阅读说明：本技术 一种高通量鲍耐高温能力的快速测评方法 (Rapid evaluation method for high-throughput abalone high-temperature resistance ) 是由 骆轩 於峰 彭文竹 游伟伟 柯才焕 于 2021-07-16 设计创作，主要内容包括：一种高通量鲍耐高温能力的快速测评方法,涉及水产生物技术领域。所述用于开展鲍耐高温能力的测评装置包括养殖箱、海水温度调节器、制氧机、蛋白分离器、温度监测仪、水泵和附着板。利用鲍拥有宽大而发达的腹足,腹足吸力极强,可牢固吸附在光滑表面的物理特性,以鲍个体在持续高温胁迫下,足肌逐渐失去吸附能力,会从附着物上脱离的时间作为指征,测评不同鲍鱼个体对高水温的耐受能力。操作简便、单次测试样本数量大、测评周期短、受试鲍鱼不会因测评工作造成损伤,完成测评后的鲍样本可用于开展后续育种工作,有助于提高鲍耐高温品种的育种效率。可避免环境对受试鲍样品造成影响,测试结果精准度高,可大量开展鲍耐高温性状的测评工作。(A rapid evaluation method for high-throughput abalone high-temperature resistance relates to the technical field of aquatic organisms. The device for evaluating the high-temperature resistance of the abalones comprises a culture box, a seawater temperature regulator, an oxygen generator, a protein separator, a temperature monitor, a water pump and an attachment plate. The physical characteristics that the abalones have wide and developed abdomens, the abdomens have strong suction force and can be firmly adsorbed on the smooth surface are utilized, the time that the foot muscles gradually lose adsorption capacity under continuous high-temperature stress of the abalones is taken as an indication, and the tolerance capacity of different abalones to high water temperature is evaluated. The method is simple and convenient to operate, large in number of single test samples, short in evaluation period, and free from damage to tested abalones due to evaluation work, and the abalone samples subjected to evaluation can be used for carrying out subsequent breeding work, so that the method is beneficial to improving the breeding efficiency of high-temperature-resistant varieties of abalones. The influence of the environment on the tested abalone sample can be avoided, the test result precision is high, and the evaluation work of the high-temperature resistance of the abalone can be carried out in a large quantity.)

1. A rapid evaluation method for high-throughput abalone high-temperature resistance is characterized by comprising the following steps:

1) selecting abalones of different types or varieties, labeling the abalones to be tested, measuring the growth parameters of the abalones samples, temporarily culturing the abalones in a culture box, setting the water temperature and the salinity, feeding fresh gracilaria, asparagus or kelp, replacing one fourth of fresh seawater every day, timely cleaning residual baits and excrement, and feeding for 5-15 days to ensure that the abalones to be tested are in a normal state;

2) when temporarily culturing is carried out till the 7 th day, the attachment plate for evaluation is placed at the bottom of the culture box, the temperature of the attachment plate for evaluation is gradually increased from 18 ℃ at the speed of 1 ℃/h, the attachment plate attached with the abalones is vertically hung in the culture seawater at the temperature of 28 ℃, and then the culture seawater is continuously increased to the final stress temperature of 30-34 ℃;

3) starting timing when the water temperature reaches the final stress temperature, and recording the time that each individual loses the adhesive capacity and falls off from the adhesive plate, namely the time for maintaining the adsorption capacity at high temperature; transferring the individuals losing the adhesive ability to fresh seawater for recovery, wherein the initial water temperature of the fresh seawater is 14-18 ℃, the recovery time is 3 days, the temperature rises by 1 ℃ every day, and the death condition within 3 days is recorded; the whole evaluation process is carried out in a dark room, and when a light source is needed for observing and moving the abalone sample, the wavelength of the used light source is 550-600 nm; statistical analysis of the data was performed using span 22, plotted using GraphPad Prism 8 and R-package.

