CN115403161A - Microalgae self-flocculating particle, preparation method and application thereof - Google Patents
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Abstract
The invention provides microalgae self-flocculating particles, a preparation method and application thereof, and belongs to the field of microalgae culture and rare earth tailing wastewater treatment. The invention inoculates the zeocin and the Cleminensis respectively in BG11 culture medium for propagation; and (3) centrifugally harvesting and pretreating the two types of microalgae after the propagation culture, putting the two types of microalgae into a new BG11 culture medium or actual wastewater to be treated according to a certain proportion, placing the two types of microalgae on a shaking table for shaking culture, and culturing for 2-4 weeks to obtain microalgae self-flocculating particles which exist stably and are not leaked basically. According to the invention, microalgae is cultured into microalgae self-flocculating particles with similar or even better flocculation effect similar to activated sludge, so that the microalgae has excellent and stable flocculation and precipitation effect, 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 microalgae self-flocculating particles have the industrial application potential of 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-culturing Zerewitinospora and Clementpur algae and a method for applying the same to wastewater treatment.
Background
In recent years, the concept of treating environmental pollution by biological methods is gradually accepted by the public, and the biological methods are widely applied to the treatment of sewage, soil, waste gas and the like, and achieve good effects.
In the traditional activated sludge method, nitrification and denitrification and anaerobic phosphorus release and aerobic phosphorus absorption are carried out by setting different conditions such as an aerobic zone, an anoxic zone, an anaerobic zone and the like, so that nitrogen and phosphorus removal is realized, the process is complex, the operation difficulty is high, the effect is general, and synchronous nitrogen and phosphorus removal is difficult to realize. Compared with the traditional activated sludge method, the microalgae has great advantages in removing nitrogen and phosphorus in sewage treatment. The mechanism of removing nitrogen and phosphorus by microalgae comprises direct absorption and conversion and indirect action. Part of microalgae cells can use various inorganic nitrogen and organic nitrogen compounds in water as nitrogen sources, inorganic phosphorus and organic phosphorus as phosphorus sources, and carbon dioxide, carbonate and organic carbon as carbon sources, so as to grow and reproduce in an autotrophic or heterotrophic way. 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, phospholipid and the like through various phosphorylation ways. 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 is energy-saving and CO-saving 2 The method has obvious advantages in the aspects of emission reduction and operation cost; meanwhile, nutrient elements in the sewage are absorbed and utilized by the algae and converted into biomass energy production raw materials, so that the cost for culturing the algae biomass is reduced. Therefore, the coupling technology based on the biological wastewater treatment and the algae biomass production has wide application prospect. But microalgae are currently locatedThe 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 granular microalgae formed by mixed culture of Zerewing and Cleminensis, which has biological activity and maintains stable flocculation precipitation effect by culturing the microalgae into flocculent granules similar to activated sludge.
The invention also provides a preparation method of the microalgae self-flocculating particle.
In addition, the invention also provides application of the microalgae self-flocculation 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:
respectively inoculating the zeocin and the kleima respectively in a BG11 culture medium for propagation;
centrifugally separating the two microalgae in the logarithmic growth phase in the propagation process, washing the microalgae for 2 to 3 times by using sterile water, and then carrying out starvation treatment for later use;
adding the 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, placing the conical flask on a shaking table for shaking culture, and obtaining microalgae self-flocculating particles.
Preferably, said alga cryptosporidium clerman: the adding proportion of the zerewitinosa is 30.
Preferably, the total adding amount of the two microalgae biomass during the shaking culture is 1-3 g/L.
Preferably, the shaking speed of the shaking table is 60-180 rpm.
Preferably, the flask water volume: the scale capacity is 1:5-3:5.
Preferably, said c.clerman is: the adding proportion of the zeuisticia is 15, the total adding amount of the two kinds of microalgae biomass is 2g/L, the shaking table oscillation speed is 130rpm, and the volume of water in the conical flask is as follows: the scale capacity is 1:2.
The microalgae self-flocculation particles are harvested by a standing, precipitating and filtering mode, and the rotation speed of centrifugal separation and enrichment of the two types of microalgae is preferably 6000r/min.
The Climan Dupri algae and Zephycus before inoculation are subjected to screening and domestication of sewage to be treated.
The composition of BG11 medium is shown in the following table:
the Climan Dupri algae is of genus Climan Dupri Oenoidosa, and the Zeylanica is of genus Zeylanica.
