CN114368886B - Biological microcapsule-based efficient black and odorous water body black and odorous substrate mud repairing agent - Google Patents
Biological microcapsule-based efficient black and odorous water body black and odorous substrate mud repairing agent Download PDFInfo
- Publication number
- CN114368886B CN114368886B CN202111428976.5A CN202111428976A CN114368886B CN 114368886 B CN114368886 B CN 114368886B CN 202111428976 A CN202111428976 A CN 202111428976A CN 114368886 B CN114368886 B CN 114368886B
- Authority
- CN
- China
- Prior art keywords
- black
- microcapsule
- odorous
- biological
- water body
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F11/00—Treatment of sludge; Devices therefor
- C02F11/02—Biological treatment
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/105—Phosphorus compounds
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Molecular 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)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
Abstract
The invention relates to a biological microcapsule-based high-efficiency black and odorous water body black and odorous substrate mud repairing agent, which is characterized by comprising capsules, biological microcapsules and porous materials, wherein the biological microcapsules are loaded in the porous materials to form porous functional materials, and the capsules and the porous functional materials undergo a crosslinking reaction to form the biological microcapsule-based high-efficiency black and odorous water body black and odorous substrate mud repairing agent; the preparation method comprises the following steps of firstly, preparing capsules; preparing biological microcapsules; step three, preparing a porous functional material; preparing a high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on biological microcapsules; the method has the advantages that the method is not easy to block, microorganisms can survive for a long time in a severe environment, the action time is long, the method is efficient and durable, COD, total phosphorus and ammonia nitrogen concentration in the water body are effectively reduced, sulfide and ammonia nitrogen concentration in the sediment have positive effects on ecological restoration of the water body for a long time, the purification efficiency is high, and the release area of the water body is large.
Description
Technical Field
The invention relates to a biological microcapsule-based efficient black and odorous water body black and odorous substrate mud repairing agent.
Background
Most of river channels in China have black and odorous phenomena with different degrees, the life of residents is seriously influenced, the social development is restricted, and the river channels are increasingly focused and focused. The polluted bottom mud is one of main pollution sources of the river, is a 'sedimentation reservoir' for receiving and decomposing pollutants in the river water body, is a 'source' for upward water covering to release nutrient salt and organic pollutants, and has great influence on the river water environment. Because of long-term exogenous input, a large amount of pollutants such as nutrition, salt elements, refractory organic matters, heavy metals and the like in the river water body and silt in the river are accumulated in the river sediment through long-term physical and chemical reactions, the pollutants in the sediment are unstable, and the pollutants can be released from the sediment again under proper conditions such as hydraulic flushing, external disturbance and the like, so that the water body is subjected to secondary pollution. Along with the effective control of exogenous pollution and the promotion of river water body treatment, the endogenous pollution problem caused by polluted sediment is increasingly prominent, and the sediment of the river needs to be repaired and the release of sediment pollutants is controlled.
The currently common black and odorous substrate sludge repairing method mainly comprises chemical, physical and biological repairing technologies. The physical repair technology is mainly to treat the river by means of measures such as sewage interception and dredging, water diversion and flushing, dredging, sediment aeration and the like, and although the sewage interception of a pollution source can be controlled from the source, non-point sources and other pollution sources still exist in the river, and the river is not easy to cut off in the treatment process. The water diversion scouring technology has the technical advantages of quick response and obvious treatment effect. Chemical restoration technology is generally to add chemical agents such as ferric salt, aluminum salt and the like into river polluted bottom mud. The problem of black and odorous river is solved by using a chemical restoration technology, and the chemical agent can be used as an emergency in emergency, so that the generation of sludge in the treatment process is increased, and meanwhile, certain toxicity is generated. The bioremediation method is a scientific, environment-friendly, green and effective method.
The microbial agent, such as EM bacteria, bacillus subtilis, compound bacteria and the like, has the effects of promoting plant growth, resisting diseases and environmental stress, improving soil microecology, improving soil fertility and the like, and is an agricultural green biological agent with very good application prospect (to be determined). Patent CN1736901a proposes a method for treating wastewater containing ammonia nitrogen, which adopts microorganism strains such as ammonia oxidizing bacteria, nitrous acid oxidizing bacteria, etc., so as to effectively maintain higher biological activity of the water treatment system, thereby ensuring excellent ammonia nitrogen treatment efficiency of the system. However, the factors such as the water inflow, the water inflow quality, the dissolved oxygen concentration and the pH value of the system are required to be strictly controlled, and the influence of each condition on the microbial activity is great. Therefore, the microbial agent has the problems of low stability of thalli, low viable count, poor lasting effect and the like in the application process. The microbial microencapsulation technology, namely the biological microcapsule, embeds or encapsulates the thalli into the microcapsule, realizes the protection of the functional strain, and can effectively improve the survival activity, the stability and the slow release property of the thalli.
