CN109251866B - Chlamydomonas strain and application thereof in biogas slurry purification - Google Patents
Chlamydomonas strain and application thereof in biogas slurry purification Download PDFInfo
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Abstract
The invention provides a chlamydomonas strain, which has a preservation number of: CGMCC NO. 15497. The invention also provides application of the chlamydomonas strain in biogas slurry purification. The chlamydomonas with the preservation number of CGMCC NO.15497 can grow in the pig manure biogas slurry under a proper culture condition, the pollutant ammonia nitrogen in the pig manure biogas slurry is used as a self-growing nitrogen source, and the pollutant total phosphorus in the pig manure biogas slurry is used as a self-growing phosphorus source, so that the pig manure biogas slurry is purified by nitrogen and phosphorus removal. Compared with chlamydomonas strains purchased from other commercial algae libraries, the chlamydomonas with the preservation number of CGMCC NO.15497 has obvious advantages in biomass accumulation and nitrogen and phosphorus removal effects on pig manure biogas slurry.
Description
Technical Field
The invention belongs to the field of microorganisms, and particularly relates to chlamydomonas and application thereof in biogas slurry purification.
Background
The method adopts the biogas engineering technology to treat the livestock waste, realizes pollution treatment while harvesting energy, can obtain multiple benefits such as environmental protection, economy and the like, and is an effective means for solving the problem of livestock manure pollution. However, the development of large and medium-sized biogas engineering also faces a plurality of environmental problems, and the problem of how to treat and utilize biogas slurry generated after anaerobic fermentation of livestock and poultry feces is particularly obvious. The biogas slurry produced by large and medium-sized biogas projects every day is considerable, and the biogas slurry subjected to anaerobic treatment still contains a large amount of nitrogen, phosphorus and other nutrient components, and is easy to cause eutrophication and secondary pollution when directly discharged into a water body. Meanwhile, most of large and medium-sized biogas projects are built in suburbs, local consumption of biogas slurry has certain difficulty, one-time consumption far exceeds the general rule of crop fertilization, and the problems of high energy consumption and high cost exist if the biogas slurry is transported in a long distance. Therefore, how to carry out low-cost and deep treatment on biogas slurry in situ is one of the main problems to be solved urgently in the development of large and medium-sized biogas projects.
The biogas slurry is residual liquid after anaerobic fermentation, mainly comprises organic and inorganic salts decomposed and released in the fermentation process, such as soluble substances of ammonium salt, potassium salt, phosphate and the like, and the total solid content is less than 1%. Compared with biogas residues, the nutrients in the biogas slurry are mainly quick-acting nutrients, and researches show that the livestock and poultry manure biogas slurry not only contains rich medium and trace elements such as nitrogen, phosphorus, potassium, calcium, magnesium, iron, manganese and the like, but also contains plant growth regulating substances such as indoleacetic acid, cytokinin, gibberellin and the like and bioactive components such as quinolinone, saccharides, vitamins, polyamines and the like, so that the biogas slurry is a wastewater, but if reasonably utilized, the biogas slurry is also a resource with abundant nutritional components.
The microalgae is a photoautotrophic aquatic microorganism capable of rapidly growing and propagating, has high photosynthetic efficiency, simple cell structure, strong environment adaptation capability and high utilization rate of nutrient elements in the environment, and does not occupy the cultivated land area for culturing the microalgae. The microalgae biomass can be used as a raw material for refining biofuel oil and biological natural pigments (carotenoid, astaxanthin, phycocyanin and the like), can be processed into health food for human beings, animal feed and aquatic feed, and can also be used as an organic slow-release fertilizer to be applied to soil to promote the growth of crops, so that the microalgae biomass is a microbial raw material with high added value and wide application. The artificial culture of microalgae requires a liquid culture medium containing sufficient nutrient elements such as carbon, nitrogen, phosphorus and the like, and the nutrient components in some organic wastewater are similar to those of a microalgae culture medium, so that the culture of microalgae by using wastewater quickly becomes a hotspot in the research of microalgae biotechnology. Besides a large amount of nutrient salts, microalgae culture also needs to consume a large amount of water resources, and the cost of the microalgae culture medium accounts for 30-60% of the total cost of microalgae culture. The microalgae is cultured by utilizing the biogas slurry, and can absorb nutrient elements such as carbon, nitrogen, phosphorus and the like in the biogas slurry to promote the growth of the microalgae, so that the biogas slurry is purified in the process.
