Heterotrophic culture method for isochrysis galbana
By establishing a monitoring timeline and analyzing growth, the heterotrophic culture method of phlebroscopic is able to monitor abnormal conditions in real time and adjust the pH value, solving the problems of slow response speed and difficulty in optimization in traditional culture methods, and improving the culture efficiency and success rate.
Patent Information
- Application Number
- CN202510271862.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-03-10
- Publication Date
- 2025-04-11
- Estimated Expiration
- Not applicable · inactive patent
AI Technical Summary
Traditional phlebol-like whip algae culture methods are difficult to monitor abnormal situations in real time, cannot improve the response speed to problems, and cannot optimize culture management and improve culture efficiency and success rate.
By establishing a monitoring timeline, comprehensively analyzing the growth situation, calculating the abnormal proportion and culture progress ratio, generating analysis signals, and processing them based on the pH culture data, adjusting the pH value to optimize the culture conditions.
The monitoring efficiency and problem response speed of the culture process are improved, the impact of pH on the growth of saccharomyceae is accurately identified, and the culture quality and success rate are significantly improved by adjusting the pH value.
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Figure CN119799499A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of algae cultivation, and in particular to a heterotrophic cultivation method of Isochrysis galbana. Background Art
[0002] In the field of Isochrysis galbana cultivation, traditional cultivation methods often have difficulty in timely detecting abnormal conditions during the cultivation process, cannot monitor changes in abnormal conditions in real time, cannot improve the response speed to problems, and cannot optimize cultivation management content. At the same time, it is impossible to accurately detect dynamic changes in the cultivation process of Isochrysis galbana, cannot accurately identify the key factors that cause abnormal cell growth, and cannot timely observe and adjust the algae growth during the cultivation cycle, making it difficult to optimize the cultivation conditions in a targeted manner, and cannot improve the cultivation efficiency and success rate. It also faces many obstacles in large-scale cultivation applications.
[0003] Based on this, a heterotrophic culture method of Isochrysis galbana was proposed. Summary of the invention
[0004] The object of the present invention is to provide a heterotrophic culture method of Isochrysis galbana to solve at least one of the above-mentioned problems of the prior art.
[0005] The present invention provides a heterotrophic culture method of Isochrysis galbana, comprising the following steps: The culture cycle is preset, and the abnormal proportion and the culture progress ratio are obtained based on the combined analysis of the monitoring time axis and the growth situation. The abnormal proportion and the culture progress ratio are summed to obtain the growth abnormality value; If the growth abnormality value is greater than or equal to the growth abnormality threshold, an analysis signal is generated; According to the analysis signal, the pH culture data is processed to obtain the pH value deviation value. If the pH value deviation value is greater than the pH value deviation threshold, the sub-period is marked as a pH value abnormal sub-period, and the number of pH value abnormal sub-periods is processed by ratio processing with the number of growth abnormal sub-periods to obtain the pH value abnormality ratio. If the pH value abnormality ratio is greater than or equal to the pH value abnormality ratio threshold, a suspected abnormality signal is generated; Among them, pH culture data include the ratio of suitable culture time and the ratio of pH value exceeding area; Based on the suspected abnormal signal, the deviation data is analyzed and processed to obtain the deviation correlation value based on pH and cell density, and the deviation correlation value is processed to obtain the deviation correlation variance; Among them, the deviation data include cell density deviation and pH value deviation; If the deviation from the correlation variance is less than the deviation from the correlation variance threshold, it means that the pH value has an impact on the heterotrophic growth of Isochrysis galbana and the pH value is adjusted.
[0006] As a further solution of the present invention: the abnormality ratio is obtained in the following manner: The cultivation cycle is divided into several sub-cycles, and the division of the sub-cycles is matched to the monitoring time axis to obtain the growth conditions of Isochrysis globosum in the sub-cycles; According to the monitoring time axis sequence, the growth of Isochrysis spherica in any sub-period is analyzed to obtain the cell density of Isochrysis spherica in the sub-period, and the difference between the cell density of the sub-period and the standard value of the cell density of the sub-period is processed and the absolute value is taken to obtain the cell density difference of the sub-period; The sub-cycle cell density difference is compared with the sub-cycle cell density standard value. The specific comparison process is as follows: If the difference in sub-cycle cell density is less than or equal to the standard value of sub-cycle cell density, no other treatment is performed; If the cell density of a sub-cycle is greater than the standard value of the cell density of a sub-cycle, the sub-cycle is marked as an abnormal growth sub-cycle; The number of abnormal growth sub-cycles was counted, and the ratio of the number of abnormal growth sub-cycles to the number of cultured sub-cycles was processed to obtain the abnormal proportion.
[0007] As a further solution of the present invention: the method for obtaining the culture progress ratio is: The number of sub-cycles that have been cultivated is compared with the number of sub-cycles in the cultivation period to obtain a cultivation progress ratio; The abnormal proportion and the culture progress ratio are summed to obtain the growth abnormality value, which reflects a comprehensive indicator for evaluating the growth situation during the culture cycle.
