CN117625397A - Vermicelli wastewater treatment process based on microalgae - Google Patents
Vermicelli wastewater treatment process based on microalgae Download PDFInfo
- Publication number
- CN117625397A CN117625397A CN202311634536.4A CN202311634536A CN117625397A CN 117625397 A CN117625397 A CN 117625397A CN 202311634536 A CN202311634536 A CN 202311634536A CN 117625397 A CN117625397 A CN 117625397A
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- CN
- China
- Prior art keywords
- vermicelli
- wastewater
- chlorella
- algae
- microalgae
- 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.)
- Granted
Links
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/12—Unicellular algae; Culture media therefor
- C12N1/125—Unicellular algae isolates
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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
- C02F3/32—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae
- C02F3/322—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae use of algae
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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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Abstract
The invention relates to a vermicelli wastewater treatment process based on microalgae, and belongs to the technical field of microbial wastewater treatment. According to the invention, the chlorella (Chlorella sorokiniana) NZ1 and the Monogimplemented algae (Monogimplemented sp.) NZ4 are screened out, and analysis of experimental results shows that the biomass in the wastewater reaches the highest level when the chlorella NZ1 and the Monogimplemented algae NZ4 are mixed and cultured. Meanwhile, the removal rate of the water quality index is improved most obviously. Specifically, removal rates for COD, TN, ammonia nitrogen, and TP can be as high as 83.4%, 91.19%, 81.51%, and 78.71%, respectively. Thus, it can be concluded that: the mixed culture of algae NZ1 and NZ4 is the most effective vermicelli wastewater treatment method.
Description
Technical Field
The invention relates to a vermicelli wastewater treatment process based on microalgae, and belongs to the technical field of microbial wastewater treatment.
Background
At present, enterprises in China for producing vermicelli by using beans as raw materials have formed a huge scale. However, waste water produced in the vermicelli production process is nontoxic but has certain harmful effects. Every 1 ton of vermicelli is produced, 12-15 tons of high-concentration organic wastewater is produced, the organic wastewater is rich in various nutrients such as soluble proteins, starch, dietary fibers, oligosaccharides and the like, and meanwhile, indexes such as COD (chemical oxygen demand), SS (suspended solids) and the like are extremely high. The vermicelli wastewater discharge amount is large, the concentration is high, if the vermicelli wastewater is directly discharged, the water body is blackened and smelly, the environment is worsened, and even the underground water source is polluted. Therefore, the vermicelli industry is vigorously developed, and the corresponding wastewater treatment problem is also accompanied. Aiming at the problems of high treatment difficulty, high input cost, serious resource waste and the like in the treatment of the wastewater. There is an urgent need to find a green, economical, environment-friendly and sustainable vermicelli wastewater treatment mode.
Chinese patent document CN103508621A (application No. 201210219902.5) discloses a treatment system for vermicelli production wastewater, wherein the removal rate of COD (chemical oxygen demand) of vermicelli wastewater after anaerobic reaction reaches 90%. Chinese patent document CN110550832A (application No. 201910989225.7) discloses a system and a method for recycling corn starch wastewater, wherein the wastewater is treated to realize the complete recovery of water, carbon, nitrogen and phosphorus resources, the water resources are efficiently recovered, and the water recycling rate is more than 55%; the carbon resource is efficiently recovered, and the methane recovery rate can reach 85%; the nitrogen and phosphorus resources are efficiently recovered, the nitrogen recovery rate is more than 85 percent, and the phosphorus recovery rate is more than 82 percent. Chinese patent document CN114014505A (application No. 202111458549.1) discloses a method for treating corn starch processing wastewater by utilizing microalgae, wherein the chemical oxygen demand removal rate of the wastewater after chlorella HQ treatment can reach 91.28%; the total phosphorus removal rate can reach 78.33%; and the total nitrogen removal rate and the ammonia nitrogen removal rate respectively reach 65.51 percent and 67.33 percent.
The vermicelli waste water contains abundant soluble proteins, starch, dietary fibers, oligosaccharides and other nutrient substances, and if the substances are utilized while the waste water is treated, certain economic benefits can be generated. At present, the research on the treatment of vermicelli wastewater and further conversion into byproducts with economic value is not very much, and the produced substances are single and are mostly protein products. Chinese patent document CN110305831A (application number 201910619486. X) discloses a method for producing growth hormone protein for fish by using vermicelli waste water and application thereof, and bacterial liquid after vermicelli waste water fermentation can be mixed with feed to feed fish, and can also be directly mixed into the feed to feed fish during feed processing, thereby having obvious growth promotion effect on fish growth and obvious yield increase effect. Chinese patent document CN105859357A (application No. 201610289845.6) discloses a method for preparing microbial fertilizer by using byproducts generated in the production process of vermicelli, wherein the byproducts generated in the production process of vermicelli are used as microbial seed culture solution after wastewater treatment, the vermicelli residues and black powder are used as microbial fertilizer carriers, and valuable components such as starch, protein, amino acid and the like in the byproducts are fully utilized by microbial fermentation to realize comprehensive utilization of resources. Chinese patent document CN101302062A (application number 200810138006. X) discloses a process for treating sewage of low pollution vermicelli production technology by a natural method, wherein protein in starch production wastewater is extracted by an acid precipitation method, a compound coagulant rapid strengthening method and a protein curing recovery unit curing recovery method after vermicelli wastewater treatment, and is used for preparing protein feed.
At present, the vermicelli wastewater treatment has the problems of high treatment difficulty, high input cost, complex equipment and the like. The sewage treatment process of the low-pollution vermicelli production process disclosed in the Chinese patent document CN101302062A (application number 200810138006. X) is a sewage treatment process by a natural method, wherein vermicelli wastewater needs to enter a processor such as an anaerobic tank with an upflow anaerobic sludge blanket, an aerobic tank with an aerobic bioreactor, a biological filter, an oxidation pond, a sand filtering tank and the like after passing through a grating and settling; chinese patent document CN103508621a (application No. 201210219902.5) discloses a treatment system for vermicelli production wastewater, wherein the wastewater is discharged after passing through a first sedimentation tank, an acidification adjusting tank, a lifting water pump, an anaerobic reactor, a second sedimentation tank and an aerobic treatment tank. The treatment modes have more equipment and higher input cost. Therefore, it is important to find a green and environment-friendly sustainable vermicelli wastewater treatment mode.
