CN115403161B - Microalgae self-flocculating particle, preparation method and application thereof - Google Patents
Microalgae self-flocculating particle, preparation method and application thereof Download PDFInfo
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- C02F3/00—Biological treatment of water, waste water, or sewage
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Abstract
The invention provides microalgae self-flocculating particles, a preparation method and application thereof, and belongs to the fields of microalgae culture and rare earth tailing wastewater treatment. According to the invention, the euphorbia helioscopia and the Criman Goodyear algae are respectively inoculated in a BG11 culture medium for expansion culture; and (3) harvesting the two microalgae after the expansion culture by centrifugation, pretreating, putting a new BG11 culture medium or actual wastewater to be treated into the culture medium according to a certain proportion, and placing the culture medium on a shaking table for shake culture, and culturing for 2-4 weeks to obtain microalgae self-flocculating particles which are stable and basically not leaked. According to the invention, microalgae are cultured into the microalgae self-flocculating particles with similar or even better flocculation effect than activated sludge, so that the microalgae have excellent and stable flocculation precipitation effect, and therefore, the problems of difficult microalgae separation, high recovery cost, low microalgae density and the like are solved, a powerful technical support is provided for the application of the microalgae in sewage treatment, and the method has industrial application potential for realizing the combination of sewage treatment and microalgae culture.
Description
Technical Field
The invention relates to the field of microalgae, in particular to microalgae particles prepared by co-culture of Zostera Marina and Classification Goodyear algae, and a method for applying the same to wastewater treatment.
Background
In recent years, the concept of treating environmental pollution by adopting a biological method is accepted by the public, and the method is widely applied to treatment of sewage, soil, waste gas and the like, and good effects are obtained.
The traditional activated sludge method realizes denitrification and dephosphorization by setting different conditions such as an aerobic zone, an anoxic zone, an anaerobic zone and the like to perform nitrification and denitrification and anaerobic phosphorus release and aerobic phosphorus absorption, and has the advantages of complex process and difficult operationThe degree is large, the effect is general, and synchronous denitrification and dephosphorization are difficult to realize. Compared with the traditional activated sludge method, the microalgae has great advantage of removing nitrogen and phosphorus in sewage treatment. Mechanisms of microalgae for removing nitrogen and phosphorus include direct absorption conversion and indirect action. And part of microalgae cells can utilize various inorganic nitrogen and organic nitrogen compounds in the water body as nitrogen sources, inorganic phosphorus and organic phosphorus as phosphorus sources and carbon dioxide, carbonate and organic carbon as carbon sources, so that autotrophic or heterotrophic growth and propagation can be realized. Nitrate, nitrite and ammonium salt absorbed by algae cells can be used for synthesizing substances such as amino acid, protein and the like; the phosphorus in the water can be directly absorbed by algae cells and converted into organic matters such as ATP, phospholipids and the like through various phosphorylation pathways. And the microalgae has the advantages of high photosynthetic rate, rapid propagation, strong environmental adaptability, high treatment efficiency and the like. In addition, compared with the traditional sewage biological treatment technology, the method has the advantages of energy conservation and CO 2 The method has obvious advantages in the aspects of emission reduction and running cost; meanwhile, nutrient elements in the sewage are absorbed and utilized by the algae and are converted into biomass energy production raw materials, and the cost for culturing the algae biomass is reduced. Therefore, the coupling technology based on wastewater biological treatment and algae biomass production has wide application prospect. However, the existing microalgae wastewater treatment technology still has the problems of high microalgae separation and recovery cost, low density, poor wastewater treatment effect and the like.
Disclosure of Invention
The invention provides a self-flocculating particle microalgae formed by mixed culture of Zostera Marinae and Classification Goodyear algae, which is enabled to have biological activity and maintain stable flocculating sedimentation effect by culturing the microalgae into flocculent particles similar to activated sludge.
The invention also provides a preparation method of the microalgae self-flocculating particles.