[0008] As a further solution of the present invention: the pH value deviation degree value is obtained in the following manner: The duration of the pH value in the appropriate pH value range is processed by ratio to the total time of the pH value change analysis curve to obtain the appropriate culture time ratio, and the pH value excess area is processed by ratio to the pH value change analysis curve area to obtain the pH value excess area ratio; The pH deviation degree value was obtained by performing ratio processing on the appropriate culture time ratio and the pH value exceeding area ratio.
[0009] As a further solution of the present invention: the method for obtaining the suitable culture time ratio is: According to the monitoring time axis, the pH value change of the sub-period is obtained to obtain the pH value change curve, and according to the pH value change curve, the pH value of the sub-period is combined with the appropriate pH value range for analysis; The pH value change curve is divided according to the number of sub-periods to obtain a number of pH value change analysis curves; Analyze the curve based on any pH value change; Mark the lowest range value of the suitable pH value and the highest range value of the suitable pH value in the coordinate system, draw a straight line parallel to the X-axis based on the marked points, mark the two straight lines as the lower limit of the suitable pH value and the upper limit of the suitable pH value, respectively, and the part between the lower limit of the suitable pH value and the upper limit of the suitable pH value is the suitable pH value range; The duration of the pH value in the appropriate pH range was counted and the ratio was processed with the total time of the pH change analysis curve to obtain the appropriate culture time ratio.
[0010] As a further solution of the present invention: the pH value exceeding the area ratio is obtained by: The area of the figure formed by the upper limit of the suitable pH value, the Y axis and the horizontal asymptote is calculated using a mathematical method integral formula to obtain the area exceeding the upper limit; Similarly, the area of the figure formed by the lower limit of the suitable pH value, the Y axis and the horizontal asymptote is calculated using the mathematical method integration formula to obtain the area exceeding the lower limit; The horizontal asymptote represents a straight line passing through the final value of the X-axis and parallel to the Y-axis; The pH value excess area is obtained by summing the upper bound excess area and the lower bound excess area; The pH value excess area and the pH value change analysis curve area are ratio-processed to obtain the pH value excess area ratio.
[0011] As a further solution of the present invention: the deviation correlation value is obtained in the following manner: The cell density difference is processed by ratio processing with the cell density standard value to obtain the cell density deviation, and the pH value lower limit deviation is summed with the pH value upper limit deviation to obtain the pH value deviation; The deviation-related value was obtained by calculating the ratio of the cell density deviation to the pH value deviation.
[0012] As a further solution of the present invention: the cell density deviation is obtained by: The cell density is processed by difference with the standard value of cell density to obtain the cell density difference, and the cell density difference is processed by ratio with the standard value of cell density to obtain the cell density deviation.
[0013] As a further solution of the present invention: the pH value deviation is obtained by: The lowest pH value range difference is compared with the lowest pH value range value, and the pH lower limit deviation is obtained. The highest pH value range difference is compared with the highest pH value range value, and the pH upper limit deviation is obtained. The pH value deviation is obtained by summing the pH value lower limit deviation and the pH value upper limit deviation.
[0014] As a further solution of the present invention: the method for obtaining the pH value lower limit deviation and the pH value upper limit deviation is: According to the pH value change curve, analysis is carried out in combination with the monitoring time axis; Specifically, the pH value lower than the lowest range value of the appropriate pH value is subjected to difference processing with the lowest range value of the appropriate pH value, and the absolute value is taken to obtain the pH value lowest range difference, and the pH value lowest range difference is subjected to ratio processing with the lowest range value of the appropriate pH value to obtain the pH value lower limit deviation; Similarly, the pH value higher than the highest range value of the suitable pH value is subjected to difference processing with the highest range value of the suitable pH value, and the absolute value is taken to obtain the pH value highest range difference, and the pH value highest range difference is subjected to ratio processing with the highest range value of the suitable pH value to obtain the pH value upper limit deviation.
[0015] Beneficial effects of the present invention: 1. The present invention establishes a monitoring time axis according to the culture cycle, and comprehensively analyzes the monitoring time axis and the growth situation to obtain the abnormal proportion and the culture progress ratio, and then calculates the growth abnormal value. The growth abnormal value can be used to determine whether an abnormality occurs and generate an analysis signal, which effectively improves the monitoring efficiency and problem response speed of the culture process. According to the analysis signal, the pH culture data is analyzed to obtain the pH value deviation degree value. By comparing the pH value deviation degree value with the pH value deviation degree threshold, the pH problem of the culturing environment of Isochrysis globosum can be timely prompted, and a suspected abnormal signal is generated, which has a significant beneficial effect on optimizing culture management and ensuring culture quality; 2. The present invention obtains and analyzes deviation data based on suspected abnormal signals to obtain deviation related values, obtains deviation related variances by further calculating the deviation related variances, and determines whether the pH value is the cause of the growth by comparing the deviation related variances with the deviation related variance threshold, thereby improving monitoring efficiency and culture success rate. The present invention can accurately identify that the pH value is a key factor affecting cell growth, and adjust the pH value accordingly to optimize the culture conditions. BRIEF DESCRIPTION OF THE DRAWINGS
[0016] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.
