Method for preventing contamination by zooplankton in cultivation of chrysophytes

Abstract

The present application relates to the field of chrysophyte culture, and discloses a method for preventing and treating zooplankton pollution in chrysophyte culture, which comprises the following steps: when the number of zooplankton in chrysophyte algae liquid is 20 pieces / mL or more, controlling CO2 flow and adding ammonium bicarbonate into the algae liquid to increase the pH of the algae liquid to 8-9, increasing the temperature of the algae liquid to 30-40 DEG C, and maintaining for 0.5-12 hours for prevention and treatment; wherein the adding amount of the ammonium bicarbonate is such that the content of ammonium bicarbonate in the chrysophyte algae liquid is 50-500 mg / L. Through the method, the zooplankton pollution in large-scale production of chrysophyte can be effectively inhibited, and the yield reduction of chrysophyte caused by zooplankton pollution can be reduced; the method has little influence on the activity of algae cells and has no significant influence on the growth rate of chrysophyte, effectively solving the problem of poor recovery of chrysophyte cell activity. The method has the advantages of simple operation, low cost and short time consumption.

Description

Technical Field

[0001] This invention relates to the field of golden algae cultivation, and more specifically to a method for preventing zooplankton contamination during golden algae cultivation. Background Technology

[0002] Zooplankton such as flagellates, ciliates, and amoebas can reach very high concentrations in golden algae culture media, reducing microalgae biomass by more than 50% and sometimes causing complete failure of golden algae cultivation.

[0003] Currently, large-scale cultivation of golden algae generally employs acidification or sodium hypochlorite treatment methods. Acidification involves adding hydrochloric acid to the algal solution to lower the pH to below 3, treating for 4 hours, and then adding NaOH to restore the pH to normal cultivation levels. For large-scale cultivation, the reactor volume for a single algae culture is typically several tons to tens of tons. After acidification, it takes several hours for the algal solution pH to return to normal, which can be time-consuming and may lead to prolonged periods of excessively low pH in certain areas, often resulting in golden algal cells failing to regain activity after treatment. Sodium hypochlorite treatment involves adding 15-40 ppm of sodium hypochlorite to the algal solution for 10-12 hours, followed by neutralizing residual chlorine with sodium thiosulfate to control zooplankton. However, after sodium hypochlorite treatment, sodium thiosulfate needs to be added to neutralize the residual chlorine. Often, this results in incomplete removal of residual chlorine or an excess of sodium thiosulfate. Both residual chlorine and sodium thiosulfate significantly inhibit the growth of golden algae. While adding small amounts of sodium thiosulfate multiple times can partially solve the problem, it is time-consuming and cumbersome. Therefore, there is an urgent need for a method that can effectively suppress zooplankton pollution while simultaneously alleviating the inhibition of golden algae growth. Summary of the Invention

[0004] The purpose of this invention is to provide a method for preventing zooplankton contamination in the cultivation of golden algae. The method described in this invention can effectively inhibit zooplankton contamination in the large-scale production of golden algae, reduce the reduction of golden algae production caused by zooplankton contamination, and effectively solve the problem of poor recovery of golden algae cell activity. In addition, the method of this invention also has the advantages of simple operation and short time consumption.

[0005] To achieve the above objectives, the present invention provides a method for preventing zooplankton pollution in the cultivation of golden algae. The method includes: when the number of zooplankton in the golden algae solution is more than 20 per mL, controlling the CO2 flow rate and adding ammonium bicarbonate to the algae solution to raise the pH of the algae solution to 8-9, while raising the temperature of the algae solution to 30-40°C and maintaining it for 0.5-12 hours for prevention and control treatment. The amount of ammonium bicarbonate added is such that the content of ammonium bicarbonate in the algal solution is 50-500 mg / L.

[0006] Preferably, during the prevention and treatment process, the temperature is 35-40℃ and the pH is 8.5-9.

