CN115010265B - Method for removing antibiotics and resistance genes in water body by using plants and attached biological films thereof - Google Patents
Method for removing antibiotics and resistance genes in water body by using plants and attached biological films thereof Download PDFInfo
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- CN115010265B CN115010265B CN202210769126.XA CN202210769126A CN115010265B CN 115010265 B CN115010265 B CN 115010265B CN 202210769126 A CN202210769126 A CN 202210769126A CN 115010265 B CN115010265 B CN 115010265B
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 84
- 239000003242 anti bacterial agent Substances 0.000 title claims abstract description 59
- 229940088710 antibiotic agent Drugs 0.000 title claims abstract description 59
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 43
- 238000000034 method Methods 0.000 title claims abstract description 24
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- 239000004098 Tetracycline Substances 0.000 claims description 17
- 235000019364 tetracycline Nutrition 0.000 claims description 17
- 150000003522 tetracyclines Chemical class 0.000 claims description 17
- 229960004099 azithromycin Drugs 0.000 claims description 15
- MQTOSJVFKKJCRP-BICOPXKESA-N azithromycin Chemical compound O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)N(C)C[C@H](C)C[C@@](C)(O)[C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 MQTOSJVFKKJCRP-BICOPXKESA-N 0.000 claims description 15
- 101100424890 Butyrivibrio fibrisolvens tetW gene Proteins 0.000 claims description 7
- 101100206300 Escherichia coli tetC gene Proteins 0.000 claims description 7
- 101100445523 Bacteroides fragilis ermF gene Proteins 0.000 claims description 6
- 101100119095 Enterococcus faecalis (strain ATCC 700802 / V583) ermB gene Proteins 0.000 claims description 6
- 241000195493 Cryptophyta Species 0.000 claims description 5
- -1 ermX Proteins 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 5
- 229940040944 tetracyclines Drugs 0.000 claims description 5
- 241000252229 Carassius auratus Species 0.000 claims description 3
- 229960004306 sulfadiazine Drugs 0.000 claims description 2
- SEEPANYCNGTZFQ-UHFFFAOYSA-N sulfadiazine Chemical compound C1=CC(N)=CC=C1S(=O)(=O)NC1=NC=CC=N1 SEEPANYCNGTZFQ-UHFFFAOYSA-N 0.000 claims description 2
- 239000006228 supernatant Substances 0.000 claims description 2
- 238000003912 environmental pollution Methods 0.000 abstract description 2
- 241000196324 Embryophyta Species 0.000 description 45
- 239000000243 solution Substances 0.000 description 14
- 229960002180 tetracycline Drugs 0.000 description 12
- 229930101283 tetracycline Natural products 0.000 description 12
- 230000003115 biocidal effect Effects 0.000 description 8
- 244000005700 microbiome Species 0.000 description 7
- 244000025254 Cannabis sativa Species 0.000 description 6
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- ZCCIPPOKBCJFDN-UHFFFAOYSA-N calcium nitrate Chemical compound [Ca+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O ZCCIPPOKBCJFDN-UHFFFAOYSA-N 0.000 description 6
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 6
- 239000012984 antibiotic solution Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 241000282414 Homo sapiens Species 0.000 description 4
- 239000002131 composite material Substances 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 4
- SFOKDWPZOYRZFF-UHFFFAOYSA-H 2,3-dihydroxybutanedioate;iron(3+) Chemical compound [Fe+3].[Fe+3].[O-]C(=O)C(O)C(O)C([O-])=O.[O-]C(=O)C(O)C(O)C([O-])=O.[O-]C(=O)C(O)C(O)C([O-])=O SFOKDWPZOYRZFF-UHFFFAOYSA-H 0.000 description 3
- 238000009825 accumulation Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000003344 environmental pollutant Substances 0.000 description 3
- 239000010410 layer Substances 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000003120 macrolide antibiotic agent Substances 0.000 description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 3
- 235000019341 magnesium sulphate Nutrition 0.000 description 3
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 3
- 235000019796 monopotassium phosphate Nutrition 0.000 description 3
- 231100000719 pollutant Toxicity 0.000 description 3
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 3
- 235000010333 potassium nitrate Nutrition 0.000 description 3
- 239000004323 potassium nitrate Substances 0.000 description 3
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 238000003753 real-time PCR Methods 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 2
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 2
- 238000011529 RT qPCR Methods 0.000 description 2
- 230000001186 cumulative effect Effects 0.000 description 2
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- 235000019253 formic acid Nutrition 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 2
- 229940041033 macrolides Drugs 0.000 description 2
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- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 108030001722 Tentoxilysin Proteins 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/32—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2305/00—Use of specific compounds during water treatment