2. The method for rapidly evaluating the high-throughput abalone temperature resistance according to claim 1, wherein in step 1), the water temperature and salinity are set, the water temperature is controlled at 20 ℃, the salinity is controlled at 30-35, the dissolved oxygen concentration is not lower than 6mg/L, and the pH is not lower than 8.0.

3. The method for rapidly evaluating the high-throughput abalone temperature resistance according to claim 1, wherein in step 1), the cultivation tank is a conventional cultivation aquarium sold in the market, and different sizes are selected according to the number of samples of abalones to be evaluated.

4. The method for rapidly evaluating the high-throughput abalone temperature resistance according to claim 1, wherein in step 2), the attachment plate is made of acrylic plastic plate, and the surface of the plate is smooth.

5. The method for rapidly evaluating the high-throughput abalone temperature resistance according to claim 1, wherein in step 2), the final stress temperature is 32 ℃.

6. The device for evaluating the high-temperature resistance of the abalones is characterized by comprising a culture box, a seawater temperature regulator, an oxygen generator, a protein separator, a temperature monitor, a water pump and an attachment plate; the culture box is connected with the protein separator, the water pump and the seawater temperature regulator through PVC water pipes; an oxygen generator is arranged between the protein separator and the water pump and is used for filling sufficient oxygen into the culture seawater; the device comprises a culture box, a water pump, a protein separator, a temperature detector, a temperature probe and a temperature sensor, wherein seawater is filled in the culture box and used for culturing abalones to be tested, the seawater is pushed by the water pump to circulate, the outflow seawater is firstly treated by the protein separator and then regulated to the temporary culture water temperature by the seawater temperature controller and then flows into the culture box again, the temperature detector is connected with one end of the temperature probe, and the other end of the temperature probe is arranged below the seawater level and used for monitoring the temperature in the culture box; at least two lifting ropes for fixing the attachment plates are arranged at the top of the culture box and used for pulling the attachment plates to be vertical from the plane.

7. The apparatus for evaluating high temperature resistance of an abalone as recited in claim 6, wherein the attachment plate has a through hole at the end for inserting a hanging rope for hanging.

Technical Field

The invention relates to an evaluation technology for high-temperature resistance of abalones in the technical field of aquatic organisms, in particular to a high-throughput quick evaluation method for high-temperature resistance of abalones, which distinguishes different varieties or different culture groups of abalones.

Background

China is the first major country of abalone culture in the world, the culture yield accounts for more than 85% of the global yield, in 2019, the abalone yield in China exceeds 18 million tons, wherein Fujian is taken as the main culture area of abalones in China, the annual yield of the abalones in Fujian province accounts for 82.7% of the national yield, the annual yield value is about 200 million yuan, the abalone culture has been developed into the first mariculture variety of the output value in the Fujian province, and the main culture variety is the haliotis discus hannai originally produced from Liaoning and Shandong coasts. The haliotis discus hannai native place is in a yellow Bohai sea area of a temperate zone, and is a temperate zone species, the high temperature resistance of the haliotis discus hannai is improved after genetic improvement, and the haliotis discus hannai is moved to the south for culture, so that the industry of the haliotis discus hannai is rapidly developed, but the marine culture is limited by the original ecological habits of the haliotis discus hannai, global warming and other reasons, the phenomenon that the haliotis discus hannai is violently killed due to high temperature intolerance in part of the marine culture every summer high temperature period is generated, and the marine culture becomes one of the main bottlenecks which troubles the healthy development of the marine culture industry at present. Therefore, the cultivation of new species (line) of high temperature resistant abalone is very important, and how to quickly and accurately evaluate the high temperature resistance of the cultivated line in the cultivation process of the high temperature resistant variety is very important.