Microalgae are single-cell microorganisms, can grow in various water areas, and are favored by virtue of the characteristics of short growth cycle, high light energy utilization efficiency and the like. Microalgae can grow by using carbon dioxide and nutrients (such as nitrogen and phosphorus) in water, and are therefore considered as a promising sewage treatment mode. They can grow and reproduce even in many harsh water environments, such as wastewater, brackish water, and even high salinity waters. Compared with the traditional sewage treatment method, the microalgae can remarkably improve the removal efficiency of nutrient substances, organic pollutants, heavy metals and pathogens in sewage, has a certain capability of tolerating toxic substances, and has a very wide treatment range. In addition, microalgae is rich in carbohydrate, protein, lipid and other rich biomasses, and the metabolites of the microalgae can be used for preparing various products such as animal feeds, medical supplies, foods, renewable energy sources and the like. Microalgae also have CO absorption 2 Can accumulate biomass by utilizing various nutrient substances such as nitrogen, phosphorus and the like in various waste water, and convert the biomass into protein, fatty acid and other cell components. Research has shown that the oil content of certain microalgae can even reach more than 50% of its dry weight.
Microalgae are rich in fatty acids, of which linoleic acid and alpha-linolenic acid are two very important components. Both play an indispensable role in human and animal health, but are often lacking in the daily diet, requiring supplementation urgently. It is particularly worth mentioning that linoleic acid has multiple benefits, including reducing blood lipid, softening blood vessels, lowering blood pressure, promoting microcirculation, etc., and can effectively prevent or reduce the incidence of cardiovascular diseases. Especially has great benefit for preventing and treating diseases such as hypertension, hyperlipidemia, angina pectoris, coronary heart disease, atherosclerosis, senile obesity and the like, and can effectively prevent the deposition of serum cholesterol of a human body on the wall of a blood vessel, so the blood vessel cleaner is known as a blood vessel scavenger, and has remarkable health care effect in preventing and treating atherosclerosis and cardiovascular diseases. Although the production of fatty acids using microalgae has proven to be viable at present, there are still relatively few patents related to the production of essential fatty acid oils using microalgae.
At present, microalgae have certain effects on the treatment of different waste water, such as industrial waste water, municipal waste water, agricultural waste water, livestock breeding waste water and the like, but the research on food processing waste water and the like still needs to be enhanced. Such as screening of microalgae strains, optimization of culture conditions, and the like. In the future, the application of microalgae in the vermicelli wastewater treatment field can be promoted by further intensive research and combination with modern biotechnology means, so that the efficient utilization of resources and the sustainable development of the environment are realized. Aiming at the characteristics of vermicelli wastewater, proper microalgae strains are selected for culture, so that the concentration of nutrient substances in the wastewater can be effectively reduced, the water quality is improved, and the pressure on the environment is reduced. Meanwhile, abundant biomass can be produced in the growth process of the microalgae, and the microalgae has a certain resource utilization value. The invention mainly researches the treatment of vermicelli processing wastewater by using microalgae, cultures the microalgae by using vermicelli wastewater, not only performs harmless treatment on vermicelli wastewater, but also utilizes biomass in the microalgae for recycling, thus realizing the combination of vermicelli wastewater treatment process and production process.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a vermicelli wastewater treatment process based on microalgae. According to the invention, two target algae strains, namely chlorella (Chlorella sorokiniana) NZ1 and Monognathus (Monognathus sp.) NZ4, are obtained through screening, and compared with the culture of a single algae strain, the combined culture of the two algae strains shows higher vermicelli wastewater treatment efficiency and vermicelli wastewater adaptability, and can more effectively remove nutrient substances in vermicelli wastewater.
The technical scheme of the invention is as follows:
a vermicelli waste water treatment process based on microalgae comprises the following steps:
(1) Amplifying and culturing Chlorella NZ1 and Monopodium NZ4 in BG-11 liquid culture medium to obtain Chlorella NZ1 seed solution and Monopodium NZ4 seed solution;
(2) Combining the chlorella NZ1 seed liquid and the Monopodium NZ4 seed liquid in the step (1) and then inoculating the combined chlorella NZ1 seed liquid and the Monopodium NZ4 seed liquid into vermicelli wastewater, wherein the initial inoculation amount is OD 680 =0.1-0.2, and the vermicelli wastewater is treated by light culture.
According to the present invention, in the step (1), the chlorella NZ1 is preserved in the China Center for Type Culture Collection (CCTCC) at a preservation address of 2023, 3 and 13: the China center for type culture collection of Wuhan university, eight-way Wuhan in Wuhan district, hubei province, with the preservation number: cctccc NO: m2023313.
According to the present invention, preferably, the single needle algae NZ4 in step (1) is preserved in China Center for Type Culture Collection (CCTCC) at a preservation address of 2023, 10 months and 20 days: the China center for type culture collection of Wuhan university, eight-way Wuhan in Wuhan district, hubei province, with the preservation number: cctccc NO: m20231949.
According to a preferred embodiment of the invention, the cultivation is expanded to the logarithmic phase in step (1).
According to a preferred embodiment of the present invention, the conditions for the expansion culture in the step (1) are: the temperature is 25-30deg.C, and the illumination intensity is 40-45 μmol photons.m -2 ·s -1 And (5) culturing.
According to a preferred embodiment of the invention, the BG-11 liquid medium in step (1) comprises the following per liter:
10mL of 100 XBG-11 solution, 1mL of ferric ammonium citrate solution with the concentration of 6mg/mL and Na with the concentration of 20mg/mL 2 CO 3 1mL of solution with a concentration of 30.5mg/mL K 2 HPO 4 1mL of solution, and the balance of water;
wherein the 100 XBG-11 solution comprises the following components per liter:
NaNO 3 149.6g，MgSO 4 ·7H 2 O 7.5g，CaCl 2 ·2H 2 o3.6 g, citric acid 0.6g, pH8.0,0.25 MNA 2 EDTA1.12mL, trace elements 100mL; the trace elements comprise the following components in per liter: h 3 BO 3 2.86g，ZnSO 4 ·7H 2 O 0.22g，MnCl 2 ·4H 2 O 1.81g，Na 2 MoO 4 ·2H 2 O 0.39g，CuSO 4 ·5H 2 O 0.079g，Co(NO 3 ) 2 ·6H 2 O 0.049g。
According to the invention, preferably, the volume ratio of the chlorella NZ1 seed solution and the Mononegae NZ4 seed solution in the step (2) is 1: (0.5-3); further preferably 1:1.
According to a preferred embodiment of the present invention, the initial inoculum size in step (2) is OD 680 ＝0.1。
According to the present invention, preferably, the conditions for the light culture in the step (2) are: the culture temperature is 25-30deg.C, and the illumination intensity is 40-45 μmol photons.m -2 ·s -1 Culturing for 10-15 days; further preferably, the incubation time is 12 days.