[0007] Preferably, the amount of ammonium bicarbonate added is such that the content of ammonium bicarbonate in the algal solution is 100-300 mg / L.

[0008] Preferably, the method further includes controlling the CO2 flow rate and adding ammonium bicarbonate to the algal solution when the number of zooplankton in the algal solution is less than 20 per mL, thereby raising the pH of the algal solution to 8.5-9 and maintaining this pH for 4-6 hours for pretreatment.

[0009] The method described in this invention can effectively suppress zooplankton pollution in the large-scale production of golden algae, greatly reduce the reproduction rate of zooplankton, and reduce the reduction in golden algae production caused by zooplankton pollution.

[0010] After using the method described in this invention to control zooplankton, the algal culture medium quickly returns to normal conditions, with minimal impact on algal cell activity and no significant impact on the growth rate of golden algae, effectively solving the problem of poor recovery of golden algal cell activity.

[0011] In this invention, ammonium bicarbonate can be added in the form of a mother liquor, which is simple to operate; moreover, by adjusting the CO2 flow rate, the pH value of the algal solution can be quickly adjusted so that the pH of the algal solution can reach the target pH or return to the normal level as soon as possible; when cultivating outdoors, the temperature of the algal solution can be quickly raised by turning off the cooling facilities. In summary, the method of this invention has the advantages of simple operation, low cost and short time consumption.

[0012] Under the preferred conditions of this invention, the control effect on zooplankton pollution can be further improved, the treatment time can be further shortened, and the impact on golden algae biomass can be reduced. Detailed Implementation

[0013] The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0014] This invention provides a method for preventing zooplankton pollution in the cultivation of golden algae. The method includes: when the number of zooplankton in the golden algae solution is more than 20 per mL, controlling the CO2 flow rate and adding ammonium bicarbonate to the algae solution to raise the pH of the algae solution to 8-9, while raising the temperature of the algae solution to 30-40°C and maintaining it for 0.5-12 hours for prevention and control treatment. The amount of ammonium bicarbonate added is such that the content of ammonium bicarbonate in the algal solution is 50-500 mg / L.

[0015] It should be understood that after the prevention and control treatment is completed, normal cultivation conditions should be restored to allow the golden algae to grow normally. Specifically, for example, the CO2 flow rate can be adjusted to restore the pH of the algal solution to a normal level, and the temperature of the algal solution can be restored to a normal level through heat exchange.

[0016] Preferably, the culture conditions include: a culture temperature not exceeding 30°C, preferably 15-28°C; a pH of 6.5-8, preferably 7.2-7.8; and a light intensity not exceeding 800 μmol / m². -2 s -1 Preferably not exceeding 600 μmol / m -2 s -1 .

[0017] Generally, temperature control during outdoor cultivation can be achieved through cooling devices, such as a sprinkler system. Light intensity during cultivation can be controlled using shading devices (such as shade nets).

[0018] Preferably, during the culture process, the CO2 inlet flow rate is 20-2000 mL / min, based on the volume of CO2. Those skilled in the art can adjust the CO2 inlet flow rate according to actual circumstances.

[0019] Preferably, the CO2 exists in the form of a CO2-containing gas, such as a CO2 / N2 mixture. Preferably, the CO2 content in the CO2-containing gas is 99% by volume or more. The CO2-containing gas is commercially available.

[0020] In this invention, there are no particular limitations on the scale of golden algae cultivation or the location of cultivation; it can be carried out indoors or outdoors. However, since zooplankton contamination is more severe when golden algae are produced on a large scale outdoors, this method is preferably used in the process of cultivating golden algae outdoors, especially in large-scale production.

[0021] The cultivation of golden algae can be carried out in reactors conventionally used in the field, including but not limited to membrane bag photobioreactors, tubular photobioreactors, flat plate photobioreactors, column photobioreactors or bottle photobioreactors, preferably membrane bag photobioreactors or tubular photobioreactors.

