CN107555608B - Method and system for removing microcystis and toxins in water body by strengthening constructed wetland - Google Patents
Method and system for removing microcystis and toxins in water body by strengthening constructed wetland Download PDFInfo
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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
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Microbiology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Hydrology & Water Resources (AREA)
- Engineering & Computer Science (AREA)
- Environmental & Geological Engineering (AREA)
- Water Supply & Treatment (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
Abstract
The application discloses a method and a system for removing microcystis and toxins in a water body by strengthening an artificial wetland. The application enhances the synergistic effect of physical, chemical and biological of the matrix in the wetland system by adding the exogenous degradation bacterial liquid, and further carries out harmless treatment on the microcystin, the application can strengthen the efficiency of treating the microcystin in sewage by the artificial wetland system, accelerate drainage to reach national sanitary standard of drinking water, lead the total nitrogen removal rate to be more than 67.5%, lead the ammoniacal nitrogen removal rate to be more than 52.6%, lead the nitrate nitrogen removal rate to be more than 91.6%, lead the total phosphorus removal rate to be more than 76.0%, lead the toxic algae cell removal rate to be up to 95.4% and lead the microcystin removal rate to be up to 98.4%. The method enhances the purification effect of the wetland system on sewage, has no secondary pollution, can effectively reduce construction and operation cost, has stable water treatment effect, and can adapt to the impact of different microcystin pollution loads.
Description
Technical Field
The application relates to the technical field of water treatment, in particular to a treatment method for removing toxic algae and microcystins in water by strengthening an artificial wetland, which is suitable for efficient and simple treatment of microcystins and toxins in drinking water sources or eutrophication water bodies.
Background
The problem of toxic cyanobacterial bloom caused by eutrophication and climate change is increasingly prominent, the toxic cyanobacterial bloom not only seriously damages the ecological system of water, but also releases algae toxins to threaten the health of aquatic animals, plants and humans, wherein Microcystins (MCs) are the most common, one of the most serious to cause damage, microcystins are generally produced and exist in algae cells, but under certain conditions, the algae cells are ruptured, and the intracellular algae toxins (IMCs) are released into the water to become dissolved extracellular toxins (EMC), which are commonly existing in algal bloom water and are often detected in various surface water bodies, especially drinking water sources, at present, the Microcystins have found more than 90 configurations, and the most common Microcystins are the three types of MC-LR, MC-RR and MC-YR which have the greatest influence. The continuous existence of the algae toxins increases the load of the sewage treatment plant, also causes the drinking risk of the effluent of the water works, and can cause symptoms such as liver cancer, kidney injury and the like when long-term drinking of the water containing the algae toxins.
At present, the treatment methods for algae mainly comprise artificial fishing, flocculation, chemical agent method, air floatation, chemical oxidation method and the like, and the removal methods for microcystins can be roughly classified into a physical method, a chemical method and a biological method 3 according to the removal principle, and are more common such as air floatation, activated carbon adsorption, ozone oxidation, fenton oxidation and the like. Most of the physical methods remove the intracellular algae toxins (IMC) by removing the microcystis cells, but the removal effect on the extracellular toxins (EMC) in the water body is not ideal, the microcystis cells are easily broken, and the extracellular toxins in the water body are increased. The chemical method mainly damages the structure of MCs through oxidation, and most of the chemical method has better treatment effect on intracellular toxins and extracellular toxins, but a large amount of chemical agents are easy to produce toxic and harmful byproducts, bring secondary pollution, and have extremely high treatment cost and are difficult to apply on a large scale.
At present, no related technical report of applying the constructed wetland treatment technology to remove the microcystis and toxins in the water body is seen.
Disclosure of Invention
The application aims to overcome the defects of the prior treatment technology and provides a method for removing microcystis and toxins in water by utilizing exogenous microcystin degrading bacteria to strengthen an artificial wetland.
Another technical problem to be solved by the application is to provide an artificial wetland system for realizing the method.
