Method and use of a diaphragm vacuum pump and a microporous filter membrane for counting small numbers of brine shrimps

Abstract

The application discloses a method for counting a small amount of brine shrimps by using a diaphragm vacuum pump and a microporous filter film, and comprises the following steps: placing the microporous filter film above a filter head of the diaphragm vacuum pump, pouring hatching liquid into a filter cup of the diaphragm vacuum pump for filtration, and filtering the hatched brine shrimps on the microporous filter film by using the diaphragm vacuum pump, so that the hatched brine shrimp larvae and empty shells can be clearly distinguished on the microporous filter film, and the brine shrimps can be accurately counted under naked eyes or a dissecting microscope. The method greatly reduces the loss of the number of brine shrimps in the process of hatching the brine shrimps in a laboratory, reduces experimental data errors, more accurately completes counting, greatly improves the counting accuracy, and obtains more accurate hatching rates of brine shrimp eggs. The method is simple in steps and easy to operate, provides a better method for accurately calculating the hatching rate of the brine shrimps, and lays an important foundation for experiments related to the brine shrimps.

Description

Technical Field

[0001] The invention belongs to the technical field of aquaculture, and in particular relates to a method for counting a small amount of Artemia by using a diaphragm vacuum pump and a microporous filter membrane and its application. Background Art

[0002] Artemia, also known as brine shrimp, hatches from Artemia eggs. Due to their palatability and high nutritional value, the nauplii hatched from Artemia eggs are suitable for use as bait and as a live carrier of trace elements. They are currently widely used in the cultivation of fish, shrimp, and crab seedlings. Research shows that over 85% of marine aquaculture species worldwide rely on Artemia as a primary source of larval food. Artemia not only improves the survival rate of fish and shrimp fry but also serves as a carrier for the prevention and treatment of fish and shrimp diseases. Artemia is also widely used as an important experimental material in biotechnology research. In recent years, rampant overfishing has severely damaged the ecological environment and Artemia resources. With the rapid development of the aquaculture industry, the demand for Artemia is increasing. Natural Artemia cannot meet market demand, making artificial cultivation a necessity. When studying the optimal habitat for Artemia cultivation, the most important and initial consideration is the hatching and survival rates of Artemia under different environments. Therefore, calculating hatching and survival rates requires counting methods.

[0003] At present, the method of counting a small amount of Artemia using a separation filter and a rubber-tipped dropper is generally used to calculate the hatching rate. The main steps of this method are as follows:

[0004] like Figure 1 and Figure 2 As shown, the separation filter consists of a thin mesh and a dense mesh. The mesh pocket is made of nylon. The dense mesh typically has a pore size of 180 mesh, while the thin mesh has a pore size of 90-100 mesh. When in use, the thin mesh is placed on top, with the dense mesh below. The thin mesh pocket is approximately 11-15 cm deep, while the dense mesh pocket is approximately 14-18 cm deep. The distance between the two meshes is generally 3-4 cm. The mesh openings are typically made of durable stainless steel, with a diameter typically ranging from 75 mm to 110 mm. They have silicone or wooden handles approximately 8 cm long. The separation process involves removing hatched Artemia eggs from the separation filter and flushing them with water. Sinking eggs or empty shells are trapped in the thin mesh on top, while pure brine shrimp larvae remain in the dense mesh on the bottom. The separation filter is then disassembled, and the larvae trapped in the dense mesh are aspirated with a rubber-tipped pipette and placed in a Petri dish. The larvae are then counted visually in the dish. This technique is primarily used in laboratory experiments to calculate Artemia hatchability.

[0005] When using the common method in the prior art, a large amount of larvae are lost when using the separation filter. The specific reasons are as follows:

[0006] 1. The larvae are relatively small. When most of the Artemia gathered together in the dense net are sucked up with a rubber-tipped dropper, many scattered larvae will stick to the net and are not easy to be sucked up.

[0007] 2. When the Artemia sucked out with a rubber-tipped dropper is placed in a culture dish for counting, since the newly hatched Artemia are alive and have the characteristics of flexible swimming, it is difficult to control the counting with the naked eye or under a dissecting microscope, resulting in inaccurate counting results.

[0008] Therefore, one or more new related methods are urgently needed. Summary of the Invention

[0009] The purpose of the present invention is to overcome the problems existing in the prior art and provide a method and application for counting a small amount of Artemia using a diaphragm vacuum pump and a microporous filter membrane.