At present, the preparation method of the sediment repairing agent mostly adopts chemical or physical formulas, often adopts a formula combination of an oxygen releasing agent, an adsorbent, an embedding agent and the like, the research method of biological formulas is relatively few, and only patent (patent number CN 202010861371.4) develops a double-layer sediment repairing capsule which is divided into an outer bacterial liquid coating layer and an inner adsorption material layer, wherein the outer bacterial liquid coating layer provides nutrition for the growth of microorganisms by degrading sediment pollutant substances, and the inner layer releases oxygen gas layer to provide oxygen for the growth of microorganisms. However, this method has some drawbacks: (1) As the growth of microorganisms is affected and limited by various factors, the contents of nitrogen and phosphorus in urban sewage are continuously increased, and the high-concentration nitrogen and phosphorus pollutants and heavy metal toxic substances can inhibit the vital activities of traditional microorganisms, so that the degradation rate of the pollutants by the microorganisms is reduced, and even a large number of the microorganisms die. (2) The microorganism immobilization method is adopted, so that microorganisms can be effectively protected, but the sodium alginate serving as a coating material is easily degraded in a short time, and the growth of the microorganisms can be influenced in the degradation process, so that the action time is short, and the action effect is influenced. (3) The contact surface between the immobilization method of the large capsule and the water body is small, so that the action effect is affected. Therefore, the important reason why the biological ecological method is not popularized at present is mainly that a proper method which can adapt microorganisms to various black and odorous water body treatments is not developed yet.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide the high-efficiency black and odorous substrate sludge repairing agent for the black and odorous water body based on the biological microcapsule, aiming at the difficulties and problems of the prior art in repairing the sewage water body, the defects of the prior art are overcome, microorganisms can survive for a long time in a severe environment, the action time is long, the effect is high and durable, the COD, the total phosphorus and the ammonia nitrogen concentration in the water body are effectively reduced, the sulfide and the ammonia nitrogen concentration in the substrate sludge have positive effects on the ecological restoration of the water body for a long time, the purification efficiency is high, and the contact area with the water body is large.
In order to achieve the above purpose, the invention is realized by the method, which is a high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on biological microcapsules, comprising capsules, biological microcapsules and porous materials, wherein the biological microcapsules are uniformly loaded in the porous materials to form porous functional materials, the capsules and the porous functional materials undergo a crosslinking reaction to form the high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on the biological microcapsules, the loading rate of the biological microcapsules loaded on the porous materials is 5% -60%, and the mass ratio of the capsules, the biological microcapsules and the porous materials is 30:20:5, a step of;
the preparation method of the high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on the biological microcapsule comprises the following steps:
step one, preparing capsules;
(1) Dissolving 1-3 parts of sodium alginate in 75 parts of water thoroughly to form flocculent colloid solution, slowly adding 6-8 parts of calcium peroxide, 6-8 parts of diatomite and 8-10 parts of fine sand after dissolving and cooling, stirring and uniformly mixing to form inner flocculent bodies, and putting the inner flocculent mixed bodies into 3% -5% of calcium chloride solution for crosslinking reaction to form capsules;
preparing biological microcapsules;
(1) Adding 5-10ml of microbial liquid seeds and 10-20ml of nutrient solution into 1-5% sodium alginate aqueous solution, dripping into a coagulating bath of 1-5% calcium chloride solution through a needle syringe at normal temperature, stabilizing for a period of time to form a microcapsule embryonic form, performing secondary calcification in 1-5% calcium chloride solution, performing secondary complex coacervation reaction in 0.5-5% chitosan solution for 20-30 minutes, and performing soaking and proliferation culture by using 100-150ml of sterile physiological saline with the concentration of 0.85-0.9% to form a biological microcapsule;
step three, preparing a porous functional material;
(1) Taking volcanic rock with the particle size of 1-3mm, wherein the weight part is 30-50 parts, immersing the volcanic rock into a device filled with 100-150ml of biological microcapsules in the second step for adsorption, and forming a porous functional material with biological microcapsules;
preparing a high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on biological microcapsules;
(1) 3-5 parts of sodium alginate is thoroughly dissolved in 75 parts of water, the porous functional material with biological microcapsules in the third step is added after dissolution and cooling, uniformly stirred to form floccules, the capsules in the first step are wrapped in the floccules and are put into a 3% -5% calcium chloride solution crosslinking reagent for crosslinking reaction, and the high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on the biological microcapsules is formed.
In the technical scheme, the microbial thallus seeds comprise EM bacteria or bacillus subtilis or composite bacteria, and the nutrient solution is suitable for the growth of the EM bacteria or the bacillus subtilis or the composite bacteria.
In the technical scheme, the diameter of the microcapsule is 0.06-0.72mm.
In the technical scheme, the mass ratio of the high-efficiency black and odorous sediment repairing agent for the black and odorous water body based on the biological microcapsules to the sediment in the water body is 1:40.
In the present technical solution, in the first step, the time for the crosslinking reaction is 10 to 20 minutes.
In the technical scheme, in the second step, the stabilization time of the coagulating bath in which the needle injector is dripped into the calcium chloride mixed solution is 20-30 minutes, so as to form a microcapsule embryonic form.
In the technical scheme, in the third step, the volcanic rock is immersed in the device filled with the biological microcapsules of the second step for adsorption for 16-24 hours. In the technical scheme, in the fourth step, the capsule in the first step is wrapped in the floccule, and the floccule is put into a crosslinking reagent for crosslinking reaction for 15-20 minutes.
In the technical scheme, in the third step, the diameter of the volcanic rock is 1-2mm.
In the technical scheme, the concentration of the sodium alginate aqueous solution in the step II is 3% -5%, the dropping speed of the needle head is 40-60 drops/min, the pH value of the calcium chloride solution is 6.8-7.2, the concentration of the chitosan solution is 3% -5%, and the proliferation culture time is 2-4h.
Compared with the prior art, the invention has the advantages that: the microbial agent is not easy to block, microorganisms can survive for a long time in a severe environment, the action time is long, the microbial agent is efficient and durable, COD, total phosphorus and ammonia nitrogen concentration in water are effectively reduced, sulfide and ammonia nitrogen concentration in bottom mud have positive effects on ecological restoration of the water for a long time, the purification efficiency is high, and the removal area of the microbial agent and the water is large.