[0017] Figure 1 This is a flow chart of a heterotrophic culture method of Isochrysis galbana provided in Example 1 of the present invention; Figure 2 This is a flow chart of a heterotrophic culture method of Isochrysis galbana provided in Example 2 of the present invention; Figure 3 This is a flow chart of a heterotrophic culture method of Isochrysis galbana provided in Example 3 of the present invention; Figure 4 This is a flow chart of a heterotrophic culture method of Isochrysis galbana provided in Example 4 of the present invention; Figure 5 It is a schematic diagram of the structure of a heterotrophic culture system of Isochrysis galbana provided in Example 5 of the present invention. DETAILED DESCRIPTION
[0018] In order to enable those skilled in the art to better understand the scheme of the present invention, the technical scheme in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.
[0019] Embodiment 1, Figure 1 The flowchart of a heterotrophic culture method of Isochrysis galbana provided in the first embodiment of the present invention is applicable to the situation of culture requirements under specific environment. The heterotrophic culture method of Isochrysis galbana can be performed by a heterotrophic culture system of Isochrysis galbana, which can be implemented by software and / or hardware, and the heterotrophic culture system of Isochrysis galbana can be configured in a heterotrophic culture device of Isochrysis galbana. Optionally, the heterotrophic culture device of Isochrysis galbana can be an electronic device, which can be a notebook, a desktop computer, a smart tablet, etc., and the embodiment of the present invention does not limit this.
[0020] like Figure 1 As shown, a heterotrophic culture method of Isochrysis galbana provided by an embodiment of the present invention specifically comprises the following steps: Step 1: The heterotrophic culture process of Isochrysis galbana is as follows: S1, using KS (G) heterotrophic medium as the basic medium to meet the growth requirements of algae; The concentration of glucose in the culture medium is 40 g / L, urea is 1.0 g / L, and KH2PO4 is 13.6 mg / L; seawater preparation: 10.0 g of glucose, 1.0 g of NaNO3, 0.12 g of NaH2PO4, 0.2 g of Na2SiO3, 10 mg of trace elements Na2EDTA, 10 mg of H3BO3, 2.0 mg of MnCl2, 0.1 mg of CuSO4, 0.05 mg of CoCl2, 10 mg of FeCl3, and 0.1 mg of ZnSO4 are added to each liter of seawater; The prepared medium was adjusted to pH 7.0 with 0.5 mol sodium hydroxide solution, sterilized at 121°C for 30 min, and after cooling, vitamin B12 0.05 mg / L was added for later use; S2. Add 200 mL of sterile heterotrophic culture medium into a 500 mL triangular flask, inoculate 50 mL of golden algae seed solution, place in a MQD-B3R incubator and culture at 30°C and 160 r / min in the dark. Stop the experiment when the algae cells enter the stable growth period. The present invention uses Isochrysis galbana 3011, which is provided by the Microalgae Germplasm Bank of Ocean University of China (MACC); S3. Regularly monitor the growth of algae, further observe the pH value of the culture medium based on the monitoring results, and adjust the culture conditions in a timely manner; S4. When the algae cells in the culture medium have completed their growth, they can be harvested. The harvesting methods include centrifugation, filtration, etc. It should be noted that the stable period means that the growth rate of algal cells slows down significantly or the net number of cells does not fluctuate greatly; S5, storing the harvested algae; In some embodiments, S3 is analyzed; Step 2: During the culture period of Isochrysis galbana, the culture period is divided into several sub-periods, and the growth of Isochrysis galbana is monitored every day, and the growth condition includes cell density; In some embodiments, the specific operation of monitoring the cell density of Isochrysis galbana by flow cytometry is: Collect Isochrysis galbana liquid from the culture medium, remove impurities and cell clumps by filtration or centrifugation to obtain a single cell suspension, then fix the cells with a fixative (such as paraformaldehyde), use phycoerythrin (PE) to stain Isochrysis galbana, mix phycoerythrin (PE) with the cell suspension at 3-5 μg / mL, incubate for 15-20 minutes under light-proof conditions to allow phycoerythrin (PE) to fully enter the cells and bind to the target substance, incubate under light-proof conditions to allow phycoerythrin (PE) to fully bind, then wash the cells with a buffer such as phosphate buffered saline (PBS) to remove unbound dyes and impurities, prepare the flow cytometer and debug and calibrate it, then resuspend the washed cells with a buffer to an appropriate concentration to obtain the Isochrysis galbana to be tested. Golden algae cell suspension, adding fluorescently labeled standard microspheres of known quantity and concentration to the test Isochrysis galbana cell suspension, fully mixing the cells with Isochrysis galbana cells, passing the cells one by one through the detection area, and recording the fluorescence and scattered light signals by the instrument, finally analyzing the data with professional software, calculating the cell density by the standard curve method, the ratio method or an