One important technical difficulty in purifying livestock and poultry biogas slurry by using microalgae is to find a microalgae strain with good tolerance to specific biogas slurry, wherein the strain can grow in the specific biogas slurry within a certain concentration range, takes nitrogen and phosphorus pollutants in the biogas slurry as a nutrient source of the strain, and has tolerance to toxic and harmful substances such as ammonia nitrogen, pathogenic microorganisms, aquaculture feed drug residues and the like in the biogas slurry.
Disclosure of Invention
In order to solve the problems mentioned in the background, the wild microalgae is obtained by separating and purifying the soil environment polluted by the pig manure biogas slurry, can grow in the pig manure biogas slurry, and can absorb pollutants such as nitrogen, phosphorus and the like in the pig manure biogas slurry to serve as a nitrogen and phosphorus nutrition source of the wild microalgae so as to promote the growth of algae cells, and can play a good purification role in the pig manure biogas slurry. Meanwhile, the additive has certain tolerance on toxic and harmful substances in the pig manure biogas slurry; namely, the first object of the present invention is to propose a strain of Chlamydomonas.
The second purpose of the invention is to provide the application of the chlamydomonas strain.
The technical scheme for realizing the purpose of the invention is as follows:
a Chlamydomonas (Chlamydomonas sp.) strain with the preservation number: CGMCC NO. 15497.
The Chlamydomonas strain was deposited in China general microbiological culture Collection center (CGMCC) of the institute of microbiology, China (institute of microbiological culture Collection, China, No. 3, No.1 Hospital, North Chen, Kogyang, Beijing), at 29 months 3 and 2018.
The chlamydomonas strain is applied to biogas slurry purification.
A biogas slurry purification method adopts the Chlamydomonas strain and comprises the following steps:
inoculating a culture solution of the chlamydomonas strain into biogas slurry by an inoculation amount of 0.01-0.1 g/L of dry weight of algae seeds, wherein COD of the biogas slurry is 200-800 mg/L, ammonia nitrogen is 40-150 mg/L, total phosphorus is 10-50 mg/L, suspended solids are 0-500 mg/L, and the pH value is 7.5-8.5.
In the inoculation amount, g refers to the dry weight of the microalgae, and can be measured by a drying specific gravity method; l is the culture volume of the microalgae, the liquid component is mainly biogas slurry, and the microalgae cells can be regarded as solid particles.
Further, inoculating the culture solution of the chlamydomonas strain into the biogas slurry in an inoculation amount of 0.02-0.05 g/L of the dry weight of the algae seeds.
If the microalgae are subjected to pure culture in an artificial culture medium, the inoculation amount is generally about 0.02-0.03 g/L in order to obtain much biomass as possible. However, when the microalgae is inoculated into the biogas slurry, the inoculation amount needs to be properly increased in consideration of the impact of various complex substances in the biogas slurry on the microalgae, otherwise, if the inoculation amount is the same as that of pure culture, the microalgae is likely to die without tolerating the biogas slurry. The optimized value determined after multiple experiments is about 0.03-0.05 g/L.
Wherein the culture solution of the chlamydomonas strain is obtained by culturing the chlamydomonas strain in a liquid culture medium to logarithmic growth phase; the inoculation amount is 0.02-0.025 g/L of liquid culture medium; and/or the liquid culture medium is BG 11.
Wherein the conditions for culturing the chlamydomonas strain in the liquid culture medium are as follows: the intensity of irradiation is 100 to 300. mu. mol/m2The photoperiod is the ratio of light to dark (12-18): 6-12), and the temperature is 15-30 ℃.
Preferably, the conditions for culturing the chlamydomonas strain in the liquid culture medium are as follows: the illumination intensity is 170-190 mu mol/square meter/s, the light-dark ratio is 18:6, and the temperature is 26 +/-0.5 ℃.
Wherein when the chlamydomonas strain is cultured in the liquid culture medium, the culture container is shaken 2-4 times every day.
One preferable technical scheme of the invention is that the method comprises the following steps:
1) pretreating the pig manure biogas slurry to reach the conditions of COD (chemical oxygen demand) 200-800 mg/L, ammonia nitrogen 40-150 mg/L, total phosphorus 10-50 mg/L, suspended solid 0-500 mg/L and pH value 7.5-8.5;
2) adding a culture solution of the chlamydomonas strain for growth, wherein the growth conditions are as follows: the illumination intensity is 100-200 mu mol/square meter/s, and the light cycle is the light-dark ratio (12-20): (4-12) the temperature is 24-28 ℃.