The traditional evaluation method for the high-temperature resistance of the abalones is mostly based on summer culture data or a laboratory evaluation method represented by methods such as semi-lethal temperature and the like, the summer culture data in the natural sea area is used as an index, the evaluation period is long, the hydrological conditions in the natural sea area are complex, the evaluation result is easily influenced by other factors, accurate data cannot be provided, and the reliability is low; the laboratory evaluation method represented by methods such as a semi-lethal temperature and the like generally needs to change the water temperature in a short time and finish the evaluation work by taking the survival rate of the abalones as an index, but the method has great damage to the abalones, individuals with strong tolerance often cannot survive after the evaluation is finished, so that the experimental abalones die, and although relevant evaluation data is obtained, breeding individuals cannot be obtained for carrying out subsequent breeding work.

Chinese patent CN 107372273A discloses a method for rapidly distinguishing temperature resistance of abalone, which comprises temporarily culturing abalone samples to be evaluated under a proper water temperature condition, adopting a non-invasive method, fixing an infrared sensor on abalone shells corresponding to the accurate position of abalone heart by glue, measuring the abalone heart rate measured in a non-invasive manner, recording the abalone heart rate under different temperature conditions in real time, then obtaining Arrhenius curve, further calculating the ABT value of each sample, thereby measuring the difference of heat tolerance of each sample, compared with the common method, the method has the advantages of convenient operation, small damage to abalone, individual measurement and the like, but because the accurate position of abalone heart needs to be searched by adopting the infrared sensor during measurement, simultaneously, the obtained signal strength needs to be ensured not less than 1V, the sensor needs to be fixed on the abalone shell by glue after positioning, the position of the heart of the abalone is found, the operation that the infrared sensor is fixed on the shell of the abalone to accurately reflect the heart rate of the abalone has higher requirement on the professional degree of an operator, and the risk of inaccurate heart rate measurement exists. In addition, when the operation steps are carried out, the tested abalone sample needs to be separated from the originally adapted culture seawater environment, when the operation is improper, the sample is frightened by operations such as prying and the like, or the activity of the tested abalone is influenced due to exposure, and if the subsequent clean seawater recovery process is not proper, the measured result is very easy to deviate. In addition, the method needs to use infrared sensors to be adhered to abalone shells, the number of samples which can be tested by the same operator each time is limited by the number of the sensors, the adhering proficiency, the time for putting back into clean seawater to recover and the like, only dozens of samples can be tested each time, and the evaluation work of one group needs about 10-20 days. The operation process is still more complicated, error evaluation results are easy to occur, and the large-scale breeding work of the high-temperature resistant abalone variety is not facilitated. Therefore, how to rapidly, accurately and high-flux evaluate the high-temperature resistance of the abalone in a laboratory becomes one of the problems that the abalone breeding workers need to solve urgently.

The abalone possesses wide and developed ventral foot, the foot muscle structure makes it capable of being adsorbed firmly to smooth rock surface in deep underwater place, and the adsorption force has high strength. Under the continuous high-temperature stress, the foot muscles gradually lose the adsorption capacity and are separated from attachments, so the attachment capacity of the abalones is taken as an important index for judging the health state of the abalones.

Disclosure of Invention

The invention aims to provide the evaluation device for developing the high-temperature resistance of the abalones, which has the advantages of simple and convenient operation, large number of single test samples, short evaluation period, no damage to tested abalones caused by evaluation work, capability of developing subsequent breeding work by using the evaluated abalones samples and the like.

The invention also aims to provide a rapid evaluation method for high-throughput abalone high-temperature resistance, which is beneficial to improving the breeding efficiency of high-temperature resistant varieties of abalones.

The device for evaluating the high-temperature resistance of the abalones comprises a culture box, a seawater temperature regulator, an oxygen generator, a protein separator, a temperature monitor, a water pump and an attachment plate; the culture box is connected with the protein separator, the water pump and the seawater temperature regulator through PVC water pipes; an oxygen generator is arranged between the protein separator and the water pump and is used for filling sufficient oxygen into the culture seawater; the device comprises a culture box, a water pump, a protein separator, a temperature controller, a temperature detector, a temperature probe and a temperature sensor, wherein seawater is filled in the culture box and used for culturing abalones to be tested, the seawater is pushed by the water pump to circulate, the flowing-out seawater is firstly treated by the protein separator and then regulated to temporary culture water temperature by the seawater temperature controller to flow into the culture box again, the temperature detector is connected with one end of the temperature probe, and the other end of the temperature probe is arranged below the seawater level and used for monitoring the temperature in the culture box. At least two lifting ropes for fixing the attachment plates are arranged at the top of the culture box and used for pulling the attachment plates to be vertical from the plane.

