CN111039415A - Preparation method of immobilized slow-release microorganism for in-situ restoration of water body - Google Patents
Preparation method of immobilized slow-release microorganism for in-situ restoration of water body Download PDFInfo
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- 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
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/28—Treatment of water, waste water, or sewage by sorption
- C02F1/281—Treatment of water, waste water, or sewage by sorption using inorganic sorbents
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- 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
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- 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
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/007—Contaminated open waterways, rivers, lakes or ponds
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2305/00—Use of specific compounds during water treatment
- C02F2305/06—Nutrients for stimulating the growth of microorganisms
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Abstract
The invention belongs to the technical field of water treatment, and particularly relates to a preparation method of immobilized slow-release microorganisms for in-situ restoration of a water body. Compared with the prior art, the immobilized slow-release microorganism particles prepared by the preparation method disclosed by the invention have higher stability, can purify a water body stably for a long time, prolong the service life of the immobilized slow-release microorganism and avoid secondary pollution of the water body.
Description
Technical Field
The invention belongs to the technical field of water treatment, and particularly relates to a preparation method of immobilized slow-release microorganisms for in-situ restoration of a water body.
Background
The area of 50000 multiple drainage basins in China is more than 100km2Most of the rivers are polluted to various degrees. Although the microorganism with specific functions can effectively repair the water body and improve the water quality, the microorganism is in a free stateThe ecological microbial inoculum has the problems of easy loss of thalli, large adding amount, easy influence by water conservancy conditions and the like.
The immobilized slow-release microbial technology can effectively solve the problems of free biological agents. The embedding method is a common microorganism immobilization method at present, can effectively maintain the activity of microorganisms, commonly used materials comprise polyvinyl alcohol, sodium alginate, polyethylene glycol and the like, and the novel embedding method is added with powdered activated carbon, attapulgite, calcium carbonate and the like on the basis to prepare immobilized pellets. However, due to the limitation of the material, the immobilized beads are difficult to maintain stability when facing complicated and variable river water, and are very easy to swell and even break after being soaked in the river water for a long time, and even cause secondary pollution to water bodies.
In view of the above, it is necessary to provide a method for improving the stability and the service life of the immobilized slow-release microorganism particles.
Disclosure of Invention
The invention aims to: aiming at the defects of the prior art, the preparation method of the immobilized slow-release microorganism for in-situ restoration of the water body is provided, the immobilized slow-release microorganism particles for purifying the river water have higher stability, can purify the water body stably for a long time, prolong the service life of the immobilized slow-release microorganism, and avoid the secondary pollution of the water body.
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of immobilized slow-release microorganisms for in-situ restoration of water bodies comprises the following steps:
s1, microbial adsorption: washing the molecular sieve particles, washing away floating ash on the surface, then immersing the molecular sieve particles into the prepared denitrified functional bacteria liquid for 4-7 days, and maintaining the temperature at 25 ℃;
s2, preparing a chitosan solution: preparing a glacial acetic acid solution with the volume ratio of 1%, adding chitosan with the deacetylation degree of more than or equal to 90% into the glacial acetic acid solution with the concentration of 1.5%, heating, stirring and dissolving to prepare a chitosan solution;
s3, coating chitosan: removing adsorbed microorganisms from the bacterial liquidImmersing the molecular sieve carrier in chitosan solution for coating, coating for 20min, taking out the molecular sieve carrier, and adding into 0.25mol/LNa2SO4Fixing in the solution for 20min, and cleaning with ultrapure water to obtain immobilized slow-release microorganism.