In addition, the invention also provides application of the microalgae self-flocculating particles in sewage treatment.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a preparation method of microalgae self-flocculating particles, which comprises the following steps:
inoculating the Zostera Marina and the Cladophora claima respectively in a BG11 culture medium for expansion culture;
centrifuging two microalgae in logarithmic growth phase in the expanding culture, cleaning with sterile water for 2-3 times, and starving for later use;
adding two kinds of microalgae into a conical flask filled with a new BG11 culture medium or sewage to be treated according to a certain proportion, and placing the conical flask on a shaking table for shaking culture to obtain microalgae self-flocculating particles.
Preferably, the claiman imperial algae: the adding proportion of the zernia alga is 30:1-10:1.
Preferably, the total adding amount of the two microalgae biomass is 1-3 g/L during the shaking culture.
Preferably, the shaking speed of the shaking table is 60-180 rpm.
Preferably, the erlenmeyer flask water volume: the scale capacity is 1:5-3:5.
Preferably, the claiman imperial algae: the adding ratio of the zernia alga is 15:1, the total adding amount of the two microalgae biomass is 2g/L, the shaking table shaking speed is 130rpm, and the volume of water in the conical flask is: the scale capacity is 1:2.
The microalgae self-flocculating particles are harvested by a standing precipitation filtration mode, and the rotation speed of centrifugal separation and enrichment of the two microalgae is preferably 6000r/min.
The Clayman algae and the zernia algae are subjected to screening and domestication of sewage to be treated before inoculation.
The composition of the BG11 medium is shown in the following table:
the Clay algae is Clay algae of Clay oil surface, the Zea mays is of the genus Zea.
Preferably, the environment humidity of the shake culture is 55-80%, the illumination is LED full-day illumination, the intensity is 8000-10000 Lx, and the time is 2-28 days.
The microalgae self-flocculating particles prepared by the method have the particle size range of 0.05-1.5 mm.
The application of microalgae self-flocculating particles in sewage treatment.
The high-concentration sewage is rare earth tailing wastewater.
When the method is used for primarily treating high-concentration sewage, the adding amount of the microalgae self-flocculating particles is 1-3 g/L.
The invention provides a method for preparing microalgae self-flocculating particles applicable to rare earth tailing wastewater treatment by co-culturing of euphorbia helioscopia and claiman imperial algae, which comprises the following steps: inoculating the Zostera Marina and the Cladophora claima respectively in a BG11 culture medium for expansion culture; centrifuging two microalgae in logarithmic growth phase in the expanding culture, cleaning with sterile water for 2-3 times, and starving for later use; the two microalgae are added into a conical flask filled with new BG11 culture medium or actual wastewater (such as rare earth mine tail water) according to a proportion and placed on a shaking table for shaking culture. The shaking culture time will be determined according to the desired microalgae particle size. Harvesting the cultured microalgae particles by a standing precipitation filtration mode, and cleaning with sterile water for 3 times to obtain the microalgae particles which can be used for subsequent treatment of rare earth tailing wastewater.
The microalgae particles obtained by the method have the polyculture capability (autotrophy and heterotrophy), and can be suitable for high-concentration wastewater and low-concentration wastewater. The pollutants can be synchronously removed in the high-concentration wastewater containing substances such as COD, N, P and the like, N, P can be efficiently removed in the wastewater lacking carbon sources, and the increase of COD is limited. The method can ensure that the COD is not increased (not exceeding the standard) while the N, P in the water meets the III type requirement of the quality standard of the surface water environment after the treatment of the low-concentration wastewater with low nutrition, and provides a new prospect for the application of microalgae in sewage treatment and water quality improvement.