According to the invention, the water quality index of the vermicelli wastewater in the step (2) is preferably as follows: the chemical oxygen demand is 15800mg/L, the ammonia nitrogen content is 128.82mg/L, the total nitrogen content is 270.48mg/L, the total phosphorus content is 105.90mg/L, and the pH value is 3.3-3.8.
Chlorella (Chlorella sorokiniana) NZ1 was deposited at the China Center for Type Culture Collection (CCTCC) at 13/2023 at the following deposit: the China center for type culture collection of Wuhan university, eight-way Wuhan in Wuhan district, hubei province, with the preservation number: cctccc NO: m2023313.
Mononegae (monospora sp.) NZ4 was deposited at the China Center for Type Culture Collection (CCTCC) at 10 and 20 days 2023 with the deposit address: the China center for type culture collection of Wuhan university, eight-way Wuhan in Wuhan district, hubei province, with the preservation number: cctccc NO: m20231949.
The beneficial effects are that:
1. according to the invention, the chlorella (Chlorella sorokiniana) NZ1 and the Monogimplemented algae (Monogimplemented sp.) NZ4 are screened out, and analysis of experimental results shows that the biomass in the wastewater reaches the highest level when the chlorella NZ1 and the Monogimplemented algae NZ4 are mixed and cultured. Meanwhile, the removal rate of the water quality index is improved most obviously. Specifically, removal rates for COD, TN, ammonia nitrogen, and TP can be as high as 83.4%, 91.19%, 81.51%, and 78.71%, respectively. Thus, it can be concluded that: the mixed culture of algae NZ1 and NZ4 is the most effective vermicelli wastewater treatment method.
2. Mixed culture of algae strain NZ1 and NZ4 in vermicelli waste water and OD thereof 680 The chlorophyll a content and the carotenoid content reach the highest level. Specifically, at the end of the culture, OD 680 Can reach 6.664, chlorophyll a content of 25.91mg/L and carotenoid content of 8.44mg/L. In addition, the mixed culture of the algae strains NZ1 and NZ4 also obtains the maximum value of the oil yield and reaches 256.79mg/L.
3. The chlorella (Chlorella sorokiniana) NZ1 and Monoguanium sp) NZ4 mixed culture can be used for treating vermicelli wastewater and simultaneously producing grease. The production of the biomass of the chlorella (Chlorella sorokiniana) NZ1 and the Monoguanium sp) NZ4 and the purification treatment of the vermicelli wastewater are combined, so that a foundation is laid for the clean treatment and the recycling of the vermicelli wastewater, and a thought is provided for the mass production of microalgae grease.
Drawings
FIG. 1 is a microscopic image of algal strains NZ1 and NZ4.
FIG. 2 is a phylogenetic tree diagram of algal strains NZ1 and NZ4 constructed based on 18S rDNA sequences.
FIG. 3 shows the wastewater quality index removal condition of the original vermicelli wastewater under different ventilation amounts.
FIG. 4 shows the growth of different algal strains in vermicelli wastewater.
FIG. 5 shows the variation of chlorophyll a content (A) and carotenoid content (B) when different algal strain combinations are cultivated in vermicelli wastewater.
FIG. 6 is a bar graph of oil yield and oil content for different algal strain combinations cultured in vermicelli wastewater.
FIG. 7 shows a graph (A) showing the change in COD in the case of culturing different algal strains in vermicelli wastewater and a column (B) showing the removal rate of COD in the wastewater.
FIG. 8 is a graph (A) showing the change of ammonia nitrogen in wastewater and a bar graph (B) showing the removal rate of ammonia nitrogen in wastewater when different algae strains are combined and cultured in vermicelli wastewater.
FIG. 9 is a graph (A) showing the change of total nitrogen in wastewater and a histogram (B) showing the removal rate of total nitrogen in wastewater when different algal strain combinations are cultured in vermicelli wastewater.
FIG. 10 is a graph (A) showing the change of total phosphorus in wastewater and a histogram (B) showing the removal rate of total phosphorus in wastewater when different algal strain combinations are cultured in vermicelli wastewater.
Detailed Description
The technical scheme of the invention is further described below with reference to the drawings and the embodiments of the invention, but the scope of the invention is not limited thereto.
The experimental materials used in the examples were derived as follows:
the strain NZ1 and strain NZ4 were isolated and purified from Shandong Jiyu vermicelli wastewater and wastewater treatment sludge, and strain SP1 was purchased from Shanghai optical and biological technologies, inc., under the product number (GY-D19 Chlorella SP.).
The vermicelli wastewater is from pea vermicelli processing wastewater of a vermicelli factory in Qingyuan city of Kangtai of Shandong province, and is used for culturing microalgae. The water quality index of the vermicelli wastewater is as follows: the Chemical Oxygen Demand (COD) is 15800mg/L, the ammonia nitrogen content is 128.82mg/L, the Total Nitrogen (TN) content is 270.48mg/L, the Total Phosphorus (TP) content is 105.90mg/L, and the pH value is 3.3-3.8.
The corn starch wastewater is from corn starch processing wastewater of a starch factory in Shandong province and is used for culturing microalgae. The water quality index of the corn starch wastewater is as follows: the Chemical Oxygen Demand (COD) is 9093.33mg/L, the ammonia nitrogen content is 45.81mg/L, the Total Nitrogen (TN) content is 757.69mg/L, the Total Phosphorus (TP) content is 18.30mg/L, and the pH value is 3.5-4.5.
The composition of the media involved in the examples is as follows:
the BG-11 liquid medium had the following composition per liter:
100 XBG-11 solution (free of Fe, phosphate, carbonate) 10mL, ferric ammonium citrate solution at a concentration of 6mg/mL 1mL, na at a concentration of 20mg/mL 2 CO 3 1mL of solution with a concentration of 30.5mg/mL K 2 HPO 4 1mL of solution, and the balance of water.
The BG-11 solid culture medium is prepared by adding the following components in each liter based on the BG-11 liquid culture medium:
3g of sodium thiosulfate, 1mol/L NaOH solution for adjusting the pH to 8.2, 10mL of mol/L trimethylol methyl amino ethane sulfonic acid solution and 15g of agar powder.