In order to solve the technical problems, the application adopts the following technical scheme:
the method for strengthening the artificial wetland to remove the microcystis and toxins in the water body is to apply the combined action of exogenous microcystis toxin degrading bacteria and the biological membrane indigenous microorganisms of the artificial wetland to improve the removal efficiency of the artificial wetland to the algae and the toxins; the method for applying exogenous microcystin degrading bacteria is to enrich and increase the concentration of microcystin in water body with microcystin bloom, periodically detect degrading bacteria gene mlrA during enrichment, confirm the culturing time of degrading bacteria, and culture until the concentration of microcystin degrading bacteria in microcystin degrading bacteria liquid is 1×10 6 ～2×10 6 Cell/m 2 The bacterial liquid is applied to the constructed wetland after the above steps.
Preferably, the artificial wetland adopts a subsurface flow artificial wetland, the wetland matrix is divided into a three-layer structure, the three-layer structure comprises a bottom layer, a middle layer and a surface layer from bottom to top, the bottom layer is zeolite, the middle layer is crushed gravel, the surface layer is high-organic matter peat soil, and perennial aquatic herbaceous plants are planted.
Preferably, the application of the exogenous microcystin degrading bacteria method enriches and domesticates microcystin degrading bacteria in a laboratory by 1×10 6 ～2×10 6 Cell/m 2 The above concentration is uniformly spread on the surface layer of the artificial wetland, and the artificial wetland is turned over and cultivated, and is used after being empty. Preferably, the time for the blank is about 1 day.
Preferably, the method further comprises inputting the microcystin-containing wastewater into the constructed wetland. Preferably by drip irrigation means.
Preferably, the thickness of the bottom layer is 5-9 cm; further preferably, the zeolite has a particle size of 1 to 2. 2 cm. Preferably, the thickness of the intermediate layer is 15-20 cm; further preferably, the crushed gravel has a particle size of 0.5 to 1cm.
Preferably, the thickness of the skin layer is 5 to 10 a cm a. The surface layer adopts peat soil with high organic matter, which is beneficial to microorganisms
Fixing and breeding.
Preferably, the perennial aquatic herb is Acorus calamus or the like.
Further preferably, the method comprises the steps of:
s1, constructing an artificial wetland system; the substrate three-layer structure of the system sequentially comprises a bottom layer, a middle layer and a surface layer from bottom to top, wherein the bottom layer is zeolite, the middle layer is crushed gravel, the surface layer is high-organic peat soil, and perennial herbaceous plants are planted and covered;
s2, membrane hanging treatment of eutrophic lake water: before the artificial wetland system operates, the wetland is subjected to membrane formation through eutrophic lake water so as to enrich indigenous microorganisms in the wetland, accelerate the formation of a biological membrane and facilitate the fixation of exogenous degradation bacteria;
s3, drip irrigation of sewage with high and low algae toxin loads:
the Gao Zao toxin loading sewage drip irrigation method comprises the following steps:
s31, screening microcystin degrading bacteria from a water body in which microcystin bloom is generated for a long time, enriching the microcystin serving as a unique carbon and nitrogen source, and gradually increasing the concentration of the microcystin; periodically detecting and measuring a degrading bacterium gene mlrA in the enrichment process so as to confirm the culture time of the degrading bacterium;
s32, mixing the enriched and domesticated microcystin degrading bacterial liquid with a ratio of 1 multiplied by 10 6 ～2×10 6 Cell/m 2 Uniformly spreading the above concentration on the surface layer of the artificial wetland matrix, turning over, and standing in the air for later use (planting plants);
the low algae toxin load sewage drip irrigation method is that
S33, inputting the sewage containing the microcystin into an artificial wetland system, staying for a period of time, fully contacting with a wetland substrate, and discharging the sewage out of the artificial wetland system as effluent;
s4, taking out the water sample at regular time, and analyzing the concentration of nutritive salt, the number of microcystis cells and the toxin content in the water sample; if the effluent does not reach the standard, returning to the step S3 after the effluent is subjected to the stay treatment; and discharging when the standard is reached.