- C02F2305/06—Nutrients for stimulating the growth of microorganisms
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W10/00—Technologies for wastewater treatment
- Y02W10/10—Biological treatment of water, waste water, or sewage
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Hydrology & Water Resources (AREA)
- Environmental & Geological Engineering (AREA)
- Water Supply & Treatment (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a method for removing antibiotics and resistance genes in a water body by using plants and attached biological films thereof, belonging to the field of environmental pollution restoration. The method comprises the following steps: (1) Placing the aquatic plants in a pretreated water body, adding a nutrient solution into the pretreated water body, and culturing for 10-30d to obtain the aquatic plants and a biological membrane system thereof; (2) Planting the aquatic plants and the biological film system thereof in the polluted water body so as to remove antibiotics and resistance genes in the water body. The method can remove antibiotics and resistance genes in the water body under the condition of not damaging a natural ecological system, the removal rate of the coexisting antibiotics is more than 90 percent, and the method is simple to operate, low in running cost and friendly to the ecology. Has important guiding significance for removing antibiotics and resistance genes and purifying water bodies.
Description
Technical Field
The invention relates to the field of environmental pollution restoration, in particular to a method for removing antibiotics and resistance genes in a water body by using plants and attached biological films thereof.
Background
Antibiotics are released into aquatic and terrestrial environments as an emerging contaminant and threaten the ecosystem. The wastewater from the chemical and pharmaceutical industries and industrial aquaculture is a major source of antibiotic contamination in the environment. With the widespread use of antibiotics, antibiotic resistance genes spread and are widely distributed in the natural ecosystem and pose a significant risk to the health of animals and individuals.
Recently, a variety of antibiotics such as tetracyclines, sulfonamides and macrolides have been frequently observed by students in various environments. Of these, tetracyclines and macrolides are the most important antibiotics for human and veterinary use, and the usage rate in the European region of 2016-2018 is relatively high. Azithromycin is one of the most commonly used macrolide drugs in human disease, whereas high levels of known and novel Azithromycin resistance genes are found in contaminated surface water and sediments (msr, mph, mef, erm). Tetracyclines can affect the growth of food crops, aquatic and terrestrial organisms, and ultimately affect human health through accumulation in the food chain. The widespread use of tetracyclines has led to the diffusion of their related antibiotic resistance genes (tetA, tetC, tetW and tetX) in aquatic environments. The constructed wetland is used as an ecological friendly and sustainable water treatment technology, and can compensate the negative influence on human beings and an ecological system.
However, artificial wetland treatment wastewater is mainly directed to traditional pollutants such as nitrogen, phosphorus, COD and the like, and few researches and designs are directed to antibiotics in polluted water bodies. The treatment technology disclosed in the prior art mostly adopts a film material to adsorb and remove single antibiotics, which not only increases the treatment cost and damages the ecological environment, but also can not meet the requirement of removing various antibiotics in the actual aquatic ecological system. Therefore, in order to solve the technical problems, the invention aims to study a method for removing different antibiotics and resistance genes in water by using plants and attached biological films thereof.
Disclosure of Invention
The invention aims to provide a method for removing antibiotics and resistance genes in a water body by using plants and attached biological films thereof, so as to solve the problems of the prior art.
In order to achieve the above object, the present invention provides the following solutions:
the invention provides a method for removing antibiotics and resistance genes in a water body by using plants and attached biological films thereof, which comprises the following steps:
(1) Placing the aquatic plants in a pretreated water body, adding a nutrient solution into the pretreated water body, and culturing for 10-30d to obtain the aquatic plants and a biological membrane system thereof;
(2) Planting the aquatic plants and the biological film system thereof in the polluted water body so as to remove antibiotics and resistance genes in the polluted water body.
Further, in step (1), the nutrient solution is Hoagland nutrient solution, the concentration of which is 10% w/v.
Further, the pretreatment water body is obtained by the following steps: filtering the water body polluted by the antibiotics, standing for 24-48h, and collecting supernatant to obtain the pretreated water body.
