Determination method for abalone foot muscle adsorption force

Abstract

The invention discloses a method for measuring abalone foot muscle adsorption capacity. The method comprises the following steps: S1, placing abalone to be measured in an inflated seawater tank to be measured; s2, the adsorption force measuring device is connected with a control computer, software is started, the height of the claw hook is adjusted to a proper position through the software or a device controller, a tensile experiment scheme is selected in the software, and force and displacement readings are zeroed; s3, taking out the abalone from the seawater tank, putting the abalone into a transparent acrylic tank filled with fresh seawater, enabling the abalone to move freely, after the feet are completely unfolded, touching the antennae of the abalone to stop moving and adsorb the abalone, taking a picture of the feet by using a camera component, hooking the abalone shell by using a claw hook, and starting an adsorption capacity measuring device to measure the adsorption capacity; s4, after the measurement is finished, exporting the obtained force measurement data and the sole picture, calculating the sole area of abalone by using ImageJ software, and calculating the abalone foot muscle adsorption strength; the method realizes low-cost, high-efficiency and standardized measurement of the abalone foot muscle adsorption capacity.

Description

Technical Field

[0001] The invention belongs to the technical field of gastropod quality detection, and in particular relates to a method for measuring the adsorption force of abalone muscles. Background Art

[0002] In the process of breeding new stress-resistant varieties, accurate evaluation of stress resistance traits is crucial and is the key basis for the success of new variety breeding. Accurate evaluation of abalone stress resistance traits has always been the main technical problem that plagues the breeding of stress-resistant varieties. At present, a large number of studies use traditional classic evaluation methods, that is, measurements are made under high stress conditions, including survival rate, half-lethal temperature (Lethal temperature 50%, LT50), etc. These methods are time-consuming and labor-intensive and cannot guarantee the survival of the measured parental materials, which is quite unfavorable for genetic breeding research on stress resistance traits. Therefore, it is urgent to explore new trait evaluation methods to help the breeding work proceed smoothly and efficiently.

[0003] Abalone is a benthic organism that lives in a non-permanent attachment state. It attaches to reefs through muscle contraction to form an adsorption effect. Adsorption is an important behavioral characteristic of abalone, and it has extremely strong adsorption capacity. Many reports have shown that the adsorption capacity of abalone is strongly correlated with its tolerance to various environments. Based on this, the adsorption capacity of abalone has been indirectly applied in various forms to the evaluation of its stress resistance traits, such as high temperature resistance, low oxygen resistance, and low salt resistance. The application forms include CTM (Critical thermal maximum) value, HAD (Heat adhesion duration) and HYAD (hypoxia adhesion duration). Compared with traditional methods, these indirect indicators related to adsorption can accurately obtain stress resistance phenotypic values ​​in high throughput and retain most of the evaluated parents, which can greatly assist the breeding of stress resistance traits. However, these adsorption-related indicators are still associated with other traits through indirect means, lacking direct use of adsorption phenotypes.

[0004] In addition, another behavior related to the adsorption force (self-righting behavior) is widely used as a parameter for evaluating the overall health of abalones. This indicator has become an important consideration when purchasing abalone fry, and it is believed that the faster this posture is adopted, the higher the likelihood of survival. In short, the adsorption force phenotype can reflect the overall health status of abalones and their tolerance to various environmental limiting conditions. Therefore, in addition to being used as an evaluation index for a single stress-resistant trait, it also shows the potential to be used as a robust trait. In the future, selective breeding targeting the adsorption force is expected to improve the robustness of abalones. However, whether directly using the adsorption force phenotype to assist in the selection and breeding of stress-resistant traits, or using the adsorption force trait as a comprehensive robust trait of abalones, it is first necessary to achieve direct and accurate measurement of the adsorption force trait. Currently, some scholars have conducted some related basic research on the adsorption force of abalones. They used a tensile testing machine commonly used in materials mechanics research as the equipment for measuring the adsorption force of abalones and tried to directly measure it. However, it has poor adaptability to the measurement of the adsorption force of abalones, the measurement process is cumbersome, and the error is large because it is impossible to judge the unfolding and adsorption of the abalone's foot sole. Therefore, this application proposes a new method for efficiently and accurately measuring the adsorption force of abalone foot muscles. Summary of the Invention

[0005] To solve the above problems, the present invention proposes a method for measuring the adsorption force of abalone foot muscles. By improving the existing equipment of the abalone foot muscle adsorption force measurement system, a standardized measurement method for the abalone foot muscle adsorption force index is established, realizing low-cost, high-efficiency, and standardized measurement of the abalone foot muscle adsorption force, providing a reliable technical means for subsequent abalone breeding work.