[0022] The golden algae ( Isochrysis sp. ) can be common marine golden algae in this field, such as Isochrysis galbana ( Isochrysis galbana), Zhanjiang isochoria ( Isochrysis zhanjiangensis ), Zhanjiang Forked Whip Algae ( Dicrateria zhanjiangensis It is one of the golden algae such as ).

[0023] Preferably, the number of golden algae cells in the golden algae solution is 500,000 or more per mL.

[0024] The culture medium used for cultivating golden algae can be a conventional seawater culture medium in the art, such as f / 2 medium. The f / 2 medium preferably contains: NaNO3 20-30 mg / L, NaH2PO4 2-6 mg / L, Na2EDTA·2H2O 0.001-0.01 mg / L, FeCl3·6H2O 0.001-0.005 mg / L, MnCl2·4H2O 0.1-0.3 mg / L, ZnSO4·7H2O 0.01-0.05 mg / L, CuSO4·5H2O 0.001-0.003 mg / L, CoCl2·6H2O 0.005-0.02 mg / L, Na2MO4·2H2O 0.001-0.01 mg / L, biotin 0.01-0.2 μg / L, vitamin B12 0.1-1 μg / L, and vitamin B1... 0.1-1 μg / L.

[0025] In this invention, the golden algae can be cultured continuously, semi-continuously, or in batches. Continuous culture means harvesting a certain volume of golden algae solution and replenishing it with a certain volume of fresh culture medium every day until the algae begin to degenerate and the growth rate slows down, at which point the culture is stopped. Semi-continuous culture means harvesting half of the algae solution when it reaches the required concentration in the reactor and replenishing half of the culture medium, and then culturing until the end, at which point the culture is stopped and the algae are harvested. Batch culture means harvesting all of the algae solution when it reaches the required concentration in the reactor.

[0026] In this invention, when the number of zooplankton in the algal solution is more than 20 per mL, the CO2 flow rate is controlled and ammonium bicarbonate is added to the algal solution to raise the pH of the algal solution to 8-9. At the same time, the temperature of the algal solution is raised to 30-40°C and maintained for 0.5-12 hours for prevention and control treatment.

[0027] One way to control the CO2 flow rate is to reduce the CO2 flow rate or stop the CO2 supply. As long as this is combined with the addition of ammonium bicarbonate, the pH of the algal solution can be raised to 8-9.

[0028] The amount of ammonium bicarbonate added is such that the content of ammonium bicarbonate in the algal solution is 50-500 mg / L, for example, it can be 50, 100, 150, 200, 250, 300, 350, 400, 450, 500 mg / L and any range between any two values, preferably 100-300 mg / L.

[0029] Preferably, the ammonium bicarbonate is added in the form of ammonium bicarbonate mother liquor, and the concentration of ammonium bicarbonate in the ammonium bicarbonate mother liquor is 100-200 g / L.

[0030] In this invention, the temperature during the prevention and control treatment process is 30-40℃, for example, it can be 30, 32, 34, 36, 38, 40℃ or any range between any two values; the pH is 8-9, for example, it can be 8, 8.2, 8.4, 8.6, 8.8, 9 or any range between any two values. Preferably, during the prevention and control treatment process, the temperature is 35-40℃ and the pH is 8.5-9. In the preferred case, the prevention and control effect can be further improved and the prevention and control time can be shortened.

[0031] It should be understood that temperature and pH will fluctuate normally during the actual cultivation process. As long as the temperature remains within the above range most of the time, it is acceptable.

[0032] When producing outdoors, prevention and control measures can be carried out at midday on sunny days. The temperature of the algae solution can be controlled by adjusting the use of the spraying device (such as turning off the spray water or reducing the spray volume).

[0033] In this invention, algal solutions with little or no zooplankton can also be pretreated. Preferably, the method further includes controlling the CO2 flow rate and adding ammonium bicarbonate to the algal solution when the number of zooplankton in the golden algae solution is less than 20 cells / mL, thereby raising the pH of the algal solution to 8.5-9 and maintaining this state for 4-6 hours for pretreatment. This pretreatment can, for example, inhibit the growth of zooplankton eggs, reduce egg activity, or even kill the eggs, thereby preventing explosive growth of zooplankton and improving the control effect of zooplankton pollution.

