A method and device for simulating the cultivation of phytoplankton environmental conditions

Abstract

The present invention discloses a device and method for simulating the cultivation of phytoplankton under environmental conditions. The device includes a glass container, a support plate, a first synchronous wheel, a synchronous belt, a second synchronous wheel, a bracket, a stepper motor, a temperature probe, a salinity detector, a first upper water level probe water adding device, a second upper water level probe water adding device and a lower water level probe water pumping device. The cultivation method comprises the steps of collecting the environmental conditions for the growth of the phytoplankton, including light, temperature, salinity, etc.; using multi-condition control intelligent software to separate or synthesize the three conditions of light-temperature-salinity, obtain the frequency and range of environmental condition changes, and cultivate the phytoplankton. The present invention can analyze and judge the adaptability and optimal growth conditions of phytoplankton to light, temperature, and salinity through one experiment, thereby screening out the optimal environmental conditions for phytoplankton, which is conducive to the rapid development of relevant physiological experiments on phytoplankton and the mechanism of red tide outbreak, saving a lot of manpower and material resources.

Description

Technical Field

[0001] The invention relates to the field of marine science and technology, and in particular to a method and device for simulating the cultivation of phytoplankton under environmental conditions. Background Art

[0002] Phytoplankton growth is affected by many biological and non-biological environmental factors, including predators, light, temperature, salinity, nutrients and water flow speed, among which light, temperature and salinity are three key factors. According to the growth and distribution characteristics of phytoplankton, the frequency and range of changes in the three environmental conditions of light, temperature and salinity can be intelligently controlled to simulate real environmental conditions and study the growth of phytoplankton under real environmental conditions.

[0003] Phytoplankton, also known as microalgae, have chromatophores like land plants and need sunlight to grow and reproduce. Most phytoplankton float in the upper layer of seawater where sunlight can penetrate. And the growth of phytoplankton also requires CO 2 , nutrients, temperature, salinity, water depth and wind, etc. When conditions are right, phytoplankton populations grow in an explosive manner, which will form red tides.

[0004] In order to study the effect of environmental conditions on the growth of phytoplankton, especially the formation of red tides, the method of simulating environmental conditions is often used to study plankton. The defects of current experimental devices and experimental methods are that the environmental conditions of light, temperature and salinity are mostly set separately and cannot be automatically and intelligently controlled and monitored over time. They cannot simulate the real environmental conditions of continuous changes and complex types, and cannot be applied and reacted to the complex mechanism research of offshore phytoplankton growth and proliferation and red tide outbreaks. Therefore, an experimental method and device is invented that can analyze and judge the three environmental conditions of light, temperature and salinity of phytoplankton in one experiment, and intelligently control them on the same experimental object at the same time, which is conducive to the laboratory research and practical application of the physiological mechanism of phytoplankton environmental condition simulation experiment and growth. Summary of the invention

[0005] The technical problem to be solved by the embodiments of the present invention is that, in the current experimental methods for studying the effects of environmental conditions on the growth of phytoplankton, the environmental conditions of light, temperature and salinity are mostly set individually and cannot be automatically and intelligently controlled and monitored over time, and cannot simulate the continuously changing complex real environmental conditions, and cannot be applied and responded to the complex mechanism research of the growth and proliferation of offshore phytoplankton and the outbreak of red tides. A method for simulating the cultivation of phytoplankton environmental conditions is proposed.

