CN112931311B - Flounder health condition evaluation method - Google Patents
Flounder health condition evaluation method Download PDFInfo
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- CN112931311B CN112931311B CN202110178423.2A CN202110178423A CN112931311B CN 112931311 B CN112931311 B CN 112931311B CN 202110178423 A CN202110178423 A CN 202110178423A CN 112931311 B CN112931311 B CN 112931311B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K29/00—Other apparatus for animal husbandry
- A01K29/005—Monitoring or measuring activity, e.g. detecting heat or mating
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K63/00—Receptacles for live fish, e.g. aquaria; Terraria
- A01K63/04—Arrangements for treating water specially adapted to receptacles for live fish
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/347—Use of yeasts or fungi
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/20—Nature of the water, waste water, sewage or sludge to be treated from animal husbandry
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
Abstract
The invention discloses a flounder health condition evaluation method, which comprises the following steps: after the early preparation of a culture system and the monitoring of a culture environment, flounder to be evaluated is placed into the culture system, and health evaluation is performed on the flounder to be evaluated through capture and anti-capture behaviors, metabolic indexes, metabolic changes after acute stress, appetite recovery behaviors after stress autism, sand diving behaviors and circadian rhythm behaviors in sequence. According to individual differences of physiological and behavioral characteristics of flounder, multiple evaluations and grading evaluations are adopted, a health evaluation standard is established, the health conditions of flounder groups in artificial breeding environment and natural environment are comprehensively evaluated, breeding technology is improved, and flounder breeding management and healthy and continuous development of breeding industry are promoted.
Description
Technical Field
The invention relates to an aquaculture and marine organism health evaluation method, in particular to a flounder health evaluation method.
Background
With global extreme climate change and aggravation of the influence of human activities on the natural environment, the environment on which organisms in the nature live is greatly changed, and the survival of the organisms is greatly threatened. In addition, the aquaculture management and breeding theory in China is relatively lagged, so that the water area environment is deteriorated, the seed resources are declined, and the diseases are frequent. In the normal breeding process of flounders, various environmental changes and artificial stresses are inevitably encountered, weak or sub-healthy individuals in breeding groups suffer from anorexia and attack diseases first, and the whole group is infected, thereby causing large-scale disasters. Therefore, health assessment of the artificial breeding environment and flounders in nature is beneficial to environmental monitoring, breeding disasters can be avoided, breeding rate of the cultured flounders is improved, breeding cost is reduced, healthy continuous development of flounders breeding industry in China is promoted, marine flounders resources are protected, and the method has important significance for biological effect early warning and ecological damage assessment of marine environmental change.
Disclosure of Invention
In view of the above problems, the present invention aims to provide a method for evaluating health status of flounder.
In order to achieve the purpose, the invention adopts the following technical scheme:
a flounder fish health condition evaluation method, the individual to be evaluated can be temporarily cultured in running water or circulating water system, or can be temporarily cultured in the sea water of net cage and pond during the health evaluation period, the health evaluation includes the following steps:
s1, disinfecting a culture system by bleaching powder, washing by fresh water, soaking, and injecting filtered natural seawater or diluted concentrated seawater with salinity of 25-35 per mill; yeast extract powder and photosynthetic bacteria are required to be put into the circulating water system, the circulating water system is operated for a plurality of days, and the biomembrane in the filter tank is cultured;
s2, monitoring the culture environment to enable the culture environment to reach a preset environmental index;
s3, after the environmental index is detected to be qualified, the flatfish to be introduced into the culture system is soaked in seawater containing povidone iodine, then is soaked in aerated seawater, and is then put into the culture system;
s4, carrying out first-round evaluation on flounder to be evaluated by using a fishing net catching method and an air exposure method, respectively recording the individual behaviors of immobility (inactivity) and escape (liveness), fishing out the individuals to be evaluated one by one, observing the breaking-off condition of the individuals, respectively recording behavior indexes, and recovering the individuals for a plurality of hours;
s5, utilizing a full-automatic intermittent flow respiratory motion meter (AutoResp) TM Logo Systems) for health evaluation of flounder, before use and after disinfecting the equipment to be used every two days with mild bleaching liquid, rinsing with fresh water and soaking, and injecting filtered natural seawater with salinity of 25-35 per mill;
monitoring the measuring environment to enable the measuring environment to reach a preset environment index;
setting a circulation period for the water body flow in the respiration monitoring chamber, wherein each period is divided into three stages, the first stage is a process of slowly adding new water into the monitoring chamber by an external water storage tank, the second stage is waiting time, and the third stage is a process of monitoring the change time of dissolved oxygen in the water body by the monitoring chamber and calculating the oxygen consumption value of the dissolved oxygen; before the experiment begins, the system is not provided with fish and at least carries out a plurality of cycles to monitor the background oxygen consumption value in the water body; then, measuring the maximum metabolism rate, the standard metabolism rate and the absolute aerobic range of the flounder to be evaluated, and respectively recording according to the maximum metabolism rate, the standard metabolism rate and the absolute aerobic range;
s6, in the step S5, at the stage close to the end of the experiment, in the third stage of the system, no new water injection period exists, a respiration monitoring experiment under acute stimulation is carried out, the index is used as a respiration selection strategy index under the condition of evaluating the acute stress of the flounder, the oxygen consumption change of the flounder is recorded, and then individuals to be evaluated are placed into independent culture cylinders one by one to wait for the evaluation of the condition of food intake recovery in the next step;
s7, evaluating the appetite recovery condition of flounder to be evaluated after further stress solitary, evaluating the individuals to be evaluated one by bait fishing, recording the individuals respectively, and recovering the individuals for a plurality of hours;
s8, evaluating flounder to be evaluated by using a sand diving behavior, recording the sand coverage area of an individual to be evaluated, and recovering for a plurality of hours;
s9, performing five-round evaluation on flounder to be evaluated by using a circadian rhythm behavior detector (bigbother 3, actimetrics), flushing and soaking clean by fresh water after equipment to be used is disinfected by mild bleaching solution before and during use, injecting filtered natural seawater with the salinity of 25-35 per mill, gently placing the flounder to be evaluated in a monitoring room, keeping a normal photoperiod, continuously monitoring each fish for 24 hours, recording day and night movement conditions of each fish, and recovering the fish for several hours;
s10, analyzing and mapping the results of the capture-anti-capture behavior, the appetite recovery behavior after stress orphan, the sand-diving behavior and the circadian rhythm behavior by using a principal component analysis method in XLSAT 2014 to obtain correlation distribution maps of different individuals and the behavior test indexes, obtaining a set of three individuals with different health conditions, and obtaining the health conditions of the different individuals by combining metabolism results.