Preferably, the microcystin degrading bacterial liquid after acclimation in step S4 is cultured in LB medium (composition: peptone 5.0. 5.0 g, yeast extract 2.5 g, naCl 5.0 g, sterilized ultra pure water 500 mL, pH adjusted to 7.0) and resuspended in normal saline for sprinkling.
The application also provides an artificial wetland system for realizing the method, which comprises a subsurface flow constructed wetland, a microcystin degrading bacterial liquid conveying device and a microcystin-containing sewage conveying device, wherein the subsurface flow constructed wetland comprises a wetland main body, a water inlet device and a water discharge device, the wetland main body is internally filled with a wetland substrate, the water inlet device is arranged on the side wall of the wetland main body, and the water discharge device is arranged at the bottom of the wetland main body; the water inlet device is respectively connected with a conveying device of sewage containing microcystin and a conveying device of microcystin degrading bacterial liquid.
Preferably, the system further comprises a water outlet sampling device and a sewage retention treatment device, wherein the water outlet sampling device is connected with the drainage device, one end of the sewage retention treatment device is connected with the drainage device, and the other end of the sewage retention treatment device is connected with the wetland main body.
Preferably, the water inlet device adopts a drip irrigation mode.
Preferably, the wetland substrate is divided into a three-layer structure, and comprises a bottom layer, a middle layer and a surface layer from bottom to top in sequence, wherein the bottom layer is zeolite, the middle layer is crushed gravel, the surface layer is high-organic matter peat soil, and perennial aquatic herbaceous plants are planted.
Preferably, the thickness of the bottom layer is 5-9 cm; further preferably, the zeolite has a particle size of 1 to 2. 2 cm. Preferably, the thickness of the intermediate layer is 15-20 cm; further preferably, the crushed gravel has a particle size of
0.5～1cm。
Preferably, the thickness of the skin layer is 5 to 10 a cm a. The surface layer adopts peat soil with high organic matter, which is beneficial to microorganisms
Fixing and breeding.
Preferably, the perennial aquatic herb is Acorus calamus or the like.
Preferably, the wetland body is of a cylindrical structure.
Further preferably, the diameter of the cylindrical structure is 22cm, the height is 35cm, and the total height of the wet foundation is 30cm.
Compared with the prior art, the application has the following beneficial effects:
the inventor has summarized through a large number of researches for a long time, overcomes the defects and limitations of the prior art, combines the combined action of exogenous microcystin degrading bacteria and artificial wetland biological membrane indigenous microorganisms with the advantages of the artificial wetland, creatively applies the artificial wetland treatment technology to remove microcystin and toxins in water, exerts the advantages of the ecological treatment technology, does not cause secondary pollution, has low operation cost and rapid treatment, has better removal potential on extracellular toxins and intracellular toxins, can be well docked with the existing water treatment technology, and has good application prospect.
A large number of experiments prove that the method has low running cost and high treatment efficiency, the total nitrogen removal rate reaches more than 67.5%, the ammonia nitrogen removal rate reaches more than 52.6%, the nitrate nitrogen removal rate reaches more than 91.6%, the total phosphorus removal rate reaches more than 76.0%, the toxic algae cell removal rate reaches 95.4%, the microcystin removal rate reaches 98.4%, and the ecological risk of water body algae toxins caused by cyanobacteria bloom is effectively reduced.
According to the method, the microcystin degrading bacterial liquid is additionally added for treatment, so that the synergistic effect of physical, chemical and biological functions of a matrix in a wetland system is exerted, the microcystin is further subjected to harmless treatment, the effect of treating the microcystin in the wetland can be accelerated, the water body residence time of the microcystin in the wetland is reduced, the removal load of the microcystin treated in the wetland in unit time is improved, the removal effect of toxigenic algae cells and toxins is good, the removal rate of the toxigenic algae cells is up to 95.4%, and the removal rate of the microcystin is up to 98.4%. The microcystin treatment operation is flexible, the reaction time and the addition amount of exogenous degradation bacteria can be flexibly selected according to the content of residual microcystin in the water body, the practicability is strong, and the microcystin treatment process can be well docked with the existing water treatment process.