Drawings
FIG. 1 is a schematic structural diagram of a biological microcapsule-based efficient black and odorous water body black and odorous substrate mud repairing agent;
FIG. 2 is a schematic representation of a biological microcapsule of the invention;
FIG. 3 is a schematic diagram of the preparation method of the biological microcapsule in the second step of the present invention;
FIG. 4 is a plot of ammonia nitrogen content in a body of water according to one embodiment of the present invention;
FIG. 5 is a graph showing the variation line of COD content in water according to the first embodiment of the present invention;
FIG. 6 is a plot of the total phosphorus content of a body of water according to one embodiment of the present invention;
FIG. 7 is a bar graph of sulfide content before and after sediment treatment in accordance with an embodiment of the present invention;
FIG. 8 is a bar graph of ammonia nitrogen content before and after sediment treatment in accordance with an embodiment of the present invention;
FIG. 9 is a plot of ammonia nitrogen content in water according to a second embodiment of the present invention;
FIG. 10 is a plot of variation of COD content in water according to a second embodiment of the invention;
FIG. 11 is a plot of the total phosphorus content of a body of water according to a second embodiment of the present invention;
FIG. 12 is a bar graph of two sulfide levels for an embodiment of the present invention;
FIG. 13 is a bar graph of ammonia nitrogen content before and after sediment treatment in accordance with the second embodiment of the present invention;
FIG. 14 is a plot of ammonia nitrogen content in a body of water according to a third embodiment of the present invention;
FIG. 15 is a plot of variation of COD content of a body of water according to a third embodiment of the present invention;
FIG. 16 is a plot of total phosphorus content of a body of water according to embodiment III of the present invention;
FIG. 17 is a bar graph of three sulfide levels for an embodiment of the present invention;
FIG. 18 is a bar graph of ammonia nitrogen content before and after sediment treatment in accordance with example III of the present invention.
Detailed Description
In order that the invention may be understood more fully, a more particular description of the invention will be rendered by reference to specific embodiments thereof which are illustrated in the appended claims. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. These embodiments are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, 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. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.
All percentages, fractions and ratios are by weight of the total composition of the present invention, unless otherwise specified. All weights as they pertain to listed ingredients are to the active level and, therefore, do not include solvents or by-products that may be included in commercially available materials, unless otherwise specified. The term "weight content" is used herein to denote in accordance with "%".
All formulations and tests herein occur in a room temperature environment unless otherwise indicated.
Herein, "comprising," "including," "containing," "having," or other variations thereof are intended to cover non-closed-form inclusion, as do not distinguish between such terms, but rather the term "comprising" refers to the addition of other steps and components that do not affect the end result. The term "comprising" also includes the terms "consisting of …" and "consisting essentially of …". The compositions and methods/processes of the present invention can comprise, consist of, and consist essentially of the essential elements and limitations described herein, as well as additional or optional ingredients, compositions, steps, or limitations of any of the embodiments described herein.
The novel embodiments of the present invention will be further described with reference to the accompanying drawings. The description of these embodiments is provided to assist understanding of the novel form of the present invention, but is not intended to limit the novel form of the present invention. In addition, the technical features of the novel embodiments of the present invention described below may be combined with each other as long as they do not collide with each other.
Example 1
As shown in fig. 1 to 8, the high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on biological microcapsules comprises capsules, biological microcapsules and a porous material, wherein the biological microcapsules are uniformly loaded in the porous material to form a porous functional material, the capsules and the porous functional material undergo a crosslinking reaction to form the high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on the biological microcapsules, the loading rate of the biological microcapsules loaded on the porous material is 5%, the preferable loading rate is 7%, and the mass ratio of the capsules, the biological microcapsules and the porous material is 30:20:5, a step of;
the preparation method of the high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on the biological microcapsule comprises the following steps:
step one, preparing capsules;
(1) 1 part of sodium alginate is thoroughly dissolved into flocculent colloid solution in 75 parts of water, 6 parts of calcium peroxide, 6 parts of diatomite and 8 parts of fine sand are slowly added after dissolution and cooling, and are stirred and mixed uniformly to form inner flocculent bodies, and the inner flocculent bodies are put into 3% calcium chloride solution for crosslinking reaction to form capsules;
preparing biological microcapsules;
(1) Adding 5ml of microorganism bacterium liquid seeds and 10ml of nutrient solution into 1% sodium alginate aqueous solution, dripping into coagulating bath of 1% calcium chloride solution through a needle syringe at normal temperature, stabilizing for a period of time to form microcapsule embryonic form, performing secondary calcification in 1% calcium chloride solution, performing secondary complex coacervation reaction in 0.5% chitosan solution for 20 minutes, and performing soaking and proliferation culture by using 100ml of sterile physiological saline with concentration of 0.85% to form biological microcapsules;
step three, preparing a porous functional material;
(1) Taking volcanic rock with the particle size of 1mm, wherein the weight part of the volcanic rock is 30 parts, immersing the volcanic rock into a device filled with 100ml of biological microcapsules in the second step for adsorption, and forming a porous functional material with biological microcapsules;
preparing a high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on biological microcapsules;
(1) And 3 parts of sodium alginate is thoroughly dissolved in 75 parts of water, the porous functional material with biological microcapsules in the third step is added after dissolution and cooling, uniformly stirred to form floccules, the capsules in the first step are wrapped in the floccules and are put into a 3% calcium chloride solution crosslinking reagent for crosslinking reaction, and the high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on the biological microcapsules is formed.
Biological microcapsule
The microcapsule technology is a technology of wrapping trace substances in a high polymer film, is a miniature packaging technology for storing solid, liquid and gas, and mainly comprises a wrapped core material and a wrapped wall material, and is shown in the following figure. The biological microcapsule adopts a microencapsulation technology to embed or encapsulate microbial thalli such as EM bacteria, bacillus subtilis, composite bacteria and the like as core materials into wall material calcium alginate and the like of the microcapsule, so as to protect functional strains and improve the survival activity, stability and slow release property of the thalli.
