, pH 7.0~7.2, hydraulic retention time 48h, to promote the adsorption, fixation and enzymatic degradation of eARGs.

[0057] (4) Material regeneration and recycling: After the treatment of poultry and livestock manure wastewater is completed, the composite material is recovered by solid-liquid separation. After washing with PBS buffer, it is regenerated by incubation at pH 5.0 for 30 min.

[0058] As shown in Table 1-4, the eARGs removal rate reached 85% after treatment, and the material maintained an 81% removal rate even after three regenerations.

[0059] The composite material can be recycled up to 5 times, and the removal rate of eARGs remains above 80%. The by-product liquid generated during the regeneration process can be further treated or utilized as a resource.

[0060] Example 2

[0061] In this embodiment, the waste plastic used is waste PP, and the remaining steps are the same as in Embodiment 1.

[0062] As shown in Table 1-4, the removal rate of eARGs after treatment reached 89%, and the material maintained a removal rate of 84% even after three regenerations.

[0063] The composite material can be recycled up to 5 times, and the removal rate of eARGs remains above 80%. The by-product liquid generated during the regeneration process can be further treated or utilized as a resource.

[0064] Example 3

[0065] In this embodiment, the waste plastic used is waste PET, and the remaining steps are the same as in Embodiment 1.

[0066] As shown in Table 1-4, the eARGs removal rate reached 87% after treatment, and the material maintained an 83% removal rate even after three regenerations.

[0067] The composite material can be recycled up to 5 times, and the removal rate of eARGs remains above 80%. The by-product liquid generated during the regeneration process can be further treated or utilized as a resource.

[0068] Table 1. Removal rate of eARGs from pig manure wastewater by a single type of waste plastic microporous carbon / nuclease-activated bacteria composite material.

[0069]

[0070] Table 2. Removal rate of eARGs from pig manure wastewater by the first regeneration of a single type of waste plastic microporous carbon / nuclease-active bacteria composite material.

[0071]

[0072]

[0073] Table 3. Removal rate of eARGs from pig manure wastewater by the second regeneration of a single type of waste plastic microporous carbon / nuclease-active bacteria composite material.

[0074]

[0075] Table 4. Removal rate of eARGs from pig manure wastewater by the third regeneration of a single type of waste plastic microporous carbon / nuclease-active bacteria composite material.

[0076]

[0077] Example 4

[0078] Composite microporous carbon particles were prepared by pyrolyzing a mixture of waste PE, PP, and PET plastics and activating the mixture with FeCl3. Nuclease-expressing strains (Bacillus licheniformis and B. pumilus, Pseudomonas P. putida, Streptomyces S. coelicolor, and Xanthomonas X. campestris) screened from soil samples were loaded onto the material. 1000 mg of the composite microporous carbon loaded with these strains was added to a 5 L anaerobic reactor to treat chicken manure wastewater at 35℃ and a hydraulic retention time (HRT) of 72 hours. The eARG removal rate exceeded 90%, and the degradation efficiency of the composite material remained above 85% even after five uses.

[0079] (1) Preparation of microporous carbon: Under nitrogen atmosphere, waste PE, PP and PET are mixed in equal proportion and pyrolyzed at 500℃ for 2h to obtain the initial product. The initial product is mixed with activator FeCl3 at a mass ratio of 1:3 and activated at 700℃ for 60min to obtain microporous carbon particles. The surface of the microporous carbon is modified with imine to make it positively charged. The specific method is as follows.

[0080] 1) Surface grafting: Preparation of 4-formylphenyl diazonium salt and grafting onto carbon

[0081] ① Under ice bath conditions, 1 g of 4-aminobenzaldehyde was dissolved in 20 mL of 1 M water / EtOH mixed solvent to obtain an acidic solution containing 4-aminobenzaldehyde;

[0082] Add 1.0 g of well-dispersed microporous carbon to 50 mL of water / EtOH (the carbon was first ultrasonically dispersed for 10 min) to form a carbon suspension, and stir in an ice bath (0 °C);

[0083] ② In another flask, dissolve NaNO2 (equimolar with 4-aminobenzaldehyde) in a small amount of water (5 mL), and add it dropwise to an acidic solution containing 4-aminobenzaldehyde. At 0°C, a diazonium salt is formed (the temperature is controlled at 0°C during the dropwise addition process, and the reaction takes 10 min).

[0084] ③ The generated diazonium salt solution is added dropwise to the carbon suspension, stirred at 0°C for 30 min, then heated to room temperature and stirred for 1.5 h. During this process, aryl radicals or diazonium compounds will graft onto the carbon surface;

[0085] ④ After the reaction is complete, centrifuge to separate the components, wash repeatedly with water and ethanol until the washing solution is colorless / free of impurities, and vacuum dry to obtain CHO-functionalized carbon (C-CHO);

[0086] 2) Imine (Schiff base) — Condensation of surface-CHO with DMAPA

[0087] ① Suspend 1.0g of C-CHO in 40mL of ethanol, add DMAPA (excess, 1.2mL) and a small amount of acetic acid as a catalyst (0.2mL);

[0088] ② Stirring at room temperature or under reflux for 8 hours will condense to form a surface -C=N-(CH2)3-N(CH3)2 (i.e., the surface imine is connected to a tertiary amine at one end);

[0089] ③ After the reaction, wash away the free amine with a filter / centrifugation, wash with ethanol and water until there is no amine odor or o-benzaldehyde residue in the washing solution, and dry to obtain C-Schiff-N(CH3)2 (containing surface imine and terminal tertiary amine);

[0090] 3) Alkylation → Quaternization (to obtain a positive charge)

[0091] ① Place the material obtained in the previous step into a dry anhydrous acetonitrile (20 mL), add excess methyl iodine (MeI), and stir at 50 °C for 18 h (protected from light). The reaction is: RN(CH3)2 + MeI → RN^+(CH3)3I^- (surface quaternary ammonium).