Firstly, the invention effectively removes pollutants in river water by using denitrifying functional bacteria, and takes the molecular sieve which can observe special porous structure under an electron microscope as an adsorption inner core, thereby being beneficial to adsorbing microorganisms. Secondly, the chitosan is insoluble in water and is non-toxic and harmless, and the invention adopts the chitosan coated film to stabilize the microorganisms adsorbed on the surface of the molecular sieve and achieve the slow release effect. Thirdly, the microorganisms adsorbed by the molecular sieve can continuously propagate and grow on the surface of the molecular sieve by utilizing nutrient substances in river water and are released into the water body, so that nitrogen pollutants and organic pollutants in the water body are effectively degraded. Meanwhile, the molecular sieve is used as an excellent adsorption material and can adsorb pollutants in a water body, and the pollutants are used as nutrient substances of microorganisms to ensure that the microorganisms continuously grow and reproduce on the surface of the molecular sieve carrier, so that the river water is effectively purified for a long time. In conclusion, the immobilized slow-release microorganisms prepared by the method have higher stability, can purify water stably for a long time, prolong the service life of the immobilized slow-release microorganisms and avoid secondary pollution of the water.
The improvement of the preparation method of the immobilized slow-release microorganism for in-situ restoration of the water body further comprises the step S4 of activating the immobilized slow-release microorganism after chitosan coating: before the immobilized slow-release microorganism particles are used for in-situ restoration of a water body, the immobilized slow-release microorganism particles are activated and cultured for 1 day by a denitrifying bacteria culture medium at the temperature of 25-30 ℃, so that microorganisms in the immobilized particles are fully recovered and activated.
As an improvement of the preparation method of the immobilized slow-release microorganism for in-situ remediation of water according to the present invention, in step S1, the denitrification functional bacteria are obtained by enrichment screening of microorganisms in the bottom sediment of the actual river.
As an improvement of the preparation method of the immobilized slow-release microorganism for in-situ remediation of water body, in step S1, the molecular sieve particles are spherical particles, and the particle size of the molecular sieve particles is 3-4 mm.
As an improvement of the preparation method of the immobilized slow-release microorganism for in-situ remediation of water body, before the step S2 is performed, the step S1 is repeated 3 times.
As an improvement of the preparation method of the immobilized slow-release microorganism for in-situ restoration of the water body, in step S2, chitosan is magnetically stirred in a constant-temperature water bath heating pot with the temperature of 35 ℃ at the rotating speed of 500r/min until the chitosan is completely dissolved, and the mixture is cooled to the room temperature.
As an improvement of the preparation method of the immobilized slow-release microorganism for in-situ remediation of water body, in step S3, the volume ratio of the chitosan solution to the molecular sieve carrier is greater than 2: 1.
As an improvement of the preparation method of the immobilized slow-release microorganism for in-situ restoration of the water body, in the step S3, in the film covering process, the molecular sieve carrier is stirred by a glass rod at intervals of 5 min.
As an improvement of the preparation method of the immobilized slow-release microorganism for in-situ remediation of water body, in step S4, the formulation of the culture medium for denitrifying bacteria is as follows: 6g sodium citrate, 1g (NH)4)2SO4、2gKNO3、1gK2HPO4、0.8gKH2PO4、0.2gMgSO4·7H2O, distilled water was supplemented to 1L, pH 7.0.
Compared with the prior art, the invention at least has the following beneficial effects:
(1) the preparation method can better keep the activity of the microorganism, and the microorganism is attached to the carrier for generation propagation and is slowly released in the water body to purify the river water; and the carrier also has good physical and chemical stability and strong impact resistance, and the adsorption performance of the carrier is more favorable for denitrification of river water and degradation of organic matters. Therefore, the invention has good purification effect on black and odorous water, surface water of inferior class V and class V.
(2) The method takes cheap and efficient molecular sieve particles as an immobilized carrier and is assisted by chitosan coating, so that the material cost is low and the chitosan coating is easy to obtain, and the method for immobilizing the slow-release microorganisms has the cost advantage.
(3) The materials adopted in the preparation method are nontoxic and harmless, no negative influence is brought to the environment, and the prepared immobilized slow-release microbial particles are convenient to recover and have no secondary pollution problem.