Preferably, the environmental humidity of the shaking culture is 55-80%, the illumination is LED all-day illumination, the intensity is 8000-10000 Lx, and the time is 2-28 days.
The microalgae self-flocculating granule prepared by the method has the grain size range of 0.05-1.5 mm.
The application of the microalgae self-flocculation particles in sewage treatment.
The high-concentration sewage is the rare earth tailing wastewater.
When the application is used for primarily treating high-concentration sewage, the adding amount of the microalgae self-flocculation 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 Zerewitinospora and Cleminn Dudi algae, which comprises the following steps: respectively inoculating the zeocin and the kleima respectively in a BG11 culture medium for propagation; centrifugally separating the two microalgae in the logarithmic growth phase in the propagation process, washing the microalgae for 2 to 3 times by using sterile water, and then carrying out starvation treatment for later use; adding the two kinds of microalgae into a conical flask filled with a new BG11 culture medium or actual wastewater (such as rare earth mine tail water) according to a proportion, and placing the conical flask on a shaking table for shaking culture. The shaking culture time is determined according to the size of the needed microalgae particles. And harvesting the cultured microalgae particles in a standing, precipitating and filtering mode, and cleaning for 3 times by using sterile water to be used for subsequent rare earth tailing wastewater treatment.
The microalgae particles obtained by the method have mixed culture capability (autotrophy and heterotrophy), and can adapt to high-concentration wastewater and low-concentration wastewater. The pollutants can be synchronously removed in high-concentration wastewater simultaneously containing COD, N, P and other substances, N, P can be efficiently removed in the wastewater lacking carbon sources, and the increase of COD is limited. For the oligotrophic low-concentration wastewater, N, P in water can meet the requirements of surface water environmental quality standard class III after treatment, and meanwhile, COD is not increased (not exceeding the standard), and a new prospect is provided for the application of microalgae in sewage treatment and water quality improvement.
In the invention, the Zerewitinospora and the Cleminensis Dunaliella can be co-cultured, and can be combined and gradually form stable microalgae particles under the action of self and water force. Wherein the size of the mature individual of the selected Climandi cleamana is about 20 μm, and the mature individual has the characteristic of forming a sub-assembly by aggregation and growth, as shown in figure 1; the mature Zealand algae can reach 200-400 μm, are interwoven with each other and have certain swimming capacity, as shown in figure 2, so that under the action of extracellular secretion and external force of the Zealand algae, the two kinds of algae are combined together to form the main body of the Cleman Duddick algae, and the Zealand algae is used as a net rack to gradually form stable microalgae self-flocculation particles as shown in figures 3 and 4. The diameter of the microalgae particles can be controlled by regulating and controlling culture parameters, and the size range of the self-flocculation particle size of the microalgae is between 0.05 and 1.5 mm. Meanwhile, the close combination of the two types of microalgae also improves the tolerance degree of the microalgae to the toxicity of some pollutants, the microalgae is verified to have good tolerance to the toxicity of high ammonia nitrogen in the rare earth tailing wastewater and can be effectively removed, and meanwhile, the screened and domesticated Clemenmenda dupuipiensis has mixed nutrition, and can directly convert some organic matters in the wastewater into self cell substances including organic matters such as saccharides, organic acids, amino acids, alcohols and the like.
The method provided by the invention realizes co-culture of two kinds of microalgae into microalgae self-flocculating particles with similar activated sludge and even better flocculation and precipitation effects, so that the microalgae has excellent and stable flocculation and precipitation effects, and the problems of difficult separation, high recovery cost, low microalgae density and the like of the microalgae 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 algae biomass can be obtained, and economic benefits are brought to sewage treatment engineering.