Further, in the step (1), the aquatic plants cultured for 10-30d are taken out from the pretreated water body, and the aquatic plants and the biological membrane system thereof are obtained.
Further, in the step (2), the density of the planting is 50-200g/m 2 。
Further, the aquatic plant is a submerged plant and/or a floating plant.
Further, the submerged plant comprises kusnezoff monkshood, black algae or goldfish algae; the floating plant comprises a large float.
Further, in step (2), the antibiotics include tetracycline, sulfadiazine, and azithromycin; the resistance genes include ermF, ermX, ermB, tetA, tetC and tetW.
The invention discloses the following technical effects:
the invention discloses a method for removing antibiotics and resistance genes in water by using plants and attached biological films thereof, which is characterized in that the plants are placed into a polluted water containing microorganisms, and nutrient solution is added, so that the plants can grow better, the biological films are formed on the surfaces of the plants faster, and the plants and the attached biological films thereof can remove the antibiotics and the resistance genes in the water under the condition of not damaging a natural ecological system, thereby having remarkable removal capability for single and composite antibiotics. Meanwhile, the method is simple to operate, low in running cost, friendly to the environment, easy to popularize and apply and has important guiding significance for removing antibiotics and resistance genes and purifying water bodies.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a flow chart of an embodiment of the method for removing various antibiotics and resistance genes in a water body by using plants and attached biological films according to the invention;
FIG. 2 shows the concentration and removal efficiency of two antibiotics in water when treating a body of water with multiple antibiotics and resistance genes using a kucao and its attached biofilm system;
FIG. 3 is a graph showing the cumulative amount of ku grass and its attached biofilm system to single and mixed antibiotic pollutants;
FIG. 4 shows the enrichment of resistance genes for two different antibiotics in the bitter grass biofilm and bitter grass plants.
Detailed Description
Various exemplary embodiments of the invention will now be described in detail, which should not be considered as limiting the invention, but rather as more detailed descriptions of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples of the present invention are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.
FIG. 1 is a flow chart of a method for removing various antibiotics and resistance genes in a water body by using plants and attached biological films thereof, which is mainly implemented by selecting aquatic plants, constructing an aquatic plant and attached biological film system capable of removing the antibiotics and the resistance genes by using a polluted water body, and planting the cultivated plants and attached biological films thereof into the polluted water body for treatment.
Example 1
(1) Preparing a composite polluted water body:
and collecting water body polluted by antibiotics, filtering by using a filter screen, removing most of silt, phytoplankton and the like, standing for 24 hours, and collecting upper-layer liquid to obtain the water body containing microorganisms and polluted.
(2) Preparation of plant-attached biofilm system:
pre-culturing submerged plant herba Sonchi Oleracei in the water body containing microorganism and polluted in step (1), adding 10% w/v Hoagland nutrient solution, and culturing for 14d to form stable biomembrane on plant surface. Taking out the cultured plant to obtain a plant-attached biological film system;
the microbiota is taken as an important organism in the water body, can absorb antibiotics, and then is gathered and attached on the surface layer of the submerged plant to form an attached biological film. The Hoagland nutrient solution with the concentration of 10% w/v is added into the polluted water body for culture, so that the growth of the biological film can be further promoted.
The Hoagland water formula is as follows: 0.821 g of calcium nitrate, 0.506 g of potassium nitrate, 0.136 g of monopotassium phosphate, 0.120 g of magnesium sulfate and 0.005 g of ferric tartrate.
(3) Removal of antibiotics and resistance genes from water body by plant-attached biofilm system
The plant-biofilm system prepared in the step (2) is carried out at the speed of 125g/m 2 Biomass was grown in water contaminated with antibiotics and resistance genes (wherein the antibiotics were azithromycin and tetracycline, and the resistance genes were mainly ermF, ermX, ermB, tetA, tetC, tetW), and the water and plants were analyzed for azithromycin and tetracycline concentrations after 14 days.
Example 2
(1) Preparing a composite polluted water body:
and collecting water body polluted by antibiotics, filtering by using a filter screen, removing most of silt, phytoplankton and the like, standing for 32h, and collecting upper-layer liquid to obtain the water body containing microorganisms and polluted.