Wherein 100 XBG-11 (free of Fe, phosphate, carbonate) comprises the following per liter components:
NaNO 3 149.6g，MgSO 4 ·7H 2 O 7.5g，CaCl 2 ·2H 2 o3.6 g, citric acid 0.6g, na 2 EDTA (pH 8.0, 0.25M) 1.12mL, trace elements 100mL; the trace elements comprise the following components in per liter: h 3 BO 3 2.86g，ZnSO 4 ·7H 2 O0.22g，MnCl 2 ·4H 2 O 1.81g，Na 2 MoO 4 ·2H 2 O 0.39g，CuSO 4 ·5H 2 O 0.079g，Co(NO 3 ) 2 ·6H 2 O 0.049g；
Ferric ammonium citrate solution (6 mg/mL,1000×): 0.6g of ferric ammonium citrate is weighed and dissolved in 100mL of ddH 2 O, storing in a refrigerator at 4 ℃ for standby.
Na 2 CO 3 Solution (20 mg/mL, 1000X): weigh 2g Na 2 CO 3 Dissolved in 100mL ddH 2 O, storing in a refrigerator at 4 ℃ for standby.
K 2 HPO 4 Solution (30.5 mg/mL, 1000X)): weigh 3.05g K 2 HPO 4 Dissolved in 100mL ddH 2 O, storing in a refrigerator at 4 ℃ for standby.
L mol/L solution of trimethylol methyl amino ethane sulfonic acid: 114.62g of trimethylolethane sulfonic acid (TES) was weighed and dissolved in 500mL of ddH 2 O, regulating the pH value to 8.2,4 ℃ by using 1mol/L NaOH solution, and storing in a refrigerator for standby.
The expansion culture of the algae strain is carried out in BG-11 medium.
Example 1: screening and identification of algae strain NZ1 and algae strain NZ4
Separating microalgae from waste water and waste water treatment sludge samples obtained from Shandong Jiyun vermicelli by adopting a plate separation technology, and performing plate culture by utilizing a BG-11 solid culture medium, wherein the method comprises the following specific steps of:
the sludge sample is preliminarily diluted by sterile water, plankton and larger suspended particles are removed by gauze filtration, and then the diluted suspension is subjected to gradient dilution by sterile water to be diluted to 10 -1 、10 -2 、10 -3 、10 -4 Adding 0.5mL of the diluted solution absorbed by each concentration to the surface of BG-11 solid medium, uniformly coating by a sterile coater, and then inverting the mixture at the temperature of 30 ℃ under the illumination of 40 mu mol photons m -2 ·s -1 Is cultured in an illumination incubator. And then adopting a plate separation technology to pick out monoclonal algae with different forms, sizes and colors, placing the monoclonal algae on a BG-11 solid culture medium, streaking, purifying and culturing, and repeating the operation for at least 3-5 times after the monoclonal algae appear again until the monoclonal algae are purified into single algae species. Inoculating single algae into 100mL conical flask containing 50mL BG-11 liquid culture medium, shake culturing on shaking table under the same culture condition to accelerate microalgae enrichment speed, observing whether microalgae morphology and appearance are purified by microscopic examination after enrichment, and simultaneously, coating algae liquid onto LB solid culture medium to verify whether mixed bacteria grow or not, and confirming whether purification is completed.
According to the size, shape, color and the like of the algae, 2 algae species are primarily purified, and are named as an algae strain NZ1 and an algae strain NZ4 respectively. Then, the culture in the BG-11 liquid culture medium is observed by a microscope, preliminary judgment and differentiation are carried out according to the shape, the color, the size and the like (figure 1), and the genome DNA of the microalgae is extracted by adopting a modified CTAB method. 18S-F with the 18S rDNA sequence primer pair: 5'-CCTGGTTGATCCTGCCAGTAG-3', 18S-R:5'-TTGATCCTTCTGCAGGTTCA-3' PCR amplification to obtain 18S rDNA sequence, purifying PCR product by 1% agarose gel electrophoresis, TA cloning, and sequencing by Qingdao qing department Biotechnology Co. Sequencing to obtain 18S rDNA sequence of algae strain NZ1 shown in SEQ ID NO.1, and 18S rDNA sequence of algae strain NZ4 shown in SEQ ID NO. 2. Sequence comparison is carried out on the sequencing result through GenBank Blast, and sequence analysis is carried out on the sequencing result by using MEGA4.0, so that a phylogenetic tree is constructed, and the phylogenetic tree is shown in figure 2.
Comprehensively analyzing the algae fall characteristics, cell morphology and 18S rDNA sequencing result of the separated and purified algae strain, and judging the species classification status of the separated and purified algae strain. Observing by microscopic examination, observing that the strain NZ1 is a spherical single-cell alga according to morphological characteristics of microalgae, wherein the size is 5-10 mu m, the strain is dark green, the pigment is obvious, and the splitting phenomenon, flagella-free and similar to the morphology of the chlorella are observed; the strain NZ4 is elliptic unicellular algae with one end being big and the other end being small, the size is 2-8 mu m, and the strain is flagella-free and is similar to elliptic chlorella. Through homology comparison of phylogenetic trees, the affinity of the algae strain NZ1 and chlorella (Chlorella sorokiniana) can be seen to be relatively close; the strain NZ4 is closely related to monospora sp.
Chlorella (Chlorella sorokiniana) NZ1 was preserved in China Center for Type Culture Collection (CCTCC) at 13/2023 with a preservation address of: the China center for type culture collection of Wuhan university, eight-way Wuhan in Wuhan district, hubei province, with the preservation number: cctccc NO: m2023313;
mononegae (Monofacium sp.) NZ4 was preserved in China Center for Type Culture Collection (CCTCC) at 10 and 20 days 2023 with the preservation addresses: the China center for type culture collection of Wuhan university, eight-way Wuhan in Wuhan district, hubei province, with the preservation number: cctccc NO: m20231949.
Example 2: influence of different ventilation on vermicelli wastewater quality index
The influence of different ventilation amounts on the water quality index of the vermicelli wastewater is studied, and the method comprises the following steps:
(1) Carrying out solid-liquid separation on vermicelli wastewater, and filtering through 8 layers of gauze to separate fine solids;
(2) 1.5L of the wastewater from the step (1) is placed in a 3L conical flask, an electromagnetic air compression pump is used for aeration, and a glass rotameter is used for controlling the flow. Two ventilation volumes were set: 1L/min and 2L/min, and simultaneously setting a non-ventilation condition as a control group; sampling every 24 hours and measuring water quality indexes such as COD, total nitrogen, ammonia nitrogen, total phosphorus and the like.