instrument using the absolute counting function, performing flow cytometry detection on the mixed suspension, respectively obtaining the fluorescence signals of Isochrysis galbana cells and the standard microspheres, distinguishing and counting the number or fluorescence intensity of Isochrysis galbana cells and the standard microspheres by gate setting and other operations, and calculating the cell density of Isochrysis galbana according to the known number of standard microspheres and the ratio of the number of detected Isochrysis galbana cells to the standard microspheres; For example, add the cell suspension of Isochrysis sphericalensis to be tested / mL fluorescent microspheres, the number of Isochrysis galbana cells detected is 5 times the number of standard microspheres, so the cell density of Isochrysis galbana is pcs / mL= Pieces / mL; In some embodiments, the first embodiment is described in detail; Embodiment 2, as Figure 2 As shown, based on the first embodiment, the growth of Isochrysis galbana in the culture medium is monitored. The heterotrophic culture method of Isochrysis galbana described in the embodiment of the present invention includes: Step 3: According to the culture cycle, a monitoring timeline is established, and the abnormal proportion and the culture progress ratio are obtained by combining the monitoring timeline and the growth situation. The abnormal proportion and the culture progress ratio are summed to obtain the growth abnormality value; Specifically, the culture cycle is divided into several sub-cycles, and the division of the sub-cycles corresponds to the monitoring time axis to obtain the growth status of Isochrysis globosum in the sub-cycles; According to the monitoring time axis sequence, the growth of Isochrysis spherica in any sub-period is analyzed to obtain the cell density of Isochrysis spherica in the sub-period, and the difference between the cell density of the sub-period and the standard value of the cell density of the sub-period is processed and the absolute value is taken to obtain the cell density difference of the sub-period; The sub-cycle cell density difference is compared with the sub-cycle cell density standard value. The specific comparison process is as follows: If the difference in sub-cycle cell density is less than or equal to the standard value of sub-cycle cell density, no other treatment is performed; If the cell density of a sub-cycle is greater than the standard value of the cell density of a sub-cycle, the sub-cycle is marked as an abnormal growth sub-cycle; Count the number of abnormal growth sub-cycles, and perform ratio processing on the number of abnormal growth sub-cycles and the number of cultured sub-cycles to obtain the abnormal proportion; The number of sub-cycles that have been cultivated is compared with the number of sub-cycles in the cultivation period to obtain a cultivation progress ratio; The abnormal proportion and the cultivation progress ratio are summed to obtain the growth abnormality value; It should be noted that the growth abnormality value reflects a comprehensive indicator for evaluating the growth situation during the culture period. The larger the growth abnormality value, the worse the growth situation during the culture period. Conversely, the smaller the growth abnormality value, the better the growth situation during the culture period. Step 4: Compare the growth abnormality value with the growth abnormality threshold. If the growth abnormality value is greater than or equal to the growth abnormality threshold, generate an analysis signal. If the growth abnormality value is less than the growth abnormality threshold, no processing is performed. Specifically, the growth abnormality value is compared with the growth abnormality threshold; If the growth abnormality value is greater than or equal to the growth abnormality threshold, an analysis signal is generated; If the growth abnormality value is less than the growth abnormality threshold, no processing is performed; The technical solution of the embodiment of the present invention is: according to the culture cycle, a monitoring timeline is established, and according to the combined analysis of the monitoring timeline and the growth situation, the abnormality ratio and the culture progress ratio are obtained, the abnormality ratio and the culture progress ratio are summed to obtain the growth abnormality value, and the growth abnormality value is compared with the growth abnormality threshold. If the growth abnormality value is greater than or equal to the growth abnormality threshold, an analysis signal is generated; if the growth abnormality value is less than the growth abnormality threshold, no processing is performed. The present invention establishes a monitoring timeline in combination with the culture cycle, and comprehensively analyzes the abnormality ratio and the culture progress ratio according to the growth situation, and then calculates the growth abnormality value, and compares it with its threshold, so as to timely discover and generate analysis signals, effectively improve the monitoring efficiency and problem response speed of the culture process, and has significant beneficial effects on optimizing culture management and ensuring culture quality.