Further, in step S1, the disinfectant is bleaching powder, the available residual chlorine in the disinfectant should be greater than 30ppm, the disinfectant is washed with fresh water for 3 times and soaked for 1 day, 0.02g/L yeast extract powder and 0.2ml/L photosynthetic bacteria are added into the circulating water system, the newly used circulating water system is operated for at least 30 days, and the system which has been used is operated for one week.
Further, in step S2, the preset environmental index is: the water temperature in the culture system is 17-20 ℃, the illumination time and the dark time are respectively 12 hours, the pH value is 7.5-8.5, the dissolved oxygen concentration is more than or equal to 6.8mg/L, the ammonia nitrogen concentration is less than or equal to 0.1mg/L, the nitrite nitrogen concentration is less than or equal to 0.1mg/L, and the sulfide concentration is less than or equal to 0.1mg/L.
Further, in step S3, the disinfection tank is filtered or diluted seawater containing 0.006% povidone iodine (the specific dosage is determined according to the purchased drug parameters), the salinity is 25% o to 35% o, and the dissolved oxygen concentration is more than or equal to 6.8mg/L; if the throwing density of the flounder seedlings introduced into the culture system is 5-10kg/m 2 10-15kg/m of adult flounder 2 And aerating the sterilized water sufficiently by using an air pump, and soaking for about 1h.
Further, in step S4, continuing to stop feeding food before evaluation, starting preliminary evaluation at about 9 a.m. after the pool flow is stable and the fish school is quiet, and when performing a first round of evaluation on flounder, firstly sliding a fishing net nearby to perform simulated capture, wherein the behavior of a fish escaping rapidly in the process that the fishing net approaches slowly is marked as 1, the behavior of a fish escaping after the fishing net contacts the fish body is marked as 0, and the behavior of a fish which still does not respond when the fishing net contacts the fish body during simulated capture is marked as-1; then further fishing net grabbing evaluation is carried out, flounder to be evaluated is quickly fished into the net one by using a fishing net, the water surface is fished out for 70 seconds, the time from the fish grazing into the fishing net to the beginning of struggling and the struggling times within 70 seconds are respectively recorded, the struggling time is arranged from short to long, 1 minute is taken for the first third flounder, 0 minute is taken for the middle third flounder, and-1 minute is taken for the last third flounder; the struggling times are that the first third of flounder is counted for 1 point, the middle third of flounder is counted for 0 point and the last third of flounder is counted for-1 point according to the sequence from high to low, two groups of fishes with the same data are placed in the same system, and after the first round of evaluation, the fish school is recovered from stress for 12-24 hours;
further, in step S5, a fully automatic intermittent flow respiratory motion meter (AutoResp) is used TM Logo Systems) to perform a second round of health evaluation on flounder to be evaluated, after disinfecting equipment to be used every two days before starting use by using a mild bleaching solution, washing the equipment for 3 times by using fresh water and soaking and washing the equipment clean, injecting filtered natural seawater with full aeration and salinity of 25-35 per mill, and performing temperature control by using TMP-REG temperature monitoring and controlling equipment (logo Systems);
the preset environmental indexes are as follows: the water temperature in the monitoring system is 18 +/-0.1 ℃, the pH value is 7.5-8.5, the dissolved oxygen concentration is more than or equal to 6.8mg/L, the ammonia nitrogen is less than or equal to 0.1mg/L, the nitrite nitrogen is less than or equal to 0.1mg/L, and the sulfide is less than or equal to 0.1mg/L;
the dissolved oxygen monitoring chamber is 48cm × 28cm × 4cm (length × width × height), can hold 6.3L of water volume, and is arranged in the monitoring systemThe flow of the water body is controlled by LDAQ-4 (logo Systems), dissolved oxygen is continuously monitored by using an electro-oxygen probe (logo Systems), a monitoring period is set to be 10 minutes as a cycle, wherein 0-4 minutes is a process that an external water storage tank slowly adds new water into a monitoring chamber so as to ensure that the dissolved oxygen of the water body of the monitoring chamber reaches 100 percent, 5 minutes is the time for waiting for the data of a breathing chamber to be stable, 6-10 minutes is the time for monitoring the change of the dissolved oxygen in the water body of the monitoring chamber, and an automatic resp is used TM software (logo Systems) collects temperature and dissolved oxygen data, corresponding respiration values are calculated, at least 3 cycles are carried out before experiments are started to monitor background oxygen consumption values caused by bacteria in a water body without putting fishes in the system, and the background oxygen consumption values are controlled to be within 5% of the maximum metabolism rate value of adult fishes to be detected;
firstly, carrying out a test for measuring the maximum metabolism rate, the standard metabolism rate and the absolute aerobic range of flounder, wherein the index is used as the maximum aerobic respiration capacity index of the flounder, before 8 am, putting the flounder to be evaluated into a rectangular temporary rearing chamber (70 cm multiplied by 50cm multiplied by 30cm, length multiplied by width multiplied by height) with preset environmental indexes from a rearing pond, then continuously catching for 3 minutes by using a fishing net, if the fish to be evaluated decelerates or stops swimming in the process, touching the tail part of the fish by using the fishing net to stimulate the fish to continuously carry out explosive swimming, after 3 minutes, completely exhausting all physical strength of all individuals, then immediately putting the fish out of the water surface by using the fishing net for 1 minute, then putting the fish into a monitoring chamber within 30 seconds, discharging and locking all gas in the monitoring chamber, covering the surface of the monitoring chamber by using a black baffle plate, reducing the external disturbance to the minimum in a recovery stage, carrying out the test for 24 hours, wherein the maximum respiration rate is the maximum metabolism rate, when the 1 st-2 nd cycle appears, the absolute respiration rate is used as the maximum aerobic metabolism rate and the absolute aerobic metabolism range is subtracted as the standard metabolism rate and the absolute aerobic metabolism range to be used as the maximum metabolism rate of each fish to be evaluated; respectively taking the first third of fishes to mark 1 minute, the middle third of fishes to mark 0 minute and the last third of fishes to mark-1 minute according to the arrangement of the maximum metabolism rate, the standard metabolism rate and the absolute aerobic range from high to low;