(4) The preparation method is simple and convenient, has few steps, short time consumption and strong practicability, and has the potential advantages of large-scale production and use.
Drawings
FIG. 1 is a graph showing the concentration change in test 1 according to the present invention.
FIG. 2 is a graph showing the concentration change obtained in test 2 of the present invention.
FIG. 3 is one of the concentration profiles obtained in test No. 3 of the present invention.
FIG. 4 is a second graph showing the concentration variation obtained in test 3 of the present invention.
Detailed Description
The present invention will be described in further detail with reference to the following detailed description and the accompanying drawings, but the embodiments of the invention are not limited thereto.
Example 1
And (3) adsorbing the microorganisms: washing the molecular sieve particles with ultrapure water, soaking the molecular sieve particles in the prepared denitrified functional bacteria liquid for 3 periods, wherein each period is 5 days, and the temperature is maintained at 25 ℃ for 15 days.
Preparing a chitosan solution: preparing a glacial acetic acid solution with the volume ratio of 1%, weighing a certain amount of chitosan (the deacetylation degree is more than or equal to 90%) and adding the chitosan into the glacial acetic acid solution with the volume ratio of 1% to ensure that the mass concentration of the chitosan is 1.5%, carrying out water bath at 35 ℃ for 20min, and stirring to dissolve the chitosan to prepare the chitosan solution.
Coating with chitosan: taking out the molecular sieve carrier with adsorbed microorganism from the bacteria liquid, soaking in chitosan solution for coating, after coating for 20min, taking out the carrier, adding 0.25mol/L Na2SO4In solution, fixing for 20min, and collectingAnd (4) discharging, washing with ultrapure water for 3 times, and refrigerating for standby.
Activation of the immobilized slow-release microorganisms: activating the prepared immobilized slow-release microorganism particles in a denitrification culture medium for one day, wherein the denitrification culture medium comprises the following specific formula: 6g sodium citrate, 1g (NH)4)2SO4、2gKNO3、1gK2HPO4、0.8gKH2PO4、0.2gMgSO4·7H2O, distilled water was supplemented to 1L, pH 7.0.
Performance testing
Test 1: 10g of the activated immobilized slow-release microorganisms obtained in example 1 were added to 2L of river water, and a certain amount of ammonium sulfate, potassium nitrate and sodium citrate was added to the river water to deteriorate nitrogen pollutants and COD index of the river water quality into inferior V-class simulated river water, and two sets of parallel tests were set. The test results are shown in fig. 1.
And (3) testing 2: 5g of the activated immobilized slow-release microorganisms of example 1 were added to 2L of actual river water (river water obtained from the second river of Tianjin City), and two sets of parallel tests were set. The test results are shown in fig. 2.
And (3) testing: the activated immobilized slow-release microorganisms of example 1 were put into a simulated riverway pilot plant at an amount of 2g/L, and the purification effect of the immobilized slow-release microorganisms on river water was examined using the pilot plant. The length of the water tank of the pilot plant of the simulated riverway is 1.8m, the height is 1m, the width is 0.5m, the height of the water level of the river water after the river water is injected is 0.8m, the effective volume is 0.72m3, and the river water is taken from a second-level riverway in Tianjin city. The basin bottom is equipped with the high bed mud baffle of 10cm, the thick bed mud of 8cm of tiling in the middle of two baffles, and the bed mud is got from same river course. The concentration of dissolved oxygen in the two simulated riverways is kept at 6-9mg/L, and the simulated riverway without the immobilized slow-release microorganism is set as a control group. The test time is 60 days in total, and the purification capacity of the immobilized slow-release microorganisms on river water under the condition of sufficient dissolved oxygen is investigated in the first 20 days; and (3) adding pollutants on the 21 st day, and inspecting the emergency purifying capacity of the immobilized slow-release microorganisms after the river is influenced by sudden pollution. And taking the pollutants from the sludge entering a digestion tank of a sludge plant, diluting and uniformly stirring the pollutants after sterilizing the pollutants by a high-pressure steam sterilization pot, and adding the pollutants into a simulated river channel pilot plant. The results are shown in FIGS. 3 to 4.