[0021] Embodiment three, as Figure 3 As shown, on the basis of Example 2, based on further analysis of the comparison between the growth abnormality value and the growth abnormality threshold, a heterotrophic culture method of Isochrysis galbana described in an embodiment of the present invention includes: Step 5: According to the analysis signal, pH culture data is obtained, the pH culture data includes a suitable culture time ratio and a pH value exceeding area ratio, the suitable culture time ratio and the pH value exceeding area ratio are processed to obtain a pH value deviation value, the pH value deviation value is compared with a pH value deviation degree threshold, if the pH value deviation degree value is less than or equal to the pH value deviation degree threshold, no other processing is performed, if the pH value deviation degree value is greater than the pH value deviation degree threshold, the sub-period number is marked as the number of pH value abnormal sub-periods; It should be noted that the suitable pH range for the cultivation of Isochrysis galbana is 7.0-8.1, of which 7.0 is the lowest range value of the suitable pH value and 8.1 is the highest range value of the suitable pH value; Specifically, according to the monitoring time axis, the pH value change of the sub-period is obtained, and an XY coordinate system is established, wherein the X axis is the monitoring time and the Y axis is the pH value. The pH value change of the sub-period is marked in the coordinate system, and the data points are connected to obtain a pH value change curve; According to the pH value change curve, the pH value of the sub-period is combined with the appropriate pH value range for analysis; Specifically, the pH value change curve is divided according to the number of sub-periods to obtain a number of pH value change analysis curves; Analyze the curve based on any pH value change; Mark the lowest range value of the suitable pH value and the highest range value of the suitable pH value in the coordinate system, draw a straight line parallel to the X-axis based on the marked points, mark the two straight lines as the lower limit of the suitable pH value and the upper limit of the suitable pH value, respectively, and the part between the lower limit of the suitable pH value and the upper limit of the suitable pH value is the suitable pH value range; The duration of the pH value in the appropriate pH range is counted, and the ratio is processed with the total time of the pH value change analysis curve to obtain the appropriate culture time ratio; In some embodiments, a mathematical method integral formula is used to calculate the area of a graph formed by the upper limit of the suitable pH value, the Y axis and the horizontal asymptote to obtain an area exceeding the upper limit; Similarly, the area of the figure formed by the lower limit of the suitable pH value, the Y axis and the horizontal asymptote is calculated using the mathematical method integration formula to obtain the area exceeding the lower limit; It should be noted that the meaning of the horizontal asymptote is: the straight line passing through the final value of the X-axis and parallel to the Y-axis is called a horizontal asymptote; The pH value excess area is obtained by summing the upper bound excess area and the lower bound excess area; The pH value excess area and the pH value change analysis curve area are processed by ratio to obtain the pH value excess area ratio; The appropriate culture time ratio and the pH value exceeding area ratio are processed to obtain the pH value deviation degree value; The pH value deviation value is compared with the pH value deviation threshold value. The specific comparison process is as follows: If the pH value deviation value is less than or equal to the pH value deviation threshold, no other processing is performed; If the pH value deviation value is greater than the pH value deviation threshold, the sub-period number is marked as the pH value abnormal sub-period number; Step 6: Based on the comparison between the pH value deviation degree value and the pH value deviation degree threshold, the number of pH value abnormal sub-cycles and the number of growth abnormal sub-cycles are ratio-processed to obtain the pH value abnormality ratio, and the pH value abnormality ratio is compared with the pH value abnormality ratio threshold. If the pH value abnormality ratio is greater than or equal to the pH value abnormality ratio threshold, a suspected abnormality signal is generated. If the pH value abnormality ratio is less than the pH value abnormality ratio threshold, no other processing is performed. Specifically, the number of abnormal pH value sub-cycles within the culture cycle is counted, and the number of abnormal pH value sub-cycles is ratioed to the number of abnormal growth sub-cycles to obtain the abnormal pH value ratio; The pH value abnormality ratio is compared with the pH value abnormality ratio threshold. The specific comparison process is as follows: If the pH value abnormality ratio is greater than or equal to the pH value abnormality ratio threshold, a suspected abnormality signal is generated; If the pH value abnormality ratio is less than the pH value abnormality ratio threshold, no other processing is performed; The technical scheme of the embodiment of the present invention is as follows: according to the analysis signal, pH culture data is obtained, the pH culture data includes the suitable culture time ratio and the pH value exceeding area ratio, the suitable culture time ratio and the pH value exceeding area ratio are processed to obtain the pH value deviation degree value, the pH value deviation degree value is compared with the pH value deviation degree threshold, if the pH value deviation degree value is greater than the pH value deviation degree threshold, the number of sub-cycles is marked as the number of pH value abnormal sub-cycles, based on the comparison of the pH value deviation degree value and the pH value deviation degree threshold, the number of pH value abnormal sub-cycles and the number of growth abnormal sub-cycles are processed by ratio to obtain the pH value abnormality ratio, the pH value abnormality ratio is compared with the pH value abnormality ratio threshold, if the pH value abnormality ratio is greater than or equal to the pH value abnormality ratio threshold, a suspected abnormal signal is generated, and the present invention can effectively identify and generate suspected abnormal signals, timely prompt the pH problem of the culturing environment of Isochrysis galbana, and improve the monitoring efficiency and the culturing success rate.