further, in step S6, after the 24-hour experiment in S5 is finished, in the 6 th to 10 th minute dissolved oxygen change monitoring period of the system, performing an evaluation of oxygen consumption change under acute stimulation for 5 minutes, wherein the evaluation index is used as a metabolism change index under acute stress conditions of flounder, removing a black baffle on the surface of a monitoring room before the experiment starts, simulating predators to disturb the fish by using shadows generated by hands under light, wherein 30 seconds is a cycle, 0 th to 15 th second testers continuously move hands above the monitoring room, the shadows caused by the test personnel simulate predators under natural conditions, stopping the disturbance for 16 th to 30 th seconds, and then continuously performing for 5 minutes, wherein the process always monitors the change of dissolved oxygen in the water body, records the oxygen consumption value of the flounder to be evaluated when disturbed, and records 1 minute when the oxygen consumption obviously rises according to the oxygen consumption change during the disturbance, 0 minute when the oxygen consumption does not change, and records-1 minute when the oxygen consumption drops; putting flounder to be evaluated into independent culture barrels one by one, covering a half barrel mouth of each culture barrel with a cover, and placing the culture barrels in a semi-light-shading mode, wherein each culture barrel is connected with a water supply system of a culture pond, so that the water quality conditions in all the culture barrels are ensured to be the same, and the fishes are recovered from the second round of evaluation stress in the culture barrels for 12-24 hours;
further, in step S7, after fasting, feeding reaction health evaluation is carried out on flounder to be evaluated, feeding is carried out on one side of the uncovered barrel opening of the breeding barrel, and the illumination intensity in the barrel is about 40LX, so that adult fishes can observe 'prey'; preparing a fishing rod, replacing a fishhook at one end of a fishing line with a small wooden clamp, tying the other section of the fishing rod on the fishing rod, clamping a small fish to be fed on the wooden clamp, moving the fishing rod to drag the small fish to swim on the water surface, observing the reaction of the adult fish to be detected at the bottom of the pool, recording the reaction as-1 if the adult fish pounding the hunting to be started in 30 seconds, recording the reaction as-0 if the adult fish pounding the hunting to be started in 2 minutes, and recording the reaction as-1 if the reaction time exceeds 2 minutes; the method can be used in a culture barrel and a fish pond, and the illumination visibility of the bottom of the fish pond is adjusted by an artificial light source, so that the illumination intensity is 40LX.
Further, in step S8, in order to avoid predators and strategic predation in nature, sand diving behavior of flatfish occurs, and the time for starting the sand diving behavior of flatfish is recorded; arranging the time for starting sand-burying from short to long, taking the first third of flounder to count 1, the middle third of flounder to count 0 and the last third of flounder to count 1; and then further evaluating the sand coverage area, wherein the sand coverage area is gradually reduced, the first third of the marks of the flounder are taken for 1 point, the middle third of the marks of the flounder are taken for 0 point, and the last third of the marks of the flounder are taken for-1 point.
Further, in step S9, in order to avoid predation by predators in the daytime in nature, activities and predation are performed at night, five rounds of health evaluation are performed on flounder to be evaluated by using a circadian rhythm behavior detector (bigbother 3, actimetrics), equipment to be used the day before use is started is sterilized by a mild bleaching solution, and then washed by fresh water and soaked cleanly, and each monitoring room is a light color culture tank with the size of 70cm × 50cm × 30cm (length × width × height); before 8 am, respectively injecting 52L of filtered natural seawater with sufficient aeration and salinity of 25-35 per mill into each monitoring chamber, then gently placing flounder to be evaluated into the monitoring chambers one by one, wherein the light cycle time is 12 hours and the light is dark for 12 hours, a camera tracks the swimming distance of each monitored flounder, the swimming distance of each fish is reported once per minute, each fish is continuously monitored for 24 hours, and then the fish is fished out and placed back into a corresponding culture system to recover for 12-24 hours in stress; taking whether obvious movement exists in the daytime and the total distance of the movement at night as scoring indexes, recording 1 score for no movement in the daytime, recording 0 score for no obvious movement and recording-1 score for obvious movement; the movement distance at night is arranged from high to low, the first third of flounder is counted for 1 point, the middle third of flounder is counted for 0 point, and the last third of flounder is counted for 1 point.
Further, in step S10, the results of the capture-anti-capture behavior, the metabolic index, the metabolic change after acute stress, the appetite recovery behavior after stress autism, the sand-diving behavior, and the circadian rhythm behavior are analyzed and plotted by using a principal component analysis method in XLSTAT 2014, so as to obtain correlation distribution maps of different individuals and the behavior test indexes, and obtain a set of three different health conditions: the fish with the characteristic value of 2 or above is active fish, the fish with the characteristic value of-2 to 2 is intermediate fish, and the fish with the characteristic value of-2 or below is inactive fish; then the metabolism results are integrated to further obtain the health conditions of different individuals, and according to the score from high to low, the fish with the integrated score of 9 or more are live fish; fish of intermediate type at-9 to 9; inactive fish in-9 cents or below.