The method of the application is economical and feasible, does not need to add chemical reagents, has low and universal price of the required materials, does not obviously increase the cost of water treatment, has simpler and more convenient filler replacement, and does not cause secondary pollution.
The method of the application further optimizes the constructed wetland system to better realize the application.
Drawings
FIG. 1 is a schematic flow chart of the method of the present application.
FIG. 2 is a schematic diagram of a system for removing microcystis and toxins in a water body according to the present application.
Wherein 11 is a water inlet device, 121 is a substrate surface layer, 122 is a substrate middle layer, 123 is a substrate bottom layer, 14 is aquatic plants, 15 is a water discharge device (water discharge port), 2 is a microcystin degrading bacterial liquid conveying device, and 3 is a microcystin-containing sewage conveying device.
FIG. 3 is a diagram showing the mlrA electrophoresis of the exogenous microcystin degrading bacterium gene.
Fig. 4 is a schematic diagram of the change of microcystins in the water body treated by the constructed wetland.
The specific embodiment is as follows:
the application will be further illustrated with reference to specific examples. The following examples are illustrative only and are not to be construed as limiting the application. Unless otherwise indicated, the reagents used in the examples which follow were those conventionally commercially available or commercially available, and the methods and apparatus used in the examples which follow were those conventionally used in the art unless otherwise indicated.
Example 1:
the embodiment provides a method for strengthening the artificial wetland to remove water microcystis and toxins, which uses the combined action of exogenous microcystin degrading bacteria and artificial wetland biomembrane indigenous microorganisms to improve the removal efficiency of algae and toxins in the artificial wetland; the method for degrading the microcystin by using the exogenous microcystin is to enrich and increase the concentration of the microcystin in water bodies in which microcystin bloom is generated, periodically detecting a degrading bacterium gene mlrA in the enrichment process, confirming the culturing time of the degrading bacterium, and culturing until the microcystin in the microcystin degrading bacterium liquid is degradedThe concentration of the degerming bacteria is 1 multiplied by 10 6 ～2×10 6 Cell/m 2 The bacterial liquid is applied to the constructed wetland after the above steps.
The artificial wetland adopts a subsurface flow artificial wetland, the wetland matrix is divided into a three-layer structure, the three-layer structure comprises a bottom layer, a middle layer and a surface layer from bottom to top, the bottom layer is zeolite, the middle layer is crushed gravel, the surface layer is high-organic matter peat soil, and perennial aquatic herbaceous plants are planted.
The flow of the method is shown in figure 1. The method comprises the following steps:
s1, constructing an artificial wetland system; the substrate three-layer structure of the system sequentially comprises a bottom layer, a middle layer and a surface layer from bottom to top, wherein the bottom layer is zeolite, the middle layer is crushed gravel, the surface layer is high-organic peat soil, and perennial herbaceous plants are planted and covered;
s2, membrane hanging treatment of eutrophic lake water: before the artificial wetland system operates, the wetland is subjected to membrane formation through eutrophic lake water so as to enrich indigenous microorganisms in the wetland, accelerate the formation of a biological membrane and facilitate the fixation of exogenous degradation bacteria;
s3, drip irrigation of sewage with high and low algae toxin loads:
the Gao Zao toxin loading sewage drip irrigation method comprises the following steps:
s31, screening microcystin degrading bacteria from a water body in which microcystin bloom is generated for a long time, enriching the microcystin serving as a unique carbon and nitrogen source, and gradually increasing the concentration of the microcystin; periodically detecting and measuring a degrading bacterium gene mlrA in the enrichment process so as to confirm the culture time of the degrading bacterium;
s32, mixing the enriched and domesticated microcystin degrading bacterial liquid with a ratio of 1 multiplied by 10 6 ～2×10 6 Cell/m 2 Uniformly spreading the above concentration on the surface layer of the artificial wetland matrix, turning over, and standing in the air for later use (planting plants);
the low algae toxin load sewage drip irrigation method is that
S33, inputting the sewage containing the microcystin into an artificial wetland system, staying for a period of time, fully contacting with a wetland substrate, and discharging the sewage out of the artificial wetland system as effluent;
s4, taking out the water sample at regular time, and analyzing the concentration of nutritive salt, the number of microcystis cells and the toxin content in the water sample; if the effluent does not reach the standard, returning to the step S3 after the effluent is subjected to the stay treatment; and discharging when the standard is reached.
