CN111792719A - Oxidation method for removing antibiotic resistance genes and resistant bacteria in sewage - Google Patents

Oxidation method for removing antibiotic resistance genes and resistant bacteria in sewage Download PDF

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Publication number
CN111792719A
CN111792719A CN201910694816.1A CN201910694816A CN111792719A CN 111792719 A CN111792719 A CN 111792719A CN 201910694816 A CN201910694816 A CN 201910694816A CN 111792719 A CN111792719 A CN 111792719A
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China
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sewage
antibiotic resistance
resistant bacteria
resistance genes
oxidation method
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CN201910694816.1A
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Inventor
曹广丽
周春爽
武继文
刘冰峰
武秀坤
王琪
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Harbin Institute of Technology
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Harbin Institute of Technology
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    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F1/00Treatment of water, waste water, or sewage
    • C02F1/72Treatment of water, waste water, or sewage by oxidation
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F1/00Treatment of water, waste water, or sewage
    • C02F1/30Treatment of water, waste water, or sewage by irradiation
    • C02F1/32Treatment of water, waste water, or sewage by irradiation with ultraviolet light
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00Nature of the contaminant
    • C02F2101/30Organic compounds

Abstract

The invention discloses an oxidation method for removing antibiotic resistance genes and resistant bacteria in sewage, relating to the technical field of sewage advanced treatment, and the technical scheme is as follows: adding oxidant into sewage, and ultraviolet irradiating. The invention adopts a novel advanced oxidation process, utilizes UV to activate peroxymonosulfate/peroxydisulfate to generate sulfate radical (SO)4 ·–) And a hydroxyl radical (C·OH), and then remove ARGs and ARB by free radicals with strong oxidizing ability and UV damage to bacteria. The advanced oxidation process is environment-friendly, has strong oxidation capacity, does not generate intermediate pollutants, and has better removal effect on ARB and ARGs.