[0006] To achieve the above object, the present invention adopts the following technical solutions:

[0007] A method for measuring the adsorption force of abalone foot muscles, comprising the following steps:

[0008] S1. Preparation before abalone testing: Place the abalone to be tested in an aerated seawater tank for testing.

[0009] S2. Equipment connection and debugging: Connect the adsorption force measurement device to the control computer, turn on the software, adjust the height of the grab hook to a suitable position through the software or the device controller, select the tensile test scheme in the software, and zero the force and displacement readings.

[0010] S3. Abalone adsorption force detection: Take the abalone out of the seawater tank and place it in a transparent acrylic tank filled with fresh seawater, let the abalone move freely, after the foot is fully unfolded, touch the abalone's antenna to make the abalone stop moving and adsorb, use the camera component to take a photo of the foot, and hook the grab hook onto the abalone shell, then turn on the adsorption force measurement device to measure the adsorption force.

[0011] S4. Adsorption force parameter analysis: After the measurement is completed, the obtained force measurement data and the sole photos of the abalone are exported. The bottom foot area of the abalone is calculated using ImageJ software, and the adsorption strength of the abalone foot muscle is calculated.

[0012] Preferably, the test speed set by the adsorption force measuring device in step S2 is 50 mm / min.

[0013] Preferably, the formula for calculating the adsorption strength of the abalone foot muscle in step S4 is:

[0014] Fs = Fn / S,

[0015] where Fs is the adsorption strength of the abalone foot muscle, with the unit of Kpa; Fn is the adsorption force of the abalone foot muscle, that is, the maximum pull-off force measured by the adsorption force measuring device, with the unit of N; S is the bottom foot area of the abalone, with the unit of cm 2 .

[0016] Preferably, the adsorption force measuring device includes a material testing machine, a steering bracket, an eagle beak buckle, two hooks, and a test platform; the material testing machine is connected to a control computer for transmitting control signals and mechanical test data; the material testing machine includes a lifting platform and a lifting controller, and the lifting of the lifting platform is adjusted through the lifting controller; the steering bracket is fixed to the lower end of the material testing machine, and a fixed pulley is provided on the steering bracket. The upper end of the first pulling rope is fixedly connected to the lifting platform, and the lower end of the first pulling rope passes through the two fixed pulleys and is fixedly connected to the eagle beak buckle; the two hooks are fixedly connected by a second pulling rope, and the second pulling rope is arranged through the lower end of the eagle beak buckle; the test platform includes a transparent acrylic cylinder, a camera assembly, and a platform frame; the transparent acrylic cylinder is fixed to the upper end of the platform frame, and the camera assembly is arranged on the platform frame below the transparent acrylic cylinder to take photos of the abalone's foot.

[0017] Preferably, a supplementary light is fixed on the platform frame at the lower end of the transparent acrylic cylinder to improve the lighting effect of the abalone's sole.

[0018] Preferably, the camera assembly includes a camera module, a radiator, and a USB multi-port docking station; the supplementary light is connected to the USB multi-port docking station through a USB cable to receive power supply, and the camera module and the radiator are respectively connected to the USB multi-port docking station through USB cables to receive power supply and transmit data; the USB multi-port docking station is connected to the control computer through a USB cable to receive power supply and transmit data.

[0019] After adopting the above technical solution, the present invention has the following beneficial effects:

[0020] 1. The measurement method of the present invention improves the existing equipment for measuring the adsorption force of abalone foot muscle, and establishes a standardized measurement method for the adsorption force index of abalone foot muscle, realizing the low-cost, high-efficiency and standardized measurement of the adsorption force of abalone foot muscle, and providing a reliable technical means for the subsequent breeding work of abalone.