The aquaculture tank can adopt a conventional aquaculture aquarium, and different sizes can be selected according to the number of samples of the abalones to be evaluated.

The attachment plate can be made of an acrylic plastic plate, and the surface of the plate is smooth; the tail part of the attachment plate is provided with a through hole for the insertion of the lifting rope so as to be beneficial to the suspension of the lifting rope.

A method for rapidly evaluating high-throughput abalone high-temperature resistance comprises the following steps:

1) selecting abalones of different types or varieties, labeling the abalones to be tested, measuring the growth parameters of the abalones samples, temporarily culturing the abalones in a culture box, setting the water temperature and the salinity, feeding fresh gracilaria, asparagus or kelp, replacing one fourth of fresh seawater every day, timely cleaning residual baits and excrement, and feeding for 5-15 days to ensure that the abalones to be tested are in a normal state;

in the step 1), setting water temperature and salinity, wherein the water temperature is controlled to be 18-22 ℃, the salinity is 30-35, the concentration of dissolved oxygen is not lower than 6mg/L, and the pH is not lower than 8.0; the water temperature is preferably 20 ℃.

2) When temporarily culturing is carried out till the 7 th day, the attachment plate for evaluation is placed at the bottom of the culture box, the temperature of the attachment plate for evaluation is gradually increased from 18 ℃ at the speed of 1 ℃/h, the attachment plate attached with the abalones is vertically hung in the culture seawater at the temperature of 28 ℃, and then the culture seawater is continuously increased to the final stress temperature of 30-34 ℃;

in step 2), the final stress temperature is preferably 32 ℃.

3) Starting timing when the water temperature reaches the final stress temperature, and recording the time that each individual loses the adhesive capacity and falls off from the adhesive plate, namely the time for maintaining the adsorption capacity at high temperature; transferring the individuals losing the adhesive ability to fresh seawater for recovery, wherein the initial water temperature of the fresh seawater is 14-18 ℃, the recovery time is 3 days, the temperature rises by 1 ℃ every day, and the death condition within 3 days is recorded; the whole evaluation process is carried out in a dark room, and when a light source is needed for observing and moving the abalone sample, the wavelength of the used light source is 550-600 nm; statistical analysis of the data was performed using span 22, plotted using GraphPad Prism 8 and R-package.

The invention discloses a high-flux and convenient-to-operate method for evaluating the high-temperature resistance of abalones based on the specific attachment capacity of the abalones different from other main-cultured shellfish, which utilizes the physical characteristics that the abalones have wide and developed ventral legs, have strong suction force of the ventral legs and can be firmly adsorbed on a smooth surface, and evaluates the high-temperature resistance of different abalones by taking the time that the foot muscles gradually lose the adsorption capacity and can be separated from attachments as an indication under the continuous high-temperature stress of the abalones. The method has the advantages of simple and convenient operation, large number of single test samples, short evaluation period, no damage to tested abalones due to evaluation work, capability of developing subsequent breeding work of the abalones samples after evaluation, contribution to improving the breeding efficiency of high-temperature resistant varieties of the abalones, and the like. The method can avoid the influence of the environment on the tested abalone sample, has high precision of the test result, and can carry out the evaluation work of the high temperature resistance of the abalone in a large quantity.

Drawings

FIG. 1 is a schematic structural view of a device for evaluating the resistance of a fire-resistant abalone.

FIG. 2 is a graph comparing the duration of heat stress attachment (HAD) of the southern population of Haliotis discus hannai and the northern population.