[0010] The technical solution adopted by the present invention to solve the technical problem is:

[0011] A method for counting a small amount of Artemia using a diaphragm vacuum pump and a microporous filter membrane comprises the following steps:

[0012] Place the microporous filter membrane above the filter head of the diaphragm vacuum pump, and pour the hatching liquid into the filter cup of the diaphragm vacuum pump for filtration. The diaphragm vacuum pump uses a common negative pressure type with a pumping rate of 20L / min, a limit vacuum of 0.08Mpa, and a motor power of 160W. There is no requirement for filtration time. When there is no water in the filter cup, unplug the silicone tube on the filter cup, turn off the vacuum pump, and use the diaphragm vacuum pump to filter the hatched Artemia onto the microporous filter membrane. Finally, the hatched Artemia larvae and empty shells can be clearly distinguished on the microporous filter membrane, and can be accurately counted with the naked eye or under a dissecting microscope.

[0013] Furthermore, the Artemia larvae are orange-red teardrop-shaped, and the empty shells or sunken eggs are brown spherical.

[0014] Furthermore, the microporous filtration membrane is a water-based microporous filtration membrane with a diameter of 50 mm and a pore size of 0.45 μm. The material of the filtration membrane is a mixed cellulose MCE membrane.

[0015] Furthermore, the specific steps are as follows:

[0016] The diaphragm vacuum pump equipped with microporous filter membrane is energized for standby use, the incubator equipped with Artemia larvae is placed in the upper filter cup mouth of the vacuum pump, the incubator bottle is slowly opened, the salt solution with a salinity of 30 ‰ slowly flows into the upper filter cup of the vacuum pump, the incubator is rinsed, and it is guaranteed that there is no residue in the incubator, the vacuum pump is opened for filtration, and when the salt solution is about to leak, the upper filter cup wall of the vacuum pump is rinsed to ensure that there is no residue on the filter cup wall, and final Artemia larvae, ovules and sunken eggs are completely sunken on the microporous filter membrane, the vacuum pump is turned off, the microporous filter membrane is clamped by tweezers, and counting is started;

[0017] Among them, the pH of salt water is between 7-8.

[0018] Furthermore, the microporous filtration membrane is nylon 66, polyethersulfone PSE, polytetrafluoroethylene PTFE, polyvinylidene fluoride PVDF or polypropylene PP.

[0019] Application of the above method in Artemia counting.

[0020] The beneficial effects achieved by the present invention are:

[0021] 1. The method of the present invention greatly reduces the loss of Artemia during laboratory hatching, reduces experimental data errors, completes counting more accurately, greatly improves counting accuracy, and obtains a more accurate Artemia egg hatch rate. The method is simple and easy to operate, providing a more high-quality method for accurately calculating Artemia hatch rates and laying an important foundation for Artemia-related experiments.

[0022] 2. The method of the present invention uses a microporous filtration membrane, which is a special filtration membrane for water systems. The diameters of the microporous filtration membrane are generally 25mm, 47mm, 50mm and 60mm, and the pore sizes are 0.2μm, 0.45μm, and 0.8μm. Artemia eggs are generally between 200μm and 250μm, and newly hatched Artemia larvae are generally between 400μm and 800μm. The above pore sizes are much smaller than the diameters of Artemia eggs and Artemia larvae, and microporous filtration membranes of different specifications can be freely selected. The filter membrane is made of mixed cellulose MCE membrane, which has a smooth surface, a light texture, a high porosity, good retention effect, good hydrophilicity, a large water flux, a uniform microporous structure, and an extremely low adsorption rate. It can tolerate pH values ​​of 2-9 and is suitable for filtering aqueous drugs or other aqueous solvents. It is commonly used in laboratories, removing particles, sterilizing and filtering culture media and culture fluids, and determining bacterial flora in water bodies.

[0023] 3. The method of the present invention uses a microporous filtration membrane, which is made of nylon 66, which has good hydrophilicity, high strength, high flow rate, high porosity, and is suitable for high flux of most organic solvents and water solvents. It is resistant to high temperatures, acidic water solvents and organic solvents, has high surface quality and is compatible with a wide range of drugs.