[0021] 2. The measurement method of the present invention does not require sacrificing the test parents, and will not cause damage and great stress to the test samples. At the same time, the test efficiency is high, the test speed can reach 1 min / abalone, and the test speed can be adjusted according to needs. The lifting speed of the lifting platform can reach 50 mm / min. In addition, the adsorption force measurement device provided by the present invention is easy to operate, automatically controlled, accurately measured, and has a low learning cost, and can be widely used.

[0022] 3. Compared with other evaluation methods for the stress resistance of abalone, the measurement method of the present invention can retain the measurement parents to a greater extent for subsequent breeding use, and takes less time, has higher efficiency, and has lower requirements for the test site and environment. The improvement of equipment such as the steering bracket and camera assembly of the present invention realizes the more accurate and rapid measurement of the adsorption force. In addition, the present invention realizes the direct measurement of the adsorption force trait of abalone, and can provide the required technical support for the future breeding of abalone adsorption force trait. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 It is a schematic structural diagram of the adsorption force measurement device of the present invention;

[0024] Figure 2 It is a diagram showing the display of the adsorption force numerical data after the adsorption force measurement device of the present invention completes the test;

[0025] Figure 3 It is a diagram showing the display of the adsorption force image data after the adsorption force measurement device of the present invention completes the test;

[0026] Figure 4 It is a schematic diagram of measuring the bottom area of abalone foot using ImageJ software in the present invention;

[0027] Figure 5 It is a box plot of the difference in abalone adsorption force under different conditions in the present invention;

[0028] Figure 6 It is a scatter plot of the difference in abalone adsorption force at different measurement time intervals in the present invention;

[0029] Figure 7 It is a scatter plot of the change in the adsorption force of repeated measurement of abalone adsorption force in the present invention.

[0030] The reference numerals in the figure are shown as:

[0031] 1. Lifting platform; 2. Material testing machine; 3. Steering bracket; 4. Lifting controller; 5. Eagle beak buckle; 6. Grappling hook; 7. Transparent acrylic cylinder; 8. Supplementary light; 9. Camera assembly; 10. Camera module; 11. Radiator; 12. USB multi-port docking station; 13. Platform frame; 14. Test platform. Detailed implementation manner

[0032] In order to make the objectives, technical solutions and advantages of the present invention clearer and more understandable, the present invention will be further described in detail below in conjunction with embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0033] As Figures 1 to 7 shown, a method for measuring the adsorption force of abalone foot muscle includes the following steps:

[0034] S1. Preparation before abalone test: Place the abalone to be tested in an aerated seawater tank for testing;

[0035] S2. Equipment connection and debugging: Connect the adsorption force measuring device to the control computer, turn on the software, adjust the height of the grappling hook 6 to a suitable position through the software or the device controller, select the tensile test plan in the software, and zero the force and displacement readings;

[0036] S3. Abalone adsorption force detection: Take the abalone out of the seawater tank and place it in the transparent acrylic cylinder 7 filled with fresh seawater, let the abalone move freely, after the foot is fully extended, touch the abalone antenna to make the abalone stop moving and adsorb, use the camera assembly 9 to take a photo of the foot, and hook the grappling hook 6 to the abalone shell, and turn on the adsorption force measuring device to measure the adsorption force;

[0037] S4. Adsorption force parameter analysis: After the measurement, export the obtained force measurement data and the sole photo, use the ImageJ software to calculate the bottom foot area of the abalone, and calculate the adsorption strength of the abalone foot muscle.

[0038] As Figure 1 shown, the test speed set by the adsorption force measuring device in step S2 is 50 mm / min.

[0039] As Figure 1 shown, the formula for calculating the adsorption strength of the abalone foot muscle in step S4 is:

[0040] Fs = Fn / S,

[0041] where Fs is the adsorption strength of the abalone foot muscle, with the unit of Kpa; Fn is the adsorption force of the abalone foot muscle, that is, the maximum pull-off force measured by the adsorption force measuring device, with the unit of N; S is the bottom foot area of the abalone, with the unit of cm 2 .