Culturing the microcystin degrading bacterial liquid after domestication in the step S4 with LB culture medium (composition: peptone 5.0 g, yeast extract 2.5 g, naCl 5.0 g, sterilized ultra-pure water 500 mL, pH value adjusted to 7.0), and re-suspending with normal saline for spraying.
Wherein, colony sequencing: collecting water samples on the surface of a water body in a lake with microcystis water bloom in a long-term burst, extracting DNA of the water samples by using a Powerwater DNA Isolation Kit (MO-BIO, USA) extraction kit, and then carrying out PCR amplification.
The conditions for amplification were: (1) The reaction system (25. Mu.L) comprises 2.0. Mu.L of DNA template, 2.5. Mu.L of 10 XPCR buffer, mgCl 2 (25. Mu.M) 2.0. Mu.L, dNTPs (2.5. 2.5 mM) 2.0. Mu.L, PCR forward and reverse primers (10. Mu. Mol/L) 0.5. Mu.L each, tap DNA polymerase (5U/. Mu.L) 0.2. Mu.L, and made up with sterile deionized water. The primer information used is shown in Table 1.
(2) The reaction system was subjected to PCR amplification by a 96-well C100TM thermal cycler PCR instrument (Bio-Rad, USA). PCR reaction conditions: (1) 95 ℃ for 3min; (2) 95℃30s,62℃1min,72℃1min,35 cycles; (3) 7min at 72℃and forever at 4 ℃.
TABLE 1mlrAPrimer sequences for gene PCR and qPCR reactions
After amplification, electrophoresis was performed on a 1.5% agarose gel containing 1 XGelRed nucleic acid dye, as shown in FIG. 3. And observing with gel imager (Bio-Rad, USA), and observing microcystin degrading bacteriamlrAThe target band was cut with gel recovery and purification using AxyPrep DNA gel recovery kit (AxyGEN), and the purified product was further checked for accuracy by PCR while sequencing the purified product.
The bacterial liquid enrichment and culture operation comprises the following steps: confirming the bacterial sourceAfter the microcystin degrading bacteria, the crude microcystin extracted in a laboratory is used as a unique carbon and nitrogen source for enrichment, the concentration of the microcystin is gradually increased, and the degrading bacteria gene mlrA is detected periodically in the enrichment process to confirm that the degrading bacteria can be cultured. The method comprises the following specific steps: firstly, a small amount of water bloom lake water is inoculated into LB culture solution to be cultured for 1-2 d, then 1.0 mL is taken from enriched bacterial liquid to be inoculated into an inorganic salt culture medium (composition: cuCl) containing microcystin extracted in a laboratory 2 ，FeSO 4 、ZnCl 2 、MnCl 2 0.0005 g each of CaCl 2 0.02 g，MgSO 4 •7H 2 0 1.0 g，NaCl 1.0 g，KH 2 PO 4 0.5 g，K 2 HPO 4 4.0 g, 1000mL of sterilized ultrapure water, pH 7.5, and shaking at constant temperature (120 r/min,30 ℃ C.). Inoculating into new inorganic salt culture medium at 10% inoculation amount every 3 days, gradually increasing the concentration of microcystin in the selective culture medium (1-20 mg/L) to gradually enrich microcystin degrading bacteria, centrifuging to collect thallus after the final enrichment, cleaning with buffer solution, suspending thallus with physiological saline (9%o), and detecting by real-time fluorescence quantitative PCR methodmlrAThe gene copy number is then put into the wetland.