The pretreatment is flocculation and air flotation treatment, and the pig manure biogas slurry is pretreated to reach the conditions of COD (chemical oxygen demand) 300-450 mg/L, ammonia nitrogen 840-120 mg/L, total phosphorus 20-30 mg/L, suspended solids 0-300 mg/L and pH value 8.0-8.5.
The flocculation and air flotation treatment can adopt the treatment mode existing in the field.
The invention has the beneficial effects that:
the chlamydomonas with the preservation number of CGMCC NO.15497 can grow in the pig manure biogas slurry under a proper culture condition, the pollutant ammonia nitrogen in the pig manure biogas slurry is used as a self-growing nitrogen source, and the pollutant total phosphorus in the pig manure biogas slurry is used as a self-growing phosphorus source, so that the pig manure biogas slurry is purified by nitrogen and phosphorus removal. Compared with chlamydomonas strains purchased from other commercial algae libraries, the chlamydomonas with the preservation number of CGMCC NO.15497 has obvious advantages in biomass accumulation and nitrogen and phosphorus removal effects on pig manure biogas slurry.
Drawings
FIG. 1 shows the cell morphology (10 times of ocular lens x 40 times of objective lens) of Chlamydomonas strain CGMCC NO.15497 under an optical microscope.
FIG. 2 shows a phylogenetic tree constructed based on Chlamydomonas strain CGMCC NO.15497 in the present invention.
FIG. 3 is a comparison graph of the growth of Chlamydomonas strain CGMCC NO.15497 separated and purified by the invention and the growth of other two Chlamydomonas reinhardtii strain FACHB-265 and Chlamydomonas strain FACHB-1935 randomly purchased from commercial algae seed bank in pig manure biogas slurry within 20 days.
FIG. 4 shows the comparison of Chlamydomonas strain CGMCC NO.15497 obtained by separation and purification of the invention and the removal of ammonia nitrogen in pig manure biogas slurry within 20 days by two other strains of Chlamydomonas reinhardtii strain FACHB-265 and Chlamydomonas strain FACHB-1935 randomly purchased from commercial algae seed banks.
FIG. 5 is a comparison of Chlamydomonas strain CGMCC NO.15497 separated and purified by the present invention and another two randomly purchased Chlamydomonas reinhardtii strain FACHB-265 and Chlamydomonas strain FACHB-1935 in pig manure biogas slurry for removing total phosphorus within 20 days.
Detailed Description
The following detailed description of embodiments of the present invention is provided in connection with the accompanying drawings and examples.
It will be appreciated by those skilled in the art that the following examples are illustrative only and are not intended to limit the scope of the present invention.
Example 1: separation, purification and preservation of algal strains
(1) Sample collection
The algae strain is obtained by sampling and separating the soil polluted by biogas slurry through pig manure anaerobic digestion in a biogas service station of Daxing village, town, west cypress shop in the Pinggu district of Beijing. Collecting soil sample from soil surface layer of selected place, placing in sealed bag, and storing in refrigerator at 4 deg.C for use.
(2) Algae strain separation, purification and algae strain culture
Taking about 0.5g of soil sample in a 24-well tissue culture dish, adding BG11 liquid culture medium (the specific composition of the BG11 culture medium is shown in Table 1) and placing the culture medium in an illumination incubator with adjustable environmental parameters for algae seed enrichment culture, wherein the color of the culture medium can be obviously changed from colorless to light green after 3-4 days of culture. Then separating and purifying by using a plate-scribing method on a BG11 solid plate culture medium (prepared by adding 1.5-2% of agar powder into a sterilized BG11 liquid culture medium) until algae colonies in a plate are single, then selecting a sterile single algae colony, inoculating the single algae colony into a liquid BG11 culture medium, and placing the single algae colony in an illumination incubator for culture. And observing whether the cell forms of the cultured microalgae strains are consistent by using an optical microscope, if so, achieving the purpose of separation, and if not, repeating the plate scribing work until the cell forms are single.
The culture container used in the algae strain culture stage is 100mL conical flask (effective culture volume is 50mL) and BG11 liquid culture medium, and the culture conditions are 28 + -0.5 deg.C, and illumination intensity is 150 μmol/m2The lighting period is 12:12, artificial work is carried out every dayShake flasks three times.