The loading rate of the biological microcapsules on the geopolymer porous material is calculated as follows:
load ratio = (bio-microcapsule load)/(bio-microcapsule load + porous functional material) ×100%
Wherein, bio-microcapsule loading = bio-microcapsule total amount-bio-microcapsule remaining in solution
In the invention, the calcium peroxide of the capsule is used as an oxygenation agent to change the anaerobic environment of the sediment into an aerobic environment, and the diatomite is used as an adsorption material to effectively adsorb pollutants in the sediment and can provide a carrier for biological growth.
Microorganisms are indispensable components in water and soil environment restoration, and play an indispensable role. The microbial technology has the characteristics of high efficiency, low cost, no secondary pollution and the like in the process of repairing water pollution. Compared with the characteristic that thalli are easy to inactivate in the environment, the method adopts a microcapsule form to wrap thalli, namely the biological microcapsule is formed, so that the thalli can be effectively prevented from being damaged by external bad conditions, the stability of the thalli is improved, and the storage time of the thalli is prolonged. Some cells have a low survival rate when subjected to adverse environmental influences (ultraviolet rays, pH, etc.) in the environment. The microbial cells can be isolated from the bad environment by the microcapsule technology, so that the effect of improving the stability of the microbial cells is achieved. In the biological microcapsule in the project, the microcapsule not only provides nutrient substances for the growth of microorganisms, but also can protect the microorganisms from the external environment, thereby greatly improving the capability of the microorganisms for adapting to the environment; different from the traditional bottom mud repairing powdery medicament, the biological microcapsule loaded on the degradable porous material is not influenced by water flow and cannot run off along with the water flow; the outer wall coating also has slow release effect, and can prolong the action time of microorganisms in the microcapsule and reduce degradation time.
In this embodiment, the microbial cell seed includes EM bacteria or bacillus subtilis or a complex bacteria, and the nutrient solution is a nutrient solution suitable for growth of the EM bacteria or bacillus subtilis or complex bacteria.
In this example, the diameter of the microcapsules is 0.06mm.
In the embodiment, the mass ratio of the high-efficiency black and odorous sediment repairing agent based on the biological microcapsules to the sediment in the water body is 1:40.
In this example, in the first step, the time for the crosslinking reaction was 10 minutes.
In this example, in the second step, the stabilization time of the coagulation bath in which the needle syringe was dropped into the calcium chloride mixed solution was 20 minutes to form a microcapsule prototype.
In this example, in the third step, the volcanic rock is immersed in the device filled with the biological microcapsule of the second step for adsorption for 16 hours. In this embodiment, in the fourth step, the capsule in the first step is wrapped in the flock and placed in a crosslinking reagent for a crosslinking reaction for 15 minutes.
In this embodiment, in the third step, the diameter of the volcanic rock is 1.1mm.
In the embodiment, the concentration of the sodium alginate aqueous solution in the step II is 3%, the dropping speed of the needle is 40 drops/min, the pH value of the calcium chloride solution is 6.8, the concentration of the chitosan solution is 3%, and the proliferation culture time is 2 hours.
Respectively taking 300g of sediment from 2 samples of river, removing all rubbles, branches and other garbage in the acquired sediment, uniformly mixing for later use, uniformly placing the sediment in a glass jar, wherein the thickness of the sediment in each jar is 10cm, and injecting water body sewage in the same river with the depth of 20 cm. The mass ratio of the sediment to the capsules is 40:1, the capsules of the invention are put into the sediment and stirred uniformly, 4 groups of experiments are arranged, and the arrangement scheme is shown in table 1.
Table 1 experimental grouping and different treatment modes of black and odorous substrate sludge
As shown in the figure, after the novel ecological black and odorous sediment remediation agent reacts for 3 days, the concentration of pollutants in two river surges is reduced by more than 40%, the concentration of ammonia nitrogen is reduced by 50% on average, and after 7 days, the concentration indexes of pollutants in water quality can reach the V-class water level of the national surface water standard, the treatment rate of ammonia nitrogen and sulfide of the sediment reaches more than 85%, so that the effect of thoroughly eliminating black and odorous is realized.
Example two
As shown in fig. 1 to 3 and fig. 9 to 13, the high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on biological microcapsules comprises capsules, biological microcapsules and porous materials, wherein the biological microcapsules are uniformly loaded in the porous materials to form porous functional materials, the capsules and the porous functional materials undergo a crosslinking reaction to form the high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on the biological microcapsules, the loading rate of the biological microcapsules loaded in the porous functional materials is 32.5%, the preferable loading rate is 18.5%, and the mass part ratio of the capsules, the biological microcapsules and the porous materials is 30:20:5, a step of;
the preparation method of the high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on the biological microcapsule comprises the following steps:
step one, preparing capsules;
(1) 2 parts of sodium alginate is thoroughly dissolved into flocculent colloid solution in 75 parts of water, 7 parts of calcium peroxide, 7 parts of diatomite and 9 parts of fine sand are slowly added after dissolution and cooling, and are stirred and mixed uniformly to form inner flocculent bodies, and the inner flocculent mixed bodies are put into 4% calcium chloride solution for crosslinking reaction to form capsules;
preparing biological microcapsules;
(1) Adding 7.5ml of microorganism bacterium liquid seeds and 15ml of nutrient solution into a 3% sodium alginate aqueous solution, dripping the solution into a coagulating bath of a 3% calcium chloride mixed solution through a needle syringe at normal temperature, stabilizing for a period of time to form microcapsule embryonic form, performing secondary calcification in the 3% calcium chloride solution, performing secondary complex coacervation reaction in a 2.75% chitosan solution for 25 minutes, and performing soaking and proliferation culture by using 125ml of sterile physiological saline with the concentration of 0.875% to form a biological microcapsule;
step three, preparing a porous functional material;
(1) Taking volcanic rock with the particle size of 2mm, wherein the weight part of the volcanic rock is 40 parts, immersing the volcanic rock into a device filled with 125ml of biological microcapsules in the second step for adsorption, and forming a porous functional material with biological microcapsules;
preparing a high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on biological microcapsules;
(1) And (3) thoroughly dissolving 4 parts of sodium alginate in 75 parts of water, adding the porous functional material with the biological microcapsules in the third step after dissolving and cooling, uniformly stirring to form floccules, wrapping the capsules in the first step in the floccules, and putting the capsules in a 4% calcium chloride solution crosslinking reagent for crosslinking reaction to form the high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on the biological microcapsules.