Sequencing and comparison: sequencing the mlrA of the screened colony, comparing with BLAST, finding that the similarity of the gene fragment of the degrading strain and the reported mlrA gene of Sphingomonas USTB-05 is up to 99%, further confirming that the degrading strain of microcystin is screened, and sequencing the result is shown in SEQ ID NO: 1.
The embodiment also provides a preferred constructed wetland system, as shown in figure 2. The sewage treatment device comprises a subsurface flow constructed wetland, a microcystin degrading bacterial liquid conveying device 2 and a microcystin-containing sewage conveying device 3, wherein the subsurface flow constructed wetland comprises a wetland main body, a water inlet device 11 and a drainage device 15, a wetland substrate is filled in the wetland main body, and the water inlet device 11 is arranged on the side wall of the wetland main body, preferably on the top of the side wall. The drainage device 15 is arranged at the bottom of the wetland main body; the water inlet device is respectively connected with a conveying device 3 of sewage containing microcystin and a conveying device 2 of microcystin degradation bacterial liquid.
The system also comprises a water outlet sampling device and a sewage retention treatment device, wherein the water outlet sampling device is connected with a drainage device, one end of the sewage retention treatment device is connected with the drainage device, and the other end of the sewage retention treatment device is connected with a wetland main body (not labeled in fig. 2).
The water inlet device 11 adopts a drip irrigation mode.
The wetland substrate is divided into a three-layer structure, namely a surface layer 121, a middle layer 122 and a bottom layer 123 from bottom to top, wherein the bottom layer 123 is zeolite, the middle layer 122 is crushed gravel, the surface layer 121 is high-organic matter peat soil, and perennial aquatic herbaceous plants 14 are planted.
The thickness of the bottom layer 123 is 5-9 cm; the particle size of the zeolite is 1-2 cm.
The thickness of the intermediate layer 122 is 15-20 cm; the particle size of the crushed gravel is 0.5-1 cm.
The thickness of the surface layer is 5-10 cm. The surface layer adopts peat soil with high organic matter, which is beneficial to microorganisms
Fixing and breeding.
The perennial aquatic herb is rhizoma Acori Graminei, etc.
The wetland main body can adopt a cylindrical structure.
Example 2 test of the removal effect of nutrient salts such as Nitrogen and phosphorus according to the application
The undercurrent constructed wetland system and method described in example 1 were constructed and utilized, a constructed wetland with an effective volume of 2.5L was constructed by using a PVC plastic bucket of 10L, a filling matrix was 5cm soil layers, 15 cm gravel (particle size 5 mm) layers in this order from top to bottom, the gravel was washed clean with tap water before filling, and the bottom layer was 5cm large-particle zeolite (20 mm) filled to prevent water blocking, and 3 yellow sweetflag plants were uniformly planted in each device.
The device operates: after the device is built, the eutrophic lake water body is used for irrigation culture for 1 year, nutrient salts are artificially added, and the concentration of the nutrient salts such as nitrogen, phosphorus and the like in the inlet water is shown in a table 2:
TABLE 2 quality of incoming water
Nitrogen and phosphorus removal effect: the constructed wetland system has stable effect of removing nitrogen and phosphorus nutritive salts. The average removal rate of the TN of the system is 67.5 to 75.3 percent, and NH is calculated 4 + The average removal rate of the-N is 52.6 to 66.3 percent, and the NO 3 - The average removal rate of the N is 91.6-94.2%, and the average removal rate of the TP is 76.0-92.8%.