(2) Preparation of plant-attached biofilm system:
the floating plants are pre-cultured in the water body containing microorganisms and polluted in the step (1), 10% w/v Hoagland nutrient solution is added, and the culture time is 30d, so that stable biological films are formed on the surfaces of the plants. Taking out the cultured plant to obtain a plant-attached biological film system;
the Hoagland water formula is as follows: 0.821 g of calcium nitrate, 0.506 g of potassium nitrate, 0.136 g of monopotassium phosphate, 0.120 g of magnesium sulfate and 0.005 g of ferric tartrate.
(3) Removal of antibiotics and resistance genes from water body by plant-attached biofilm system
The plant-biofilm system prepared in the step (2) is mixed with 50g/m 2 Is planted in water body polluted by antibiotics and resistance genes (wherein the antibiotics comprise azithromycin and tetracycline, and the resistance genesMainly comprises ermF, ermX, ermB, tetA, tetC and tetW, and the concentration of azithromycin and tetracycline in water and plants is analyzed after 14 d.
Example 3
(1) Preparing a composite polluted water body:
and collecting water body polluted by antibiotics, filtering by using a filter screen, removing most of silt, phytoplankton and the like, standing for 48 hours, and collecting upper-layer liquid to obtain the water body containing microorganisms and polluted.
(2) Preparation of plant-attached biofilm system:
pre-culturing submerged plant Goldfish algae in the water body containing microorganisms and polluted in the step (1), adding 10% w/v Hoagland nutrient solution, and culturing for 10d to form stable biological film on the plant surface. Taking out the cultured plant to obtain a plant-attached biological film system;
the Hoagland water formula is as follows: 0.821 g of calcium nitrate, 0.506 g of potassium nitrate, 0.136 g of monopotassium phosphate, 0.120 g of magnesium sulfate and 0.005 g of ferric tartrate.
(3) Removal of antibiotics and resistance genes from water body by plant-attached biofilm system
The plant-biofilm system prepared in the step (2) is carried out at a speed of 200g/m 2 Biomass was grown in water contaminated with antibiotics and resistance genes (where the antibiotics azithromycin and tetracycline, the resistance genes were mainly ermF, ermX, ermB, tetA, tetC, tetW) and the water and plants were analyzed for azithromycin and tetracycline concentrations after 14 d.
Effect verification
Azithromycin single antibiotic solution (concentration is 0.10, 0.50 and 1.00 mg/L), tetracycline single antibiotic solution (concentration is 0.10, 0.50 and 1.00 mg/L) and mixed antibiotic solution (total concentration of Azithromycin and tetracycline is 0.10, 0.50 and 1.00 mg/L) are respectively prepared as water samples polluted by antibiotics and resistance genes, the water samples with different concentrations are respectively treated according to the method of the embodiment 1, and the water samples and the concentration of the Azithromycin and the tetracycline in plants are analyzed after 14 d.
The sample is enriched by Solid Phase Extraction (SPE). The water sample seeped through the solid filling device at a flow rate of 5mL/min, and the filling device was then washed with 5mL deionized water. The eluate was collected, extracted 2 times with 4mL of methanol, and the solvent was evaporated in a stream of nitrogen. Finally, the extract was dissolved in 0.5mL of mobile phase a (0.1% formic acid in water). Azithromycin and tetracycline concentrations in water and plants were analyzed by liquid chromatography-mass spectrometry (LC-MS: H-Class UPLC-Xevo TQD, waters Corporation, USA). The column was Acquity UPLC BEH C (100 mm. Times.2.1 mm, particle size 1.7 μm, waters, USA). Sample injection amount: 20. Mu.L, the optimal separation technique was determined using gradient elution. The mobile phase was solvent A (1% formic acid in water) and solvent B (methanol) at a flow rate of 0.3mL/min. The MS conditions were: TSQ Quantum Ultra triple quadrupole mass spectrometer + electrospray source, ESI (Thermo Fisher Scientific, san Jose, CA), ion source polarity positive ion mode, sheath gas pressure (N2): 40units, ion transport tube temperature: collision air pressure at 350 ℃): 1.0mTorr (argon), Q1 resolution: 0.2FWHM, q3 resolution 0.7FWHM, dwell time: 0.2s, scan type: and SRM.