Biological microcapsule
The microcapsule technology is a technology of wrapping trace substances in a high polymer film, is a miniature packaging technology for storing solid, liquid and gas, and mainly comprises a wrapped core material and a wrapped wall material, and is shown in the following figure. The biological microcapsule adopts a microencapsulation technology to embed or encapsulate microbial thalli such as EM bacteria, bacillus subtilis, composite bacteria and the like as core materials into wall material calcium alginate and the like of the microcapsule, so as to protect functional strains and improve the survival activity, stability and slow release property of the thalli.
The loading rate of the biological microcapsules on the geopolymer porous material is calculated as follows:
load ratio = (bio-microcapsule load)/(bio-microcapsule load + porous functional material) ×100%
Wherein, bio-microcapsule loading = bio-microcapsule total amount-bio-microcapsule remaining in solution
In the invention, the calcium peroxide of the capsule is used as an oxygenation agent to change the anaerobic environment of the sediment into an aerobic environment, and the diatomite is used as an adsorption material to effectively adsorb pollutants in the sediment and can provide a carrier for biological growth.
Microorganisms are indispensable components in water and soil environment restoration, and play an indispensable role. The microbial technology has the characteristics of high efficiency, low cost, no secondary pollution and the like in the process of repairing water pollution. Compared with the characteristic that thalli are easy to inactivate in the environment, the method adopts a microcapsule form to wrap thalli, namely the biological microcapsule is formed, so that the thalli can be effectively prevented from being damaged by external bad conditions, the stability of the thalli is improved, and the storage time of the thalli is prolonged. Some cells have a low survival rate when subjected to adverse environmental influences (ultraviolet rays, pH, etc.) in the environment. The microbial cells can be isolated from the bad environment by the microcapsule technology, so that the effect of improving the stability of the microbial cells is achieved. In the biological microcapsule in the project, the microcapsule not only provides nutrient substances for the growth of microorganisms, but also can protect the microorganisms from the external environment, thereby greatly improving the capability of the microorganisms for adapting to the environment; different from the traditional bottom mud repairing powdery medicament, the biological microcapsule loaded on the degradable porous material is not influenced by water flow and cannot run off along with the water flow; the outer wall coating also has slow release effect, and can prolong the action time of microorganisms in the microcapsule and reduce degradation time.
In this embodiment, the microbial cell seed includes EM bacteria or bacillus subtilis or a complex bacteria, and the nutrient solution is a nutrient solution suitable for growth of the EM bacteria or bacillus subtilis or complex bacteria.
In this embodiment, the diameter of the bio-microcapsule is 0.53mm, preferably the diameter of the bio-microcapsule is 0.39mm.
In the embodiment, the mass ratio of the high-efficiency black and odorous sediment repairing agent based on the biological microcapsules to the sediment in the water body is 1:40.
In this example, in the first step, the time of the crosslinking reaction was 15 minutes.
In this example, in the second step, the stabilization time of the coagulation bath in which the needle syringe was dropped into the calcium chloride mixed solution was 25 minutes to form a microcapsule prototype.
In this example, in the third step, the volcanic rock is immersed in the device filled with the biological microcapsule of the second step for adsorption for 20 hours. In this embodiment, in the fourth step, the capsule in the first step is wrapped in the flock and placed in a crosslinking reagent for a crosslinking reaction for 17.5 minutes.
In this example, in the third step, the diameter of the volcanic rock is 1.5mm.
In this example, the concentration of the sodium alginate aqueous solution in the step II is 4%, the dropping speed of the needle is 50 drops/min, the pH value of the calcium chloride solution is 7, the concentration of the chitosan solution is 4%, and the proliferation culture time is 3 hours.
The invention can reduce COD, total phosphorus, ammonia nitrogen, and bottom mud ammonia nitrogen concentration and sulfide in the water body.
The invention is used for sampling test
Respectively taking 300g of sediment from 2 samples of river, removing all rubbles, branches and other garbage in the acquired sediment, uniformly mixing for later use, uniformly placing the sediment in a glass jar, wherein the thickness of the sediment in each jar is 10cm, and injecting water body sewage in the same river with the depth of 20 cm. The mass ratio of the sediment to the capsules is 40:1, the capsules of the invention are put into the sediment and stirred uniformly, 4 groups of experiments are arranged, and the arrangement scheme is shown in table 1.
TABLE 2 Experimental grouping and different treatment modes of black and odorous substrate sludge
| Group number | Treatment mode | Ratio of addition |
| 1 | River 1 | 40：1 |
| 2 | River 2 | 40：1 |
As shown in the figure, after the novel ecological black and odorous sediment remediation agent reacts for 3 days, the concentration of pollutants in two river surges is reduced by more than 40%, the concentration of ammonia nitrogen is reduced by 50% on average, and after 7 days, the concentration indexes of pollutants in water quality can reach the V-class water level of the national surface water standard, the treatment rate of ammonia nitrogen and sulfide of the sediment reaches more than 85%, so that the effect of thoroughly eliminating black and odorous is realized.