[0034] Preferably, during the pretreatment process, the amount of ammonium bicarbonate added is such that the content of ammonium bicarbonate in the algal solution is 50-150 mg / L, for example, it can be 50, 60, 80, 100, 120, 140, 150 mg / L or any range between any two values.

[0035] Preferably, the method further includes monitoring zooplankton pollution after the prevention and control treatment, and repeating the prevention and control treatment according to the pollution situation.

[0036] Preferably, the frequency of the prevention and control treatment is no more than once every two days, for example, once every two days, once every three days, once every four days, etc.

[0037] In this invention, the zooplankton can be zooplankton that appear during the cultivation of golden algae, such as flagellates, ciliates, amoebas, rotifers, Sarcodina, and scale-forming organisms. Arcella sp. ), sand beetles ( Difflugia sp. ), Dyspodum ( Gromia sp. At least one of the following, preferably flagellates and / or ciliates, can further improve the control effect of zooplankton pollution and increase the biomass of golden algae.

[0038] It should be understood that different species of zooplankton are suppressed or killed to varying degrees after treatment.

[0039] The present invention will be described in detail below through embodiments.

[0040] In the following examples, the golden algae used is *Isochrysis galbana* (…). Isochrysis galbana (), preserved in the algal strain bank of the SDIC Microalgae Biotechnology Center of SDIC Biotechnology Investment Co., Ltd.

[0041] The number of golden algae and zooplankton in the algal solution was determined by counting.

[0042] The f / 2 medium contains: NaNO3 25 mg / L, NaH2PO4 4 mg / L, Na2EDTA·2H2O 0.004 mg / L, FeCl3·6H2O 0.003 mg / L, MnCl2·4H2O 0.18 mg / L, ZnSO4·7H2O 0.022 mg / L, CuSO4·5H2O 0.0098 mg / L, CoCl2·6H2O 0.01 mg / L, Na2MO4·2H2O 0.006 mg / L, biotin 0.1 μg / L, vitamin B12 0.5 μg / L, and vitamin B1 0.5 μg / L, with a pH of 7-7.5. CO2 gas is introduced during the culture process.

[0043] Ammonium bicarbonate mother liquor: The concentration of ammonium bicarbonate is 100 g / L.

[0044] The large-scale production of golden algae was carried out in a tubular photobioreactor (hereinafter referred to as a tubular PBR) with a diameter of 5 cm and a length of 15 m. Each reactor had a cultivation system of 2.5 tons, with a total volume of 20 tons, and a production scale of 1000 t / year. A semi-continuous cultivation mode was adopted. The experiment was conducted in August. During normal cultivation, the temperature was controlled to not exceed 30℃ and the light intensity to not exceed 800 μmol / m² by controlling the spray system and shading nets. -2 s -1 .

[0045] Example 1 This embodiment illustrates the method for controlling zooplankton in the cultivation of golden algae according to the present invention.

[0046] Golden algae were cultured in an outdoor pipe-type PBR at a temperature of 25-28℃, a pH of approximately 7.5, and a light intensity not exceeding 500 μmol / m². -2 s -1 The number of zooplankton is counted under a microscope every day. When the number of zooplankton exceeds 20 per mL, pollution control measures are implemented.

[0047] On a sunny midday (ambient temperature 35±2℃), stop introducing carbon dioxide into the algal solution and add ammonium bicarbonate stock solution (to a final concentration of 250 mg / L) to raise the pH of the algal solution to approximately 8.8. Stop the cooling spray and allow the temperature of the algal solution to rise to 38-40℃, maintaining this temperature for 2 hours. After treatment, introduce carbon dioxide into the algal solution to restore the pH to approximately 7.5. Resume the cooling spray to restore the temperature to 25-28℃ and begin normal cultivation. After 3-5 days of cultivation, end the cultivation process.