FIG. 3 is a graph showing a comparison of heat stress attachment duration (HAD) of Haliotis discus hannai, and Haliotis discus hannai.

Detailed Description

The following examples will further illustrate and describe the technical solutions of the present invention with reference to the accompanying drawings. Commercially available products may be used unless otherwise specified.

As shown in figure 1, the evaluation device for carrying out the high temperature resistance of the abalone comprises a culture box (12), a seawater temperature regulator (1), an oxygen generator (2), a protein separator (3), a temperature monitor (5), a water pump (13) and an attachment plate (8); at least two lifting ropes (7) for fixing the attachment plates are arranged at the top of the culture box (12) and used for pulling the attachment plates to be vertical from the plane; the culture box (12) is connected with the protein separator (3), the water pump (13) and the seawater temperature regulator (1) through a PVC water pipe (11); an oxygen generator (2) is arranged between the protein separator (3) and the water pump (13) and is used for filling sufficient oxygen into the culture seawater; the aquaculture device is characterized in that seawater (10) is filled in the aquaculture box (12) and used for aquaculture of abalones to be tested and evaluated (9), a water pump (13) pushes the seawater to circulate, the flowing-out seawater (10) is firstly treated by a protein separator (3) and then regulated to a temporary aquaculture water temperature by a seawater temperature controller (1) and then flows into the aquaculture box (12) again, the temperature detector (5) is connected with one end of a temperature probe (4), and the other end of the temperature probe (4) is arranged below a seawater liquid level (6) and used for monitoring the temperature in the aquaculture box.

The breeding tank (12) can adopt a conventional breeding aquarium, and different sizes can be selected according to the number of samples of the abalones to be evaluated.

The attachment plate (8) can be made of an acrylic plastic plate, and the surface of the plate is smooth. The tail part of the attachment plate (8) is correspondingly provided with a through hole for the insertion of the lifting rope (7) so as to facilitate the suspension of the lifting rope.

The embodiment of the rapid evaluation method for the high-temperature resistance of the high-flux abalone comprises the following steps:

1) when carrying out the high temperature resistance evaluation of the abalones, selecting different types or varieties of abalones, attaching labels to samples of the abalones to be evaluated (9), measuring growth parameters of the samples of the abalones, temporarily culturing the samples in a culture tank, wherein the culture tank is a conventional culture aquarium, and different sizes can be selected according to the number of the samples of the abalones to be evaluated. Controlling the water temperature in the culture box at 20 ℃, controlling the salinity at 30-35, filling sufficient oxygen into the culture seawater by an oxygen generator (2), ensuring that the concentration of dissolved oxygen is not lower than 6mg/L and the pH is not lower than 8.0, feeding fresh gracilaria, asparagus or kelp, replacing one fourth of fresh seawater every day, timely cleaning residual bait and excrement, feeding for 5-15 days, and ensuring that the tested abalone is in a normal state;

the method comprises the steps that an abalone to be tested is cultured in a culture box (12), seawater (10) is filled in the culture box, the seawater is pushed by a water pump to circulate, the flowing-out seawater (10) is firstly treated by a protein separator (3), is regulated to temporary culture water temperature by a seawater temperature controller (1) and then flows into the culture box (12) again, the temporary culture water temperature is controlled at 20 ℃, the salinity is 30-35, the concentration of dissolved oxygen is not lower than 6mg/L, the pH is not lower than 8.0, fresh gracilaria, asparagus or kelp are fed, one fourth of fresh seawater is replaced every day, residual bait and excrement are cleaned in time, and the abalone is cultured for 5-15 days to ensure that the tested abalone is in a normal state.

The protein separator can be selected from the manufacturers and models comprising: ansu ger environmental protection equipment ltd, model: GEER-DB 5.

The oxygen generator can select manufacturers: shandong Yuanxi environmental technologies, Inc., model number: YT-9000-10P.