Example III
As shown in fig. 1 to 3 and fig. 14 to 18, the high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on biological microcapsules comprises capsules, biological microcapsules and porous materials, wherein the biological microcapsules are uniformly loaded in the porous materials to form porous functional materials, the capsules and the porous functional materials undergo a crosslinking reaction to form the high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on the biological microcapsules, the loading rate of the biological microcapsules on the porous functional materials is 60%, the preferable loading rate is 30%, and the mass ratio of the capsules, the biological microcapsules and the porous materials is 30:20:5, a step of;
the preparation method of the high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on the biological microcapsule comprises the following steps:
step one, preparing capsules;
(1) 3 parts of sodium alginate is thoroughly dissolved into flocculent colloid solution in 75 parts of water, 8 parts of calcium peroxide, 8 parts of diatomite and 10 parts of fine sand are slowly added after dissolution and cooling, and are stirred and mixed uniformly to form inner flocculent bodies, and the inner flocculent mixed bodies are put into 5% calcium chloride solution for crosslinking reaction to form capsules;
preparing biological microcapsules;
(1) Adding 10ml of microbial liquid seeds and 20ml of nutrient solution into 5% sodium alginate aqueous solution, dripping into coagulating bath of 5% calcium chloride mixed solution through a needle syringe at normal temperature, stabilizing for a period of time to form microcapsule embryonic form, performing secondary calcification in 5% calcium chloride solution, performing secondary complex coacervation reaction in 5% chitosan solution for 30 min, and performing soaking and proliferation culture by using 150ml of sterile physiological saline with concentration of 0.9% to form biological microcapsule;
step three, preparing a porous functional material;
(1) Taking volcanic rock with the particle size of 3mm, wherein the weight part of the volcanic rock is 50 parts, immersing the volcanic rock into a device filled with 150ml of biological microcapsules in the second step for adsorption, and forming a porous functional material with biological microcapsules;
preparing a high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on biological microcapsules;
(1) And 5 parts of sodium alginate is thoroughly dissolved in 75 parts of water, the porous functional material with biological microcapsules in the third step is added after dissolution and cooling, uniformly stirred to form floccules, the capsules in the first step are wrapped in the floccules and are put into a 5% calcium chloride solution crosslinking reagent for crosslinking reaction, and the high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on the biological microcapsules is formed.
Biological microcapsule
The microcapsule technology is a technology of wrapping trace substances in a high polymer film, is a miniature packaging technology for storing solid, liquid and gas, and mainly comprises a wrapped core material and a wrapped wall material, and is shown in the following figure. The biological microcapsule adopts a microencapsulation technology to embed or encapsulate microbial thalli such as EM bacteria, bacillus subtilis, composite bacteria and the like as core materials into wall material calcium alginate and the like of the microcapsule, so as to protect functional strains and improve the survival activity, stability and slow release property of the thalli.
The loading rate of the biological microcapsules on the geopolymer porous material is calculated as follows:
load ratio = (bio-microcapsule load)/(bio-microcapsule load + porous functional material) ×100%
Wherein, bio-microcapsule loading = bio-microcapsule total amount-bio-microcapsule remaining in solution
In the invention, the calcium peroxide of the capsule is used as an oxygenation agent to change the anaerobic environment of the sediment into an aerobic environment, and the diatomite is used as an adsorption material to effectively adsorb pollutants in the sediment and can provide a carrier for biological growth.
Microorganisms are indispensable components in water and soil environment restoration, and play an indispensable role. The microbial technology has the characteristics of high efficiency, low cost, no secondary pollution and the like in the process of repairing water pollution. Compared with the characteristic that thalli are easy to inactivate in the environment, the method adopts a microcapsule form to wrap thalli, namely the biological microcapsule is formed, so that the thalli can be effectively prevented from being damaged by external bad conditions, the stability of the thalli is improved, and the storage time of the thalli is prolonged. Some cells have a low survival rate when subjected to adverse environmental influences (ultraviolet rays, pH, etc.) in the environment. The microbial cells can be isolated from the bad environment by the microcapsule technology, so that the effect of improving the stability of the microbial cells is achieved. In the biological microcapsule in the project, the microcapsule not only provides nutrient substances for the growth of microorganisms, but also can protect the microorganisms from the external environment, thereby greatly improving the capability of the microorganisms for adapting to the environment; different from the traditional bottom mud repairing powdery medicament, the biological microcapsule loaded on the degradable porous material is not influenced by water flow and cannot run off along with the water flow; the outer wall coating also has slow release effect, and can prolong the action time of microorganisms in the microcapsule and reduce degradation time.
In this embodiment, the microbial cell seed includes EM bacteria or bacillus subtilis or a complex bacteria, and the nutrient solution is a nutrient solution suitable for growth of the EM bacteria or bacillus subtilis or complex bacteria.
In this example, the diameter of the bio-microcapsules is 0.72mm.
In this embodiment, the preferred diameter of the bio-microcapsule is 0.6mm.
In the embodiment, the mass ratio of the high-efficiency black and odorous sediment repairing agent based on the biological microcapsules to the sediment in the water body is 1:40.
In this example, in the first step, the time of the crosslinking reaction was 20 minutes.
In this example, in the second step, the stabilization time of the coagulation bath in which the needle syringe was dropped into the calcium chloride mixed solution was 30 minutes to form a microcapsule prototype.
In this example, in the third step, the volcanic rock is immersed in the device filled with the biological microcapsule of the second step for adsorption for 24 hours. In this embodiment, in the fourth step, the capsule in the first step is wrapped in the flock and placed in a crosslinking reagent for a crosslinking reaction for 20 minutes.
In this embodiment, in the third step, the diameter of the volcanic rock is 2mm.