The level of abundance of the resistance gene in the DNA of the different samples was detected using Real-time qPCR. All real-time qPCR assays were performed by CFX96 Touch TM 2 XUItra SYBR mix (CWBIO, china) on real-time PCR detection system (Bio-Rad, USA) was performed in 3 replicates to detect the abundance of the resistance gene. DNA extraction Using a kit (Omega, cat: M5635-02) was performed according to the instructions, standards were prepared and standard curves were constructed, SYRB and primer mixtureA were prepared as shown in Table 1:
TABLE 1SYBR and primer mix A System
The configuration of the on-line system is shown in a table II:
table 2 upper body system
Real-time PCR reaction the PCR reaction solution prepared according to the reaction system is placed on a Real-time PCR instrument to carry out PCR reaction, and the reaction procedure is as follows:
the primers for PCR were:
tetA F:GCTACATCCTGCTTGCCTTC
R:CATAGATCGCCGTGAAGAGG
tetC F:CTTGAGAGCCTTCAACCCAG
R:ATGGTCGTCATCTACCTGCC
tetW F:GAGAGCCTGCTATATGCCAGC
R:GGGCGTATCCACAATGTTAAC
ermF F:CCACCGCCAACTGTCAAATC
R:TTTCAGGGACAACTTCCAGCA
ermX F:GATAGGACCAGGAAGCGGTG
R:AGAATGGCAGTGGTGAGGTG
ermB F:AGGGTTGCTCTTGCACACTC
R:CTGTGGTATGGCGGGTAAGT
16S rDNA F:CAATGGACGAAAGTCTGACG
R:ACGTAGTTAGCCGTGGCTTT
and calculating the adsorption removal capacity of the plant-biological film system on the antibiotics according to the difference value between the initial concentration and the final concentration of the antibiotics in the solution.
The calculation formula is as follows: removal rate (%) = (initial concentration-final concentration)/initial concentration, wherein the antibiotic concentration unit is mg/L.
The experimental results are shown in fig. 2-4:
FIG. 2 shows the concentration and removal efficiency of two antibiotics in water when treating a body of water containing multiple antibiotics and resistance genes using the grass and its attached biofilm system. According to the graph, the water body is treated through the biological film system, the removal efficiency of the azithromycin and the tetracycline in the water is 93.06% -99.80% and 73.35% -97.74%, respectively, and the plant-biological film system has strong capability of removing various antibiotic pollution in the water. And the antibiotic removal rate in the mixed antibiotic solution is higher than that of the single antibiotic solution.
FIG. 3 is the cumulative amount of ku cao and its attached biofilm system to single and mixed antibiotic pollutants. With the increase of the initial content of antibiotics, the accumulation of two antibiotics in the plant body is increased, and the accumulation of the antibiotics in the plant reaches the highest in the highest concentration solution. Therefore, the plant and the attached biological film system have better adsorption and removal effects on the water body polluted by the high-concentration antibiotics.
FIG. 4 is an enriched number (gene copy number) of resistance genes for two different antibiotics in a bitter grass biofilm and bitter grass plants. The presence of various antibiotics can obviously increase the number of resistance genes in water, plants and biological films can transfer the resistance genes in water, and the enrichment of the resistance genes of the biological films is higher than that of the plants.
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.
Claims (5)
1. A method for removing antibiotics and resistance genes in water by using plants and attached biological films thereof, which is characterized by comprising the following steps:
(1) Placing the aquatic plants in a pretreatment water body, adding a nutrient solution into the pretreatment water body, culturing for 10-30d, and then taking out the aquatic plants cultured for 10-30d from the pretreatment water body to obtain the aquatic plants and a biological membrane system thereof;
(2) Planting the aquatic plants and the biological film system thereof in a polluted water body to remove antibiotics and resistance genes in the polluted water body;
the pretreatment water body is obtained by the following steps: filtering the water body polluted by antibiotics, standing for 24-48 hours, and collecting supernatant to obtain a pretreated water body;
the antibiotics include tetracyclines, sulfadiazine and azithromycin; the resistance genes include ermF, ermX, ermB, tetA, tetC and tetW.
2. The method of claim 1, wherein in step (1) the nutrient solution is a Hoagland nutrient solution at a concentration of 10% w/v.
3. The method according to claim 1, wherein in step (2), the density of the planting is 50-200g/m 2 。
4. The method of claim 1, wherein the aquatic plant is a submerged plant and/or a floating plant.
5. The method of claim 4, wherein the submerged plant comprises kusnezoff, black algae, or goldfish algae; the floating plant comprises a large float.
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