[0006] In order to solve the above technical problems, the present invention provides a phytoplankton environmental condition simulation culture device, which comprises: a glass container, a support plate, a first synchronous wheel, a synchronous belt, a second synchronous wheel, a bracket, a stepper motor, a temperature probe, a salinity detector, a first upper water level probe water adding device, a second upper water level probe water adding device and a lower water level probe water pumping device;

[0007] The glass container is arranged on a support plate, a first synchronous wheel is installed at the bottom of the support plate, the first synchronous wheel is cooperatively installed on the upper end surface of the bracket, a second synchronous wheel drives the first synchronous wheel to rotate through a synchronous belt, the second synchronous wheel is fixedly arranged on the top of the stepper motor, the stepper motor is fixedly installed on the inner side of the bottom of the bracket, the bottom of the bracket is fixedly supported by a support plate, and a support bottom plate is fixedly installed at the bottom of the support plate;

[0008] A temperature heating controller is installed on the inner side of the bottom of the glass container, and a light tube is installed on the inner side of the top. A quantitative culture medium nutrient liquid is filled in the glass container. A temperature probe, a salinity detector, a first upper water level probe water adding device, a second upper water level probe water adding device and a lower water level probe water pumping device are all inserted in the glass container, and at least part of them are arranged in the nutrient solution.

[0009] The temperature probe is used to detect the temperature of the nutrient solution, the salinity detector is used to detect the brine concentration of the nutrient solution, the first upper water level probe water adding device is used to add nutrient solution with a good concentration, the second upper water level probe water adding device is used to add distilled water to adjust the brine concentration, and the lower water level probe pumping device is used to extract the nutrient solution.

[0010] Among them, the stepper motor, temperature heating controller, light tube, temperature probe, salinity detector, first upper water level probe water adding device, second upper water level probe water adding device and lower water level probe water pumping device are all electrically connected to the computer host or mobile phone.

[0011] Among them, the support plate is made of plastic material, including a base plate and multiple straight plates that are evenly spaced and vertically connected to the base plate. There is a gap of 1-2mm between the inner side of the straight plate and the outer side of the glass container. The support plate is tilted with an inclination angle of 5-10 degrees. The support plate is connected to the first synchronous wheel through multiple bearings.

[0012] Among them, the temperature probe, salinity detector, the first upper water level probe water adding device, the second upper water level probe water adding device and the lower water level probe pumping device are clamped by plastic brackets and inserted into the glass container, and the bottom of the plastic bracket is fixed on the workbench outside the phytoplankton environmental condition simulation culture device.

[0013] Wherein, a protruding circular frame is arranged on the upper end surface of the bracket, and the first synchronous wheel is inserted into the circular frame.

[0014] The inner side of the bottom of the bracket is connected to the upper surface of the body of the stepper motor by screws.

[0015] The lower end surface of the bracket is connected to the upper end surface of the support plate by screws, and the lower end surface of the support plate is connected to the upper end surface of the support base plate by screws.

[0016] In order to solve the above technical problems, the present invention also provides a method for simulating the cultivation of phytoplankton environmental conditions, comprising the following steps:

[0017] Step 1: Investigate and collect the growth environment conditions of phytoplankton used in the experiment: including geographical distribution, whether red tides are formed, natural environment records of the three growth conditions of light, temperature, and salinity, and screening of experimental conditions;

[0018] Step 2: Cultivate the phytoplankton required for the experiment, and perform laboratory pre-cultivation using culture medium and an algae cultivation room with controlled temperature and light;

[0019] Step 3, adding culture medium nutrient solution to the phytoplankton environmental condition simulation culture device, and then setting the environmental conditions of the phytoplankton environmental condition simulation culture device according to the natural environment record of the growth conditions in step 1 on a computer or mobile phone, including the frequency and range of changes in light, temperature and salinity and the duration of the experiment;

[0020] Step 4: Place the phytoplankton obtained by the experimental culture into a phytoplankton environmental condition simulation culture device according to a certain density ratio, and start the culture;

[0021] Step 5, taking a quantitative phytoplankton sample from the phytoplankton environmental condition simulation culture device at a certain interval;

[0022] Step 6: Add a fixative to a portion of the taken phytoplankton samples, and count the cells of the sample with the fixative; and detect and record the chlorophyll fluorescence Fv / Fm of the other portion of the samples;

[0023] Step 7: Based on the data obtained in step 6, analyze the growth trend and changing rules of phytoplankton.