And (4) testing: and analyzing the propagation condition of the microorganisms in the river water added with the immobilized slow-release microorganism particles by using a flow cytometer.
Test results
As can be seen from FIG. 1, NH in simulated river water4 +-N、NO3 -Initial concentrations of-N, TN and COD were 8.12mg/L, 12.77mg/L, 22.95mg/L, 103mg/L, NH after 4 days4 +-N、NO3 -The average concentrations of-N, TN and COD are respectively reduced to 1.60mg/L, 0.17mg/L, 1.94mg/L and 21mg/L, the removal rates are respectively 80.32%, 98.66%, 91.54% and 79.61%, and the concentrations of nitrogen pollutants and COD reach the standard of surface water class V.
As can be seen from FIG. 2, river NH4 +-N、NO3 -Initial concentrations of-N, TN and COD were 3.52mg/L, 0.72mg/L, 4.70mg/L, 45mg/L, NH after 3 days4 +-N、NO3 -The concentrations of N, TN and COD are respectively reduced to 0.68mg/L, 0.07mg/L, 1.43mg/L and 6mg/L, the removal rates are respectively 80.68%, 90.04%, 65.04% and 86.67%, and nitrogen pollutants and COD indexes are promoted to IV standards of surface water from V classes of surface water.
As can be seen from FIGS. 3 to 4, the initial NH of river water in the test group4 +-N、NO3 --N、NO2 -The concentrations of-N, TN and COD were 3.00mg/L, 0.21mg/L, 0.005mg/L, 3.15mg/L and 46mg/L, respectively. After 20 days of the test, NH4 +-N、NO3 --N、NO2 -the-N, TN and COD are respectively reduced to 0.45mg/L, 0.06mg/L, 0.001mg/L, 0.61mg/L and 28mg/L, wherein NH4 +-N、NO3 -The removal rates of-N, TN and COD are 85.16%, 72.30%, 82.56% and 39.13% respectively, NO NO is generated2 --N accumulation.
In addition, as can be seen from FIGS. 3 to 4, NH of the test group was added after exogenous contamination was added4 +-N、NO3 -the-N, TN and COD are respectively increased to 3.72mg/L, 1.73mg/L, 6.32mg/L and 64mg/L,the immobilized slow-release microorganisms help the denitrification of the river water and degrade COD, and NH is added at the end of the test on day 604 +-N、NO3 -The concentration of-N, TN and COD are respectively 0.50mg/L, 0.22mg/L, 0.94mg/L and 26mg/L, and the removal rate is respectively 86.56%, 87.20%, 85.21% and 59.38%. NO2 -N is always less than 0.10 mg/L. As can be seen by comparison, the removal rate and the removal rate of the pollutants in the test group are higher than those in the control group. That is, the denitrification process of river water can be accelerated by adding the immobilized slow-release microorganisms, and the immobilized slow-release microorganisms prepared by the preparation method have better impact resistance to external pollution. In conclusion, in the process of purifying the river water for a long time, particularly after exogenous pollutants are added, the immobilized slow-release microorganisms have obvious removal effect on nitrogen and organic matters in the river water, which shows that the activity of the denitrification functional bacteria is well maintained, and the denitrification and organic matter degradation effects are continuously exerted in the river water.
Finally, the results of test 4 show that the number of living cells in the river water increased only 91.60% and the amount of growth was less than doubled after the river water purification test was carried out for 15 days. Therefore, the immobilized slow-release microbial particles slowly release thalli and have the microbial slow-release capacity.