[0022] Embodiment 4, as Figure 4 As shown, according to the comparison between the pH value deviation value and the pH value deviation threshold, the cell density and pH value are combined and analyzed to determine whether the pH value is the cause of the growth. If so, the pH value is adjusted according to the analysis result. The heterotrophic culture method of Isochrysis galbana described in the embodiment of the present invention includes: Step 7: Based on the suspected abnormal signal, the deviation data is obtained, and the deviation data includes the cell density deviation and the pH value deviation. The deviation correlation value is obtained according to the analysis of the deviation data. The deviation correlation value is processed to obtain the deviation correlation variance. The deviation correlation variance is compared with the deviation correlation variance threshold. If the deviation correlation variance is less than the deviation correlation variance threshold, the deviation correlation value data fluctuates little, indicating that the pH value is the reason affecting the growth. If the deviation correlation variance is greater than or equal to the deviation correlation variance threshold, the deviation correlation value data fluctuates greatly, indicating that other factors cause the growth to fail to meet expectations during the culture period, and no other processing is performed. Specifically, based on the monitoring time axis, any sub-period is analyzed; Performing difference processing on the cell density and the standard value of cell density to obtain the cell density difference, and performing ratio processing on the cell density difference and the standard value of cell density to obtain the cell density deviation; According to the pH value change curve, analysis is carried out in combination with the monitoring time axis; Specifically, the pH value lower than the lowest range value of the appropriate pH value is subjected to difference processing with the lowest range value of the appropriate pH value, and the absolute value is taken to obtain the pH value lowest range difference, and the pH value lowest range difference is subjected to ratio processing with the lowest range value of the appropriate pH value to obtain the pH value lower limit deviation; Similarly, the pH value higher than the highest range value of the appropriate pH value is processed with the highest range value of the appropriate pH value, and the absolute value is taken to obtain the highest range difference of the pH value, and the highest range difference of the pH value is processed with the highest range value of the appropriate pH value to obtain the deviation of the upper limit of the pH value; The pH value deviation is obtained by summing the pH value lower limit deviation and the pH value upper limit deviation; The deviation of cell density and pH value are calculated by ratio, and the deviation correlation value is obtained; The deviation correlation value is calculated using the mathematical variance formula to obtain the deviation correlation variance; The deviation from the correlation variance is compared with the deviation from the correlation variance threshold. The specific comparison process is as follows: If the deviation correlation variance is less than the deviation correlation variance threshold, the deviation correlation value data fluctuates little, indicating that pH value is the cause of the growth condition; If the deviation correlation variance is greater than or equal to the deviation correlation variance threshold, the deviation correlation value data fluctuates greatly, indicating that other factors have caused the growth situation within the culture cycle to fail to meet expectations, and no other treatment is performed; For example, if the first sub-cycle and the second sub-cycle in the culture cycle are both abnormal sub-cycle numbers, the cell density deviation of the first sub-cycle in the culture cycle is 2, and the pH value deviation is 1, then the deviation correlation value is 2, and the deviation correlation value of the second sub-cycle in the culture cycle is also 2.01, and the difference between the deviation correlation values of the first sub-cycle and the second sub-cycle in the culture cycle is small, it means that the pH value in the first two sub-cycles of the culture cycle is the reason that affects the growth. Otherwise, it means that other factors cause the growth in the first two days of the culture cycle to fail to meet expectations; Step 8: Based on the fact that pH is the factor that affects growth, make adjustments according to changes in pH; Specifically, when the pH value is greater than the maximum range of the appropriate pH value, an acidic substance such as dilute hydrochloric acid may be added in an appropriate amount to lower the pH value, and the pH value change curve is observed in real time until the pH value reaches the appropriate pH value range; When the pH value is lower than the lowest value in the appropriate pH range, alkaline substances such as sodium hydroxide can be added to increase the pH value. However, during the addition process, it is necessary to add slowly and stir thoroughly to prevent the local pH value from changing too much and causing impact on the algae until the pH value reaches the appropriate pH range. The technical solution of the embodiment of the present invention is: based on the suspected abnormal signal, deviation data is obtained, the deviation data includes the cell density deviation and the pH value deviation, the deviation correlation value is obtained according to the analysis of the deviation data, the deviation correlation value is subjected to data processing to obtain the deviation correlation variance, the deviation correlation variance is compared with the deviation correlation variance threshold, if the deviation correlation variance is less than the deviation correlation variance threshold, the deviation correlation value data fluctuates little, indicating that the pH value is the reason affecting the growth condition, based on the fact that the pH value is the reason affecting the growth condition, the pH value is adjusted according to the change of the pH value, the present invention can accurately identify that the pH value is a key factor affecting cell growth, and adjust the pH value accordingly to optimize the culture conditions.