[0048] A control group was set up, with the same culture conditions, but without zooplankton control treatment.

[0049] Samples were taken once a day at regular intervals, and the number of zooplankton in the field of view was counted by microscopic examination. The number of zooplankton was obtained by counting with a zooplankton counting frame, and the concentration of golden algae was counted using a hemocytometer. The specific results are shown in Table 1.

[0050] Example 2 This embodiment illustrates the method for controlling zooplankton in the cultivation of golden algae according to the present invention.

[0051] The procedure was performed according to the method described in Example 1, except that the final concentration of ammonium bicarbonate in the algal solution was 150 mg / L.

[0052] Samples were taken once a day at a set time to determine the number of zooplankton and the number of cells in golden algae. The specific results are shown in Table 1.

[0053] Example 3 This embodiment illustrates the method for controlling zooplankton in the cultivation of golden algae according to the present invention.

[0054] The procedure was performed according to the method described in Example 1, except that the final concentration of ammonium bicarbonate in the algal solution was 300 mg / L.

[0055] Samples were taken once a day at a set time to determine the number of zooplankton and the number of cells in golden algae. The specific results are shown in Table 1.

[0056] Example 4 This embodiment illustrates the method for controlling zooplankton in the cultivation of golden algae according to the present invention.

[0057] The procedure was performed according to the method described in Example 1, except that the final concentration of ammonium bicarbonate in the algal solution was 500 mg / L.

[0058] Samples were taken once a day at a set time to determine the number of zooplankton and the number of cells in golden algae. The specific results are shown in Table 1.

[0059] Example 5 This embodiment illustrates the method for controlling zooplankton in the cultivation of golden algae according to the present invention.

[0060] The procedure was performed according to the method described in Example 1, except that the final concentration of ammonium bicarbonate in the algal solution was 80 mg / L.

[0061] Samples were taken once a day at a set time to determine the number of zooplankton and the number of cells in golden algae. The specific results are shown in Table 1.

[0062] Example 6 This embodiment illustrates the method for controlling zooplankton in the cultivation of golden algae according to the present invention.

[0063] The procedure was carried out according to the method described in Example 1, except that the temperature during the prevention and control treatment was 36°C and the treatment time was 4 hours.

[0064] Samples were taken once a day at a set time to determine the number of zooplankton and the number of cells in golden algae. The specific results are shown in Table 1.

[0065] Example 7 This embodiment illustrates the method for controlling zooplankton in the cultivation of golden algae according to the present invention.

[0066] The procedure was carried out according to the method described in Example 1, except that the temperature during the prevention and control treatment was 32°C and the treatment time was 6 hours.

[0067] Samples were taken once a day at a set time to determine the number of zooplankton and the number of cells in golden algae. The specific results are shown in Table 1.

[0068] Example 8 This embodiment illustrates the method for controlling zooplankton in the cultivation of golden algae according to the present invention.

[0069] The procedure was carried out according to the method described in Example 1, except that the CO2 flow rate was reduced and the pH of the algae solution was controlled at around 8.3 during the prevention and treatment process.

[0070] Samples were taken once a day at a set time to determine the number of zooplankton and the number of cells in golden algae. The specific results are shown in Table 1.

[0071] Comparative Example 1 This comparative example is used to illustrate a method for preventing zooplankton contamination in the cultivation of reference golden algae.

[0072] The golden algae were cultured according to the method described in Example 1, except that the prevention and control methods were different. The treatment method was as follows: the treatment was carried out in the evening, and 1% sodium hypochlorite dilution was added to make the effective chlorine content in the algal solution reach 10 ppm. After treatment for 10 hours, sodium thiosulfate was used for reduction.

[0073] Samples were taken once a day at a set time to determine the number of zooplankton and the number of cells in golden algae. The specific results are shown in Table 1.