Variations and modifications to the above-described embodiments may also occur to those skilled in the art, which fall within the scope of the invention as disclosed and taught herein. Therefore, the present invention is not limited to the above-mentioned embodiments, and any obvious improvement, replacement or modification made by those skilled in the art based on the present invention is within the protection scope of the present invention. Furthermore, although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.
Claims (9)
1. A preparation method of immobilized slow-release microorganisms for in-situ restoration of water bodies is characterized by comprising the following steps:
s1, microbial adsorption: washing the molecular sieve particles, washing away floating ash on the surface, then immersing the molecular sieve particles into the prepared denitrified functional bacteria liquid for 4-7 days, and maintaining the temperature at 25 ℃;
s2, preparing a chitosan solution: preparing a glacial acetic acid solution with the volume ratio of 1%, adding chitosan with the deacetylation degree of more than or equal to 90% into the glacial acetic acid solution with the concentration of 1.5%, heating, stirring and dissolving to prepare a chitosan solution;
s3, coating chitosan: taking out the molecular sieve carrier with adsorbed microorganism from the bacterial liquid, soaking in chitosan solution for coating, coating for 20min, taking out the molecular sieve carrier, and adding into 0.25mol/LNa2SO4Fixing in the solution for 20min, and cleaning with ultrapure water to obtain immobilized slow-release microorganism.
2. The method for preparing the immobilized slow-release microorganism for in-situ restoration of the water body according to claim 1, further comprising step S4, activation of the immobilized slow-release microorganism after chitosan coating: before the immobilized slow-release microorganism particles are used for in-situ restoration of a water body, the immobilized slow-release microorganism particles are activated and cultured for 1 day by a denitrifying bacteria culture medium at the temperature of 25-30 ℃, so that microorganisms in the immobilized particles are fully recovered and activated.
3. The method for preparing immobilized slow-release microorganisms for in situ remediation of water body as claimed in claim 1, wherein in step S1, the denitrogenating functional bacteria are selected from the microorganism enrichment in the bottom sediment of actual river channel.
4. The method for preparing the immobilized slow-release microorganism for in-situ restoration of water body according to claim 1, wherein in step S1, the molecular sieve particles are spherical particles, and the particle size of the molecular sieve particles is 3-4 mm.
5. The method for preparing immobilized slow-release microorganisms for in-situ remediation of water body according to claim 1, wherein step S1 is repeated 3 times before step S2 is performed.
6. The method for preparing the immobilized slow-release microorganism capable of being used for in-situ restoration of the water body according to claim 1, wherein in step S2, the chitosan is magnetically stirred in a constant-temperature water bath heating pot with the temperature of 35 ℃ at the rotating speed of 500r/min until the chitosan is completely dissolved, and then the mixture is cooled to room temperature.
7. The method for preparing the immobilized slow-release microorganism for in-situ restoration of water body according to claim 1, wherein in step S3, the volume ratio of the chitosan solution to the molecular sieve carrier is greater than 2: 1.
8. The method for preparing the immobilized slow-release microorganism for in-situ restoration of the water body according to claim 1, wherein in step S3, the molecular sieve carrier is stirred by a glass rod at intervals of 5min during the film covering process.
9. The method for preparing immobilized slow-release microorganisms for in situ remediation of water body as claimed in claim 2, wherein in step S4, the formulation of the culture medium for denitrifying bacteria is: 6g sodium citrate, 1g (NH)4)2SO4、2gKNO3、1gK2HPO4、0.8gKH2PO4、0.2gMgSO4·7H2O, distilled water was supplemented to 1L, pH 7.0.
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Cited By (2)
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| CN111646580A (en) * | 2020-06-17 | 2020-09-11 | 江南大学 | Preparation method and application of slow-release material for treating water pollution |
| CN112063568A (en) * | 2020-09-30 | 2020-12-11 | 天津大学 | Heterotrophic nitrification-aerobic denitrification bacterial agent and application thereof in natural water purification |
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Application publication date: 20200421 |