[0024] Wherein, the culture medium in step 2 and step 3 is f2 culture medium; the pre-culture temperature in step 2 is 20 degrees, and the light intensity is 1500 lux.

[0025] Among them, the density of phytoplankton in step 4 is 105-106 cells / L; the interval time in step 5 is 2 days; in step 6, 2 ml of phytoplankton sample is taken out, a fixative is added, the cells are counted, a growth curve is drawn, and then 2 ml of phytoplankton sample is taken out, the chlorophyll fluorescence Fv / Fm is detected and recorded, and the maximum growth rate is determined.

[0026] The implementation of the present invention has the following beneficial effects: the present invention can analyze and determine the adaptability and optimal growth conditions of phytoplankton to light, temperature, and salinity through a single experiment, thereby screening out the optimal environmental conditions for phytoplankton, which is beneficial to the rapid development of relevant physiological experiments on phytoplankton and the mechanism of red tide outbreaks, saving a lot of manpower and material resources. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. 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 paying creative work.

[0028] Figure 1 A schematic diagram of the three-dimensional structure of the phytoplankton environmental condition simulation culture device provided by the present invention;

[0029] Figure 2 A schematic side view of the structure of the phytoplankton environmental condition simulation culture device provided by the present invention;

[0030] Figure 3 This is a schematic structural diagram of a glass container provided by the present invention.

[0031] In the figure: glass container 1, support plate 2, first synchronous wheel 3, bracket 4, second synchronous wheel 5, synchronous belt 6, stepper motor 7, support plate 8, support bottom plate 9, lighting lamp 10, temperature probe 11, salinity detector 12, first upper water level probe water adding device 13, second upper water level probe water adding device 14, lower water level probe water pumping device 15, bottom plate 16, straight plate 17 and circular frame 18. DETAILED DESCRIPTION

[0032] The following will be combined with the accompanying drawings in the present invention to clearly and completely describe the technical solutions 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 are within the scope of protection of the present invention.

[0033] See also Figure 1-3 , Figure 1 A schematic diagram of the three-dimensional structure of the phytoplankton environmental condition simulation culture device provided by the present invention; Figure 2 A schematic side view of the structure of the phytoplankton environmental condition simulation culture device provided by the present invention; Figure 3 This is a schematic structural diagram of a glass container provided by the present invention.

[0034] The present invention provides a phytoplankton environmental condition simulation culture device, which comprises: a glass container 1 is arranged on a support plate 2, the support plate 2 is made of plastic material, comprises a bottom plate 16 and a plurality of straight plates 17 evenly spaced and vertically connected to the bottom plate 16, and there is a gap of 1-2 mm between the inner side of the straight plate 17 and the outer side of the glass container 1. The support plate 2 is inclined at an angle of 5-10 degrees, and the inclined setting of the support plate 2 further causes the glass container 1 to be inclined at 5-10 degrees.

[0035] The support plate 2 is connected with the first synchronous wheel 3 at the bottom thereof through a plurality of bearings. A protruding circular frame 18 is provided on the upper end surface of the bracket 4, the first synchronous wheel 3 is inserted on the circular frame 18, the second synchronous wheel 5 drives the first synchronous wheel 3 to rotate through the synchronous belt 6, the second synchronous wheel 5 is fixedly provided on the top of the stepping motor 7 and connected with the stepping motor 7, the inner side of the bottom of the bracket 4 is connected with the upper end surface of the body of the stepping motor 7 by screws, the lower end surface of the bracket 4 is connected with the upper end surface of the support plate 8 by screws, and the lower end surface of the support plate 8 is connected with the upper end surface of the support bottom plate 9 by screws.