[0024] The microporous filtration membrane is polyethersulfone PSE, a hydrophilic membrane with high chemical and thermal stability and strong acid and alkali resistance; the microporous filtration membrane is polytetrafluoroethylene PTFE, a hydrophobic membrane with good chemical compatibility with high temperature resistance, strong acid and strong alkali organic solvents and oxidants, and is resistant to most chemical solvents;

[0025] The microporous filtration membrane is polyvinylidene fluoride (PVDF), a hydrophobic membrane that does not absorb moisture, has a very low protein adsorption rate, and is resistant to chemical corrosion and oxidation.

[0026] The microporous filtration membrane is made of polypropylene (PP), which provides deep filtration, stable physical and chemical properties, good compatibility, high porosity, large dirt holding capacity, can be backflushed and sterilized at high temperature, and has good pressure resistance.

[0027] The above materials differ in their primary purpose of filtering water quality, and all can achieve the purpose of filtering salt water and retaining Artemia larvae and Artemia eggs. However, due to their functionality, the filter membranes of the above materials have obvious fibrous structures on the surface of the membranes themselves, which will affect the Artemia counting effect to a certain extent. In addition, these highly functional filter membranes are expensive to produce. In addition, the solution involved in this experiment is ordinary salt water with a pH between 7 and 8, which does not require resistance to strong acids, strong alkalis, or corrosion. Therefore, the above filter membranes can be used instead, but the water-specific filter membrane - mixed cellulose MCE membrane is the most economical and practical for this experiment. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 A photo of a sparse net and a dense net in the prior art "a method for counting a small amount of Artemia using a separation filter and a rubber-tipped dropper";

[0029] Figure 2 A photograph of a method for counting a small amount of Artemia using a separation filter and a rubber-tipped dropper in the prior art, wherein the method involves "sucking out the larvae from the dense mesh with a rubber-tipped dropper, placing them in a culture dish, and finally completing the counting by observation in the culture dish";

[0030] Figure 3 This is a physical diagram of the connection of various parts of the diaphragm vacuum pump in the present invention;

[0031] Figure 4 This is a photo of a water-based microporous filtration membrane with a diameter of 50 mm and a pore size of 0.45 μm used in the method of the present invention;

[0032] Figure 5 This is a photo of the present invention showing the removal of the microporous filter membrane after filtration and centralized counting;

[0033] Figure 6 This is a photo of the microporous filtration membrane of the present invention just after filtration is completed on a vacuum pump;

[0034] Figure 7 These are photos of incubators No. 1 to No. 6 in the present invention;

[0035] Figure 8 This is a photo of the counting and statistics of egg shells and Artemia larvae on the filter membrane in the embodiment of the present invention;

[0036] Figure 9 A schematic diagram of a process flow of an embodiment of the present invention;

[0037] Figure 10This is a photograph showing that after the filtration is completed in the embodiment of the present invention, the Artemia larvae that have lost water and stagnated on the filter membrane can continue to survive if they are re-placed in salt water within 10 minutes;

[0038] Figure 11 This is a microscopically enlarged diagram demonstrating that Artemia larvae that have lost water and stagnated on the filter membrane after filtration in an embodiment of the present invention can continue to survive if they are re-placed in salt water within 10 minutes. DETAILED DESCRIPTION

[0039] For a better understanding of the present invention, the present invention is further described in detail below with reference to the embodiments. However, the scope of protection claimed by the present invention is not limited to the scope represented by the embodiments.

[0040] Unless otherwise specified, the raw materials used in the present invention are all conventional commercial products. Unless otherwise specified, the methods used in the present invention are all conventional methods in the art. The quality of each substance used in the present invention is the quality of conventional use.

[0041] A method for counting a small amount of Artemia using a diaphragm vacuum pump and a microporous filter membrane comprises the following steps:

[0042] Place the microporous filter membrane above the filter head of the diaphragm vacuum pump, and pour the hatching liquid into the filter cup of the diaphragm vacuum pump for filtration. The diaphragm vacuum pump uses a common negative pressure type with a pumping rate of 20L / min, a limit vacuum of 0.08Mpa, and a motor power of 160W. There is no requirement for filtration time. When there is no water in the filter cup, unplug the silicone tube on the filter cup, turn off the vacuum pump, and use the diaphragm vacuum pump to filter the hatched Artemia onto the microporous filter membrane. Finally, the hatched Artemia larvae and empty shells can be clearly distinguished on the microporous filter membrane, and can be accurately counted with the naked eye or under a dissecting microscope.