Example 3: enhanced artificial wetland removal test for algae cells and soluble microcystin LR
The subsurface flow constructed wetland system and method described in example 1 are constructed and utilized, the composite bacterial liquid of the degrading bacteria is uniformly inverted on the surface soil of the wetland, and the composite bacterial liquid and the soil matrix are uniformly mixed and domesticated for one week by adopting a ploughing mode. The initial microcystin MC-LR concentration was set to 3.61. Mu.g/L, 6.80. Mu.g/L, 16.07. Mu.g/L, respectively. After one-time water inflow, the water sample stays in the wetland for 7 days, and 1mL of water sample is collected at 0h,6h,12h,24h,48h,72h,96h,120h,144h and 168h for measuring EMC-LR.
The optimal hydraulic retention time of the constructed wetland system is 3d, under the hydraulic retention time, the removal rate of the constructed wetland system to toxic algae cells (intracellular toxins IMC-LR) is 73.5% -95.4%, and the removal rate of the constructed wetland system to extracellular microcystins (EMC-LR) is 87.5% -98.4%, so that the MC-LR content can be effectively reduced to be less than 1.0 mug/L of the drinking water safety standard; further prolonging the hydraulic retention time of the wetland system, and enhancing the removal effect of MC-LR to be lower than the detection limit (0.10 mug/L). The addition of the microcystin composite degrading bacteria into the wetland can obviously improve the removal rate of the EMC-LR by the wetland systemP<0.05 The half-life period is shortened to 22 hours, which shows that the bio-enhanced constructed wetland can more effectively treat microcystin pollution
In order to overcome the defects of the existing water microcystis and toxin treatment method, the application provides a method for strengthening the artificial wetland to remove the water microcystis and toxin by utilizing exogenous microcystin degrading bacteria, and the harmless treatment of a wetland system to the microcystin is strengthened by virtue of the biodegradation effect of indigenous microorganisms and exogenous microcystin degrading bacteria added in the wetland, so that the method has the advantages of simple operation, economy, rapidness, high efficiency, flexibility and no secondary pollution, has strong practicability, is suitable for the treatment requirements of drinking water sources and eutrophic water, and can be well docked with the existing water treatment process.
Sequence listing
<110> and university of south China
<120> method and system for removing microcystis and toxins in water body by reinforced constructed wetland
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<170> SIPOSequenceListing 1.0
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<212> DNA
<213> microcystin degrading bacterium (Microcystins)
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agaagtttga ccccctggcc gcagcggtga tcctcggcct catgtggtgg gcttggcatt 480
tgccgcgcga cttgcccacg ctgttctccg gggtacccgg tgccgcatgg ggagtgatca 540
ccaaacagtt tgttatcgtt ccagggttta ttgccagcac tatcatcgct gtctttgttt 600
gcaacaaact gggtggatcg atgtggggtg gcgtgctcat tcacgcgatc cataacgaac 660
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gggatttcat cgaattcgcc gtggccattg ggctcgtcct gagtgtgggg aggaga 776
Claims (8)
1. A method for strengthening the removal of microcystis and toxins in water body by artificial wetland is characterized in that the combined action of exogenous microcystin degrading bacteria and biological membrane indigenous microorganisms of the artificial wetland is used for improving the removal efficiency of algae and toxins in the artificial wetland; the method for applying exogenous microcystin degrading bacteria is to enrich and increase the concentration of microcystin in water body with microcystin bloom, periodically detect degrading bacteria gene mlrA during enrichment, confirm the culturing time of degrading bacteria, and culture until the concentration of microcystin degrading bacteria in microcystin degrading bacteria liquid is 1×10 6 ～2×10 6 Cell/m 2 Uniformly scattering the bacterial liquid on the surface layer of the constructed wetland, turning over, and planting plants after the bacterial liquid is empty; the membrane hanging treatment is carried out on the constructed wetland biological membrane by the eutrophic lake water, specifically, before the constructed wetland system operates, the membrane hanging is carried out on the constructed wetland by the eutrophic lake water so as to enrich the indigenous microorganisms in the constructed wetland, accelerate the formation of the biological membrane and be beneficial to the fixation of exogenous degradation bacteria; the artificial wetland adopts a subsurface flow artificial wetland, the wetland matrix is divided into a three-layer structure, the three-layer structure comprises a bottom layer, a middle layer and a surface layer from bottom to top, the bottom layer is zeolite, the middle layer is crushed gravel, the surface layer is high-organic matter peat soil, and perennial aquatic herbaceous plants are planted.