[0036] A temperature heating controller is installed on the inner side of the bottom of the glass container 1, and a light tube 10 is installed on the inner side of the top. A quantitative culture medium nutrient liquid is installed in the glass container 1, and a temperature probe 11, a salinity detector 12, a first upper water level probe water adding device 13, a second upper water level probe water adding device 14 and a lower water level probe water pumping device 15 are all inserted in the glass container 1, and at least part of them are set in the nutrient liquid. The temperature probe 11 is used to detect the temperature of the nutrient liquid, the salinity detector 12 is used to detect the salt water concentration of the nutrient liquid, the first upper water level probe water adding device 13 is used to add a nutrient liquid with a good concentration, the second upper water level probe water adding device 14 is used to add distilled water to adjust the salt water concentration, and the lower water level probe water pumping device 15 is used to extract the nutrient liquid.

[0037] The stepper motor 7, temperature heating controller, light tube 10, temperature probe 11, salinity detector 12, first upper water level probe water adding device 13, second upper water level probe water adding device 14 and lower water level probe pumping device 15 in this simulation culture device are all electrically connected to the computer host or mobile phone, and are controlled by the phytoplankton environmental condition simulation culture box automatic control system software in the computer or mobile phone.

[0038] The temperature probe 11, the salinity detector 12, the first upper water level probe water adding device 13, the second upper water level probe water adding device 14 and the lower water level probe pumping device 15 are respectively clamped by plastic brackets and inserted into the glass container 1, and the bottom of the plastic bracket is fixed on the workbench outside the phytoplankton environmental condition simulation culture device.

[0039] The present invention supports the glass container 1 through the support plate 2, and the first synchronous wheel 3 under the support plate 2 together with the bearing therein is rotated at a uniform speed by the second synchronous wheel 5 on the top of the stepping motor 7 through the synchronous belt 6, so that the liquid in the glass container 1 will flow at a uniform speed. During the rotation process, timed turbulence can also be set to simulate the flow of seawater in the marine environment.

[0040] The present invention also provides a salinity detector 12, a first upper water level probe water adding device 13 (adding nutrient solution and salt water with a good concentration), a second upper water level probe water adding device 14 (adding distilled water), and a lower water level probe water pumping device 15 in the glass container 1. When the liquid in the glass container needs to be replaced regularly, the liquid pump will extract liquid from the lower water level probe water pumping device 15, stop when the liquid is pumped to the lower water level probe at the bottom of the container, and then add the nutrient solution with a good concentration from the first upper water level probe water adding device 13 to the position of the upper water level probe and stop, and then detect the liquid concentration through the salinity detector 12, and adjust the concentration of the liquid in the glass container 1 through the first upper water level probe water adding device 13 and the second upper water level probe water adding device 14.

[0041] The present invention also provides a method for simulating the cultivation of phytoplankton under environmental conditions, comprising the following steps:

[0042] Step 1: Investigate and collect the growth environment conditions of phytoplankton used in the experiment: including geographical distribution, whether red tides are formed, natural environment records of the three growth conditions of light, temperature, and salinity, and screening of experimental conditions.

[0043] Step 2: Cultivate the phytoplankton required for the experiment, and use F2 culture medium and a temperature-controlled, light-illuminated algae culture room for laboratory pre-cultivation. The pre-cultivation temperature is 20 degrees and the light intensity is 1500 lux.

[0044] Step 3, add F2 culture medium nutrient solution to the phytoplankton environmental condition simulation culture device, and then set the environmental conditions of the phytoplankton environmental condition simulation culture device according to the natural environment record of the growth conditions in step 1 on a computer or mobile phone, including the frequency and range of changes in light, temperature and salinity and the duration of the experiment.

[0045] Step 4: Place the phytoplankton obtained from the experimental culture at a density of 105-106 cells / L into a phytoplankton environmental condition simulation culture device and start culture.

[0046] Step 5: Take a quantitative amount of phytoplankton samples from the phytoplankton environmental condition simulation culture device every 2 days.