[0042] As Figure 1As shown in the figure, the adsorption force measuring device includes a material testing machine 2, a steering bracket 3, an eagle beak buckle 5, two hooks 6 and a test platform 14; the material testing machine 2 is connected to a control computer for transmitting control signals and mechanical test data; the material testing machine 2 includes a lifting platform 1 and a lifting controller 4, and the lifting platform 1 is adjusted to rise or fall through the lifting controller 4; the steering bracket 3 is fixed to the lower end of the material testing machine 2, and a fixed pulley is arranged on the steering bracket 3. The upper end of the first pulling rope is fixedly connected to the lifting platform 1, and the lower end of the first pulling rope passes through the two fixed pulleys and is fixedly connected to the eagle beak buckle 5; the two hooks 6 are fixedly connected by a second pulling rope, and the second pulling rope is arranged through the lower end of the eagle beak buckle 5; the test platform 14 includes a transparent acrylic cylinder 7, a camera assembly 9 and a platform frame 13; the transparent acrylic cylinder 7 is fixed to the upper end of the platform frame 13, and the camera assembly 9 is arranged on the platform frame 13 below the transparent acrylic cylinder 7, and the foot photos of abalone are taken through the camera assembly 9.

[0043] As Figure 1 shown, a supplementary light 8 is fixed on the platform frame 13 at the lower end of the transparent acrylic cylinder 7, and the lighting effect of the bottom of the abalone foot is improved through the supplementary light 8.

[0044] As Figure 1 shown, the camera assembly 9 includes a camera module 10, a radiator 11 and a USB multi-port docking station 12; the supplementary light 8 is connected to the USB multi-port docking station 12 through a USB cable to receive power supply, and the camera module 10 and the radiator 11 are respectively connected to the USB multi-port docking station 12 through USB cables to receive power supply and transmit data; the USB multi-port docking station 12 is connected to the control computer through a USB cable to receive power supply and transmit data.

[0045] Example 1: Exploration of Standardized Measurement Conditions for the Adsorption Force Index of Haliotis discus hannai

[0046] Select Haliotis discus hannai at the second age from Fuda Abalone Factory in Jinjiang, Quanzhou for the standardized measurement of the adsorption force index. The measurement conditions include: acclimation temperature, starvation time, gender and gonad state. The specific steps are as follows:

[0047] (1) Select 79 second-age Haliotis discus hannai as experimental materials, and acclimate them at 18 °C, 20 °C and 22 °C for 7 days (18 °C, n = 21; 20 °C, n = 33; 22 °C, n = 25), and then conduct the adsorption force measurement.

[0048] (2) Select 111 second-age Haliotis discus hannai (nmale = 63, nfemale = 48) as experimental materials, acclimate them at a water temperature of 20 °C for 7 days, and then conduct the adsorption force measurement.

[0049] (3) Sixty-six 2-year-old Haliotis discus hannai (33 in each group) were selected as experimental materials and acclimated at a water temperature of 20 °C for 7 days before fasting. The adsorption force of 33 abalones was measured after 1 day of fasting, and the adsorption force of the other 33 abalones was measured after 2 days of fasting.

[0050] (4) One hundred and eleven 2-year-old Haliotis discus hannai (empty, Null, n = 54; discharged but not empty, discharged, n = 25; full gonad, Full, n = 32) were selected as experimental materials and acclimated at a water temperature of 20 °C for 7 days, and then the adsorption force was measured.

[0051] (5) The abalones to be measured were placed in an aerated seawater tank for measurement.

[0052] (6) Equipment connection and debugging: Connect the adsorption force measurement device to the control computer, turn on the software, adjust the height of the grab hook 6 to a suitable position through the software or the device controller, select the tensile test plan in the software, zero the force and displacement readings, and set the test speed to 50 mm / min.

[0053] (7) Take the abalone out of the seawater tank and place it in a transparent acrylic tank 7 filled with fresh seawater to let the abalone move freely. After the foot is fully extended, touch the abalone's antenna to make the abalone stop moving, take a photo of the foot using the imaging component 9, and hook the grab hook 6 onto the abalone shell, then turn on the adsorption force measurement device to measure the adsorption force.