Description

Oxidation method for removing antibiotic resistance genes and resistant bacteria in sewage
Technical Field
The invention relates to the technical field of sewage advanced treatment, in particular to a method for removing antibiotic resistance genes and resistant bacteria in sewage.
Background
As an important link for guaranteeing safe discharge of sewage, the advanced treatment process of sewage has important significance in researching the influence of different advanced treatment processes on pollutant removal. In the advanced treatment process, the disinfection technology of chlorination, UV and ozone has the capacity of damaging bacterial cells and DNA in the cells, has the potential of killing Antibiotic Resistant Bacteria (ARB) and destroying Antibiotic Resistant Genes (ARGs), but the required dosage is higher than that of the conventional sewage treatment plant, and because the ARB has stronger tolerance capacity, the dosage used by the existing disinfection method can not effectively remove the ARB and the ARGs, so that the resistant bacteria and the resistant genes enter downstream rivers and lakes along with effluent, and the diffusion of the ARGs is promoted. And the existing disinfection technology is easy to generate harmful disinfection byproducts and other toxic intermediate products, and further pollutes the environment. The existing reports about the influence of ultraviolet, chlorination and the like on the ARGs and the ARB in the disinfection technology are focused on monitoring a sewage treatment plant or treating a water sample prepared in a laboratory, the influence of specific disinfection process operation parameters on actual sewage is not systematically researched, and the existing treatment method and research are not representative and do not obtain better effect.
Disclosure of Invention
In order to solve the above problems, the present invention provides an oxidation method for removing antibiotic resistance genes and resistant bacteria in sewage, which specifically comprises: adding oxidant into sewage, and ultraviolet irradiating.
The sewage is secondary effluent of a sewage treatment plant.
Preferably, the oxidizing agent is capable of being activated by ultraviolet rays to generate radicals having a strong oxidizing power.
Further preferably, the oxidant Peroxodisulfate (PDS) is potassium peroxodisulfate.
Further preferably, the oxidant per-sulphate (PMS) is potassium hydrogen peroxymonosulphate.
Preferably, the PDS is used as an oxidizing agent, and the adding amount of the PDS is 0.1-2 mmol/L of the final concentration of the PDS in the sewage.
Preferably, the PMS is used as an oxidant, and the addition amount of the PMS is 0.1-2 mmol/L of the final concentration of the PMS in the sewage.
The ultraviolet irradiation treatment is carried out by using an ultraviolet lamp with the wavelength of 254 nm.
The ultraviolet irradiation treatment is carried out, and the UV intensity is 16-36 mu w/cm2
And the irradiation treatment lasts for 1-60 min.
Preferably, all of the above operations are performed under sterile conditions.
Advantageous effects
The invention adopts a novel advanced oxidation process, and utilizes UV activation to generate sulfate radical (SO)4 ·–) And a hydroxyl radical (C·OH), ARB and ARGs are removed by strong oxidizing power of free radicals and damage of UV to bacteria. The advanced oxidation process is environment-friendly, has strong oxidation capacity, does not generate intermediate pollutants, and has good removal effect on ARB and ARGs.
Drawings
FIG. 1 shows the inactivation of macrolide antibiotic-resistant bacteria (MRB) at different times (1-60 min) and under different treatment conditions (PDS, UV, UV/PDS). It can be seen from the figure that the UV/PDS test group achieves the best effect on MBR removal at 10 minutes of treatment, where the removal rate is as high as 96.6%, while the UV and PDS control groups have MRB removal rates of 86.5% and 65.1% at 10 minutes, respectively, which are much lower than the MRB removal results of the UV/PDS test group, indicating that the new method is far superior to the conventional UV irradiation method for ARB removal.
FIG. 2 shows the removal of ARGs after ten minutes of treatment of the secondary effluent under different conditions (PDS, UV, UV/PDS). From the figure, it can be seen that UV/PDS has a better effect on removing erm B, sul 1, qnr S, tet O resistance genes and intI 1 integron comparison ratio (PDS, UV), wherein the removal rate of erm B is as high as 83.4%, and is 14.9% higher than UV, which indicates that the method is far superior to the traditional UV irradiation method in removing ARGs.
Detailed Description
Media used in examples and comparative examples:
the LB solid culture medium contains peptone with the concentration of 5-10 g/L, yeast powder with the concentration of 5-10 g/L, sodium chloride with the concentration of 5-10 g/L and agar with the concentration of 10-20 g/L.
Sulfonamide antibiotic resistance medium: sulfadiazine is added on the basis of a common LB solid culture medium, so that the final concentration of the sulfadiazine is 400-700 mug/mL.
The macrolide antibiotic resistance culture medium is characterized in that erythromycin is added on the basis of a common LB solid culture medium, so that the final concentration of the erythromycin is 5-10 mu g/mL.
Tetracycline antibiotic-resistant medium: adding tetracycline on the basis of a common LB solid culture medium to ensure that the final concentration of the tetracycline is 10-20 mu g/mL.
Quinolone antibiotic resistance medium: adding ciprofloxacin on the basis of a common LB solid culture medium to ensure that the final concentration of the ciprofloxacin is 20-40 mug/mL.
Example 1
Under aseptic condition, placing secondary effluent of sewage treatment plant into a beaker, adding PDS to make final concentration 1mmol/L, mixing well, and UV intensity 21 μ w/cm2Under ultraviolet lamp for 10 min.
Example 2
Under aseptic condition, placing secondary effluent of sewage treatment plant into a beaker, adding PDS to make final concentration of 0.1mmol/L, mixing well, and UV intensity of 16 μ w/cm2Under ultraviolet lamp for 30 min.
Example 3