[0047] Step 6: Take 2 ml of the taken phytoplankton sample and add a fixative, and count the cells of the sample with the fixative; then take another 2 ml of the sample to detect and record the chlorophyll fluorescence Fv / Fm.

[0048] Step 7: Based on the data obtained in step 6, a growth curve is drawn to determine the maximum growth rate and analyze the growth trend and change rules of phytoplankton.

[0049] In this method, the lighting, temperature, salinity and glass container speed are set on the phytoplankton environmental condition simulation incubator automatic control system software installed on a computer or mobile phone terminal, and the real environmental conditions for phytoplankton growth are simulated by the momentary changes in lighting, temperature and salinity. The system parameters and performance are as follows: temperature setting range: 0-40℃; light intensity setting range: 0-5000LUX; platform operation speed: 0-15r / min; salinity setting range: 0-proportional liquid concentration; liquid replacement time setting range: 1-999 hours; power supply voltage: AC220V±5%, good grounding; whole machine power ≤600W. Data recording time: 30min; working environment 0-60℃, relative temperature <90%; RS485 communication is used, and the computer collects data in real time.

[0050] The composition ratio of the F2 medium in this method is shown in the following table:

[0051]

[0052] The present invention provides a method and device for simulating the cultivation of phytoplankton under environmental conditions. The method collects the environmental conditions for the growth of the phytoplankton, including light, temperature and salinity; uses multi-condition control intelligent software to separate or synthesize the three conditions of light, temperature and salinity, obtains the frequency and range of environmental condition changes, cultivates the phytoplankton, and observes the growth of the phytoplankton under the intelligent control changes of the three environmental conditions, including changes in cell morphology and density.

[0053] The present invention can analyze and determine the adaptability and optimal growth conditions of phytoplankton to light, temperature, and salinity through a single experiment, thereby screening out the optimal environmental conditions for phytoplankton, which is beneficial to the rapid development of relevant physiological experiments on phytoplankton and the mechanism of red tide outbreaks, saving a lot of manpower and material resources.

[0054] The phytoplankton environmental condition simulation culture device has a simple structure, requires a small number of instruments and equipment, and is highly usable; the phytoplankton environmental condition simulation culture method does not require manual operation for data rectification, and a set of data can be imported into the software to perform operations for continuous light, temperature and salinity changes; it is easy to operate, the experimental time is controllable, and the workload is small, and the three environmental conditions can be simulated quickly and effectively.

[0055] The experimental method and device of the present invention are applicable to most phytoplankton, and can be used to conduct experimental research on the growth conditions of various phytoplankton or carry out other related experiments.

[0056] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A phytoplankton environmental condition simulation culture device, characterized in that: include: Glass container, support plate, first synchronous wheel, synchronous belt, second synchronous wheel, bracket, stepper motor, temperature probe, salinity detector, first upper water level probe water adding device, second upper water level probe water adding device and lower water level probe water pumping device; The glass container is arranged on the support plate, the bottom of the support plate is installed with the first synchronous wheel, the first synchronous wheel is cooperatively installed on the upper end surface of the bracket, the second synchronous wheel drives the first synchronous wheel to rotate through the synchronous belt, the second synchronous wheel is fixedly arranged on the top of the stepper motor, the stepper motor is fixedly installed on the inner side of the bottom of the bracket, the bottom of the bracket is fixedly supported by a support plate, and a support bottom plate is fixedly installed on the bottom of the support plate; The support plate is made of plastic material, including a bottom plate and a plurality of straight plates evenly spaced and vertically connected to the bottom plate, a gap of 1-2 mm exists between the inner side of the straight plate and the outer side of the glass container, the support plate is tilted, and the tilt angle is 5-10 degrees, and the support plate is connected with the first synchronous wheel through a plurality of bearings; A temperature heating controller is installed on the inner side of the bottom of the glass container, and a light tube is installed on the inner side of the top. A quantitative culture medium nutrient liquid is filled in the glass container. The temperature probe, the salinity detector, the first upper water level probe water adding device, the second upper water level probe water adding device and the lower water level probe water pumping device are all inserted into the glass container, and at least partially arranged in the nutrient liquid. The temperature probe is used to detect the temperature of the nutrient solution, the salinity detector is used to detect the salt water concentration of the nutrient solution, the first upper water level probe water adding device is used to add the nutrient solution with a good concentration, the second upper water level probe water adding device is used to add distilled water to adjust the salt water concentration, and the lower water level probe water pumping device is used to extract the nutrient solution; The temperature probe, the salinity detector, the first upper water level probe water adding device, the second upper water level probe water adding device and the lower water level probe water pumping device are respectively clamped by plastic brackets and inserted into the glass container, and the bottom of the plastic bracket is fixed to the workbench outside the phytoplankton environmental condition simulation culture device.