[0054] (8) Export the obtained force measurement data and the sole photos of the abalone, calculate the bottom foot area of the abalone using ImageJ software, and calculate the adsorption strength of the abalone foot muscle; the formula for calculating the adsorption strength of the abalone foot muscle is:

[0055] Fs = Fn / S,

[0056] where Fs is the adsorption strength of the abalone foot muscle, with the unit of Kpa; Fn is the adsorption force of the abalone foot muscle, that is, the maximum pull-off force measured by the adsorption force measurement device, with the unit of N; S is the bottom foot area of the abalone, with the unit of cm 2 .

[0057] Table 1: Statistical table of adsorption force indexes of Haliotis discus hannai under different test conditions

[0058]

[0059] As can be seen from Table 1, there were no significant differences among different acclimation temperatures, different starvation times, different genders, and different gonadal states (P > 0.05). The results showed that the acclimation temperature of 18 - 22 °C did not affect the determination result of the abalone's adsorption force, and any temperature within this range could be used as the acclimation temperature for the determination of the abalone's adsorption force. When measuring the adsorption force, the gender of the selected material had no effect on the measurement result. The abalone could still have enough energy to support its life activities after fasting for 2 days, and there was no obvious effect on its movement, etc. When measuring the adsorption force, the abalone could be measured after fasting for 1 day or 2 days. The abalone's gonads develop and mature during the breeding season, and the energy consumption caused by gonad discharge does not affect the magnitude of the adsorption force. When measuring the adsorption force, the gonadal state of the abalone need not be considered.

[0060] Example 2

[0061] In order to further determine the measurement interval (or the recovery time of adsorption force measurement) of the abalone's adsorption force in the present invention, the present invention selected 40 two-year-old Haliotis discus hannai (10 in each group), acclimated them at a water temperature of 20 °C for 7 days. Subsequently, the initial adsorption force was measured. After the measurement was completed, they were placed in a plastic basin for temporary cultivation, and oxygen was continuously supplied during this period to ensure sufficient dissolved oxygen. Then, the adsorption force was measured again after 5 min, 10 min, 20 min, and 30 min respectively. The results are shown in Table 2.

[0062] Table 2: Statistical table of the adsorption force indexes of Haliotis discus hannai at different test time intervals (secondary measurement)

[0063]

[0064] The paired-sample t-test was used to conduct a difference test on the measured adsorption force values. Different letters indicate significant differences between groups (p < 0.05), and the same letter represents no significant difference (p > 0.05).

[0065] As can be seen from Table 2, after the initial measurement of the adsorption force of the abalone, the abalone did not return to the initial state within 10 min. After 20 min, the abalone could recover its adsorption force. It was verified that the test interval for the secondary measurement in the present invention should be greater than 20 min. Finally, it was set that the secondary measurement interval of 30 min was appropriate to ensure that the abalone's adsorption force was fully recovered.

[0066] Example 3

[0067] In order to further determine whether the present invention can perform multiple measurements of the adsorption force of the same batch of abalones at fixed time intervals, the inventor of the present invention selected 10 two-year-old Haliotis discus hannai as experimental materials and acclimated them at a water temperature of 20 °C for 7 days. Subsequently, the adsorption force of the abalones was continuously measured. After each measurement was completed, they were placed in a plastic basin for temporary cultivation and recovery. The adsorption force of the abalones was measured again 30 min and 60 min after the first measurement. The results are shown in Table 3.

[0068] Table 3: Statistical table of adsorption index of Abalone discus henna measured continuously at the same time interval

[0069] Initial adsorption force value Measure the adsorption force value again after 30 min Measure the adsorption force value again after 60 min 1 94.19 100.09 72.98 2 81.75 56.77 27.50 3 65.88 60.10 43.03 4 96.04 52.22 39.76 5 138.67 85.16 39.84 6 76.59 76.82 70.27 7 84.20 56.18 44.44 8 43.60 59.35 20.54 9 124.26 121.04 84.77 10 53.25 28.70 40.96 85.84±29.42a 69.64±26.63a 48.41±20.73b

[0070] As shown in Table 3, the abalone adsorption capacity recovered 30 minutes after the first measurement, but could not be recovered 30 minutes after the second measurement, indicating that the recovery time of abalone adsorption capacity increased with the number of measurements.