2) When temporarily culturing to the 7 th day, placing the attachment plate (8) for evaluation at the bottom of the culture box (12), gradually raising the temperature from 20 ℃ at a rate of 1 ℃/h after the tested abalones gradually crawl to the attachment plate for evaluation, vertically suspending the attachment plate with the abalones in the culture seawater at the temperature of 28 ℃, and then continuously raising the temperature of the culture seawater to the final stress temperature of 32 ℃.

3) Counting time from the water temperature reaching 32 ℃, and recording the time when each individual loses the adhesive capacity and falls off from the adhesive plate, namely the time for maintaining the adsorption capacity at high temperature, namely heat stress adhesion duration (HAD); the individuals lost the ability to adhere were transferred to fresh seawater for recovery at an initial water temperature of 16 ℃ for 3 days, 1 ℃ per day, and the death status within 3 days was recorded. The whole evaluation process is carried out in a dark room, and when a light source is needed for observing and moving the abalone sample, the wavelength of the used light source is 550-600 nm. Statistical analysis of the data was performed using span 22, plotted using GraphPad Prism 8 and R-package.

Specific examples are given below.

Example 1:

and (3) evaluating the high-temperature resistance of the southern Haliotis discus hannai population (code: YX) and the northern Haliotis discus hannai population (code: DL), and comparing the high-temperature resistance of the southern Haliotis discus hannai population and the northern Haliotis discus hannai population.

Taking 50 samples of the abalone in the south population of the haliotis discus hannai and 50 samples of the abalone in the north population of the haliotis discus hannai respectively, sticking a label on each abalone shell, temporarily culturing the samples in a high-temperature resistance capacity evaluation device of the abalones, controlling the average shell length of the southern population of the haliotis discus hannai to be 5.3cm, controlling the average shell length of the northern population of the haliotis discus hannai to be 5.5cm, controlling the temperature of seawater in the evaluation device to be 20 ℃, controlling the salinity of the seawater to be 33, controlling the dissolved oxygen concentration to be 6.2mg/L and controlling the pH to be 8.1, feeding fresh asparagus, replacing one fourth of the fresh seawater every day, timely cleaning residual baits and excrement, feeding for 10 days normally, and confirming that the tested abalones are in a normal state; and (3) temporarily culturing until the 7 th day, putting the attachment plate for evaluation into the bottom of the tank, gradually raising the temperature of the tested abalone from 20 ℃ at a speed of 1 ℃/h, vertically suspending the attachment plate attached with the abalone in the culture water body at 28 ℃, and then continuously raising the water temperature in the culture tank to 32 ℃.

The time from the time of reaching 32 ℃ was counted, and the time at which each individual lost the adhesive ability and dropped from the adhesive plate, that is, the time at which the adsorption ability was maintained at high temperature (HAD), was recorded. And (3) moving the falling individual losing the adhesive ability to fresh seawater at 16 ℃ in time for recovery, carrying out recovery observation on the individual in a period of 3 days, wherein the recovery time is 3 days, the temperature rises by 1 ℃ every day, and recording the death condition within 3 days. The whole evaluation process is carried out in a dark room, and when a light source is needed for observing and moving the abalone sample, the wavelength of the used light source is 550-600 nm. Statistical analysis of the data was performed using span 22, plotted using GraphPad Prism 8 and R-package.

The HAD measurements of the southern and northern haliotis discus hannai populations are shown in fig. 2. As can be seen in FIG. 2, the southern population is higher than the northern population on the HAD50 (YX:4.96 h; DL:3.49h), again 5.49 + -2.47 h is significantly higher than the northern population by 4.06 + -2.49 h on the average of the HAD (P ═ 0.019< 0.05); the north-south population has larger difference in the coefficient of variation, and the numerical value is YX: 47.85% less than DL: 68.88%, reflecting that the southern population is more uniform and more concentrated in falling after long-time southern high-temperature domestication; although the initial dropping time is within 1h, the maximum attaching duration of the southern population (0.72-11.27 h) is obviously longer than that of the northern population (0.34-10.02), which indicates that the southern population has higher high temperature resistance and has high temperature breeding potential.