The method is characterized in that only two conditions of active fish and inactive fish are usually present in a natural culture population, an intermediate individual appears in an artificial culture population influenced by culture activities, the active individual has the characteristics of high food utilization rate, strong active foraging capacity, high motion level, low stress level and the like, the inactive individual has the characteristics of high stress level, environmental stress sensitivity, high neural plasticity, low aggressivity and the like, different individual individuals are screened, and the method is favorable for distinguishing the health conditions of different individuals of flounder. In an artificial culture environment, health assessment is carried out on flounder by utilizing a catching-anti-catching behavior, a metabolism index, metabolism change after acute stress and an appetite recovery behavior after stress lonely; in a natural environment, health assessment is carried out on flounder by utilizing a catching-anti-catching behavior, a metabolism index, metabolism change after acute stress, an appetite recovery behavior after lonely, a sand diving behavior and a circadian rhythm behavior. According to the method, the evaluation indexes of the health conditions of different flounders are used for screening out healthy fish species which meet the actual culture condition from 3 types of flounders with different personalities, so that the culture management and breeding technology of the flounders is promoted.
The invention can obtain the following technical effects through the technical scheme:
1. the invention utilizes the non-specificity of stress and carries out health evaluation on flounder according to the principles of different stress expression, recovery time and metabolism of flounder with different physique and activity after being subjected to various external stimuli.
2. The method utilizes the difference of stress behavior reaction on flounder individuals with different health conditions to determine the proportion of individuals with weak constitution and individuals which are not easy to recover after stress in fish stocks to distinguish, thereby evaluating the health condition and the overall stress resistance of a test population and achieving the purposes of biological effect early warning of environmental change and ecological damage evaluation.
3. The fishing net is used for catching flounder activity and limiting air exposure stress to serve as a first and second step index of flounder health evaluation.
4. The full-automatic intermittent flow respiratory motion meter is used for measuring the maximum metabolic rate, the standard metabolic rate and the absolute aerobic range, and the respiratory selection strategy under the acute stress condition is used as the third and fourth step indexes for flounder health evaluation.
5. The reaction of flounder to food bait after the solitary strange environment and short-term hunger stress is used as the fifth index of flounder health evaluation.
6. And (4) utilizing the sand diving behavior as a sixth index of flounder health evaluation.
7. And monitoring the total movement distance for 24 hours by using a circadian rhythm behavior detector as a seventh step index of flounder health evaluation.
8. The stress resistance and the disease resistance of the flounder are closely related, and the flounder individual selected by the invention has the characteristic of strong overall stress resistance.
9. According to the method, various stresses, appetite and the number proportion of sub-healthy fishes in the group of the offspring are determined through multiple rounds of health evaluation, the improved variety rate of the offspring is determined, the improved variety screening is facilitated, and the method can be widely applied to the fields of large-scale aquaculture and improved variety culture.
In conclusion, the flounder health evaluation method provided by the invention utilizes the physiological characteristics of flounder to pursuing, air exposure stress, metabolism and appetite recovery after stress orphism, combines the sand diving behavior and the circadian rhythm behavior in the natural environment, adopts a multi-evaluation and comprehensive scoring evaluation method, distinguishes flounder individuals with different stress resistance and appetite by health evaluation of flounder populations in the early culture period, maintains healthy individuals in the early culture period, keeps healthy flounder seedling and adult fish populations, and improves the culture yield and the survival rate of filial generation.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention utilizes the non-specificity of stress, according to different metabolic indexes of flounder and flounder with different physique and activity and different circadian behaviors, and after being stimulated by various external stimuli, the principles of different stress expression and recovery time are as follows: after the fish is stimulated by the outside, the recovery time of the stress generated by the external stimulation is different due to different physique and activity of the fish. The fish with good body quality and strong activity has the advantages of maximum metabolism, standard metabolism, higher metabolic space, obvious activity in the daytime and more frequent activity at night, can quickly escape from a dangerous area after being stimulated, can violently struggle under the severe stimulation of air exposure, can quickly recover in a short time after the stimulation disappears, and starts to eat; the fish with poor constitution and weak activity has the advantages of maximum metabolism, standard metabolism, lower metabolic space, no obvious activity in the daytime and less activity at night, the fish with weak activity cannot escape from a dangerous area after being stimulated, cannot violently struggle under the severe stimulation of air exposure, cannot quickly recover in a short time after the stimulation disappears, and cannot begin to eat in a short time. However, a scientific and reasonable detection method is required for health evaluation, so that death of the fish due to overlarge strength of the evaluation method is avoided, healthy and active juvenile fish or bred parent fish can be accurately distinguished, and a reasonable health evaluation method is required. The present invention will be described in detail with reference to examples.
Example one
S1, preparing a culture system:
disinfecting the existing culture system with bleaching powder disinfectant with residual chlorine of more than 30ppm, soaking for 1 day, washing with fresh water for 3 times, and injecting natural seawater with salinity of 25-35 ‰; 0.02g/L yeast extract and 0.2ml/L nitrobacteria are required to be added into the circulating water system, the circulating water system is operated for at least 15 days, and the biomembrane in the filter tank is cultured.
S2, monitoring the culture environment, and checking whether the culture environment meets preset environmental indexes:
the water temperature in the culture system is 18-20 ℃; the light time required was 12 hours (7; the pH value is 7.5, the dissolved oxygen concentration is 6.8mg/L, the ammonia nitrogen concentration is 0.1mg/L, the nitrite nitrogen concentration is 0.1mg/L, and the sulfide concentration is 0.1mg/L.