[0071] The method for determining the adsorption capacity of abalone foot muscle of the present invention has high accuracy and strong stability. As a non-destructive, precise and rapid method for determining the adsorption capacity trait of abalone, it will provide assistance for the evaluation of abalone stress resistance traits, assist in the breeding of abalone stress resistance traits, and provide a reliable technical means and an important basis for the subsequent direct breeding work on the adsorption capacity traits of abalone.

[0072] The above is only a preferred specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed by the present invention should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A method for measuring the adsorption force of abalone foot muscle, characterized in that: The following steps are involved: S1. Preparation before abalone testing: Place the abalone to be tested in an inflated seawater tank for testing; S2. Equipment connection and debugging: connect the adsorption force measuring device to the control computer, start the software, adjust the height of the grab hook (6) to a suitable position through the software or the device controller, select the stretching test program in the software, and adjust the force and displacement readings to zero; S3, abalone adsorption force detection: the abalone is taken out of the seawater tank and placed in a transparent acrylic tank (7) filled with fresh seawater, and the abalone is allowed to move freely. After the foot is fully unfolded, the abalone tentacles are touched to stop the abalone from moving and adsorbing. A camera assembly (9) is used to take a picture of the foot, and the grappling hook (6) is hooked onto the abalone shell, and the adsorption force measuring device is turned on to measure the adsorption force; S4. Adsorption force parameter analysis: After the measurement, the force measurement data and the plantar photos were exported, and the plantar area of ​​the abalone was calculated using ImageJ software, and the adsorption strength of the abalone plantar muscle was calculated.

2. The method for measuring the adsorption force of the abalone foot muscle according to claim 1, characterized in that: The test speed of the adsorption force measuring device in step S2 is set to 50 mm / min.

3. The method for measuring the adsorption force of the abalone foot muscle according to claim 1, characterized in that: The formula for calculating the adsorption strength of the abalone foot muscle in step S4 is: Fs=Fn / S, Among them, Fs is the adsorption strength of the abalone foot muscle, in KPa; Fn is the adsorption force of the abalone foot muscle, that is, the maximum pull-off force measured by the adsorption force measurement device, in N; S is the area of ​​the abalone foot, in cm 2 .

4. The method for measuring the adsorption force of the abalone foot muscle as claimed in claim 1, characterized in that: The adsorption force measuring device comprises a material testing machine (2), a steering bracket (3), a hawk beak buckle (5), two grab hooks (6) and a test platform (14); the material testing machine (2) is connected to a control computer for transmitting control signals and mechanical test data; the material testing machine (2) comprises a lifting platform (1) and a lifting controller (4), and the lifting platform (1) is adjusted to rise or fall by the lifting controller (4); the steering bracket (3) is fixed to the lower end of the material testing machine (2), and a fixed pulley is arranged on the steering bracket (3), and the upper end of the first pulling rope is connected to the upper end of the first pulling rope. The lifting platform (1) is fixedly connected, and the lower end of the first pulling rope passes through two fixed pulleys and is fixedly connected to the hawk's beak buckle (5); the two grab hooks (6) are fixedly connected through a second pulling rope, and the second pulling rope is passed through the lower end of the hawk's beak buckle (5); the test platform (14) includes a transparent acrylic cylinder (7), a camera assembly (9) and a platform frame (13); the transparent acrylic cylinder (7) is fixed to the upper end of the platform frame (13), and the camera assembly (9) is arranged on the platform frame (13) below the transparent acrylic cylinder (7), and a picture of the abalone's foot is taken through the camera assembly (9).

5. The method for measuring the adsorption force of the abalone foot muscle as claimed in claim 4, characterized in that: A fill light (8) is fixed on the platform frame (13) at the lower end of the transparent acrylic cylinder (7), and the fill light (8) is used to improve the lighting effect of the sole of the abalone.

6. The method for measuring the adsorption force of the abalone foot muscle as claimed in claim 5, characterized in that: The camera assembly (9) comprises a camera module (10), a heat sink (11) and a USB multi-port expansion dock (12); the fill light (8) is connected to the USB multi-port expansion dock (12) via a USB cable to receive power, and the camera module (10) and the heat sink (11) are respectively connected to the USB multi-port expansion dock (12) via USB cables to receive power and transmit data; the USB multi-port expansion dock (12) is connected to a control computer via a USB cable to receive power and transmit data.

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