In this example, the concentration of the sodium alginate aqueous solution in the step II is 4.9%, the dropping speed of the needle is 60 drops/min, the pH value of the calcium chloride solution is 7.2, the concentration of the chitosan solution is 4.9%, and the proliferation culture time is 4 hours.
The invention can reduce COD, total phosphorus, ammonia nitrogen, and bottom mud ammonia nitrogen concentration and sulfide in the water body.
The invention is used for sampling test
Respectively taking 300g of sediment from 2 samples of river, removing all rubbles, branches and other garbage in the acquired sediment, uniformly mixing for later use, uniformly placing the sediment in a glass jar, wherein the thickness of the sediment in each jar is 10cm, and injecting water body sewage in the same river with the depth of 20 cm. The mass ratio of the sediment to the capsules is 40:1, the capsules of the invention are put into the sediment and stirred uniformly, 4 groups of experiments are arranged, and the arrangement scheme is shown in table 1.
TABLE 3 experiment groups and different treatment modes of black and odorous substrate sludge
| Group number | Treatment mode | Ratio of addition |
| 1 | River 1 | 40：1 |
| 2 | River 2 | 40：1 |
As shown in the figure, after the novel ecological black and odorous sediment remediation agent reacts for 3 days, the concentration of pollutants in two river surges is reduced by more than 40%, the concentration of ammonia nitrogen is reduced by 50% on average, and after 7 days, the concentration indexes of pollutants in water quality can reach the V-class water level of the national surface water standard, the treatment rate of ammonia nitrogen and sulfide of the sediment reaches more than 85%, so that the effect of thoroughly eliminating black and odorous is realized.
The embodiments of the present invention have been described in detail above with reference to the accompanying drawings, but the present invention is not limited to the described embodiments. It will be apparent to those skilled in the art that various changes, modifications, substitutions and alterations can be made hereto without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (10)
1. The high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on the biological microcapsule is characterized by comprising a capsule, the biological microcapsule and a porous material, wherein the biological microcapsule is uniformly loaded in the porous material to form a porous functional material, the capsule and the porous functional material undergo a crosslinking reaction to form the high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on the biological microcapsule, the loading rate of the biological microcapsule loaded on the porous material is 5% -60%, and the mass fraction ratio of the capsule, the biological microcapsule and the porous material is 30:20:5, a step of;
the loading rate of the biological microcapsules on the porous material is calculated as follows:
load ratio = (bio-microcapsule load)/(bio-microcapsule load + porous functional material) ×100%
Wherein, bio-microcapsule loading = bio-microcapsule total amount-bio-microcapsule remaining in solution;
the preparation method of the high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on the biological microcapsule comprises the following steps:
step one, preparing capsules;
(1) Dissolving 1-3 parts of sodium alginate in 75 parts of water thoroughly to form flocculent colloid solution, slowly adding 6-8 parts of calcium peroxide, 6-8 parts of diatomite and 8-10 parts of fine sand after dissolving and cooling, stirring and uniformly mixing to form inner flocculent bodies, and putting the inner flocculent mixed bodies into 3% -5% of calcium chloride solution for crosslinking reaction to form capsules;
preparing biological microcapsules;
(1) Adding 5-10ml of microbial liquid seeds and 10-20ml of nutrient solution into 1-5% sodium alginate aqueous solution, dripping into a coagulating bath of 1-5% calcium chloride solution through a needle syringe at normal temperature, stabilizing for a period of time to form a microcapsule embryonic form, performing secondary calcification in 1-5% calcium chloride solution, performing secondary complex coacervation reaction in 0.5-5% chitosan solution for 20-30 minutes, and performing soaking and proliferation culture by using 100-150ml of sterile physiological saline with the concentration of 0.85-0.9% to form a biological microcapsule;
step three, preparing a porous functional material;
(1) Taking volcanic rock with the particle size of 1-3mm, wherein the weight part of the volcanic rock is 30-50 parts, immersing the volcanic rock into a device filled with 100-150ml of biological microcapsules in the second step for adsorption, and forming a porous functional material with biological microcapsules;
preparing a high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on biological microcapsules;
(1) 3-5 parts of sodium alginate is thoroughly dissolved in 75 parts of water, the porous functional material with biological microcapsules in the third step is added after dissolution and cooling, uniformly stirred to form floccules, the capsules in the first step are wrapped in the floccules and are put into a 3% -5% calcium chloride solution crosslinking reagent for crosslinking reaction, and the high-efficiency black and odorous water body black and odorous substrate mud repairing agent based on the biological microcapsules is formed.
2. The biological microcapsule-based efficient black and odorous water body black and odorous substrate sludge repairing agent is characterized in that the microbial liquid seeds are EM bacteria, and the nutrient solution is suitable for the growth of the EM bacteria.
3. The biological microcapsule-based efficient black and odorous water body black and odorous substrate sludge repairing agent according to claim 1, wherein the diameter of the microcapsule is 0.06-0.72mm.
4. The biological microcapsule-based efficient black and odorous water body black and odorous substrate mud repairing agent is characterized in that the mass ratio of the biological microcapsule-based efficient black and odorous water body black and odorous substrate mud repairing agent to substrate mud in a water body is 1:40.
5. The biological microcapsule-based efficient black and odorous water body black and odorous substrate sludge repairing agent is characterized in that in the first step, the time of the crosslinking reaction is 10-20 minutes.
6. The biological microcapsule-based efficient black and odorous water body black and odorous substrate sludge repairing agent is characterized in that in the second step, the stabilization time of the coagulating bath in which a needle injector is dripped into a calcium chloride mixed solution is 20-30 minutes, so as to form a microcapsule prototype.