[0023] Embodiment 5, as Figure 5 The heterotrophic culture system of Isochrysis galbana described in the embodiment of the present invention comprises: Data acquisition module: according to the cultivation cycle, a monitoring time axis is established, and the abnormal proportion and the cultivation progress ratio are obtained by combining the monitoring time axis and the growth situation. The abnormal proportion and the cultivation progress ratio are summed to obtain the growth abnormality value; Analysis signal acquisition module: compares the growth abnormality value with the growth abnormality threshold, and generates an analysis signal if the growth abnormality value is greater than or equal to the growth abnormality threshold; Analysis module: according to the analysis signal, pH culture data is obtained, the pH culture data includes the suitable culture time ratio and the pH value exceeding area ratio, the suitable culture time ratio and the pH value exceeding area ratio are processed to obtain the pH value deviation degree value, the pH value deviation degree value is compared with the pH value deviation degree threshold, if the pH value deviation degree value is greater than the pH value deviation degree threshold, the sub-cycle number is marked as the pH value abnormal sub-cycle number; Suspected abnormal signal acquisition module: Based on the comparison between the pH value deviation degree value and the pH value deviation degree threshold, the number of pH value abnormal sub-cycles and the number of growth abnormal sub-cycles are ratio-processed to obtain the pH value abnormality ratio, and the pH value abnormality ratio is compared with the pH value abnormality ratio threshold. If the pH value abnormality ratio is greater than or equal to the pH value abnormality ratio threshold, a suspected abnormal signal is generated; Suspected abnormal signal analysis module: based on the suspected abnormal signal, the deviation data is obtained, the deviation data includes the cell density deviation and the pH value deviation, the deviation correlation value is obtained according to the analysis of the deviation data, the deviation correlation value is processed to obtain the deviation correlation variance, the deviation correlation variance is compared with the deviation correlation variance threshold, if the deviation correlation variance is less than the deviation correlation variance threshold, the deviation correlation value data fluctuates little, indicating that the pH value is the reason affecting the growth; pH adjustment module: Based on the fact that pH is the factor that affects growth conditions, adjustments are made according to changes in pH.
[0024] The above formulas are all dimensionless and numerical calculations. The formula is a formula for the most recent real situation obtained by collecting a large amount of data and performing software simulation. The preset parameters in the formula are set by technicians in this field according to actual conditions.
[0025] The above is a detailed description of an embodiment of the present invention, but the content is only a preferred embodiment of the present invention and cannot be considered to limit the scope of implementation of the present invention. All equivalent changes and improvements made within the scope of the present invention should still fall within the scope of the patent coverage of the present invention.
Claims
1. A heterotrophic culture method of Isochrysis galbana, characterized in that: The following steps are involved: The culture cycle is preset, and the abnormal proportion and the culture progress ratio are obtained based on the combined analysis of the monitoring time axis and the growth situation. The abnormal proportion and the culture progress ratio are summed to obtain the growth abnormality value; If the growth abnormality value is greater than or equal to the growth abnormality threshold, an analysis signal is generated; According to the analysis signal, the pH culture data is processed to obtain the pH value deviation value. If the pH value deviation value is greater than the pH value deviation threshold, the sub-period is marked as a pH value abnormal sub-period, and the number of pH value abnormal sub-periods is processed by ratio processing with the number of growth abnormal sub-periods to obtain the pH value abnormality ratio. If the pH value abnormality ratio is greater than or equal to the pH value abnormality ratio threshold, a suspected abnormality signal is generated; Among them, pH culture data include the ratio of suitable culture time and the ratio of pH value exceeding area; Based on the suspected abnormal signal, the deviation data is analyzed and processed to obtain the deviation correlation value based on pH and cell density, and the deviation correlation value is processed to obtain the deviation correlation variance; Among them, the deviation data include cell density deviation and pH value deviation; If the deviation from the correlation variance is less than the deviation from the correlation variance threshold, it means that the pH value has an impact on the heterotrophic growth of Isochrysis galbana and the pH value is adjusted.
2. The heterotrophic culture method of Isochrysis galbana according to claim 1, characterized in that: The method for obtaining the abnormal proportion is as follows: The cultivation cycle is divided into several sub-cycles, and the division of the sub-cycles is matched to the monitoring time axis to obtain the growth conditions of Isochrysis globosum in the sub-cycles; According to the monitoring time axis sequence, the growth of Isochrysis spherica in any sub-period is analyzed to obtain the cell density of Isochrysis spherica in the sub-period, and the difference between the cell density of the sub-period and the standard value of the cell density of the sub-period is processed and the absolute value is taken to obtain the cell density difference of the sub-period; The sub-cycle cell density difference is compared with the sub-cycle cell density standard value. The specific comparison process is as follows: If the difference in sub-cycle cell density is less than or equal to the standard value of sub-cycle cell density, no other treatment is performed; If the cell density of a sub-cycle is greater than the standard value of the cell density of a sub-cycle, the sub-cycle is marked as a growth abnormality sub-cycle; The number of abnormal growth sub-cycles was counted, and the ratio of the number of abnormal growth sub-cycles to the number of cultured sub-cycles was processed to obtain the abnormal proportion.