In the invention, the algae of the Zernia and the Cladophora can be co-cultured, and can be combined and gradually form stable microalgae particles under the action of self and water. Wherein the size of the selected mature individual of Clay algae is about 20 μm, which has the characteristic of aggregation growth to form sub-aggregate, as shown in figure 1; the mature individual of the zernia algae can reach 200-400 mu m, and the zernia algae are interwoven with each other and have certain swimming capability, as shown in figure 2, so that under the action of self extracellular secretion and external force, two algae are combined together to form the stable microalgae self-flocculating particles which take the Clayman algae as a main body and the zernia algae as a net frame, as shown in figures 3 and 4. The particle diameter of the microalgae can be controlled by regulating culture parameters, and the size of the self-flocculating particle size of the microalgae is between 0.05 and 1.5 mm. Meanwhile, the tight combination of the two microalgae also improves the tolerance degree of the microalgae to the toxicity of some pollutants, so that the strong tolerance to the high ammonia nitrogen toxicity in the rare earth tailing wastewater is verified at present, and the selected and domesticated Clay Dupu Di algae nutrition mode is mixed nutrition, so that certain organic matters in the wastewater can be directly utilized to be converted into self cell matters, including organic matters such as saccharides, organic acids, amino acids and alcohols.
The method provided by the invention realizes that two microalgae are co-cultured into the microalgae self-flocculating particles with similar activated sludge and even better flocculating and settling effects, so that the microalgae have excellent and stable flocculating and settling effects, and the problems of difficult microalgae separation, high recovery cost, low microalgae density and the like are solved. The method can couple sewage treatment with microalgae culture, so that pollutants such as carbon, nitrogen, phosphorus and the like in the sewage can be fixed and purified, carbon (sludge and CO 2) emission is reduced, valuable algal biomass can be obtained, and economic benefits are brought to sewage treatment engineering.
Drawings
FIG. 1 is a microscopic picture of Clayman diptere algae;
FIG. 2 is a microscopic photograph of Zea mays; a is a microscopic picture of a single cell, and B is a microscopic picture of an algae film formed by the zerniella;
FIG. 3 is a macroscopic picture of cultivated microalgae particles, wherein the particle size of A is about 0.5mm, and the particle size of B is about 1.2 mm;
fig. 4 is a microscopic picture of the cultivated microalgae particles, a is an optical microscopic picture, and B is a scanning electron microscope picture;
FIG. 5 is a graph showing the comparison of sedimentation rates of microalgae particles with different particle sizes in example 1;
FIG. 6 is a graph showing the ammonia nitrogen of the tail water of the rare earth ore treated by the microalgae particles in example 2 over time;
FIG. 7 is a graph showing total nitrogen, ammonia nitrogen, and total phosphorus of the microalgae particles of example 3 for treating municipal tail water over time;
FIG. 8 is a graph of TOC over time for microalgae particles treating municipal tail water in example 3;
FIG. 9 is a graph showing the COD and ammonia nitrogen of simulated wastewater from microalgae pellet treatment in example 4 over time;
FIG. 10 is a flow chart of a conventional process scheme for treating wastewater by co-culturing Zostera Marina and Classification Goodyear algae to prepare microalgae self-flocculating particles applicable to treatment of rare earth tailing wastewater.
Detailed Description
The invention provides a method for microalgae self-flocculating particles, which comprises the following steps:
inoculating the Zostera Marina and the Cladophora claima respectively in a BG11 culture medium for expansion culture;
centrifuging two microalgae in logarithmic growth phase in the expanding culture, and cleaning with sterile water for 2-3 times for starvation treatment for later use;
the two microalgae are added into a conical flask filled with a new BG11 culture medium or sewage to be treated according to a proportion and placed on a shaking table for shaking culture.
The shaking culture time will be determined according to the desired microalgae particle size.
Harvesting the cultured microalgae particles by a standing precipitation filtration mode, and cleaning with sterile water for 3 times to obtain the microalgae particles which can be used for subsequent treatment of rare earth tailing wastewater.
In the present invention, the Criman Du Di algae: the preferable adding ratio of the zernia is 30:1-10:1, and more preferable 15:1.
In the present invention, the total amount of the two microalgae biomass is preferably 1 to 3g/L, more preferably 2g/L.