2. The phytoplankton environmental condition simulation culture device according to claim 1, characterized in that: The stepper motor, the temperature heating controller, the light tube, the temperature probe, the salinity detector, the first upper water level probe water adding device, the second upper water level probe water adding device and the lower water level probe water pumping device are all electrically connected to a computer host or a mobile phone.

3. The phytoplankton environmental condition simulation culture device according to claim 1, characterized in that: A protruding circular frame is arranged on the upper end surface of the bracket, and the first synchronous wheel is inserted into the circular frame.

4. The phytoplankton environmental condition simulation culture device according to claim 1, characterized in that: The inner side of the bottom of the bracket is connected to the upper surface of the body of the stepping motor by screws.

5. The phytoplankton environmental condition simulation culture device according to claim 1, characterized in that: The lower end surface of the bracket is connected to the upper end surface of the support plate by screws, and the lower end surface of the support plate is connected to the upper end surface of the support base plate by screws.

6. A method for simulating the cultivation of phytoplankton environmental conditions, characterized in that: The phytoplankton environmental condition simulation culture method comprises the following steps: Step 1: Investigate and collect the growth environment conditions of phytoplankton used in the experiment: including geographical distribution, whether red tides are formed, natural environment records of the three growth conditions of light, temperature, and salinity, and screening of experimental conditions; Step 2: Cultivate the phytoplankton required for the experiment, and perform laboratory pre-cultivation using culture medium and an algae cultivation room with controlled temperature and light; Step 3, adding culture medium nutrient solution to the phytoplankton environmental condition simulation culture device according to claim 1, and then setting the environmental conditions of the phytoplankton environmental condition simulation culture device on a computer or mobile phone according to the natural environment record of the growth conditions in step 1, including the frequency and range of changes in light, temperature and salinity and the duration of the experiment; Step 4: Place the phytoplankton obtained by the experimental culture into a phytoplankton environmental condition simulation culture device according to a certain density ratio, and start the culture; Step 5, taking a quantitative phytoplankton sample from the phytoplankton environmental condition simulation culture device at a certain interval; Step 6: Add a fixative to a portion of the taken phytoplankton samples, and count the cells of the sample with the fixative; and detect and record the chlorophyll fluorescence Fv / Fm of the other portion of the samples; Step 7: Based on the data obtained in step 6, analyze the growth trend and changing rules of phytoplankton.

7. The method for simulating culturing phytoplankton environmental conditions according to claim 6, characterized in that: The culture medium in step 2 and step 3 is f2 culture medium; the pre-culture temperature in step 2 is 20 degrees, and the light intensity is 1500 lux.

8. The method for simulating culturing phytoplankton environmental conditions according to claim 6, characterized in that: In step 4, the density of phytoplankton is 105-106 cells / L; in step 5, the interval time is 2 days; in step 6, 2 ml of phytoplankton sample is taken out, a fixative is added, cells are counted, a growth curve is drawn, and then 2 ml of phytoplankton sample is taken out, chlorophyll fluorescence Fv / Fm is detected and recorded, and the maximum growth rate is determined.

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