3. The heterotrophic culture method of Isochrysis galbana according to claim 1, characterized in that: The method for obtaining the cultivation progress ratio is as follows: The number of sub-cycles that have been cultivated is processed by ratio with the number of sub-cycles in the cultivation period to obtain the cultivation progress ratio; The abnormal proportion and the culture progress ratio are summed to obtain the growth abnormality value, which reflects a comprehensive indicator for evaluating the growth situation during the culture cycle.
4. The heterotrophic culture method of Isochrysis galbana according to claim 1, characterized in that: The pH value deviation degree value is obtained in the following manner: The duration of the pH value in the appropriate pH value range is processed by ratio to the total time of the pH value change analysis curve to obtain the appropriate culture time ratio, and the pH value excess area is processed by ratio to the pH value change analysis curve area to obtain the pH value excess area ratio; The appropriate culture time ratio and the pH value exceeding area ratio were processed to obtain the pH value deviation degree value.
5. The heterotrophic culture method of Isochrysis galbana according to claim 4, characterized in that: The method for obtaining the suitable culture time ratio is as follows: According to the monitoring time axis, the pH value change of the sub-period is obtained to obtain the pH value change curve, and according to the pH value change curve, the pH value of the sub-period is combined with the appropriate pH value range for analysis; The pH value change curve is divided according to the number of sub-periods to obtain a number of pH value change analysis curves; Analyze the curve based on any pH value change; Mark the lowest range value of the suitable pH value and the highest range value of the suitable pH value in the coordinate system, draw a straight line parallel to the X-axis based on the marked points, mark the two straight lines as the lower limit of the suitable pH value and the upper limit of the suitable pH value, respectively, and the part between the lower limit of the suitable pH value and the upper limit of the suitable pH value is the suitable pH value range; The duration of the pH value in the appropriate pH range was counted and the ratio was processed with the total time of the pH change analysis curve to obtain the appropriate culture time ratio.
6. The heterotrophic culture method of Isochrysis galbana according to claim 4, characterized in that: The pH value exceeding the area ratio is obtained in the following manner: The area of the figure formed by the upper limit of the suitable pH value, the Y axis and the horizontal asymptote is calculated using a mathematical method integral formula to obtain the area exceeding the upper limit; Similarly, the area of the figure formed by the lower limit of the suitable pH value, the Y axis and the horizontal asymptote is calculated using the mathematical method integration formula to obtain the area exceeding the lower limit; The horizontal asymptote represents a straight line passing through the final value of the X-axis and parallel to the Y-axis; The pH value excess area is obtained by summing the upper bound excess area and the lower bound excess area; The pH value excess area and the pH value change analysis curve area are ratio-processed to obtain the pH value excess area ratio.
7. The heterotrophic culture method of Isochrysis galbana according to claim 1, characterized in that: The deviation-related value is obtained in the following manner: The cell density difference is processed by ratio processing with the cell density standard value to obtain the cell density deviation, and the pH value lower limit deviation is summed with the pH value upper limit deviation to obtain the pH value deviation; The deviation-related value was obtained by calculating the ratio of the cell density deviation to the pH value deviation.
8. The heterotrophic culture method of Isochrysis galbana according to claim 7, characterized in that: The cell density deviation is obtained by: The cell density is processed by difference with the standard value of cell density to obtain the cell density difference, and the cell density difference is processed by ratio with the standard value of cell density to obtain the cell density deviation.
9. The heterotrophic culture method of Isochrysis galbana according to claim 7, characterized in that: The pH value deviation is obtained by: The lowest pH value range difference is compared with the lowest pH value range value, and the pH lower limit deviation is obtained. The highest pH value range difference is compared with the highest pH value range value, and the pH upper limit deviation is obtained. The pH value deviation is obtained by summing the pH value lower limit deviation and the pH value upper limit deviation.
10. The heterotrophic culture method of Isochrysis galbana according to claim 9, characterized in that: The method for obtaining the pH value lower limit deviation and the pH value upper limit deviation is: According to the pH value change curve, analysis is carried out in combination with the monitoring time axis; Specifically, the pH value lower than the lowest range value of the appropriate pH value is subjected to difference processing with the lowest range value of the appropriate pH value, and the absolute value is taken to obtain the pH value lowest range difference, and the pH value lowest range difference is subjected to ratio processing with the lowest range value of the appropriate pH value to obtain the pH value lower limit deviation; Similarly, the pH value higher than the highest range value of the suitable pH value is subjected to difference processing with the highest range value of the suitable pH value, and the absolute value is taken to obtain the pH value highest range difference, and the pH value highest range difference is subjected to ratio processing with the highest range value of the suitable pH value to obtain the pH value upper limit deviation.
Citation Information
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