In the present invention, the shaking speed of the shaking table is preferably 60 to 180rpm, more preferably 130rpm.
In the present invention, the volume of water in the conical flask: the scale volume is preferably 1:5 to 3:5, more preferably 1:2.
In the invention, the microalgae are preferably subjected to the following treatments in sequence before use: screening and domesticating for half a year, co-culturing and testing, expanding culture in a culture medium, centrifugally enriching and starving. In the present invention, the rotational speed of the centrifugal enrichment is preferably 6000r/min. The specific modes of the screening domestication, co-culture test, amplification culture and starvation treatment are not particularly limited.
In the present invention, the embodiment is preferably carried out in a conical flask on a shaking table.
In the invention, the ambient humidity of the denitrification is preferably 55-80%, the illumination is preferably LED full-day illumination, the intensity is preferably 8000-10000 Lx, and the time is determined according to the effect of the experiment.
A conventional process scheme flow chart of a method for preparing microalgae self-flocculating particles applicable to rare earth tailing wastewater treatment by co-culturing Zostera Marinae and Classification algae is shown in fig. 10, microalgae particles are inoculated into pretreated rare earth tailing wastewater, a traditional biochemical process is selected as an aid according to actual conditions, and the microalgae particles can be discharged after reaching standards, and the microalgae particles can be recycled after treatment.
For further explanation of the present invention, a method for preparing microalgae self-flocculating particles applicable to the treatment of rare earth tailing wastewater by co-culturing of Zostera Marinae and Clayman Goodyear algae is provided in the present invention in the following, but they should not be construed as limiting the scope of the present invention.
The following examples all meet the following experimental conditions, except for the specific descriptions.
The two microalgae are derived from algae films soaked on corncobs in the waste water of the rare earth tailings. Adding a proper amount of blocky corncob into the alkaline rare earth tailing wastewater, inoculating a small amount of natural water sample containing mixed algae, placing the mixture in a culture chamber for culture, and growing a layer of algae film on the surface of the corncob after a period of time. The algae film of some samples in parallel experiments can maintain the clarity and the transparency of surrounding water, and has excellent solid-liquid separation effect. The algae membrane is taken for separation and identification, and the algae membrane is confirmed to be the Zostera Marinae and the Claydiphora claima. The two algae are separated and purified respectively, and are amplified and stored in BG-11 (see Table 1) medium for standby.
Table 1 BG11 solid Medium formulation (1×)
The experiments were all performed in 500mL Erlenmeyer flasks with 300mL of initial wastewater per flask.
Other conditions: the temperature is 28 ℃; humidity is 55% -80%; illumination: the LED irradiates all the day, the intensity is 8000-10000 Lx, and the speed of the shaking table is 130rpm.
Example 1: culture contrast experiment of microalgae self-flocculation particles with different particle diameters
The experimental conditions of the two comparison groups are as follows except that the culture time is different:
according to the Criman Du Di algae: adding the spirulina with the adding proportion of 15:1 and the total microalgae biomass of 2g/L into a 500ml conical flask filled with 250ml of rare earth mine tail water, placing the conical flask on a constant-temperature shaking table for shaking culture at the shaking speed of 130rpm, wherein the environment humidity of the culture is 55-80%, the illumination is LED full-day illumination, the intensity is 8000-10000 Lx, the time is two weeks and 4 weeks respectively, 150ml of wastewater is replaced every week, after the culture is successful, static precipitation filtration is carried out, a sedimentation performance comparison test is carried out after 3 times of aseptic water washing, and the wastewater is represented by measuring the turbidity index for five times and taking the average value. As can be seen from FIG. 5, the sedimentation performance of the microalgae self-flocculating particles after four weeks of culture is very excellent, the turbidity can be reduced by 99% in 45 seconds, and 160 seconds are required for culturing the microalgae self-flocculating particles only for two weeks. The average particle diameter of 100 microalgae self-flocculating particles under the microscope reaches 1.2mm, and the average particle diameter of the latter is about 0.5 mm.