Example 2:

and (3) carrying out evaluation on the high-temperature resistance properties of the haliotis discus hannai (code number: DD), the haliotis discus (code number: FF) and the haliotis discus (code number: female parent) and the haliotis discus (code number: FF), and comparing the high-temperature resistance of the haliotis discus hannai (code number: FF) and the haliotis discus (code number: female parent).

Taking 60 samples of Haliotis discus hannai, Haliotis diversicolor and Haliotis diversicolor, sticking a label on each shell of the Haliotis diversicolor and temporarily culturing in a culture box of an evaluation device for high temperature resistance of the Haliotis diversicolor, wherein the average shell length of the Haliotis diversicolor is 7.8cm, the average shell length of the Haliotis diversicolor is 8.1cm, the average shell length of the Haliotis diversicolor is 7.4cm, the temperature of seawater in the evaluation device is 20 deg.C, the salinity of the seawater is 32, the dissolved oxygen concentration is 6.5mg/L, the pH is 8.1, feeding fresh thallus Gracilariae, replacing one fourth of fresh seawater every day, cleaning up residual bait and feces in time, normally feeding for 9 days, and confirming that the abalone to be tested is in a normal state; and temporarily culturing until the 7 th day, putting the attachment plate for evaluation into the bottom of the tank, gradually raising the temperature of the tested abalone gradually at the speed of 1 ℃/h from 20 ℃, vertically suspending the attachment plate attached with the abalone in the culture water body at 28 ℃, and then continuously raising the water temperature in the culture tank to 33 ℃.

The time from when the water temperature reached 33 ℃ was counted, and the time at which each individual lost the adhesive ability and dropped from the adhesive plate, that is, the time at which the adsorption ability was maintained at high temperature (HAD), was recorded. And (3) timely transferring the individuals lost the adhesive ability to fresh seawater at 17 ℃ for recovery, carrying out recovery observation on the individuals in a period of 3 days, wherein the recovery time is 3 days, the temperature rises by 1 ℃ every day, and recording the death condition within 3 days. The whole evaluation process is carried out in a dark room, and when a light source is needed for observing and moving the abalone sample, the wavelength of the used light source is 550-600 nm. Statistical analysis of the data was performed using span 22, plotted using GraphPad Prism 8 and R-package.

The HAD of Haliotis discus hannai (DD), Haliotis Diversicolor (DF), and Haliotis diversicolor (FF) at 32 deg.C is shown in FIG. 3. DD the longest adsorption time (10.97h) was at a minimum at the initial drop time (0.42h), HAD50(4.95h), average drop time (5.11. + -. 2.40 h); DF initial falling time (0.79h), HAD50(7.47h), average adsorption time (7.26 +/-3.53 h) and coefficient of variation (0.49) are all larger than those of other two abalones; the HAD mean (6.89 ± 3.23h) and median (6.89h) of FF were close and significantly higher than DD (P ═ 0.015< 0.05).

As can be seen from fig. 3, haliotis discus hannai (DD) has the minimum of the initial falling time (0.42h), HAD50(4.95h), average falling time (5.11 ± 2.40h), and the maximum adsorption time (10.97 h); the initial falling time (0.79h) of the Haliotis Diversicolor (DF), the HAD50(7.47h), the average adsorption time (7.26 +/-3.53 h) and the coefficient of variation (0.49) are all larger than those of the other two Haliotis diversicolor; the HAD average value (6.89 +/-3.23 h) and the median (6.89h) of Haliotis diversicolor (FF) are close and are remarkably higher than those of Haliotis discus hannai (DD) (P is 0.015< 0.05). The result shows that the haliotis discus hannai (DD) has the lowest high temperature resistance, the hybrid species haliotis Discus (DF) has higher high temperature resistance, and the heterosis is shown.

The invention is not limited to the examples described above, but rather, variations and equivalent modifications are possible within the true spirit and scope of the invention.