Under aseptic condition, placing secondary effluent of sewage treatment plant into a beaker, adding PDS to make final concentration of 2mmol/L, mixing well, and UV intensity of 36 μ w/cm2Under ultraviolet lamp for 60 min.
Example 4
Under the aseptic condition, taking secondary effluent of a sewage treatment plant, putting the secondary effluent into a beaker, adding PMS (permanent magnet sludge) to ensure that the final concentration of the secondary effluent is 1mmolL, mixing well, UV intensity of 21 μ w/cm2Under ultraviolet lamp for 10 min.
Example 5
Under aseptic condition, placing secondary effluent of sewage treatment plant into a beaker, adding PMS to make final concentration of 0.1mmol/L, mixing well, and UV intensity of 16 μ w/cm2Under ultraviolet lamp for 30 min.
Example 6
Under aseptic condition, placing secondary effluent of sewage treatment plant into a beaker, adding PMS to make final concentration of 2mmol/L, mixing well, and UV intensity of 36 μ w/cm2Under ultraviolet lamp for 60 min.
Comparative example 1
The difference from example 1 is that: the ultraviolet irradiation treatment is not carried out, and specifically comprises the following steps:
under aseptic conditions, taking secondary effluent of a sewage treatment plant, putting the secondary effluent into a beaker, adding PDS to enable the final concentration to be 1mmol/L, uniformly mixing, and standing for 10 min.
Comparative example 2
The difference from example 1 is that: without PDS, specifically:
under aseptic conditions, the secondary effluent from a sewage treatment plant is taken into a beaker and treated with UV light intensity of 21 μ w/cm2Under ultraviolet lamp for 10 min.
Comparative example 3
The difference from example 4 is that: adding PMS, specifically:
under aseptic conditions, the secondary effluent from a sewage treatment plant is taken into a beaker and treated with UV light intensity of 21 μ w/cm2Under ultraviolet lamp for 10 min.
The water samples used in the above examples and comparative examples are all from the same batch of secondary effluent.
And (4) checking a processing result:
preferred validation media:
the LB solid medium contains peptone with a concentration of 10g/L, yeast powder with a concentration of 5g/L, sodium chloride with a concentration of 10g/L, and agar with a concentration of 20g/L
Sulfonamide antibiotic resistance medium: sulfadiazine is added on the basis of common LB solid culture medium to make the final concentration of the sulfadiazine be 512 mug/mL.
Macrolide antibiotic resistance medium erythromycin is added on the basis of a common LB solid medium to make the final concentration be 8 mug/mL.
Tetracycline antibiotic-resistant medium: tetracycline was added to a final concentration of 16. mu.g/mL based on the normal LB solid medium.
Quinolone antibiotic resistance medium: ciprofloxacin was added to give a final concentration of 32. mu.g/mL based on the ordinary LB solid medium.
The water samples are obtained from the secondary effluent after the treatment of the examples and the comparative examples (the raw material secondary effluent is the same batch) and the untreated secondary effluent of the same batch. Respectively coating a water sample on a common LB solid culture medium and a resistance culture medium containing various antibiotics such as sulfonamides, macrolides, tetracyclines, quinolones and the like, culturing for 24 hours at 37 ℃, and calculating the concentration (CFU/mL) of resistant bacteria in the sample under different treatment conditions to obtain the removal result of the resistant bacteria.
Filtering a water sample by using a filter membrane of 0.22 mu m, extracting DNA of bacteria on the filter membrane by using a DNA extraction kit, then carrying out fluorescence quantitative qPCR on the extracted DNA to detect the relative abundance and copy number of various ARGs (resistance genes such as macrolides, aminoglycosides, tetracyclines, quinolones, sulfonamides, beta-lactones and the like) and various movable elements (MGEs: various movable elements such as integron, transposon and the like) and comparing the relative abundance and copy number of each milliliter with the original secondary effluent to obtain the removal effect on the ARGs and the MGEs.
The results of removing MRB from resistant bacteria in example 1, comparative example 1 and comparative example 2 are shown in FIG. 1, and the results of removing ARGs are shown in FIG. 2: although the results obtained in examples other than example 1 are not shown, the results are not much different from those of example 1 and are approximately: the results of the example 1 and the example 4 for removing ARB and ARGs are the closest, and the treatment effect is the best; example 2 and example 5 are the worst effect, because the PDS dosage is too small, the generated free radical content is low, and the ARB is inactivated and the ARGs are removed only by the damage of ultraviolet to bacteria and genes, so the effect is lower than that of other examples, but the effect is also greatly improved compared with that of comparative examples; the reason why the treatment effect of example 3 is far superior to that of comparative example compared to example 1, and the treatment effect of example 6 is not much different from that of example 1, is that better sterilization effect (MRB removal rate as high as 96.6%) and ARGs removal effect (erm B removal rate as high as 83.4%) are achieved under the conditions of example 1, and thus the inactivation of ARB and the ARGs removal do not change much after the PDS concentration, the uv intensity and the treatment time are increased. By comparing the treated secondary effluent with the treated secondary effluent, the method provided by the invention has better removal effect on ARB and ARGs. Moreover, the method provided by the invention not only simply combines the ultraviolet treatment with the action of the oxidant, but also combines the physical damage of ARB and the chemical oxidation of free radicals by UV, so that the final experimental result of the UV/PDS on the removal of ARB and ARGs is far better than that of a control group and is better than that of the traditional ultraviolet method.