TABLE 1 BG11 Medium formulation and amounts
Identification of algal strains
The identification of the algae strains is carried out in two steps, firstly, the morphological initial observation is carried out, and then, the molecular biology identification is carried out. The isolated and purified microalgae strains were observed using an optical microscope and photographed (10 times eyepiece × 40 times objective lens). The cell morphology, size, structure and other characteristics of the strain were observed. The morphology of the algal strains observed by an optical microscope is shown in FIG. 1.
The biological characteristics of the microalgae obtained by separation and purification of the invention observed under a microscope are that the microalgae body is green and has single cells, the cells are spherical or oval, and the front end of the microalgae body is provided with two flagella with equal length and can swim.
The microalgae were subjected to molecular biological identification, using the extracted DNA as a template, and DNAMAN and Primer 5.0 software were used to design primers (Primer sequence forward 5'-TCCGTAGGTGAACCTGCGG-3', reverse 5'-TCCTCCGCTTATTGATATGC-3'). The ITS gene is subjected to PCR amplification, and the PCR reaction system is set up to 50 mu L. Wherein the DNA template is 1 mu L, the forward primer and the reverse primer are 1 mu L respectively, the dNTP4 mu L, the reaction buffer solution of 5 XQ 5 is 10 mu L, the Q5DNA polymerase is 0.5 mu L, and the High GC Enhancer is 5 XQ 5. mu.L. The ITS gene PCR amplification program is as follows: pre-denaturation at 94 ℃ for 30s, followed by denaturation at 98 ℃ for 5s, annealing at 52 ℃ for 30s, and extension at 68 ℃ for 75s for 30 cycles, and finally extension at 68 ℃ for 5 min. The procedure commissioned sequencing by the Biotech company, Inc., of Epischiuron, Beijing. The obtained sequence was subjected to homologous detection in GenBank database (http:// www.ncbi.nlm.nih.gov /) by BLAST, and compared with the existing algal species genes in the algal species library, and the species was determined. After the algal strain is subjected to PCR amplification, an ITS fragment is obtained. The ITS fragment obtained after sequencing has a length of 694 bp. The sequence was analyzed for homology with the NCBI database, and it was found that its affinity with Chlamydomonas sp was recent and the homology reached 99%. Phylogenetic trees were constructed using MEGA5.10 for the ITS sequences, and the results are shown in FIG. 2.
Preservation of algal strains
The chlamydomonas strain obtained and identified by separation and purification is preserved in China general microbiological culture Collection center (CGMCC) (No. 3 of Xilu No.1 of Beijing Kogyo Beicheng province) of the institute of microbiology of China academy of sciences at 29 months and 3 years in 2018, and the preservation number is CGMCC NO. 15497.
Example 2 study of optimum culture conditions for algal strains
After the microalgae species with sufficient biomass is obtained, in order to explore the suitable culture conditions, the invention carries out a series of optimization researches such as single factor test-orthogonal test-response surface test and the like on four common technical parameters of microalgae culture such as illumination intensity, photoperiod, temperature and inoculation amount, and the set parameters are the illumination intensity: 100. 150, 200, 250 and 300 (mu mol/square meter/s), photoperiod (ratio of light to dark): 12/12, 14/10, 16/8, 18/6; temperature: 15 ℃, 20 ℃, 25 ℃ and 30 ℃; pure culture inoculum size (algal seed culture medium culture): 0.015g/L, 0.020g/L, 0.025g/L, 0.030g/L, and most preferably 0.025 g/L. In the same test mode, the biogas slurry culture inoculum size (culture of algae for purifying pig manure biogas slurry) is set: 0.020g/L, 0.030g/L, 0.040g/L and 0.050g/L, and the optimal inoculation amount is 0.041g/L through tests.
The optimal culture conditions of the microalgae obtained finally are that the illumination intensity is 180 mu mol/square meter/s, the photoperiod is 18:6 (light: dark), the temperature is 26 +/-0.5 ℃, and the inoculation amount is 0.022 g/L. The culture container used in this stage is a 250mL conical flask (effective culture volume is 150mL), BG11 liquid culture medium, placed in an illumination incubator with adjustable environmental parameters, and manually shaken three times a day.
In order to explore the potential of the chlamydomonas strain CGMCC NO.15497 in purifying the pig manure biogas slurry, the invention develops experimental research of culturing the chlamydomonas strain CGMCC NO.15497 by using real pig manure biogas slurry. In this experimental study, in addition to the chlamydomonas CGMCC No.15497 strain of the present invention, the ability of two other chlamydomonas strains (comparative example 1, comparative example 2) randomly selected from commercial algae seed banks to purify pig manure biogas slurry and the growth conditions thereof in pig manure biogas slurry were studied for comparison with the strain of the present invention.