[0043] like Figure 3 As shown, the diaphragm vacuum pump generally includes a filter cup 1, a filter head 2, a fixing clamp 3, a triangular flask 5, a intercepting bottle 4, a vacuum filtration pump body 7 and a silicone tube 6. When in use, the filter head is tightly installed on the triangular flask, and then the filter cup is tightly installed on the filter head and fixed with a fixing clamp. The filter head and the negative pressure gas nozzle of the vacuum filtration pump body are tightly connected with a silicone tube. The intercepting bottle is tightly set between the filter head and the negative pressure gas nozzle of the vacuum filtration pump body to prevent liquid from flowing back into the pump body.

[0044] Preferably, the Artemia larvae are orange-red teardrop-shaped, and the empty shells or sunken eggs are brown spherical.

[0045] Preferably, if Figure 4 As shown, the microporous filtration membrane is a water-based microporous filtration membrane with a diameter of 50 mm and a pore size of 0.45 μm. The material of the membrane is a mixed cellulose MCE membrane.

[0046] Preferably, the specific steps are as follows:

[0047] The diaphragm vacuum pump equipped with microporous filter membrane is energized for standby use, the incubator equipped with Artemia larvae is placed in the upper filter cup mouth of the vacuum pump, the incubator bottle is slowly opened, the salt solution with a salinity of 30 ‰ slowly flows into the upper filter cup of the vacuum pump, the incubator is rinsed, and it is guaranteed that there is no residue in the incubator, the vacuum pump is opened for filtration, and when the salt solution is about to leak, the upper filter cup wall of the vacuum pump is rinsed to ensure that there is no residue on the filter cup wall, and final Artemia larvae, ovules and sunken eggs are completely sunken on the microporous filter membrane, the vacuum pump is turned off, the microporous filter membrane is clamped by tweezers, and counting is started;

[0048] Among them, the pH of salt water is between 7-8.

[0049] Preferably, the microporous filtration membrane is nylon 66, polyethersulfone PSE, polytetrafluoroethylene PTFE, polyvinylidene fluoride PVDF or polypropylene PP.

[0050] Application of the above method in Artemia counting.

[0051] Specifically, the relevant preparation and testing are as follows:

[0052] A method for counting a small amount of Artemia using a diaphragm vacuum pump and a microporous filter membrane comprises the following steps:

[0053] Use a diaphragm vacuum pump to filter the hatched Artemia onto a microporous filter membrane. Microporous filter membranes are specialized for water systems and typically have diameters of 25mm, 47mm, 50mm, and 60mm, with pore sizes of 0.2μm, 0.45μm, and 0.8μm. Artemia eggs typically range from 200μm to 250μm, while newly hatched Artemia larvae typically range from 400μm to 800μm. These pore sizes are much smaller than those of Artemia eggs and larvae, allowing for the selection of microporous filter membranes of varying specifications. Water-based microporous filter membranes are made of mixed cellulose MCE membranes, which offer a smooth surface, lightweight texture, high porosity, excellent retention, good hydrophilicity, high water flux, a uniform microporous structure, and extremely low adsorption. They tolerate a pH range of 2-9 and are suitable for filtering aqueous pharmaceuticals or other aqueous solvents. They are commonly used in laboratories, for particle removal, sterilization filtration of culture media and broth, and for the determination of bacterial flora in water.

[0054] This method uses the most economical and practical 50mm, 0.45μm microporous filter membrane. Place the membrane above the filter head of a diaphragm vacuum pump, pour the hatching solution into the filter cup, and filter it. The microporous filter membrane allows for clear differentiation between hatched Artemia larvae and empty shells. Artemia larvae are typically orange-red, drop-shaped, while empty shells or sunken eggs are generally brown, spherical. Accurate counting is possible with the naked eye or under a dissecting microscope.

[0055] like Figures 5 to 8As shown, 300 frozen eggs of the same species and batch of Bohai Bay superfine Artemia were taken and divided into 6 groups, 50 eggs in each group, and poured into Artemia incubators numbered 1-6 filled with 100 ml of 30‰ saline water aerated 24 hours in advance, the temperature was controlled at 25-30℃, the light was 2000 Lux, and the incubation was carried out for 24 hours.