Drawings
FIG. 1 is a micrograph of Climando priciparum;
FIG. 2 is a photomicrograph of Zerewitinosa; a is a microscopic picture of a single cell, and B is a microscopic picture of an algal membrane formed by the Zerewing algae;
FIG. 3 is a macroscopic picture of cultured microalgae particles, in which 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 cultured microalgae particles, wherein A is an optical microscopic picture and B is a scanning electron microscopic picture;
FIG. 5 is a graph showing the comparison of the sedimentation rates of microalgae particles with different particle sizes in example 1;
FIG. 6 is a graph showing the change of ammonia nitrogen with time in the treatment of rare earth mine tail water by microalgae particles in example 2;
FIG. 7 is a graph showing the change of total nitrogen, ammonia nitrogen and total phosphorus with time in the treatment of municipal tail water by microalgae particles in example 3;
FIG. 8 is a graph of the TOC of the microalgae granule treated municipal tail water of example 3 as a function of time;
FIG. 9 is a graph showing the change of COD and ammonia nitrogen with time in the simulated wastewater of microalgae particle treatment in example 4;
FIG. 10 is a flow chart of a conventional process scheme for treating wastewater using a method of co-culturing Zerewitinospora and Cleveland dipteria cloremae to prepare microalgae self-flocculating granules applicable to rare earth tailings wastewater treatment.
Detailed Description
The invention provides a method for microalgae self-flocculating particles, which comprises the following steps:
respectively inoculating the zerewitinospora and the Clementopidi glaemann in BG11 culture medium for propagation;
centrifugally separating the two microalgae in the logarithmic growth phase in the propagation process, and cleaning the microalgae with sterile water for 2-3 times of hunger treatment for later use;
adding the 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 shake culture.
The shaking culture time is determined according to the required microalgae particle size.
And harvesting the cultured microalgae particles in a standing, precipitating and filtering mode, and cleaning for 3 times by using sterile water to be used for subsequent rare earth tailing wastewater treatment.
In the present invention, the alga dippersa clelmann: the preferable adding proportion of the zerewitinosa is 30-10.
In the invention, the total amount of the two microalgae biomasses is preferably 1 to 3g/L, and more preferably 2g/L.
In the present invention, the shaking speed of the shaking table is preferably 60 to 180rpm, and more preferably 130rpm.
In the present invention, the volume of water in the flask is: the scale capacity is preferably 1:5-3:5, more preferably 1:2.
In the present invention, the microalgae are preferably treated in the following order before use: the method comprises the steps of screening and domesticating for half a year, co-culture testing, expanding culture in a culture medium, centrifugal enrichment and hunger treatment. In the present invention, the rotational speed of the centrifugal enrichment is preferably 6000r/min. The specific modes of screening and domesticating, co-culture testing, amplification culture and starvation treatment are not particularly limited in the present invention.
In the present invention, the embodiments are preferably carried out in Erlenmeyer flasks on a shaker.
In the invention, the environmental humidity of denitrification is preferably 55-80%, the illumination is preferably LED all day illumination, the intensity is preferably 8000-10000 Lx, and the time is determined according to the effect of the experiment.
A flow chart of a conventional process scheme for treating wastewater by a method for preparing microalgae self-flocculating particles applicable to rare earth tailing wastewater treatment by co-culturing Zerewing algae and Cleminn Dupri algae is shown in a figure 10, the microalgae particles are inoculated into the pretreated rare earth tailing wastewater, a traditional biochemical process is selected as an auxiliary according to actual conditions, the wastewater can reach the standard and be discharged, and the treated microalgae particles can be recycled.
To further illustrate the present invention, the following examples are provided to describe in detail a method for preparing self-flocculating micro-algae particles applicable to the treatment of rare earth tailings wastewater by co-culturing Zerewitinospora and Cleveland chrysosporium, but they should not be construed as limiting the scope of the present invention.
The following examples, unless otherwise specified, all meet the following experimental conditions.
The two obtained microalgae are derived from algae membranes on corncobs soaked in the rare earth tailing wastewater. Adding a proper amount of blocky corncobs into the alkaline rare earth tailing wastewater, inoculating a small amount of natural water sample containing mixed algae, placing the natural water sample in a culture chamber for culture, and growing a layer of algae film on the surface of the corncobs after a period of time. In parallel experiments, the algae membranes of some samples can maintain the clarity and the transparency of surrounding water, and show excellent solid-liquid separation effect. The algae membrane is taken for separation and identification, and determined to be the Zerewing algae and the Cleminensis Dunaliella. The two algae were separately purified, expanded in BG-11 (see Table 1) medium and stored for further use.
TABLE 1 BG11 solid Medium formulation (1X)
The experiments were all performed in 500mL Erlenmeyer flasks with 300mL of initial wastewater per flask.
Other conditions are as follows: the temperature is 28 ℃; humidity is 55-80%; illumination: LED all day illumination, intensity 8000-10000 Lx, table speed 130rpm.