As shown in the experimental results in FIG. 3, the water quality index of the vermicelli wastewater is obviously accelerated and the difference is extremely obvious compared with that of the vermicelli wastewater subjected to stationary culture (non-aeration) under the aeration culture condition. The analytical reasons are mainly probably because under the condition of static culture (non-aeration), gas exchange only depends on the surface of liquid, so that the absorption speed of microorganisms in vermicelli wastewater on nutrient substances is relatively slow; the ventilation operation enhances the substance transmission effect between the microorganism and the wastewater, so that the gas exchange rate is obviously improved, and the growth rate of the microorganism is promoted. The removal effect of different ventilation rates on the water quality index also shows obvious difference, and the reduction rate of the water quality index is higher than that of the ventilation rate of 1L/min under the culture condition of ventilation rate of 2L/min. This is probably because the ventilation is too low, so that the contact between the microorganisms and the air is insufficient, thereby affecting the absorption process and further affecting the effect of lowering the water quality index. In addition, the water quality index is most remarkable in the condition of aeration for 3 days. However, on aeration day 4, the water quality index (ammonia nitrogen) slightly increases, possibly due to poor growth of later microorganisms, resulting in death of some microorganisms. In conclusion, under the condition of not inoculating exogenous microorganisms, the optimal ventilation amount of the vermicelli wastewater is 2L/min, and the aeration lasts for 3 days.
Example 3: influence of mixed culture of algae strains in vermicelli wastewater on microalgae and wastewater
A vermicelli waste water treatment process based on microalgae comprises the following steps:
(1) Amplifying and culturing Chlorella NZ1, monopodium NZ4 and Chlorella vulgaris SP1 in BG-11 liquid culture medium to logarithmic phase to obtain microalgae seed liquid;
(2) Inoculating the microalgae seed liquid obtained in the step (1) into vermicelli wastewater after combining, wherein the pH value is natural, and the initial inoculum size is OD 680 The combination of algae species is shown in Table 1, the cultivation temperature is 30deg.C, and the illumination intensity is 45 μmol photons.m -2 ·s -1 The ventilation is 2L/min.
TABLE 1 combination of algae species
| Algae seed combination | Microalgae seed liquid volume ratio V NZ1 ：V NZ4 ：V SP1 |
| A | 1:0:0 |
| B | 0:1:0 |
| C | 0:0:1 |
| D | 1:1:0 |
| E | 1:0:1 |
| F | 0:1:1 |
| G | 1:1:1 |
1. Influence on microalgae growth
Samples were taken every 48 hours and OD was measured 680 The growth curves of microalgae cells in single culture and mixed culture modes are shown in fig. 4. The results show that the growth curve of the microalgae is S-shaped no matter the microalgae is cultured by single algae or mixed algae, and the three microalgae can effectively utilize nutrient substances in the wastewater to realize normal growth. Under the mixed algal strain culture mode, the biomass yield is obviously higher than that of single algal strain culture. Specifically, the mixed culture of algal strain NZ1 and algal strain NZ4 exhibited the most prominent biomass yield, which is the optimal combination. Under the condition, the strain NZ1 and strain NZ4 are mixed and cultured for 12 days to obtain OD 680 The value reached 6.664, as compared with OD when the algal strains NZ1, NZ4 and SP1 were cultured alone 680 The values were increased by 44.00%, 66.93% and 54.50%, respectively. This shows that the microalgae mixed culture in vermicelli wastewater can significantly improve the biomass yield of microalgae.
2. Influence on chlorophyll a content in microalgae
And taking the algae liquid every 48 hours to determine the chlorophyll a content. The improved chlorophyll a assay method is adopted: taking 2mL of algae liquid in a 2mL centrifuge tube, centrifuging at 10,000r/min for 10min, pouring out supernatant, adding 2mL of methanol, sucking and beating, mixing uniformly, oscillating for 5min by a vortex oscillator under the condition of avoiding light, standing for 10min, centrifuging at 10,000r/min for 10min, taking supernatant for measurement, taking methanol as a reference, measuring light absorption values at wavelengths 666nm and 653nm by an ultraviolet spectrophotometer, and measuring the content of chlorophyll a according to the following formula.
C Leaves of the plant ＝15.65×OD 666 -7.34×OD 653
Wherein:
C leaves of the plant Chlorophyll a concentration, mg/L;
OD λ is the absorbance at wavelength lambda.
From the data of panel A of FIG. 5, it is understood that the chlorophyll-a content of the algal strain tends to rise as a whole throughout the cultivation. In the early growth stage, chlorophyll a increases more rapidly and finally becomes stable. After 12 days of cultivation, the chlorophyll a contents of the algae strains NZ1, NZ4 and SP1 are respectively 9.07, 1.83 and 19.64mg/L, and it can be seen that the highest chlorophyll a content of the algae strain SP1 is 19.64mg/L under a single cultivation condition; while NZ4 has the lowest chlorophyll a content of only 1.83mg/L. Under the mixed culture condition, the content of chlorophyll a in the mixed culture of NZ1 and NZ4 is highest, the stable level is reached on the 12 th day, and the content of chlorophyll a is 25.91mg/L. However, under the condition of mixed culture of other algal species, the final chlorophyll a content in mixed culture of algal strains NZ1 and NZ4 was not reached. The reason for this analysis may be that when cultured alone, no other algae compete with it, but under the condition of mixotrophic culture, other algae are sharing the same nutrient resources, which may lead to more complex allocation of resources, which is disadvantageous for SP1 to maintain the highest chlorophyll content. While the algae strains NZ1 and NZ4 may change their growth and metabolic modes during mixed culture, thereby facilitating the accumulation of chlorophyll a.
3. Effect on carotenoid content in microalgae
The algae liquid is taken every 48 hours to determine the carotenoid content. The carotenoid determination method comprises the following steps: taking 2mL of algae liquid, centrifuging at 13,000r/min for 10min, re-suspending the precipitate with 1mL of N, N-dimethylformamide solution, sucking, beating, mixing, centrifuging at 13,000r/min for 10min, taking supernatant, and respectively measuring OD by spectrophotometer 461 And OD (optical density) 664 Values and carotenoid content was calculated according to the following formula.
C Class(s) ＝(OD 461 -0.046×OD 664 )×4
Wherein:
C class(s) Is carotenoid concentration, mg/L;
OD λ is the absorbance at wavelength lambda.