In example 3 and comparative example, 6mL of culture solution per day was sampled to determine biomass and water quality. Three replicates were set for all experimental groups.
Example 3: purification of pig manure biogas slurry by using chlamydomonas strain CGMCC NO.15497
(1) Test materials
The pig manure biogas slurry used in the experiment is taken from a biogas slurry storage tank of a biogas service station in the Xingzhuangzhen west cypress village of the Changgu district in Beijing, because the original biogas slurry in the storage tank contains ammonia nitrogen with higher concentration (more than 900mg/L) and a large amount of suspended solids (700 + 800 mg/L), the original biogas slurry is subjected to flocculation-air flotation pretreatment, so that the ammonia nitrogen concentration and the suspended solids concentration are reduced to the degree which can be tolerated by microalgae. The water quality of the pretreated pig manure biogas slurry is shown in table 2.
TABLE 2 Water quality of pretreated pig manure biogas slurry
The algae strain used in the test is the chlamydomonas strain CGMCC NO.15497 separated and purified by the invention, and is used for the test research after being cultured in BG11 culture medium to logarithmic growth phase by adopting the optimal culture condition optimized in the embodiment 2.
(2) Test method
The microalgae in the logarithmic growth phase is added into a 250mL conical flask (the effective culture volume is 150mL), pig manure biogas slurry is added as a microalgae culture medium, and the microalgae culture medium is placed in an illumination incubator with adjustable environmental parameters for culture, wherein the specific culture condition is that the inoculation amount is 0.041g/L, and the illumination intensity is 150 mu mol/m2The photoperiod is 18:6 (light: dark) and the temperature is 26. + -. 0.5 ℃. All experimental groups were cultured for 20 days in three manual shake flasks per day.
Comparative example 1
The strain used in the comparative example was Chlamydomonas reinhardtii strain FACHB-265 randomly selected and purchased from the freshwater algae bank of Wuhan institute of sciences, and the strain was cultured in BG11 medium to a logarithmic growth phase under the optimum culture conditions optimized in example 2 and used in the experimental study.
Comparative example 2
The algal strain used in this comparative example was Chlamydomonas strain FACHB-1935 randomly selected and purchased from the freshwater algal species bank of Wuhan institute of sciences in China. The optimum culture conditions optimized for example 2 were used in this experimental study after culture in BG11 medium to a logarithmic growth phase.
Test results
The growth conditions of the three microalgae in the pig manure biogas slurry are shown in figure 3. In the culture time of 20 days, the Chlamydomonas reinhardtii strain FACHB-265 and the Chlamydomonas strain CGMCC NO.15497 separated and purified by the method can grow in the pig manure biogas slurry, but the growth capacity difference is obvious. The growth effect of the Chlamydomonas strain CGMCC NO.15497 in the pig manure biogas slurry is better, the biomass can reach 0.583g/L in the 20 th day, and the biomass of the Chlamydomonas reinhardtii strain FACHB-265 can only reach 0.450g/L in the 20 th day. The Chlamydomonas strain FACHB-1935 is obviously inhibited in the pig manure biogas slurry, can slowly grow at the initial stage of culture, and gradually dies down from the 9 th day of the test, so that the biomass is reduced. The test result shows that compared with the chlamydomonas strain randomly selected and purchased from a commercial algae bank, the chlamydomonas strain obtained by separation and purification of the invention has the best tolerance in pig manure biogas slurry and the strongest capacity of accumulating biomass.
The change of ammonia nitrogen in the pig manure biogas slurry is shown in figure 4. Within the culture time of 20 days, the difference of the removal capacities of the three microalgae on the ammonia nitrogen in the pig manure biogas slurry is obvious. The chlamydomonas strain CGMCC NO.15497 separated and purified by the method has the strongest removal capability on ammonia nitrogen in the pig manure biogas slurry, and the removal rate of the ammonia nitrogen in 20 days can reach 97.79%. The removal rate of ammonia nitrogen in the pig manure biogas slurry by the chlamydomonas reinhardtii strain FACHB-265 in 20 days is 81.33%. And the chlamydomonas strain FACHB-1935 gradually dies from the 9 th day of culture, so that the removal of ammonia nitrogen in the pig manure biogas slurry is only 20.11 percent finally.