Example 1: contrast experiment for culturing self-flocculating particles of microalgae with different particle sizes
The experimental conditions of the two control groups are as follows, except for the culture time:
according to the method, the following steps are carried out: the feeding proportion of the zeotis is 15, the total amount of microalgae biomass is 2g/L, the zeotis is added into a 500ml conical flask filled with 250ml rare earth mine tail water and placed on an illumination constant temperature shaking table for oscillation culture, the shaking table oscillation speed is 130rpm, the cultured environment humidity is 55-80%, the illumination is LED all-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, the settlement performance comparison test is carried out after static precipitation filtration and sterile water washing for 3 times, and the average value is obtained by measuring five times of turbidity indexes to represent the result. Fig. 5 shows that the sedimentation performance of the microalgae self-flocculating particles cultured for four weeks is very excellent, the turbidity can be reduced by 99% in 45 seconds, and the sedimentation performance of the microalgae self-flocculating particles cultured for two weeks only needs 160 seconds. The average particle size of 100 microalgae self-flocculating particles under the microscope reaches 1.2mm, and the average particle size of the latter microalgae self-flocculating particles is about 0.5 mm.
Example 2: rare earth mine tailing water treatment experiment using microalgae particles (the average particle size of the used microalgae particles is about 0.5 mm)
The rare earth tail water is 2020.06 obtained from Gannan rare earth mine, and the rare earth tail water in the water-rich period is characterized by lower ammonia nitrogen content than in the dry period and lower COD, and the water quality is shown in Table 2.
TABLE 2 Water quality of rare earth tailings wastewater
After the experiment group appropriately supplements potassium dihydrogen phosphate, the cultured microalgae self-flocculating particles are added, the initial algae density of the experiment is 2g/L, the graph 6 is a graph showing the change of total nitrogen and ammonia nitrogen along with time, the microalgae particles obtained from the graph 6 can effectively remove the 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. As can be seen from the data in Table 3, the water quality ammonia nitrogen, COD and the like of the rare earth tailing wastewater treated by the microalgae self-flocculation particle method all meet the discharge requirements of the emission Standard of pollutants for the rare earth industry (GB 26451-2011).
TABLE 3 microalgae treated with self-flocculating particles
Example 3: experiment of treating municipal sewage with microalgae granule (average diameter of microalgae granule is about 0.5 mm)
The municipal tail water is 2022.08 which is obtained from Yunnan university, and the requirement of water outlet is that the municipal tail water meets the standard of earth surface III water and is discharged into a high-altitude lake (Erhai); the water quality is shown in Table 4.
TABLE 4 municipal tailwater quality
The same amount of cultured microalgae self-flocculating particles are added into an experimental group, the initial algae amount of the experiment is 0.2g/L, a graph of the change of total nitrogen, ammonia nitrogen and total phosphorus along with time is shown in a graph 8, substances such as total nitrogen, total phosphorus, TOC and the like in municipal tail water can be effectively removed from the microalgae particles shown in a graph 8 and a graph 9, the discharge standard can be reached in six days, an SBR mode experiment is subsequently carried out, water is changed in the sixth day, the water change amount is 50%, after two days, each water quality index reaches the standard, water is further changed, sampling and measurement shows that each index can stably descend in long-term operation, the particles can keep a main body 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 quality of ammonia nitrogen, total nitrogen and total phosphorus in the effluent after the treatment of the microalgae particles all meet the III-type standard in the environmental quality Standard for surface Water (GB 3838-2002).
TABLE 5 municipal tailwater quality after microalgae self-flocculation particle treatment
Example 4: wastewater simulation experiment for microalgae particle treatment (the average diameter of the used microalgae particles is about 0.3 mm)
The used simulated wastewater is leachate of corncobs in the rare earth tailing wastewater, 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 dry season at the time; the corncobs are placed into the rare earth tailing wastewater after high-temperature sterilization, the adding amount of the corncobs is 20g/L in dry weight, the leaching time is 3d, and the total nitrogen content of ammonia nitrogen is higher than that of the corncobs in 6 months in 2020. And 3d, taking out the corncobs, centrifuging the obtained wastewater, and taking supernatant as simulated wastewater for later use. The wastewater is characterized in that both organic matters and ammonia nitrogen are high, and the specific water quality is as shown in the following table 6.