[0074] Comparative Example 2 This comparative example is used to illustrate a method for preventing zooplankton contamination in the cultivation of reference golden algae.

[0075] The golden algae were cultured according to the method described in Example 1, except that the prevention and control methods were different. The treatment method was as follows: pure CO2 was introduced in the evening to lower the pH of the algal solution to 6. After 6 hours of treatment, sodium hydroxide was added to restore the pH to 7.5.

[0076] Samples were taken once a day at a set time to determine the number of zooplankton and the number of cells in golden algae. The specific results are shown in Table 1.

[0077] Comparative Example 3 This comparative example is used to illustrate a method for preventing zooplankton contamination in the cultivation of reference golden algae.

[0078] The operation was carried out according to the method described in Example 1, except that during the prevention and control process, the flow rate of CO2 was controlled to keep the pH of the algae solution at around 7.8, and the spray cooling was maintained to keep the temperature at 25-28℃.

[0079] Samples were taken once a day at a set time to determine the number of zooplankton and the number of cells in golden algae. The specific results are shown in Table 1.

[0080] Comparative Example 4 This comparative example is used to illustrate a method for preventing zooplankton contamination in the cultivation of reference golden algae.

[0081] The procedure was performed according to the method described in Example 1, except that the final concentration of ammonium bicarbonate in the algal solution was 30 mg / L.

[0082] Samples were taken once a day at a set time to determine the number of zooplankton and the number of cells in golden algae. The specific results are shown in Table 1.

[0083] Table 1

[0084] As can be seen from the results in Table 1, compared with the CO2 control method and the sodium hypochlorite control method, the method of the present invention can significantly inhibit the growth of zooplankton in the large-scale production of golden algae. Under the optimal ammonium bicarbonate concentration and the temperature and pH during the control process, it can further achieve a significantly better control effect.

[0085] The preferred embodiments of the present invention have been described in detail above; however, the present invention is not limited thereto. Within the scope of the inventive concept, various simple modifications can be made to the technical solutions of the present invention, including combinations of various technical features in any other suitable manner. These simple modifications and combinations should also be considered as the content disclosed in the present invention and are all within the protection scope of the present invention.

Claims

1. A method for preventing zooplankton contamination in the cultivation of golden algae, characterized in that, The method includes: when the number of zooplankton in the algal solution is more than 20 per mL, controlling the CO2 flow rate and adding ammonium bicarbonate to the algal solution to raise the pH of the algal solution to 8-9, while raising the temperature of the algal solution to 30-40℃ and maintaining it for 0.5-12 hours for prevention and control treatment. The addition of ammonium bicarbonate results in an ammonium bicarbonate content of 50-500 mg / L in the algal solution. Among them, the golden algae is *Isochrysis galbana* (a type of golden algae). Isochrysis galbana ).

2. The method according to claim 1, wherein, During the aforementioned prevention and treatment process, the temperature is 35-40℃ and the pH is 8.5-9.

3. The method according to claim 1 or 2, wherein, The addition of ammonium bicarbonate results in an ammonium bicarbonate content of 100-300 mg / L in the algal solution.

4. The method according to claim 3, wherein, The ammonium bicarbonate is added in the form of ammonium bicarbonate mother liquor, and the concentration of ammonium bicarbonate in the ammonium bicarbonate mother liquor is 100-200 g / L.

5. The method according to any one of claims 1, 2, and 4, wherein, The method also includes controlling the CO2 flow rate and adding ammonium bicarbonate to the algal solution when the number of zooplankton in the algal solution is less than 20 per mL, so as to raise the pH of the algal solution to 8.5-9 and maintain it for 4-6 hours for pretreatment.

6. The method according to claim 5, wherein, During the pretreatment process, the addition of ammonium bicarbonate resulted in an ammonium bicarbonate content of 50-150 mg / L in the algal solution.

7. The method according to claim 3, wherein, The method also includes controlling the CO2 flow rate and adding ammonium bicarbonate to the algal solution when the number of zooplankton in the algal solution is less than 20 per mL, so as to raise the pH of the algal solution to 8.5-9 and maintain it for 4-6 hours for pretreatment.