[0056] Comparative Example (counting using the existing technology): Count incubators No. 1-3 using a separation filter and a rubber-tipped dropper: Pour the salt water in incubator No. 1 into the separation filter and rinse with water. Remove the upper thin net, separate the dense net from the thin net, and use a rubber-tipped dropper to suck out the egg shells and sunken eggs in the thin net and the Artemia larvae in the dense net, respectively, and squeeze them into A1 and A2 culture dishes containing an appropriate amount of salt water with a mass concentration of 30‰. Count the egg shells, sunken eggs, and Artemia larvae in the two culture dishes. The same procedure was used for incubators No. 2 and No. 3. The results are shown in Tables 1 and 2:

[0057] Table 1 Accuracy results of counting a small amount of Artemia using separation filter and rubber dropper in the prior art

[0058] A1 Petri Dish B1 Petri Dish C1 Petri Dish Eggshell and number of eggs 36 41 38 Counting accuracy 72％ 82％ 76％

[0059] (Egg loss rate = (total number of hatched eggs - number of eggshells and sunken eggs) / total number of hatched eggs × 100%)

[0060] The average counting accuracy is: 76.67%.

[0061] Table 2 Hatching rate results of a small amount of Artemia method using separation filter and rubber dropper in the prior art

[0062] A2 Petri Dish B2 Petri Dish C2 Petri Dish Number of Artemia larvae 27 38 32 Artemia hatching rate 54％ 76％ 64％

[0063] (Artemia hatching rate = number of Artemia larvae / total number of hatched eggs × 100%)

[0064] It can be seen that the above-mentioned method in the prior art has a high egg loss rate, and the Artemia hatching rate in parallel groups varies greatly and is unstable.

[0065] Example (using the method of the present invention to count): Figure 9As shown, incubators No. 4-6 are counted using a diaphragm vacuum pump: a diaphragm vacuum pump equipped with a microporous filter membrane D (D here is the number of the microporous filter membrane used during the experiment, and has no special meaning) is powered on for standby use, incubator No. 3 is placed on the filter cup mouth at the upper end of the vacuum pump, the incubator bottle mouth is slowly opened, and saline with a salinity of 30‰ is slowly flowed into the filter cup at the upper end of the vacuum pump, the incubator is rinsed to ensure that there is no residue in the incubator, the vacuum pump is turned on for filtration, and when the saline is about to leak out, the filter cup wall at the upper end of the vacuum pump is rinsed to ensure that there is no residue on the filter cup wall, and finally the Artemia larvae, egg shells and sunken eggs are completely sunk on the microporous filter membrane, the vacuum pump is turned off, the filter membrane D is clamped with tweezers, and counting is started. The same is true for incubators No. 4 and No. 5, and the results are shown in Tables 3 and 4. Figure 8 As shown:

[0066] Table 3 Accuracy and hatchability of counting a small amount of Artemia using the diaphragm vacuum pump and microporous filter membrane of the present invention

[0067] Microporous filtration membrane D Microporous filtration membrane E Microporous filtration membrane F Eggshell and number of eggs 47 49 47 Counting accuracy 94％ 98％ 94％ Number of Artemia larvae 39 40 40 Artemia hatching rate 78％ 80％ 80％

[0068] Average counting accuracy: 95.33%

[0069] The difference in accuracy between the two solutions is: 95.33% - 76.67% = 18.66%.

[0070] By comparing the comparative examples with the examples, it can be seen that the egg counting scheme of the present invention has a very low loss rate during the experiment, a high accuracy rate, and a stable and relatively accurate hatching rate. After testing, if the counting process is faster, the filter membrane with Artemia attached is placed back into the salt water again, Artemia larvae can continue to survive.

[0071] The survival of Artemia nauplii was verified as follows:

[0072] The newly hatched Artemia larvae were divided into 9 groups, each with 30 larvae. Each group was filtered onto a microporous filter membrane using a diaphragm vacuum pump. The timer was started, and the filter membrane was picked up with tweezers according to different time periods (3min, 5min, 8min, 10min, 15min, 20min, 25min, 30min, 35min). They were placed in culture dishes filled with saline and shaken to transfer the Artemia larvae attached to the filter membrane into the culture dish. When the transfer was incomplete, the filter membrane could be rinsed with a squeeze elbow containing saline and a distilled water bottle to rinse the filter membrane until the Artemia larvae on the filter membrane were completely transferred into the water for observation of survival rate. After the test, the Artemia larvae that lost water and stagnated on the filter membrane after the filtration was completed could continue to survive if they were placed in saline again within 10min. Individual deaths occurred over 10min, and all died over 30min. The results are shown in Table 4. Figure 10 and Figure 11 .