S3, after being qualified through the detection of S2, the paralichthys olivaceus needing to be introduced into the culture system is firstly soaked in seawater containing povidone iodine with the oxygen content of 0.006 thousandth for 1 hour, and then the paralichthys olivaceus is soaked in aerated seawater and then is placed into the culture system;
wherein the seawater salinity of 0.006 per mill povidone iodine is 25 per mill-35 per mill, and the dissolved oxygen concentration is 6.8mg/L. The flounder fry introduced into the culture system is thrown at a density of 5-10kg/m 2 The throwing density of parent fish for breeding paralichthys olivaceus introduced into the culture system is 10-15kg/m 2 。
And S4, continuing to stop feeding food before evaluating the flounder to be evaluated the next day. And (4) in the morning of about 9. When the first round of evaluation is performed on the paralichthys olivaceus, the simulation catching is performed by sliding the fishing net nearby. If 1 point is recorded for the flounder which escapes rapidly in the process that the fishing net approaches slowly, 0 point is recorded for the flounder which escapes after the fishing net contacts the fish body, and-1 point is recorded for the flounder which does not react when the fishing net contacts the fish body during the simulated catching. Then further fishing net grabbing evaluation is carried out, the flounder to be evaluated is rapidly placed into the net one by using the fishing net, the water surface is fished out, the time from the fish to break away from the water surface in the grabbing of the fishing net to the beginning of struggling and the struggling times are respectively used as scoring indexes, the beginning of struggling time is arranged from short to long, the first third of the flounder is taken and recorded for 1, the middle third of the flounder is recorded for 0, and the last third of the flounder is recorded for-1; the struggle times are counted as 1 point for the first third of the flounder from high to low, 0 point for the middle third of the flounder, and-1 point for the last third of the flounder. And (3) putting the paralichthys olivaceus with the same score in the same system, and recovering the fish group from stress for 12-24 hours after the first round of evaluation.
S5, utilizing a full-automatic intermittent flow respiratory motion meter (AutoResp) TM Logo Systems) performed a second round of health assessment of the flounder to be assessed. Before the use, the equipment to be used is disinfected by mild bleaching liquid every two days, then is washed by fresh water for 3 times and soaked and washed cleanly, filtered natural seawater with the salinity of 25-35 per mill is injected, and the temperature is controlled by TMP-REG temperature monitoring and controlling equipment (logo Systems).
The preset environmental indexes are as follows: the water temperature in the monitoring system is 18 +/-0.1 ℃, the pH value is 7.5-8.5, the dissolved oxygen concentration is more than or equal to 6.8mg/L, the ammonia nitrogen is less than or equal to 0.1mg/L, the nitrite nitrogen is less than or equal to 0.1mg/L, and the sulfide is less than or equal to 0.1mg/L.
The size of the dissolved oxygen monitoring chamber is 48cm multiplied by 28cm multiplied by 4cm (length multiplied by width multiplied by height), the volume of the water body which can be accommodated is 6.3L, the water body flow in the monitoring system is controlled by LDAQ-4 (logo Systems), the dissolved oxygen is continuously monitored by using an electro-oxygen probe (logo Systems), the monitoring period is set to 10 minutes as a cycle, wherein 0-4 minutes is the process of slowly adding new water into the monitoring chamber by an external water storage tank so as to ensure that the dissolved oxygen of the water body in the monitoring chamber reaches 100 percent, 5 minutes is waiting time, 6-10 minutes is used for monitoring the dissolved oxygen change in the water body by using an AutoResp, and the automatic Resp is used for monitoring the dissolved oxygen change in the water body TM software (logo Systems) collects temperature and dissolved oxygen data and calculates corresponding respiration values. Before the experiment is started, the system is not used for placing fishes for at least 3 cycles to monitor the background oxygen consumption value caused by bacteria in the water body, and the background oxygen consumption value is controlled to be within 5 percent of the maximum metabolism rate value of the flounder to be detected.
Firstly, the measurement experiment of the maximum metabolism rate, the standard metabolism rate and the absolute aerobic range of the paralichthys olivaceus is carried out, and the index is used as the maximum aerobic respiration capacity index of the paralichthys olivaceus. Before 8 am, the flounder to be evaluated is put into a rectangular temporary rearing chamber (70 cm multiplied by 50cm multiplied by 30cm, length multiplied by width multiplied by height) reaching preset environmental indexes from a rearing pond, then the fishing net is used for continuously chasing for 3 minutes, and if the flounder to be evaluated decelerates or stops swimming in the process, the fishing net can be used for touching the fish tail part to stimulate the flounder to continuously carry out explosive swimming. After 3 minutes, all individual physical power is completely exhausted, then the flounder is immediately lifted out of the water surface for 1 minute by using the fishing net, then the fish is placed into the monitoring room in 30 seconds, all gas in the monitoring room is exhausted and locked, the surface of the monitoring room is covered by using a black baffle, the external disturbance is reduced to the minimum in the recovery stage, and the experiment is carried out for 24 hours. The maximum respiration value is the maximum metabolism rate, the maximum respiration value appears in the 1 st to 2 nd circulation, the standard metabolism rate is obtained when the respiration is stable, and the absolute aerobic range is obtained by subtracting the standard metabolism rate from the maximum metabolism rate. According to the arrangement of the maximum metabolism rate, the standard metabolism rate and the absolute aerobic range from high to low, the first third of the fishes are respectively counted for 1 minute, the middle third of the fishes are counted for 0 minute, and the last third of the fishes are counted for-1 minute.
And S6, after the 24-hour experiment in the S5 is finished, in the 6 th-10 th minute dissolved oxygen change monitoring period of the system, carrying out oxygen consumption change evaluation for 5 minutes under acute stimulation, wherein the evaluation index is used as a metabolic change index of the paralichthys olivaceus under the acute stress condition. The black baffle on the surface of the monitoring room is removed before the experiment begins, interference of predators on the paralichthys olivaceus is simulated by using shadows generated by hands under light, 30 seconds is a cycle, 0-15 seconds of testers continuously move hands above the monitoring room, the shadows caused by the testers simulate the predators under natural conditions, the interference is stopped at 16-30 seconds, then the process is continuously carried out for 5 minutes, and the change of dissolved oxygen in the water body is always monitored in the process. According to the change of oxygen consumption during the disturbance, the obvious increase of oxygen consumption is recorded as 1 point, the no change of oxygen consumption is recorded as 0 point, and the decrease of oxygen consumption is recorded as-1 point.
And then, putting the flounder to be evaluated into independent half-shading culture barrels one by one, marking, covering a half barrel opening of the culture barrels by using covers, placing in a half shading mode, ensuring that the water quality conditions in all the culture barrels are the same, and recovering the flounder from the second round of evaluation stress in the culture barrels for 12-24 hours.
And S7, after fasting, carrying out feeding reaction health evaluation on the flounder to be evaluated. The feeding is carried out at the uncovered side of the mouth of the breeding barrel, the illumination intensity in the barrel is about 40LX, so that the flounder can observe 'prey'. Preparing a fishing rod, replacing a fishhook at one end of a fishing line with a small wooden clamp, tying the other section of the fishing rod on the fishing rod, clamping a small fish to be fed on the wooden clamp, moving the fishing rod to drag the small fish to move on the water surface, and carefully observing the reaction of the adult fish to be detected at the bottom of the pool. If the lefteye flounder starting to eat within 30 seconds is scored for 1 point, if the lefteye flounder starting to eat within 2 minutes is scored for 0 point, and if the lefteye flounder starting to eat exceeds 2 minutes, the lefteye flounder which has no interest in food or no predation reaction is scored for-1 point. The method can be used in a culture barrel and a fish pond, and the illumination visibility of the bottom of the fish pond is adjusted by an artificial light source, so that the illumination intensity is 40LX.
S8, in order to avoid predators and trap predators in nature and realize strategic predation, the sand-occult behavior of the paralichthys olivaceus occurs, and the time for the paralichthys olivaceus to start the sand-occult behavior is recorded. The time for starting to submerge the sand is arranged from short to long, the first third of the paralichthys olivaceus is taken to be recorded with 1, the middle third of the paralichthys olivaceus is recorded with 0, and the last third of the paralichthys olivaceus is recorded with-1. And then, further evaluating the coverage area of the sand grains, wherein the coverage area of the sand grains is from more to less, 1 point is taken from the first third of the paralichthys olivaceus, 0 point is taken from the middle third of the paralichthys olivaceus, and-1 point is taken from the last third of the paralichthys olivaceus.
S9, carrying out five rounds of health evaluation on the flounder to be evaluated by using a circadian rhythm behavior detector (Bigbother 3, actimetrics) at night in order to avoid predation of predators in the daytime in nature, disinfecting equipment to be used the day before the beginning of use by using a mild bleaching solution, washing by using fresh water, and soaking to be clean, wherein each monitoring room is a light-colored culture tank with the size of 70cm multiplied by 50cm multiplied by 30cm (length multiplied by width multiplied by height). 8 in the morning: before 00, respectively injecting 52L of filtered natural seawater with sufficient aeration and salinity of 25-35 per mill into each monitoring chamber, then gently placing the flounder to be evaluated into the monitoring chambers one by one, wherein the photoperiod time is 12 hours, the illumination is 12 hours and the darkness is 12 hours, the camera tracks the swimming distance of each monitored flounder, the swimming distance of each flounder is reported once per minute, each fish is continuously monitored for 24 hours, then the fish is taken out and placed back into the corresponding culture system, the fish is recovered for 12-24 hours in stress, the total distance of obvious motion and night motion is used as a grading index according to the existence of the obvious motion in the daytime, the mark of no motion in the daytime is 1, the mark of 0 for the existence of the unobvious motion is 0, and the mark of-1 for the existence of the obvious motion is 1; the movement distance at night is arranged from high to low, the first third of the flounder is taken and recorded for 1, the middle third of the flounder is recorded for 0, and the last third of the flounder is recorded for-1.
From this point on, the health evaluation of the paralichthys olivaceus is finished, the active paralichthys olivaceus is obtained according to the score from high to low, the score is 9 or more, the score is from-9 to 9, the intermediate paralichthys olivaceus is obtained, and the score is from-9 and below, the inactive paralichthys olivaceus is obtained. According to the embodiment, the evaluation indexes of the health conditions of different flounders can screen out healthy fish species which accord with the actual culture condition from different individual flounders.
Example two
S1, preparing a culture system:
disinfecting the existing culture system with bleaching powder disinfectant with residual chlorine more than 30ppm, soaking for 1 day, washing with fresh water for 3 times, and injecting natural seawater with salinity of 25-35 ‰; 0.02g/L yeast extract and 0.2ml/L nitrobacteria are required to be added into the circulating water system, the circulating water system is operated for at least 15 days, and the biomembrane in the filter tank is cultured.
S2, monitoring the culture environment, and checking whether the culture environment meets preset environmental indexes:
the water temperature in the culture system is 18-20 ℃; the light time required was 12 hours (7-00-19 00), the dark time was 12 hours (19; the pH value is 7.5, the dissolved oxygen concentration is 6.8mg/L, the ammonia nitrogen concentration is 0.1mg/L, the nitrite nitrogen concentration is 0.1mg/L, and the sulfide concentration is 0.1mg/L.
S3, after being qualified by the detection of S2, the cynoglossus Gunther which needs to be introduced into the culture system is firstly put into seawater of povidone iodine with 0.006 per mill of sufficient oxygen for soaking for 1 hour, and then the cynoglossus Gunther is soaked and washed by aerated seawater and then put into the culture system;
wherein the seawater salinity of 0.006 per mill povidone iodine is 25 per mill-35 per mill, and the dissolved oxygen concentration is 6.8mg/L. The cynoglossus larva introduced into the breeding system is thrown in with the density of 5-10kg/m 2 The feeding density of the parent fish bred by the cynoglossus gunther introduced into the breeding system is 10-15kg/m 2 。
And S4, continuing to stop feeding food before evaluating the cynoglossus sollaevis to be evaluated on the next day. And 9, in the morning, about 00, and performing preliminary evaluation after the water flow of the water pool is stable and the cynoglossus gunther shoals are quiet. When carrying out the first round of evaluation on the cynoglossus larva or adult fish, firstly, a fishing net slides nearby to carry out simulated capture. If the score of the cynoglossus sole running rapidly is recorded as 1 score in the process that the fishing net slowly approaches, the score of the cynoglossus sole running after the fishing net contacts the fish body is recorded as 0 score, and the score of the cynoglossus sole which still has no response is recorded as-1 score when the fishing net contacts the fish body during the simulated capture. Then, further fishing net grabbing evaluation is carried out, the fish nets are used for rapidly enabling the cynoglossus soloniensis to be evaluated to enter the net one by one, the water surface is fished out, the time from the fish grabbing to the fish net to break away from the water surface to the beginning of struggling and the struggling times are respectively used as scoring indexes, the beginning struggling time is arranged from short to long, 1 point is recorded for the first third cynoglossus soloniensis, 0 point is recorded for the middle third cynoglossus soloniensis, and-1 point is recorded for the last third cynoglossus soloniensis; the struggle times are that the first third of the cynoglossus Gunther is counted as 1 point, the middle third of the cynoglossus Gunther is counted as 0 point, and the last third of the cynoglossus Gunther is counted as-1 point according to the arrangement from high to low. The cynoglossus gunther with the same score is put in the same system, and after the first evaluation, the fish group is recovered from the stress for 12-24 hours.
S5, utilizing a full-automatic intermittent flow respiratory motion meter (AutoResp) TM Logo Systems) performed a second round of health assessment of cynoglossus to be assessed. Before the use, the equipment to be used is disinfected by mild bleaching liquid every two days, then is washed by fresh water for 3 times and soaked and washed cleanly, filtered natural seawater with the salinity of 25-35 per mill is injected, and the temperature is controlled by TMP-REG temperature monitoring and controlling equipment (logo Systems).
The preset environmental indexes are as follows: the water temperature in the monitoring system is 18 +/-0.1 ℃, the pH value is 7.5-8.5, the dissolved oxygen concentration is more than or equal to 6.8mg/L, the ammonia nitrogen is less than or equal to 0.1mg/L, the nitrite nitrogen is less than or equal to 0.1mg/L, and the sulfide is less than or equal to 0.1mg/L.
The size of the dissolved oxygen monitoring chamber is 48cm multiplied by 28cm multiplied by 4cm (length multiplied by width multiplied by height), the volume of the water body can be accommodated is 6.3L, the water body flow in the monitoring system is controlled by LDAQ-4 (logo Systems), the dissolved oxygen is continuously monitored by using an electro-oxygen probe (logo Systems), and the monitoring period is set to be 10 minutesCirculating, wherein the process that the external water storage tank slowly adds new water into the monitoring room in 0-4 minutes to ensure that the dissolved oxygen of the water body of the monitoring room reaches 100 percent, the waiting time in 5 minutes and the dissolved oxygen change in the water body monitored in 6-10 minutes by the monitoring room, and using AutoResp TM software (logo Systems) collects temperature and dissolved oxygen data and calculates corresponding respiration values. Before the experiment begins, at least 3 cycles are carried out without fish placement in the system to monitor the background oxygen consumption value caused by bacteria in the water body, and the background oxygen consumption value is controlled to be within 5% of the maximum metabolism rate value of the cynoglossus solevis to be detected.
Firstly, carrying out a measurement experiment of the maximum metabolism rate, the standard metabolism rate and the absolute aerobic range of the cynoglossus sole, wherein the index is used as the index of the maximum oxygen breathing capacity of the cynoglossus sole. Before 8 am, the cynoglossus sollaevis to be evaluated is put into a rectangular temporary rearing chamber (70 cm multiplied by 50cm multiplied by 30cm, length multiplied by width multiplied by height) reaching preset environmental indexes from a rearing pond, then the trapping is continuously carried out for 3 minutes by using a fishing net, and if the cynoglossus sollaevis to be evaluated decelerates or stops swimming in the process, the fish net can touch the fish tail part to stimulate the cynoglossus sollaevis to continuously carry out explosive swimming. After 3 minutes, all individual physical power is completely exhausted, then the cynoglossus is lifted out of the water surface for 1 minute by using a fishing net immediately, then the cynoglossus is placed into a monitoring room in 30 seconds, all gas in the monitoring room is exhausted and locked, the surface of the monitoring room is covered by using a black baffle, the external disturbance is reduced to the minimum in the recovery stage, and the experiment is carried out for 24 hours. The maximum value of respiration is the maximum metabolism rate, generally appears in the 1 st to 2 nd cycles, the standard metabolism rate is when the respiration is stable, and the absolute aerobic range is obtained by subtracting the standard metabolism rate of the Newcastle from the maximum metabolism rate. According to the arrangement of the maximum metabolism rate, the standard metabolism rate and the absolute aerobic range from high to low, the first third of the fishes are respectively counted for 1 minute, the middle third of the fishes are counted for 0 minute, and the last third of the fishes are counted for-1 minute.
And S6, after the 24-hour experiment in S5 is finished, in the 6 th-10 th minute dissolved oxygen change monitoring period of the system, oxygen consumption change evaluation under acute stimulation for 5 minutes is carried out, and the evaluation index is used as a metabolism change index of the cynoglossus gunther under the acute stress condition. The black baffle on the surface of the monitoring chamber is removed before the experiment is started, a predator is simulated to interfere with the cynoglossus gunther by using the shadow generated by hands under the light, 30 seconds is a cycle, 0-15 second testers continuously move hands above the monitoring chamber, the shadow caused by the test personnel simulates the predator under the natural condition, the interference is stopped in 16-30 seconds, then the process is continuously carried out for 5 minutes, and the change of the dissolved oxygen in the water body is always monitored in the process. According to the change of oxygen consumption during interference, the obvious rise of oxygen consumption is marked as 1 minute, the unchanged oxygen consumption is marked as 0 minute, and the decline of oxygen consumption is marked as-1 minute;
and then putting the cynoglossus to be evaluated into independent half-shading culture barrels one by one, marking, covering a half barrel opening of the culture barrels with a cover, placing in a half-shading mode, connecting each culture barrel with a water supply system of a culture pond, ensuring that the water quality conditions in all the culture barrels are the same, and recovering the cynoglossus from the second round of evaluation stress in the culture barrels for 12-24 hours.