The change of total phosphorus in the pig manure biogas slurry is shown in fig. 5. Within the culture time of 20 days, the removal rate of the chlamydomonas strain CGMCC NO.15497 separated and purified by the invention to the total phosphorus in the pig manure biogas slurry is 90.36%, the removal rate of the chlamydomonas reinhardtii strain FACHB-265 to the total phosphorus within 20 days is 79.53%, and the chlamydomonas strain FACHB-1935 gradually dies from the 9 th day of culture, so the removal rate to the ammonia nitrogen in the pig manure biogas slurry is only 18.90%.
In conclusion, the chlamydomonas strain CGMCC NO.15497 obtained by separation and purification has good tolerance to pig manure biogas slurry with certain concentration, and the microalgae cultured by the pig manure biogas slurry can play a good purifying role to the pig manure biogas slurry, and has obvious effects of nitrogen and phosphorus removal. Compared with two chlamydomonas randomly purchased from a commercial algae bank, the wild algae strain separated from the natural environment has better growth capability in the pig manure biogas slurry, so the invention is an ideal microbial material for treating the pig manure biogas slurry and has wide application prospect.
The above examples are only for describing the preferred embodiments of the present invention, and are not intended to limit the scope of the present invention, and various modifications and improvements made to the technical solution of the present invention by those skilled in the art without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.
Sequence listing
<110> university of agriculture in China
<120> chlamydomonas strain and application thereof in biogas slurry purification
<130> KHP181115747.5
<141> 2018-10-10
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Claims (9)
1. A Chlamydomonas strain (Chlamydomonas sp.) is characterized by the following preservation number: CGMCC No. 15497.
2. The use of the chlamydomonas strain of claim 1 for biogas slurry purification.
3. A method for purifying biogas slurry by using the Chlamydomonas strain of claim 1, comprising:
inoculating a culture solution of the chlamydomonas strain into biogas slurry by an inoculation amount of 0.01-0.1 g/L of dry weight of algae seeds, wherein COD of the biogas slurry is 200-800 mg/L, ammonia nitrogen is 40-150 mg/L, total phosphorus is 10-50 mg/L, suspended solids are 0-500 mg/L, and the pH value is 7.5-8.5.
4. The method for purifying biogas slurry according to claim 3, wherein the culture solution of Chlamydomonas strain is inoculated into biogas slurry at an inoculum size of 0.02-0.05 g/L of dry weight of algal species.
5. The method for purifying biogas slurry according to claim 3, wherein the Chlamydomonas strain culture solution is obtained by inoculating Chlamydomonas strain in an inoculum size of 0.02-0.025 g/L in BG11 liquid medium and culturing to logarithmic growth phase.
6. The method for purifying biogas slurry according to claim 5, wherein the Chlamydomonas strain is cultured in a liquid medium under the following conditions: the illumination intensity is 100 to 300 mu mol/m2The photoperiod is 12-18: 6-12 of light-dark ratio, and the temperature is 15-30 ℃.
7. The method for purifying biogas slurry according to claim 6, wherein the Chlamydomonas strain is cultured in a liquid medium under the following conditions: the illumination intensity is 170-190 mu mol/square meter/s, the light-dark ratio is 18:6, and the temperature is 26 +/-0.5 ℃.
8. The method for purifying biogas slurry according to any one of claims 4 to 7, wherein the Chlamydomonas strain is cultured in the liquid medium by shaking the culture vessel 2 to 4 times per day.
9. The biogas slurry purification method according to any one of claims 3 to 5, comprising the steps of:
1) pretreating the pig manure biogas slurry to reach the conditions of COD (chemical oxygen demand) 200-800 mg/L, ammonia nitrogen 40-150 mg/L, total phosphorus 10-50 mg/L, suspended solid 0-500 mg/L and pH value 7.5-8.5;
2) adding a culture solution of the chlamydomonas strain for growth, wherein the growth conditions are as follows: the illumination intensity is 100 to 200 mu mol/m2And/s, the photoperiod is that the light-dark ratio is 12-20: 4-12 ℃ and the temperature is 24-28 ℃.
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| CN103966102A (en) * | 2014-05-26 | 2014-08-06 | 临沂大学 | Culture medium for culturing Chlamydomonas reinhardtii by urea plant wastewater and culture method of Chlamydomonas reinhardtii |
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| CN105858894A (en) * | 2016-02-19 | 2016-08-17 | 南昌大学 | Method for nitrogen abundance transformation treatment of wastewater with high ammonia nitrogen |
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