[0073] Table 4 After the filtration, the Artemia larvae that lost water and stagnated on the filter membrane continued to survive after being placed in salt water within 10 minutes.

[0074]

[0075] Disadvantage 2 of the existing methods mentioned that "because the newly hatched Artemia are alive and have the characteristics of flexible swimming, it is difficult to control and count them under the naked eye or dissecting microscope." At present, most of the solutions to this problem are to use iodine and other agents to kill Artemia larvae, and then count the Artemia floating in the culture dish after death. Table 4, Figure 10 and Figure 11 The results show that the method of the present invention can avoid such unnecessary death to a certain extent. The filter membrane itself is hydrophilic, which prevents the Artemia on the filter membrane from being completely dehydrated in a short period of time and continues to maintain its own activity. Moreover, when counting a small amount of Artemia on the filter membrane, it is relatively clear and easy to distinguish, and the counting process often does not exceed 10 minutes. Generally, the counting can be completed within 3 minutes. Therefore, the present invention also effectively avoids the unnecessary death of Artemia larvae.

[0076] In summary, the accuracy of the existing method is 76.67%, and the accuracy of the method of the present invention is 95.33%, which is an increase of 18.66%. The accuracy has reached more than 95%. It can be seen that the counting scheme designed by the present invention has an extremely low egg loss rate during the experiment and the accuracy is greatly improved. The hatching rates of the existing methods are 54%, 76%, and 64%, respectively, and the hatching rates of the method of the present invention are 78%, 80%, and 80%, respectively. It can be seen that the hatching rate of the parallel group of the present invention fluctuates less than that of the existing method, and the data is stable, which can indicate that the accuracy of the hatching rate is improved.

[0077] Although the embodiments of the present invention are disclosed for illustrative purposes, those skilled in the art will understand that various substitutions, changes and modifications are possible without departing from the spirit and scope of the present invention and the appended claims. Therefore, the scope of the present invention is not limited to the contents disclosed in the embodiments.

Claims

1. A method for counting a small amount of Artemia using a diaphragm vacuum pump and a microporous filter membrane, characterized in that: The steps include: Place the microporous filter membrane above the filter head of the diaphragm vacuum pump, pour the hatching liquid into the filter cup of the diaphragm vacuum pump for filtration. The diaphragm vacuum pump is a negative pressure type with a pumping rate of 20L / min, a limit vacuum degree of 0.08Mpa, and a motor power of 160W. When there is no water in the filter cup, unplug the silicone tube on the filter cup, turn off the vacuum pump, and use the diaphragm vacuum pump to filter the hatched Artemia onto the microporous filter membrane. Finally, the hatched Artemia larvae and empty shells can be clearly distinguished on the microporous filter membrane, and can be accurately counted with the naked eye or under a dissecting microscope. After the filtration is completed, the Artemia larvae that have lost water and stagnated on the filter membrane can continue to survive if they are placed in salt water within 10 minutes. Some of them will die after more than 10 minutes, and all of them will die after more than 30 minutes. The Artemia larvae are orange-red teardrop-shaped, and the empty shells or sunken eggs are brown spherical; The microporous filtration membrane is a water-based microporous filtration membrane with a diameter of 50 mm and a pore size of 0.45 μm. The material of the membrane is a mixed cellulose MCE membrane. The specific steps are as follows: The diaphragm vacuum pump equipped with microporous filter membrane is energized for standby use, the incubator equipped with Artemia larvae is placed in the upper filter cup mouth of the vacuum pump, the incubator bottle is slowly opened, the salt solution with a salinity of 30 ‰ slowly flows into the upper filter cup of the vacuum pump, the incubator is rinsed, and it is guaranteed that there is no residue in the incubator, the vacuum pump is opened for filtration, and when the salt solution is about to leak, the upper filter cup wall of the vacuum pump is rinsed to ensure that there is no residue on the filter cup wall, and final Artemia larvae, ovules and sunken eggs are completely sunken on the microporous filter membrane, the vacuum pump is turned off, the microporous filter membrane is clamped by tweezers, and counting is started; Among them, the pH of salt water is between 7-8; The microporous filtration membrane is nylon 66, polyethersulfone PSE, polytetrafluoroethylene PTFE, polyvinylidene fluoride PVDF or polypropylene pp; The accuracy of this method is 95.33% and the hatchability of this method is 80%.

2. Use of the method according to claim 1 in Artemia counting.

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