Claims (10)

1. An oxidation method for removing antibiotic resistance genes and resistant bacteria in sewage, which is characterized in that: the method comprises the following steps: adding oxidant into sewage, and ultraviolet irradiating.
2. The oxidation method for removing antibiotic resistance genes and resistant bacteria in sewage according to claim 1, wherein: the sewage is secondary effluent of a sewage treatment plant.
3. The oxidation method for removing antibiotic resistance genes and resistant bacteria in sewage according to claim 1, wherein: the oxidant can be activated by ultraviolet rays to generate free radicals with strong oxidizing ability.
4. The oxidation method for removing antibiotic resistance genes and resistant bacteria in sewage according to claim 3, wherein: the oxidant Peroxodisulfate (PDS) is potassium peroxodisulfate.
5. The oxidation method for removing antibiotic resistance genes and resistant bacteria in sewage according to claim 3, wherein: the oxidant per-sulfate (PMS) is potassium hydrogen peroxymonosulfate.
6. The oxidation method for removing antibiotic resistance genes and resistant bacteria in sewage according to claim 4, wherein: and the PDS is used as an oxidant, and the addition amount of the PDS is that the final concentration of the PDS in the sewage is 0.1-2 mmol/L.
7. The oxidation method for removing antibiotic resistance genes and resistant bacteria in sewage according to claim 5, wherein: the PMS is used as an oxidant, and the addition amount of the PMS is 0.1-2 mmol/L of the final concentration of the PMS in the sewage.
8. The oxidation method for removing antibiotic resistance genes and resistant bacteria in sewage according to claim 1, wherein: the ultraviolet irradiation treatment is carried out by using an ultraviolet lamp with the wavelength of 254 nm.
9. The oxidation method for removing antibiotic resistance genes and resistant bacteria in wastewater according to claim 8, wherein: the ultraviolet irradiation treatment is carried out, and the UV intensity is 16-36 uw/cm2
10. The oxidation method for removing antibiotic resistance genes and resistant bacteria in wastewater according to claim 9, wherein: and the irradiation treatment lasts for 1-60 min.
CN201910694816.1A 2019-07-30 2019-07-30 Oxidation method for removing antibiotic resistance genes and resistant bacteria in sewage Pending CN111792719A (en)

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CN112624455A (en) * 2020-12-18 2021-04-09 华东理工大学 Method for disinfecting gram-negative superbacteria with antibiotic resistance in water
CN113896276A (en) * 2021-09-29 2022-01-07 同济大学 Method for removing antibiotics and resistance genes in sewage by using ultraviolet/peroxyacetic acid
CN114477459A (en) * 2022-01-17 2022-05-13 哈尔滨工业大学 Method for reducing antibiotic resistance genes in aquaculture wastewater
CN115321637A (en) * 2022-08-24 2022-11-11 北京工业大学 Method for removing antibiotic resistance genes by vacuum ultraviolet/potassium hydrogen persulfate coupled advanced oxidation and regulation and control system
WO2024051046A1 (en) * 2022-09-09 2024-03-14 长江水利委员会长江科学院 Method for estimating abundances and distribution characteristics of antibiotic resistance genes in lake/reservoir surface sediments

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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112624455A (en) * 2020-12-18 2021-04-09 华东理工大学 Method for disinfecting gram-negative superbacteria with antibiotic resistance in water
CN113896276A (en) * 2021-09-29 2022-01-07 同济大学 Method for removing antibiotics and resistance genes in sewage by using ultraviolet/peroxyacetic acid
CN114477459A (en) * 2022-01-17 2022-05-13 哈尔滨工业大学 Method for reducing antibiotic resistance genes in aquaculture wastewater
CN114477459B (en) * 2022-01-17 2023-02-07 哈尔滨工业大学 Method for reducing antibiotic resistance genes in aquaculture wastewater
CN115321637A (en) * 2022-08-24 2022-11-11 北京工业大学 Method for removing antibiotic resistance genes by vacuum ultraviolet/potassium hydrogen persulfate coupled advanced oxidation and regulation and control system
CN115321637B (en) * 2022-08-24 2023-09-22 北京工业大学 Method and regulation and control system for removing antibiotic resistance gene by coupling vacuum ultraviolet/potassium hydrogen persulfate with advanced oxidation
WO2024051046A1 (en) * 2022-09-09 2024-03-14 长江水利委员会长江科学院 Method for estimating abundances and distribution characteristics of antibiotic resistance genes in lake/reservoir surface sediments

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