7. The bio-microcapsule-based efficient black and odorous water body black and odorous matrix restoration agent according to claim 1, wherein in the third step, the volcanic rock is immersed in the device filled with the second step bio-microcapsule for adsorption for 16-24 hours.
8. The biological microcapsule-based efficient black and odorous water body black and odorous substrate sludge repairing agent is characterized in that in the fourth step, the capsules in the first step are wrapped in floccules, and the floccules are placed in a crosslinking reagent for crosslinking reaction for 15-20 minutes.
9. The bio-microcapsule-based efficient black and odorous water body black and odorous matrix restoration agent according to claim 1, characterized in that in the third step, the diameter of volcanic rock is 1-2mm.
10. The biological microcapsule-based efficient black and odorous water body black and odorous substrate sludge repairing agent is characterized in that the concentration of the sodium alginate aqueous solution in the step II is 3% -5%, the dropping speed of a needle head is 40-60 drops/min, the pH value of a calcium chloride solution is 6.8-7.2, the concentration of a chitosan solution is 3% -5%, and the proliferation culture time is 2-4h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111428976.5A CN114368886B (en) | 2021-11-29 | 2021-11-29 | Biological microcapsule-based efficient black and odorous water body black and odorous substrate mud repairing agent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111428976.5A CN114368886B (en) | 2021-11-29 | 2021-11-29 | Biological microcapsule-based efficient black and odorous water body black and odorous substrate mud repairing agent |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114368886A CN114368886A (en) | 2022-04-19 |
| CN114368886B true CN114368886B (en) | 2023-09-29 |
Family
ID=81138769
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111428976.5A Active CN114368886B (en) | 2021-11-29 | 2021-11-29 | Biological microcapsule-based efficient black and odorous water body black and odorous substrate mud repairing agent |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114368886B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108101327A (en) * | 2017-12-29 | 2018-06-01 | 广州市环境保护工程设计院有限公司 | A kind of black and odorous water polluted bed mud in-situ remediation method based on pelelith |
| CN108217968A (en) * | 2016-12-14 | 2018-06-29 | 王迎春 | A kind of water body processing component and its preparation method and application |
| CN112028422A (en) * | 2020-08-25 | 2020-12-04 | 佛山市绿之源环保技术有限公司 | Double-layer sediment repair capsule and application thereof |
-
2021
- 2021-11-29 CN CN202111428976.5A patent/CN114368886B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108217968A (en) * | 2016-12-14 | 2018-06-29 | 王迎春 | A kind of water body processing component and its preparation method and application |
| CN108101327A (en) * | 2017-12-29 | 2018-06-01 | 广州市环境保护工程设计院有限公司 | A kind of black and odorous water polluted bed mud in-situ remediation method based on pelelith |
| CN112028422A (en) * | 2020-08-25 | 2020-12-04 | 佛山市绿之源环保技术有限公司 | Double-layer sediment repair capsule and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114368886A (en) | 2022-04-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107488620B (en) | Cadmium-polluted rice field soil microbial passivator | |
| CN110697907B (en) | Immobilized composite flora material and preparation method thereof | |
| CN101812440B (en) | Nitrifying bacteria vector for degrading ammonia nitrogen and preparation method thereof | |
| CN111517477B (en) | Sulfate reducing bacteria embedded particles for treating ionic rare earth mine wastewater, and preparation method and application thereof | |
| CN109956563B (en) | Preparation method and application of efficient aerobic denitrification phosphorus-accumulating bacteria immobilized pellet | |
| CN112028422B (en) | Double-layer sediment repair capsule and application thereof | |
| CN109734199A (en) | Immobilized microorganism structural body and preparation method thereof | |
| CN102344899A (en) | Preparation method and application for compound fungus agent for degrading organic matter | |
| CN108017793B (en) | Preparation method of slow-release polyurethane reticular carrier and application of slow-release polyurethane reticular carrier in chemical wastewater treatment | |
| CN113999836B (en) | Composite microbial agent for sewage treatment, preparation method and application thereof in river channel treatment and black and odorous water body treatment | |
| CN106479935A (en) | A kind of preparation method of use in waste water treatment denitrification denitrogenation microbial inoculum | |
| Wang et al. | Improving denitrification efficiency in constructed wetlands integrated with immobilized bacteria under high saline conditions | |
| Liu et al. | Performances and mechanisms of microbial nitrate removal coupling sediment-based biochar and nanoscale zero-valent iron | |
| CN102465101B (en) | Denitrification bacterium preparation capable of utilizing nitrite to realize denitrification and use thereof | |
| CN109912143A (en) | A kind of original position black-odor riverway mud repair material and its potting and its application | |
| CN114368886B (en) | Biological microcapsule-based efficient black and odorous water body black and odorous substrate mud repairing agent | |
| CN103045578A (en) | Preparation method of composite bacterial agent of ammonia oxidation bacteria | |
| CN111924984A (en) | Complex microbial inoculant for treating deposited pollutants in open water area and preparation method thereof | |
| CN108975494B (en) | Method for treating black smelly water by using graphene modified straw material | |
| CN110938557A (en) | Composite microbial inoculum for degrading COD (chemical oxygen demand) in wastewater and preparation method thereof | |
| CN109592793A (en) | A kind of inorganic agent and preparation method thereof for aquaculture pollutant | |
| CN114477436A (en) | Biological treatment method for denitrifying low-concentration nitrate nitrogen wastewater | |
| CN114262128A (en) | Black and odorous inland river treatment by combination of encapsulated microorganisms and composite functional minerals | |
| CN113968626A (en) | Bottom mud repairing agent for polluted bottom mud-water interface aerobic layer and preparation and application thereof | |
| CN103011417A (en) | Method for efficiently treating ammonia nitrogen sewage at low temperature |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |