CN110440969B

CN110440969B - Quick patinopecten yessoensis vitality evaluation method

Info

Publication number: CN110440969B
Application number: CN201910729669.7A
Authority: CN
Inventors: 田元勇; 刘俊荣; 刘洋; 李亚煊
Original assignee: Dalian Ocean University
Current assignee: Dalian Ocean University
Priority date: 2019-08-08
Filing date: 2019-08-08
Publication date: 2023-05-23
Anticipated expiration: 2039-08-08
Also published as: CN110440969A

Abstract

The invention discloses a quick patinopecten yessoensis vitality evaluation method, which is characterized by comprising the following steps of: the method comprises the following steps of: firstly, fixing the left shell of a living patinopecten yessoensis on a square stone by quick-drying cement, and keeping the right shell in a free moving state; then, hooking the lower end of the dynamometer at the edge of the right shell of the scallop which can freely move; the scallop mantle was stimulated with a needle at 30 s/min intervals for 12 times of co-stimulation, and the frequency of closure and the instantaneous maximum of the closing force were detected within 6 minutes by real-time recording of the data by an external computer connected to a dynamometer. Drawing a closing force waveform chart by taking the horizontal axis as detection time and the vertical axis as closing force, analyzing the scallop closing force waveform chart by using origin software, calculating the average value of the instantaneous maximum closing force peak value, and finally evaluating the activity of the scallop according to the value.

Description

Quick patinopecten yessoensis vitality evaluation method

Technical Field

The invention relates to the field of living body circulation of aquatic products, in particular to a quick evaluation method for patinopecten yessoensis vitality.

Background

Patinopecten yessoensis is one of important cultured varieties in China, and has higher economic value and nutritive value. Living products are the most popular product form in China.

The quality evaluation of living products is not only a precondition means for researching the scallop quality change mechanism, but also a judgment basis for commercial operation among various links of a supply chain. In actual production, the specification, the edge shrinkage and the fertilizer fullness are indexes which are currently used by the industry at present; in the scientific research field, the theory and technology of fish preservation are mostly used. Commonly used indicators are ATP-related compounds, muscle pH, glycogen content, free amino acids, etc. The ATP and the phosphate can better reflect the quality of living products, but the ATP and the phosphate are inconvenient to measure on the sites of shellfish fishing, transportation and consumption due to complex extraction and the need of large-scale instruments such as high-efficiency liquid in the measuring process. And the measurement process needs to take several hours, and cannot meet the real-time monitoring requirement in the whole circulation process, a method or device capable of solving the above problems is needed.

Disclosure of Invention

The invention aims to solve the defects in the prior art, and provides a patinopecten yessoensis vitality evaluation method capable of conveniently and rapidly measuring patinopecten yessoensis vitality at any time on the fishing, transportation and consumption sites.

The technical scheme of the invention is as follows: a patinopecten yessoensis vitality quick evaluation method is characterized by comprising the following steps of: the method comprises the following steps of:

firstly, fixing the left shell of the living patinopecten yessoensis on a square stone by quick-drying cement, keeping the right shell in a free moving state, standing for 30 minutes in a room temperature environment to solidify the cement so as to fix the patinopecten yessoensis,

then, hooking the lower end of the dynamometer at the edge of the right shell of the scallop which can freely move, stimulating the scallop mantle with a needle at 30 s/time intervals for 12 times of co-stimulation, recording data in real time by an external computer connected with the dynamometer, detecting the instantaneous maximum value of the closing frequency and closing force within 6 minutes, drawing a closing force waveform chart by taking the horizontal axis as the detection time and the vertical axis as the closing force,

analyzing the closing force waveform diagram, counting the scallop closing generation frequency and the instantaneous maximum peak value by using origin software, and calculating the average value of the instantaneous closing force of the scallop closing generation frequency and the instantaneous maximum peak value, wherein the average value is used for evaluating the activity of the scallop, and the calculation formula is as follows: closing force average (N) =sum of peaks/closing frequency.

Compared with the prior art, the invention has the following advantages:

the invention provides a quick patinopecten yessoensis vitality evaluation method, which aims at the problems of the traditional scallop vitality evaluation method by detecting ATP and phosphate source energy of scallops, and designs a simple evaluation method which reflects the contraction force of scallop muscles through the closing force and obtains a change waveform diagram of the closing force. Then analyzing the obtained waveform diagram, and calculating the average value of closing force as a rapid evaluation index of scallop vitality; the method has positive correlation with the result of the classical biochemical evaluation method (the traditional method for detecting scallop ATP and phosphate precursor energy by using large-scale equipment), is simple and easy to implement, and can perform real-time evaluation most importantly, so that the method has various advantages, is particularly suitable for popularization and application in the field of shellfish living goods circulation, and has very broad market prospect.

Drawings

FIG. 1 is a schematic diagram of an apparatus used in the method of the present invention.

Fig. 2 is a waveform diagram showing the variation of the closing force of scallops.

FIG. 3 is a plot of ATP content and closing force variation of scallops.

Detailed Description

The following will describe embodiments of the present invention with reference to the accompanying drawings, as shown in fig. 1, 2 and 3: a patinopecten yessoensis vitality quick evaluation method sequentially comprises the following steps:

then, when the tentacles of the scallop mantle are fully unfolded and kept in a natural opening state, the lower end of the dynamometer is hooked at the edge of the right shell of the scallop which can freely move,

stimulating scallop mantle with needle at 30 s/time for 12 times, recording data in real time by external computer connected with dynamometer, detecting the frequency of closing in 6 min and the instantaneous maximum value of closing force, drawing waveform graph of closing force with horizontal axis as detection time and vertical axis as closing force,

Specific examples:

1. selecting commercial specification scallops: the shell is about 10.5cm in height, about 10cm in length, about 2.0cm in thickness, about 120g in weight, and 60 healthy living bodies.

2. Fixing patinopecten yessoensis: the left shell of patinopecten yessoensis is fixed on square stone by quick-drying cement, and the right shell is kept free to move.

3. Patinopecten yessoensis vitality standardization: placing the fixed scallop into a flowing seawater tank for temporary culture for 24 hours, so that the scallop is fully recovered to the vitality state.

4. And (3) applying dry dew stress: the scallops recovered to the optimal state are taken out from the sea water and divided into 6 groups of 10, 5 of the groups are treated by the method, and the ATP content of the muscles of the other 5 groups is measured by a traditional method (the ATP content of the muscles is measured by the traditional method is a reference group). The 6 groups of scallops are respectively placed in a refrigerator at the temperature of 4 ℃ for 0h, 6h, 8h, 12h, 36h and 48h.

5. Measuring the closing force of scallops: the closing frequency and instantaneous maximum closing force of patinopecten yessoensis within 6 min after the stimulus (30 s/time) was applied were measured, respectively, as shown in FIG. 1. The frequency of closure is between 10 and 20 times and the closing force is between 1 and 25 and N.

And each scallop performs closing force acquisition according to the operation. The AVERAGE value of the instantaneous maximum value of the closing force (AVERAGE (N1: N5)) of each group of scallops within 6 minutes was calculated as an index of the viability of each group of scallops.

6. Determination of ATP-related compound content in adductor scallop muscle: 1.0 g scallops were cut from each scallop tested as described above and mixed with 1:10 adding 5% perchloric acid (PCA), fully mashing in ice bath, adding 2 mol/L KOH, regulating the pH to 2.0-3.5, adding ultrapure water to a constant volume of 20 mL, centrifuging for 5 min at 3000 g, taking supernatant and filtering with a 0.45 mu m filter membrane. The filtrate of 4 mL was taken and 1 mL of 0.1 mol/L phosphate buffer (pH 7.5) was added. The whole process is carried out under ice bath conditions.

High performance liquid chromatography was used for analysis. Chromatographic column: DALIITE corporation (SinoChrom ODS-BP 5 μm,4.6 mm X250 mm); a detector: a Diode Array Detector (DAD); detection wavelength: 254 nm; temperature: 35 ℃; flow rate: 0.7 mL/min; sample injection amount: 0.02 And (3) mL. Mobile phase a:0.05 mol/L K HPO4-KH2PO4 buffer (pH 6.5), mobile phase B: mobile phase a: methanol solution= (8:2).

Data analysis: the closing force and the ATP content are analyzed on the abscissa and on the ordinate, respectively. The results show that: as the dry exposure time was prolonged, the closing force of the scallop was gradually decayed from 14N to 6N. The ATP content was reduced from 5.6. Mu. Mol/g to 3.6. Mu. Mol/g. Both the closing force and the ATP content show a decreasing trend. Correlation analysis of the two indices (see Table 1) showed that the correlation coefficient of the closing force magnitude and the ATP content was 0.94, indicating that the closing force was extremely significantly correlated with the change in ATP content (P < 0.01).

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Claims

1. A patinopecten yessoensis vitality quick evaluation method is characterized by comprising the following steps of: the method comprises the following steps of:

then, the lower end of the dynamometer is hooked on the edge of the right shell of the scallop which can freely move,

analyzing the closing force waveform diagram, counting the instant maximum peak value of the closing generation frequency and the closing force of the scallop by using origin software, and calculating the average value of the instant closing force of the scallop to evaluate the vitality of the scallop, wherein the calculation formula is as follows: average closing force (N) =instantaneous maximum peak sum of closing forces/closing frequency.