TABLE 6 simulation of wastewater quality
The cultured microalgae self-flocculating particles are added into the experimental group, the initial algae density is 2g/L, FIG. 9 is a graph showing the change of COD and ammonia nitrogen with time, the microalgae particles can effectively remove ammonia nitrogen and COD from FIG. 9, wherein the ammonia nitrogen removal rate is 20.265 mg.L -1 ·d -1 COD removal rate of 728.17 mg.L -1 ·d -1 The ammonia nitrogen and COD in the effluent water after the treatment of the microalgae particles are efficiently removed, and the treated water quality is shown in Table 7.
TABLE 7 simulated water quality after microalgae self-flocculation particle treatment
According to the above 4 embodiments, the present invention provides a method for utilizing Zerewitinosa and Clermann dui algaeThe method for culturing and preparing the microalgae self-flocculating particles applicable to rare earth tailing wastewater treatment realizes the preparation of the microalgae self-flocculating particles, and the microalgae is cultured into the microalgae self-flocculating particles with similar or even better flocculation effect similar to activated sludge, so that the microalgae has excellent and stable flocculation and precipitation effect, and can be used for quickly, effectively and economically treating high ammonia nitrogen rare earth ore wastewater and high COD wastewater, thereby solving the problems of difficult microalgae separation, high recovery cost, low microalgae density and the like in the microalgae rare earth tailing wastewater treatment technology. And the microalgae can survive and remove various pollutants in the low-nutrient environment of the Yunnan municipal tail water, and all indexes of the discharged water are reduced to meet the standards of surface III, so that a powerful technical support is provided for the application of the microalgae in sewage treatment. In addition, the method also has the function of coupling sewage treatment with microalgae culture to reduce carbon (sludge and CO) 2 ) And discharging to obtain valuable algae biomass, and bringing economic benefits for sewage treatment engineering and the like so as to realize the industrial application potential of combining sewage treatment and microalgae culture.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention in any way. It should be noted that, for those skilled in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be construed as the protection scope of the present invention.
Claims (10)
1. A preparation method of microalgae self-flocculating particles comprises the following steps:
(1) Respectively inoculating the zerewitinospora and the Clementopidi glaemann in BG11 culture medium for propagation;
(2) Centrifugally separating the two microalgae in the logarithmic growth phase in the propagation process, washing the microalgae for 2 to 3 times by using sterile water, and then carrying out starvation treatment for later use;
(3) Adding the two kinds of microalgae into a volumetric flask filled with a new BG11 culture medium or sewage to be treated according to a certain proportion, placing the volumetric flask on a shaking table for shaking culture, and obtaining microalgae self-flocculating particles.
2. The method of claim 1, wherein said Clermann Impatientis: the addition proportion of the zerewitinosa is 30-10.
3. The preparation method according to claim 2, wherein the total adding amount of the two microalgae biomass during the shaking culture is 1-3 g/L, and the shaking speed of the shaking table is 60-180 rpm; volume of liquid in the volumetric flask: the scale capacity is 1:5-3:5.
4. The method of claim 3, wherein said Clementania cepharantha: the adding proportion of the zeuis wires is 15, the total adding amount of the two kinds of microalgae biomass is 2g/L, the shaking table oscillation speed is 130rpm, and the liquid volume in the volumetric flask is as follows: the scale capacity is 1:2.
5. The preparation method according to claim 1, wherein the microalgae self-flocculation particles are harvested by standing, sedimentation and filtration, and the rotation speed for centrifugal separation and enrichment of the two microalgae is preferably 6000r/min.
6. The preparation method according to claim 1, wherein the Clementopteria cloramae and the Zeylania before inoculation are subjected to screening and domestication of sewage to be treated, the Clementopteria cloramiana is in the genus Clementopteria oleifera, and the Zeylania is in the genus Zeylania.
7. The method according to any one of claims 1 to 6, wherein the shaking culture is performed under an environment of 55 to 80% humidity, under LED illumination for the whole day at an intensity of 8000 to 10000Lx for 2 to 28 days.
8. The microalgae self-flocculating granule obtained by the preparation method according to any one of claims 1 to 7, which has a grain size ranging from 0.05 to 1.5 mm.
9. Use of the microalgae self-flocculating granule of claim 8 in sewage treatment.
10. The use of claim 9, wherein the wastewater is rare earth tailings wastewater.
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