8. The method according to claim 7, wherein, During the pretreatment process, the addition of ammonium bicarbonate resulted in an ammonium bicarbonate content of 50-150 mg / L in the algal solution.

9. The method according to any one of claims 1, 2, 4, 6-8, wherein, The cultivation conditions include: a cultivation temperature not exceeding 30℃, a pH of 6.5-8, and a light intensity not exceeding 800 μmol / m². -2 s -1 .

10. The method according to claim 3, wherein, The cultivation conditions include: a cultivation temperature not exceeding 30℃, a pH of 6.5-8, and a light intensity not exceeding 800 μmol / m². -2 s -1 .

11. The method according to claim 5, wherein, The cultivation conditions include: a cultivation temperature not exceeding 30℃, a pH of 6.5-8, and a light intensity not exceeding 800 μmol / m². -2 s -1 .

12. The method according to any one of claims 1, 2, 4, 6-8, 10-11, wherein, During the culture process, the CO2 flow rate is 20-2000 mL / min, based on the volume of CO2.

13. The method according to claim 12, wherein, The CO2 exists in the form of a CO2-containing gas, wherein the CO2 content in the CO2-containing gas is 99% by volume or more.

14. The method according to claim 3, wherein, During the culture process, the CO2 flow rate is 20-2000 mL / min, based on the volume of CO2.

15. The method according to claim 14, wherein, The CO2 exists in the form of a CO2-containing gas, wherein the CO2 content in the CO2-containing gas is 99% by volume or more.

16. The method according to claim 5, wherein, During the culture process, the CO2 flow rate is 20-2000 mL / min, based on the volume of CO2.

17. The method according to claim 16, wherein, The CO2 exists in the form of a CO2-containing gas, wherein the CO2 content in the CO2-containing gas is 99% by volume or more.

18. The method according to claim 9, wherein, During the culture process, the CO2 flow rate is 20-2000 mL / min, based on the volume of CO2.

19. The method according to claim 18, wherein, The CO2 exists in the form of a CO2-containing gas, wherein the CO2 content in the CO2-containing gas is 99% by volume or more.

20. The method according to any one of claims 1, 2, 4, 6-8, 10-11, 13-19, wherein, The method also includes monitoring zooplankton pollution after treatment and repeating treatment as needed based on the pollution levels.

21. The method according to claim 20, wherein, The frequency of the prevention and control treatment shall not exceed once every two days.

22. The method according to claim 3, wherein, The method also includes monitoring zooplankton pollution after treatment and repeating treatment as needed based on the pollution levels.

23. The method according to claim 22, wherein, The frequency of the prevention and control treatment shall not exceed once every two days.

24. The method according to claim 5, wherein, The method also includes monitoring zooplankton pollution after treatment and repeating treatment as needed based on the pollution levels.

25. The method according to claim 24, wherein, The frequency of the prevention and control treatment shall not exceed once every two days.

26. The method according to claim 9, wherein, The method also includes monitoring zooplankton pollution after treatment and repeating treatment as needed based on the pollution levels.

27. The method according to claim 26, wherein, The frequency of the prevention and control treatment shall not exceed once every two days.

28. The method according to claim 12, wherein, The method also includes monitoring zooplankton pollution after treatment and repeating treatment as needed based on the pollution levels.

29. The method according to claim 28, wherein, The frequency of the prevention and control treatment shall not exceed once every two days.

30. The method according to any one of claims 1, 2, 4, 6-8, 10-11, 13-19, and 21-29, wherein, The number of golden algae cells in the algal solution is above 500,000 / mL.

31. The method according to claim 3, wherein, The number of golden algae cells in the algal solution is above 500,000 / mL.

32. The method according to claim 5, wherein, The number of golden algae cells in the algal solution is above 500,000 / mL.

33. The method according to claim 9, wherein, The number of golden algae cells in the algal solution is above 500,000 / mL.