From the data shown in panel B of FIG. 5, it is apparent that the carotenoid content of algal strains tends to increase as a whole. In the early growth stage, the carotenoid increases more rapidly and finally becomes stable. Under the mixed culture condition, the algae strains NZ1 and NZ4 have the highest carotenoid content, reach a stable level on the 12 th day, and reach 8.44mg/L carotenoid content. In contrast, in the case of cultivation alone, the carotenoid content increase tendencies of the algal strains NZ1, NZ4 and SP1 were approximately similar, and stabilization was achieved on day 12, with the final carotenoid contents of 3.95, 0.90 and 6.47mg/L, respectively. However, under the conditions of both mixed culture (algal strains NZ1 and SP1, and algal strains NZ4 and SP 1), the carotenoid content was within a predictable range, and was lower than the final carotenoid content at the time of mixed culture of algal strains NZ1 and NZ4. In summary, fig. 5B clearly shows that the carotenoid yield reaches the highest level when the strain NZ1 and NZ4 are mixed-cultured, which indicates that the mixed-culture of the strain NZ1 and NZ4 has a remarkable promotion effect on the carotenoid content, and also indicates that more carotenoids are produced when the strain NZ1 and NZ4 are mixed-treated with wastewater.
4. Influence on microalgae oil yield
After 12 days of culture, the algae liquid is taken to determine the content of grease. The content of the grease is determined by adopting a modified chloroform-methanol method: taking 30mL of algae liquid in a 50mL centrifuge tube, recording the volume of the algae liquid as V, putting the centrifuge tube into a centrifuge, centrifuging for 10min at 7,000r/min, and rapidly pouring out supernatant after the centrifugation is finished to obtain grease-extracted algae mud. Adding 500 mu L of 1mol/L hydrochloric acid solution into grease-extracted algae mud, shaking uniformly on a vortex oscillator, carrying out wall breaking treatment on microalgae, then adding chloroform/methanol extraction solution (V: V=2:1) into a fume hood for grease extraction, placing a centrifuge tube on a shaking table in a dark place, and carrying out shaking reaction for 4 hours at 150r/min to fully mix. After centrifugation, 10mL of physiological saline was added to the tube, and after shaking for 60 seconds on a vortex shaker, the tube was centrifuged at 7,000r/min for 10min. After centrifugation, the layers were separated, and the lowest mixed solution of chloroform/methanol and grease was drawn into a dried tin box (dry weight of tin box M) 0 ) And the tin foil box was placed in a fume hood until chloroform methanol was completely volatilized. Then the tinfoil box is put into a baking oven at 60 ℃ to be baked to constant weight, and the weight is M 1 . The oil yield was calculated from the following formula:
wherein: η is the content of grease,%; c is the oil yield, g/L; m is M 0 Mg is the mass of the dried tinfoil box; m is M 1 Mg is the total mass of the dried tin paper box and grease; v is the volume of the algae liquid measured and mL; m is the dry weight of microalgae, g/L.
FIG. 6 shows the oil and fat production of different algae species combined in wastewater. As shown in FIG. 6, the fat content of the mixed culture of the algae strain NZ1 and NZ4 is highest and reaches 43.21%, and the fat yield is 256.79mg/L; in the process of culturing other algae combinations in wastewater, the grease content is raised, but is lower than the grease yield and grease content of mixed culture of algae strains NZ1 and NZ4. Single algal strains are cultivated in the wastewater, and the grease yield of the algal strains is the lowest. In summary, the algal strains NZ1 and NZ4 are best used for grease production in mixed culture.
5. Microalgae fatty acid component analysis
The microalgae fatty acid component was analyzed by GC-MS. 50mL of the algae solution obtained after the mixed culture of the algae strain NZ1 and NZ4 for 12 days was taken, 7500 Xg was centrifuged for 10 minutes, the supernatant was discarded, the microalgae culture was harvested, the microalgae culture was stored at-80℃and then the stored microalgae culture was freeze-dried for 12 hours using a vacuum freeze dryer.
0.1g of microalgae culture is taken, freeze-dried powder is placed in a centrifuge tube, solid acid with the mass of 10% of that of the powder is added, 2mL of methanol is added, water bath is carried out for 40min at 60 ℃, then 2mL of KOH-methanol solution with the concentration of 4% is added, and the mixture is placed in a constant-temperature water bath kettle for water bath at 60 ℃ for 1 hour and then cooled. 1mL of n-hexane is added into the reacted solution, after ultrasonic crushing is carried out for 10min, 200 mu L of the upper layer solution is extracted and placed into a clean reagent bottle, and then 25 mu L of an internal standard substance (2 mg/mL, methyl heptadecanoate) is added into the reagent bottle to be detected. The fatty acid is measured by a gas chromatograph-mass spectrometer, and the full scanning mode is adopted, and the scanning range is 50-300amu. The parameters were set as follows, column: VF-23ms, adopting a temperature programming method: keeping at 150 ℃ for 1min, and then heating to 165 ℃ at 1 ℃/min; sample inlet temperature: 220 ℃.
As shown in Table 2, the content of C16-C18 fatty acids was 96.74% or more, and the long-chain hydrocarbon component was hardly contained. The saturated fatty acid (C16:0, C18:0) obtained in the invention accounts for 38.46 percent, and the content is lower; polyunsaturated fatty acids (C18:2, C18:3) account for up to 43.01%, well above the European standard limit of 12%. Therefore, the mixed culture of microalgae NZ1 and NZ4 in wastewater is not suitable for biodiesel. However, under the condition, the contents of linoleic acid and linolenic acid are respectively 27.80 percent and 15.21 percent, so that the cultivation of microalgae NZ1 and NZ4 by using vermicelli wastewater has positive significance.
TABLE 2 microalgae fatty acid component
| Fatty acid component | Name of the name | Duty cycle (%) |
| C16:0 | Palmitic acid | 34.71 |
| C18:0 | Stearic acid | 3.75 |
| C16:1 | Palmitoleic acid | 1.62 |
| C18:1 | Oleic acid | 13.65 |
| C18:2 | Linoleic acid | 27.80 |
| C18:3 | Linolenic acid | 15.21 |
| C16-C18 | / | 96.74 |
6. Effect on pollutant removal in wastewater
Sampling every two days in the culture process, sampling 20mL each time, placing the obtained sample into a 50mL centrifuge tube, centrifuging for 10min at 7,000r/min, filtering the supernatant through a filter membrane with the aperture of 0.45 μm to remove microalgae cells, and determining the water quality index of the sewage by using the filtrate. The determination method refers to "monitoring method of Water and wastewater" (national standard monitoring method of China), wherein TN content is determined by adopting potassium persulfate digestion ultraviolet spectrophotometry, NH 4 + The N content is determined by adopting a Nahner reagent spectrophotometry, the TP content is determined by adopting a potassium persulfate digestion ammonium molybdate colorimetric method, and the COD is determined by adopting a potassium dichromate method. TN, NH 4 + -N, TP and COD removal rate:
RR＝(C 0 -Ct)/C 0
wherein, RR: removal rate (%); c (C) 0 : initial concentration (mg/L); c (C) t : concentration at time t (mg/L).