[0092] ② After the reaction is complete, filter and wash with ethanol / water to remove free formioiodine and iodide residues, then wash with water until there is no free iodine in the washing solution (residual iodine is detected by trace amount with AgNO3), and finally vacuum dry to obtain surface modified carbon with quaternary ammonium cations (C-Imine-N^+Me3 I^-).

[0093] (2) Strains screening and fixation: The exonuclease expression strains (Bacillus licheniformis and B. pumilus, P. putida, S. coelicolor, and Xanthomonas) screened in soil samples were loaded onto the surface of a microporous carbon carrier to form a microporous carbon / nuclease active bacteria composite material;

[0094] During the preparation of the composite materials, each strain of nuclease-active bacteria underwent adaptive culture in a modified selective medium, DNAagar, at 30°C. After 48 hours of culture, each strain was prepared into a concentration of 10... 7A CFU / mL suspension was prepared and an equal volume (5 mL per strain) was taken. Then, the suspension was sprayed evenly onto the surface of the microporous carbon at a ratio of 5 mL suspension to 1 mg microporous carbon under sterile conditions at 25°C. The culture dish was then moistened at 30°C and incubated for 48 h. The bacteria formed a biofilm on the surface of the microporous carbon, thus obtaining a microporous carbon / nuclease active bacteria composite material.

[0095] (3) Treatment of poultry and livestock manure wastewater: 1000 mg of microporous carbon / nuclease active bacteria composite material was added to a treatment reactor containing 5 L of chicken manure wastewater. The operating parameters were set as follows: temperature 35℃, pH 7.0~7.2, hydraulic retention time 72h, to promote the adsorption, fixation and enzymatic degradation of eARGs.

[0096] (4) Material regeneration and recycling: After the treatment of poultry and livestock manure wastewater is completed, the composite material is recovered by solid-liquid separation. After washing with PBS buffer, it is regenerated by incubation at pH 5.0 for 30 min.

[0097] As shown in Table 5, the eARGs removal rate reached 92% after treatment, and the material maintained an 86% removal rate even after 4 regenerations.

[0098] The composite material can be recycled up to 5 times, and the removal rate of eARGs remains above 80%. The by-product liquid generated during the regeneration process can be further treated or utilized as a resource.

[0099] Table 5. Removal rate of eARGs from chicken manure wastewater by the mixed waste plastic microporous carbon / nuclease-active bacteria composite material

[0100]

Claims

1. A method for removing extracellular drug resistance genes from poultry and livestock manure based on a composite material of waste plastic microporous carbon / nuclease-active bacteria, characterized in that, Includes the following steps: (1) Preparation of microporous carbon: Under a nitrogen atmosphere, waste plastics are pyrolyzed at 480~520℃ for 1.8~2.2 h to obtain a primary product. The primary product is mixed with activator FeCl3 at a mass ratio of 1:2.8~3.2 and activated at 650~750℃ for 60~70 min to obtain microporous carbon particles. Then, the surface of the microporous carbon is modified with imine to make it positively charged. The waste plastics mentioned are one or a mixture of two or more of polyethylene, polypropylene, and polyethylene terephthalate; (2) Strain screening and fixation: After culturing strains with high nuclease activity, the strains were fixed on the surface of microporous carbon by spraying to form a microporous carbon / nuclease-active bacteria composite material. The strain with high nuclease activity is one or a mixture of two or more of the genera *Pseudomonas*, *Bacillus*, *Streptomyces*, and *Xanthomonas*. (3) Treatment of livestock and poultry manure wastewater: The microporous carbon / nuclease active bacteria composite material is added to the livestock and poultry manure wastewater treatment reactor, and the operating parameters are set as follows: temperature 30~40℃, pH 6.5~7.5, hydraulic retention time 48~72h; (4) Material regeneration and recycling: After the treatment of poultry and livestock manure wastewater, the composite material is recovered by solid-liquid separation. After washing with PBS buffer, it is regenerated by incubation at pH 5.0 for 30-40 min.

2. The method for removing extracellular drug resistance genes from poultry and livestock manure based on a composite material of waste plastic microporous carbon / nuclease-active bacteria according to claim 1, characterized in that, In step (1), the microporous carbon particles prepared have a pore size of 1~3 nm, a particle size of 2~5 mm, and a specific surface area of ​​1100~1300 m² / g.

3. The method for removing extracellular drug resistance genes from poultry and livestock manure based on a composite material of waste plastic microporous carbon / nuclease-active bacteria, as described in claim 1, is characterized in that... In step (3), the feeding ratio of the microporous carbon / nuclease active bacteria composite material to poultry and livestock manure wastewater is 900~1100mg:5L.

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