Example 2: microalgae granule treatment rare earth mine tail water experiment (the average grain diameter of the microalgae granule is about 0.5 mm)
The rare earth mine tail water is 2020.06 from a Gannan rare earth mine, and the rare earth tail water in the high water period is characterized by lower ammonia nitrogen content than in the dead water period, and also has very low COD, and the water quality is shown in Table 2.
TABLE 2 rare earth tailing wastewater quality
After the experiment group is properly supplemented with monopotassium phosphate, adding the cultured microalgae self-flocculating particles, wherein the initial algae density of the experiment is 2g/L, and the time-dependent diagram of total nitrogen and ammonia nitrogen is shown in fig. 6, so that the microalgae particles can effectively remove ammonia nitrogen and the ammonia nitrogen removal rate is 23.4 mg.L -1 ·d -1 。
The water quality after treatment is shown in Table 3. From the data in Table 3, it can be seen that the effluent quality ammonia nitrogen, COD and the like after the rare earth tailing wastewater is treated by the microalgae self-flocculation particle method all meet the emission requirements of emission standard of rare earth industrial pollutants (GB 26451-2011).
TABLE 3 Water quality after microalgae self-flocculating particle treatment
Example 3: municipal sewage treatment experiment with microalgae particles (the average particle diameter of the microalgae particles is about 0.5 mm)
The municipal tail water is 2022.08 from Yunnan university, and the requirement for the effluent is that the effluent meets the standard of surface III water and is discharged into a plateau lake (Erhai); the water quality is shown in Table 4.
Table 4 municipal tail water quality
The microalgae self-flocculating particles cultured in an equivalent amount are added into an experimental group, the initial algae amount of the experiment is 0.2g/L, fig. 8 is a time-varying graph of total nitrogen, ammonia nitrogen and total phosphorus, substances such as total nitrogen, total phosphorus and TOC in municipal tail water can be effectively removed by the microalgae particles from fig. 8 and 9, the discharge standard can be met within six days, the SBR mode experiment is carried out subsequently, water is changed in the sixth day, the water change amount is 50%, and after all water quality indexes reach the standard, water is further changed, the sample measurement shows that all indexes can be stably reduced in long-term operation, the particles can maintain the main structure for a long time, and the solid-liquid separation effect is good.
The water quality after treatment is shown in Table 5. As can be seen from the data in Table 5, the ammonia nitrogen, total nitrogen and total phosphorus in the effluent water after the microalgae particles are treated all meet the class III standard in the surface water environment quality standard (GB 3838-2002).
TABLE 5 municipal tail water quality after microalgae self-flocculating particle treatment
Example 4: microalgae granule treatment simulation wastewater experiment (the average grain diameter of the microalgae granule is about 0.3 mm)
The simulated wastewater is the leaching liquid of the corncob in the rare earth tailing wastewater, and the rare earth tailing wastewater is taken from Gannan in 2020.12 months, and the ammonia nitrogen content of the rare earth tailing wastewater is higher than that of the Gannan in the dead water period; the corncob is put into the rare earth tailing wastewater after high-temperature sterilization, the dry weight of the corncob is 20g/L, the leaching time is 3d, and the total nitrogen of ammonia nitrogen is higher than that of the corncob in the period of 6 months in 2020. And taking out the corncob after 3d, and taking supernatant after centrifuging the obtained wastewater as simulated wastewater for standby. The wastewater is characterized by higher organic matters and ammonia nitrogen, and the specific water quality is shown in the following table 6.
Table 6 simulation of wastewater quality
The microalgae self-flocculating particles cultured are added into the experimental group, the initial algae density of the experiment is 2g/L, the graph of the change of COD and ammonia nitrogen with time is shown in fig. 9, the microalgae particles can be obtained from fig. 9, ammonia nitrogen and COD can be effectively removed, wherein the ammonia nitrogen removal rate is 20.265 mg.L -1 ·d -1 COD removal rate 728.17 mg.L -1 ·d -1 The ammonia nitrogen and COD in the effluent water after the microalgae particles are treated are removed efficiently, and the treated water quality is shown in Table 7.