1) Removal of COD in wastewater
The removal of COD in the vermicelli wastewater by the combination of different algal species can be clearly observed from FIG. 7. All the algae combinations showed a certain removal effect on COD, however, in these combinations, the mixed culture of algae strains NZ1 and NZ4 had significantly better removal effect on COD in wastewater than other algae combinations. In the first 8 days, the mixed culture of the algae strain NZ1 and NZ4 can effectively remove a considerable amount of COD, and finally the COD concentration is reduced from 15800mg/L to 2622.5mg/L, and the removal rate is as high as 83.4%. In contrast, the COD removal of the water quality in other mixed culture modes is relatively poor, the removal effect of the algae strains NZ1, NZ4 and SP1 which are independently cultured in the wastewater is not greatly different, and the final COD concentration is respectively reduced to 3860mg/L, 3847.5mg/L and 3297.5mg/L. In addition, the COD change is small at the end of the treatment, probably because microalgae die due to insufficient nutrients, resulting in a small change in the COD concentration in the wastewater.
2) For NH in wastewater 4 + Removal of N
As can be seen from fig. 8, ammonia nitrogen is continuously removed when algae strains NZ1 and NZ4 are mixed and cultured in wastewater, and the final removal rate reaches 81.51%, which is the highest value of each experimental group; the ammonia nitrogen removal rate of a single algae strain in the wastewater is slower than that of a combined algae strain in the wastewater, the algae strain starts to be stable on the 10 th day, and the ammonia nitrogen removal rates of the final algae strains NZ1, NZ4 and SP1 in the independent culture are 68.08%, 33.15% and 67.99%, respectively. Besides the mixed culture of NZ1 and NZ4, the mixed culture of other algae strains has the ammonia nitrogen removal effect within a predictable range. In conclusion, the mixed culture of the algae strains NZ1 and NZ4 has the best effect of removing ammonia nitrogen in vermicelli wastewater.
3) Removal of TN in wastewater
FIG. 9 shows the total nitrogen removal from vermicelli wastewater by different algae combinations. The result shows that the triple algae strain has certain removal capability on the total nitrogen in the vermicelli wastewater, and the total nitrogen content is wholly reduced. The total nitrogen concentration of the algae strains NZ1 and NZ4 is reduced from 270.477mg/L to 23.82mg/L, the removal rate is 91.19%, and the total nitrogen removal rate is highest and is higher than that of the algae strains NZ1 and NZ4 in single culture. In conclusion, the mixed culture of the algae strains NZ1 and NZ4 has the best effect of removing total nitrogen in vermicelli wastewater.
4) Removal of TP from wastewater
As can be seen from fig. 10, the mixed culture of algae strains NZ1 and NZ4 has the capability of removing total phosphorus in vermicelli wastewater and overall has a downward trend, wherein the TP concentration is rapidly decreased in the first 10 days, the total phosphorus removal rate is 78.71% in the first 10 days of culture, and the total phosphorus removal rate is highest and is higher than that of the algae strains NZ1 and NZ4 in single culture; mixing two algae strains, culturing in wastewater, and stably removing total phosphorus in the 12-day treatment process, wherein the total phosphorus concentration is reduced from 105.90mg/L to 22.54mg/L; from day 10, the treatment efficiency gradually decreased, probably due to insufficient nutrients in the wastewater, limiting microalgae growth and slowing the decrease in phosphorus content. In conclusion, the mixed culture of the algae strains NZ1 and NZ4 has the best effect of removing the total phosphorus in the vermicelli wastewater.
Comparative example 1: corn starch wastewater treatment by combined culture of algae strains NZ1 and NZ4
A corn starch wastewater treatment process based on microalgae comprises the following steps:
(1) Amplifying and culturing Chlorella NZ1 and Monopodium NZ4 in BG-11 liquid culture medium to logarithmic phase to obtain microalgae seed liquid;
(2) The microalgae seed liquid in the step (1) is inoculated into corn starch wastewater after being combined according to the volume ratio of 1:1, the pH value is natural, and the initial inoculum size is OD 680 =0.1, the starch wastewater was treated by light culture at 30℃and light intensity of 45. Mu. Mol photons.m -2 ·s -1 The incubation time was 12 days.
Determination of culture Start microalgae OD 680 And COD, total phosphorus, total nitrogen and ammonia nitrogen of the wastewater, and the results are shown in Table 3.
TABLE 3 Water quality index of corn starch wastewater
According to the data in Table 3, algal strains NZ1 and NZ4 were mixed and cultured in corn starch wastewater, and after 12 days of growth, their OD was measured 680 The value only reaches 1.776, which is to grow OD in the vermicelli waste water 680 26.65% of the value. This suggests that the growth of these two algal strains in corn starch wastewater is not ideal, and in contrast, they are more suitable for vermicelli wastewaterAnd (5) growing. Regarding the water quality index removal rate, the removal effect of the corn starch wastewater is far less than that of the vermicelli wastewater, and COD, total phosphorus, total nitrogen and ammonia nitrogen are only 51.35%, 37.52%, 24.09% and 29.91%, which indicates that the two algae strains are limited when the corn starch wastewater is treated, and the water quality index removal effect is more remarkable in the vermicelli wastewater. Based on the above results, we analyzed that it is possible that the vermicelli wastewater may contain more abundant nutrients such as nitrogen, phosphorus, etc., which are key elements necessary for the growth and propagation of microorganisms, helping to promote the growth rate of microalgae; secondly, environmental conditions may be more suitable for the growth of algae strains NZ1 and NZ4, vermicelli waste water may contain chemicals more friendly to algae strains NZ1 and NZ4, or there may be fewer inhibitors relative to corn starch waste water. In summary, the vermicelli wastewater may have characteristics more suitable for growth of algae strains NZ1 and NZ4 compared with corn starch wastewater, so that the algae strains NZ1 and NZ4 have better treatment effect on the vermicelli wastewater. However, specific influencing factors may need to be clarified by further experimentation and analysis.