34. The method according to claim 12, wherein, The number of golden algae cells in the algal solution is above 500,000 / mL.

35. The method according to claim 20, wherein, The number of golden algae cells in the algal solution is above 500,000 / mL.

36. The method according to any one of claims 1, 2, 4, 6-8, 10-11, 13-19, 21-29, and 31-35, wherein, The zooplankton include flagellates, ciliates, amoebas, rotifers, Sarcodina, and scale insects. Arcella sp. ), sand beetles ( Difflugia sp. ), Dyspodum ( Gromia sp. At least one of the following.

37. The method according to claim 3, wherein, The zooplankton include flagellates, ciliates, amoebas, rotifers, Sarcodina, and scale insects. Arcella sp. ), sand beetles ( Difflugia sp. ), Dyspodum ( Gromia sp. At least one of the following.

38. The method according to claim 5, wherein, The zooplankton include flagellates, ciliates, amoebas, rotifers, Sarcodina, and scale insects. Arcella sp. ), sand beetles ( Difflugia sp. ), Dyspodum ( Gromia sp. At least one of the following.

39. The method according to claim 9, wherein, The zooplankton include flagellates, ciliates, amoebas, rotifers, Sarcodina, and scale insects. Arcella sp. ), sand beetles ( Difflugia sp. ), Dyspodum ( Gromia sp. At least one of the following.

40. The method according to claim 12, wherein, The zooplankton include flagellates, ciliates, amoebas, rotifers, Sarcodina, and scale insects. Arcella sp. ), sand beetles ( Difflugia sp. ), Dyspodum ( Gromia sp. At least one of the following.

41. The method according to claim 20, wherein, The zooplankton include flagellates, ciliates, amoebas, rotifers, Sarcodina, and scale insects. Arcella sp. ), sand beetles ( Difflugia sp. ), Dyspodum ( Gromia sp. At least one of the following.

42. The method according to claim 30, wherein, The zooplankton include flagellates, ciliates, amoebas, rotifers, Sarcodina, and scale insects. Arcella sp. ), sand beetles ( Difflugia sp. ), Dyspodum ( Gromia sp. At least one of the following.

43. The method according to any one of claims 1, 2, 4, 6-8, 10-11, 13-19, 21-29, 31-35, and 37-42, wherein, The culture is carried out in a membrane bag photobioreactor, a tubular photobioreactor, a flat plate photobioreactor, a column photobioreactor, or a shake flask photobioreactor.

44. The method according to claim 3, wherein, The culture is carried out in a membrane bag photobioreactor, a tubular photobioreactor, a flat plate photobioreactor, a column photobioreactor, or a shake flask photobioreactor.

45. The method according to claim 5, wherein, The culture is carried out in a membrane bag photobioreactor, a tubular photobioreactor, a flat plate photobioreactor, a column photobioreactor, or a shake flask photobioreactor.

46. ​​The method according to claim 9, wherein, The culture is carried out in a membrane bag photobioreactor, a tubular photobioreactor, a flat plate photobioreactor, a column photobioreactor, or a shake flask photobioreactor.

47. The method according to claim 12, wherein, The culture is carried out in a membrane bag photobioreactor, a tubular photobioreactor, a flat plate photobioreactor, a column photobioreactor, or a shake flask photobioreactor.

48. The method of claim 20, wherein, The culture is carried out in a membrane bag photobioreactor, a tubular photobioreactor, a flat plate photobioreactor, a column photobioreactor, or a shake flask photobioreactor.

49. The method according to claim 30, wherein, The culture is carried out in a membrane bag photobioreactor, a tubular photobioreactor, a flat plate photobioreactor, a column photobioreactor, or a shake flask photobioreactor.

50. The method of claim 36, wherein, The culture is carried out in a membrane bag photobioreactor, a tubular photobioreactor, a flat plate photobioreactor, a column photobioreactor, or a shake flask photobioreactor.

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