TABLE 7 simulation of wastewater quality after microalgae self-flocculating particle treatment
According to the 4 embodiments, the method for preparing the microalgae self-flocculating particles applicable to the treatment of the rare earth tailing wastewater by co-culturing the euphorbia helioscopia and the claiman diptere is provided, the microalgae self-flocculating particles are prepared, the microalgae are cultured into the microalgae self-flocculating particles with similar or even better flocculation effects than activated sludge, the microalgae have excellent stable flocculation precipitation effects, and the high ammonia nitrogen rare earth mine wastewater and the high COD wastewater can be rapidly, effectively and economically treated, so that the problems of difficult microalgae separation, high recovery cost, low microalgae density and the like in the microalgae rare earth tailing wastewater treatment technology are solved. And various pollutants can be survived and removed in the low-nutrition environment of the Yunnan municipal tail water, and various indexes of the effluent are reduced to meet the surface III standard, so that a powerful technical support is provided for the application of microalgae in sewage treatment. In addition, the method also has the functions of coupling sewage treatment and microalgae culture, and reducing carbon (sludge and CO) 2 ) And (3) discharging to obtain valuable algae biomass, bringing economic benefits to sewage treatment engineering and the like so as to realize the industrialized application potential of combining sewage treatment and microalgae culture.
The foregoing is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. It should be noted that modifications and adaptations to the present invention may occur to one skilled in the art without departing from the principles of the present invention and are intended to be comprehended within the scope of the present invention.
Claims (9)
1. A preparation method of microalgae self-flocculating particles comprises the following steps:
(1) Inoculating the Zostera Marina and the Cladophora claima respectively in a BG11 culture medium for expansion culture;
(2) Centrifuging two microalgae in logarithmic growth phase in the expanding culture, cleaning with sterile water for 2-3 times, and starving for later use;
(3) Adding two microalgae into a volumetric flask containing a new BG11 culture medium or sewage to be treated according to a proportion, and placing the volumetric flask on a shaking table for shaking culture to obtain microalgae self-flocculating particles;
the Clayman diptere algae: the adding proportion of the zernia alga is 30:1-10:1.
2. The preparation method of claim 1, wherein the total adding amount of the two microalgae biomass is 1-3 g/L during the shake culture, and the shake speed of a shaking table is 60-180 rpm; volume of liquid in the volumetric flask: the scale capacity is 1:5-3:5.
3. The process according to claim 2, wherein said claiman imperial algae: the adding ratio of the zernia alga is 15:1, the total adding amount of the two microalgae biomass is 2g/L, the shaking speed of the shaking table is 130rpm, and the liquid volume in the volumetric flask is: the scale capacity is 1:2.
4. The preparation method according to claim 1, wherein the microalgae self-flocculating particles are harvested by a standing precipitation filtration mode, and the rotational speed of centrifugal separation and enrichment of the two microalgae is 6000r/min.
5. The preparation method according to claim 1, wherein the Cladophora and the Zea maydis are subjected to screening and domestication of sewage to be treated before inoculation, the Cladophora is Cladophora, and the Zea maydis is the genus Cladophora.
6. The preparation method according to any one of claims 1 to 5, wherein the environmental humidity of the shaking culture is 55 to 80%, the illumination is LED full-day illumination, the intensity is 8000 to 10000Lx, and the time is 2 to 28 days.
7. The microalgae self-flocculating particle obtained by the preparation method according to any one of claims 1 to 6, wherein the particle size of the microalgae self-flocculating particle ranges from 0.05 mm to 1.5 mm.
8. Use of microalgae self-flocculating particles according to claim 7 in sewage treatment.
9. The use according to claim 8, wherein the wastewater is rare earth tailings wastewater.
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