2. The method for enhancing the removal of microcystis and toxins from a body of water in a constructed wetland of claim 1, further comprising inputting microcystin-containing wastewater into the constructed wetland.
3. The method for removing microcystis and toxins from a body of water by using an enhanced constructed wetland according to claim 1, wherein the method comprises the following steps:
s1, constructing an artificial wetland system; the substrate three-layer structure of the system sequentially comprises a bottom layer, a middle layer and a surface layer from bottom to top, wherein the bottom layer is zeolite, the middle layer is crushed gravel, the surface layer is high-organic peat soil, and perennial herbaceous plants are planted and covered;
s2, membrane hanging treatment of eutrophic lake water: before the artificial wetland system operates, the wetland is subjected to membrane formation through eutrophic lake water so as to enrich indigenous microorganisms in the wetland, accelerate the formation of a biological membrane and facilitate the fixation of exogenous degradation bacteria;
s3, drip irrigation of sewage with high and low algae toxin loads:
s31, screening microcystin degrading bacteria from a water body in which microcystin bloom is generated for a long time, enriching the microcystin serving as a unique carbon and nitrogen source, and gradually increasing the concentration of the microcystin; periodically detecting and measuring a degrading bacterium gene mlrA in the enrichment process so as to confirm the culture time of the degrading bacterium;
s32, mixing the enriched and domesticated microcystin degrading bacterial liquid with a ratio of 1 multiplied by 10 6 ～2×10 6 Cell/m 2 Uniformly spreading the above concentration on the surface layer of the artificial wetland matrix, turning over, and standing for later use;
s33, inputting the sewage containing the microcystin into an artificial wetland system, staying for a period of time, fully contacting with a wetland substrate, and discharging the sewage out of the artificial wetland system as effluent;
s4, taking out the water sample at regular time, and analyzing the concentration of nutritive salt, the number of microcystis cells and the toxin content in the water sample; if the effluent does not reach the standard, returning to the step S3 after the effluent is subjected to the stay treatment; and discharging when the standard is reached.
4. A system for realizing the method for removing microcystis and toxins in water body by the reinforced constructed wetland according to any one of claims 1 to 3, which is characterized by comprising a subsurface flow constructed wetland, a microcystin degrading bacterial liquid conveying device and a microcystin-containing sewage conveying device, wherein the subsurface flow constructed wetland comprises a wetland main body, a water inlet device and a water outlet device, the wetland main body is filled with a wetland substrate, the water inlet device is arranged on the side wall of the wetland main body, and the water outlet device is arranged at the bottom of the wetland main body; the water inlet device is respectively connected with a conveying device of sewage containing microcystin and a conveying device of microcystin degrading bacterial liquid.
5. The system of claim 4, further comprising a water sampling device and a wastewater retention treatment device, wherein the water sampling device is connected to the water drainage device, and wherein the wastewater retention treatment device is connected to the water drainage device at one end and the wetland body at the other end.
6. The system of claim 4, wherein the water inlet means is in the form of drip irrigation.
7. The system of claim 4, wherein the wetland substrate is divided into a three-layer structure, namely a bottom layer, a middle layer and a surface layer from bottom to top, the bottom layer is zeolite, the middle layer is crushed gravel, the surface layer is high organic matter peat soil, and perennial aquatic herbaceous plants are planted.
8. The system of claim 4, wherein the base layer has a thickness of 5 to 9cm; the thickness of the middle layer is 15-20 cm; the thickness of the surface layer is 5-10 cm.
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