The invention separates and purifies microalgae from waste water of Shandong Jiyu vermicelli and waste water treatment sludge to obtain algae strains NZ1 and NZ4. The two microalgae are utilized to treat vermicelli wastewater, and the algae strains NZ1 and NZ4 can be found to have the highest biomass accumulation in the vermicelli wastewater during mixed culture, and OD is at the end of culture 680 The chlorophyll a content can reach 25.91mg/L and the carotenoid content can reach 8.44mg/L when the chlorophyll a content reaches 6.664. The oil yield was the maximum (256.79 mg/L) when the algal strains NZ1 and NZ4 were cultured in a mixed manner. Meanwhile, the water quality of the wastewater is well purified, and the removal rates of COD, total nitrogen, ammonia nitrogen and total phosphorus respectively reach 83.4%, 91.19%, 81.51% and 78.71%. Compared with other reports, the method has the advantages of better treatment effect, simple treatment equipment and low cost, can produce derivative products such as grease, linoleic acid, linolenic acid, chlorophyll, carotenoid and the like, and has wide application prospect. Microalgae can also be reused for treating wastewater.
Based on the above results, we speculate that chlorella NZ1 and Monochoria NZ4 have complementary advantages in vermicelli wastewater treatment. The method has the advantages of treating different types of pollutants, and can exert the synergistic effect of the two through combined culture to improve the wastewater treatment effect; chlorella NZ1 and Monochamus NZ4 may have a strong adaptability to specific environmental conditions in vermicelli wastewater, so that they can maintain a high growth rate and bioactivity in such wastewater; metabolites of chlorella NZ1 and Monochamus NZ4 can mutually promote, so that a synergistic effect is generated in combined culture, and the treatment capacity of pollutants in wastewater is improved; the combination of different algal strains may lead to higher resource allocation efficiency, such as nutrient utilization and biomass accumulation, thereby improving the overall efficiency of wastewater treatment; the chlorella NZ1 and the single needle algae NZ4 have strong stress resistance and can keep a good growth state under the condition of relatively high-concentration wastewater, so that the wastewater is effectively treated; compared with single microalgae culture, the mixed culture can form an ecological system which is mutually promoted and mutually complemented by fusing the characteristics and advantages of different microalgae strains, thereby improving the efficiency of overall wastewater treatment and biomass yield. The effect of treating vermicelli wastewater is more excellent than other algae strain treatment methods.
The invention lays a foundation for next step of condition research for optimizing microalgae to treat vermicelli wastewater, and provides a thought for improving yield and reducing culture cost of microalgae cultured in wastewater. While the foregoing embodiments have been described in detail in connection with the embodiments of the invention, it should be understood that the foregoing embodiments are merely illustrative of the invention and are not intended to limit the invention to the particular forms disclosed, but on the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the invention.
Claims (10)
1. Chlorella (Chlorella sorokiniana) NZ1 was deposited with the China center for type culture Collection at a deposit address of 2023, 3 and 13: the China center for type culture collection of Wuhan university, eight-way Wuhan in Wuhan district, hubei province, with the preservation number: cctccc NO: m2023313.
2. Mononegae (Monofacium sp.) NZ4 was deposited at the China center for type culture Collection on 10 and 20 days of 2023 with the deposit address: the China center for type culture collection of Wuhan university, eight-way Wuhan in Wuhan district, hubei province, with the preservation number: cctccc NO: m20231949.
3. A vermicelli wastewater treatment process based on microalgae is characterized by comprising the following steps:
(1) Amplifying and culturing Chlorella NZ1 and Monopodium NZ4 in BG-11 liquid culture medium to obtain Chlorella NZ1 seed solution and Monopodium NZ4 seed solution;
(2) Combining the chlorella NZ1 seed liquid and the Monopodium NZ4 seed liquid in the step (1) and then inoculating the combined chlorella NZ1 seed liquid and the Monopodium NZ4 seed liquid into vermicelli wastewater, wherein the initial inoculation amount is OD 680 =0.1-0.2, and the vermicelli wastewater is treated by light culture.
4. The vermicelli wastewater treatment process according to claim 3, wherein in step (1), the chlorella NZ1 was deposited in the chinese collection at 2023, 3 and 13 days, and the deposit address is: the China center for type culture collection of Wuhan university, eight-way Wuhan in Wuhan district, hubei province, with the preservation number: cctccc NO: m2023313;
the single needle algae NZ4 is preserved in China center for type culture Collection (China) 10-month 20-year 2023, and the preservation address is: the China center for type culture collection of Wuhan university, eight-way Wuhan in Wuhan district, hubei province, with the preservation number: cctccc NO: m20231949.
5. A vermicelli wastewater treatment process as claimed in claim 3, characterized in that in step (1) the cultivation is expanded to logarithmic phase.
6. The vermicelli wastewater treatment process according to claim 3, wherein the conditions of the expansion culture in the step (1) are as follows: the temperature is 25-30deg.C, and the illumination intensity is 40-45 μmol photons.m -2 ·s -1 And (5) culturing.
7. The vermicelli wastewater treatment process as claimed in claim 3, wherein the BG-11 liquid medium in step (1) comprises the following components per liter:
10mL of 100 XBG-11 solution, 1mL of ferric ammonium citrate solution with the concentration of 6mg/mL and Na with the concentration of 20mg/mL 2 CO 3 1mL of solution with a concentration of 30.5mg/mL K 2 HPO 4 1mL of solution, and the balance of water;
wherein the 100 XBG-11 solution comprises the following components per liter:
NaNO 3 149.6g，MgSO 4 ·7H 2 O 7.5g，CaCl 2 ·2H 2 o3.6 g, citric acid 0.6g, pH8.0,0.25 MNA 2 EDTA1.12mL, trace elements 100mL; the trace elements comprise the following components in per liter: h 3 BO 3 2.86g，ZnSO 4 ·7H 2 O 0.22g，MnCl 2 ·4H 2 O 1.81g，Na 2 MoO 4 ·2H 2 O 0.39g，CuSO 4 ·5H 2 O 0.079g，Co(NO 3 ) 2 ·6H 2 O 0.049g。
8. The vermicelli wastewater treatment process according to claim 3, wherein the volume ratio of the chlorella NZ1 seed solution to the single needle algae NZ4 seed solution in the step (2) is 1: (0.5-3); further preferably 1:1;
preferably, the initial inoculum size in step (2) is OD 680 ＝0.1。
9. The vermicelli wastewater treatment process according to claim 3, wherein the conditions of the light culture in the step (2) are as follows: the culture temperature is 25-30deg.C, and the illumination intensity is 40-45 μmol photons.m -2 ·s -1 Culturing for 10-15 days; further preferably, the incubation time is 12 days.
10. The vermicelli wastewater treatment process as claimed in claim 3, wherein the vermicelli wastewater in the step (2) has the following water quality indexes: the chemical oxygen demand is 15800mg/L, the ammonia nitrogen content is 128.82mg/L, the total nitrogen content is 270.48mg/L, the total phosphorus content is 105.90mg/L, and the pH value is 3.3-3.8.
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