And S7, after fasting, carrying out feeding reaction health evaluation on the cynoglossus to be evaluated. The tongue sole is fed on the uncovered side of the mouth of the breeding barrel, the illumination intensity in the barrel is about 40LX, and the tongue sole can observe 'preys'. Preparing a fishing rod, replacing a fishhook at one end of a fishing line with a small wooden clamp, tying the other section of the fishing rod on the fishing rod, clamping a small fish to be fed on the wooden clamp, moving the fishing rod to drag the small fish to swim on the water surface, and carefully observing the reaction of the adult fish to be detected at the bottom of the pool. If the cynoglossus Gunther which starts to swallow the game within 30 seconds is scored as 1 point, if the cynoglossus Gunther which starts to swallow the game within 2 minutes is scored as 0 point, if the cynoglossus Gunther which does not interest food or does not have predation reaction exceeds 2 minutes is scored as-1 point. The method can be used in a culture barrel and a fish pond, and the illumination visibility of the bottom of the fish pond is adjusted by an artificial light source, so that the illumination intensity is 40LX.
S8, in order to avoid predators and trap food strategically in the nature, the cynoglossus Gunther can have sand-diving behavior, and the time when the cynoglossus Gunther starts the sand-diving behavior is recorded. The time for starting sand diving is arranged from short to long, the first third of the cynoglossus Gunther is taken to count 1, the middle third of the cynoglossus Gunther is taken to count 0, and the last third of the cynoglossus Gunther is taken to count 1. And then, further evaluating the coverage area of the sand grains, wherein the coverage area of the sand grains is from more to less, 1 score is taken for the first third of the cynoglossus sole, 0 score is taken for the middle third of the cynoglossus sole, and-1 score is taken for the last third of the cynoglossus sole.
S9, performing activities and predation at night by the cynoglossus sollaevis in the daytime in nature in order to avoid predation of predators, performing five-round health evaluation on the cynoglossus sollaevis to be evaluated by using a circadian rhythm behavior detector (bigbroth 3, actimetrics), disinfecting equipment to be used in the day before use by using mild bleaching solution, washing by fresh water and soaking to be clean, wherein each monitoring chamber is a light-colored culture tank with the size of 70cm multiplied by 50cm multiplied by 30cm (length multiplied by width multiplied by height). In the morning, 8: before 00, respectively injecting 52L of fully aerated filtered natural seawater with salinity of 25-35 per mill into each monitoring chamber, then gently placing the cynoglossus gunther to be evaluated into the monitoring chambers one by one, with the photoperiod time of 12 hours and 12 hours of illumination and darkness, tracking the swimming distance of each monitored cynoglossus gunther by a camera, reporting the swimming distance of each cynoglossus gunther once per minute, continuously monitoring each cynoglossus gunther for 24 hours, then fishing out and placing back into a corresponding culture system, recovering the cynoglossus gunther in stress for 12-24 hours, taking the total distance of the obvious motion and the night motion as a scoring index according to whether the day has the obvious motion or not, recording 1 score for the non-motion in the day, recording 0 score for the existence of the unobvious motion, and recording-1 score for the existence of the obvious motion; the movement distance at night is arranged from high to low, the first third of the cynoglossus Gunther takes 1 mark, the middle third of the cynoglossus Gunther takes 0 mark, and the last third of the cynoglossus Gunther takes-1 mark.
From this point on, the evaluation of the health of the cynoglossus sole is finished, and according to the score from high to low, the cynoglossus sole is an active cynoglossus sole with the score of 9 or more, the intermediate cynoglossus sole is a score from-9 to-9, and the inactive cynoglossus sole is a score from-9 and below. According to the embodiment, the evaluation indexes of the health conditions of different flounders can screen out healthy fish species which accord with the actual culture condition from different individual flounders.
It should be noted that, although the present invention has been described in detail with reference to the foregoing embodiments, the technical solutions described in the foregoing embodiments or some technical features of the present invention may be modified or replaced equivalently by those skilled in the art, and any modifications, equivalents, improvements and the like made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (3)
1. A flounder health condition evaluation method is characterized by comprising the following steps:
s1, after a culture system is disinfected by bleaching powder, washing and soaking the culture system by fresh water, injecting filtered natural seawater or diluted concentrated seawater with the salinity of 25-35 per mill, operating for a plurality of days after yeast extract powder and photosynthetic bacteria are put into a circulating water system, and culturing a biological membrane in a filter tank;
s2, monitoring the culture environment to enable the culture environment to reach preset environmental indexes, wherein the preset environmental indexes are as follows: the water temperature in the culture system is 17-20 ℃, the illumination time and the dark time are respectively 12 hours, the pH value is 7.5-8.5, the dissolved oxygen concentration is more than or equal to 6.8mg/L, the ammonia nitrogen concentration is less than or equal to 0.1mg/L, the nitrite nitrogen concentration is less than or equal to 0.1mg/L, and the sulfide concentration is less than or equal to 0.1mg/L;
s3, after the environmental indexes are detected to be qualified, the flatfish is firstly put into a disinfection pond containing seawater containing povidone iodine for soaking, then is soaked and washed by aerated seawater, and is put into the culture system;
s4, performing first round evaluation on the flounder in the step S3 by using a fishing net catching method and an air exposure method: continuing to stop feeding food before evaluation, starting preliminary evaluation after water flow in the pool is stable and fish school is quiet 9 morning, and when performing first round of evaluation on flounder, firstly using a fishing net to slide nearby for simulated capture, marking the behavior of the fish escaping rapidly in the process that the fishing net approaches slowly as 1, marking the behavior of the fish escaping after the fishing net contacts a fish body as 0, and marking the behavior of the fish which still has no response when the fishing net contacts the fish body as-1 during simulated capture; then carrying out fishing net grabbing evaluation, rapidly fishing flounder into the net one by using a fishing net, fishing out the water surface for 70 seconds, respectively recording the time from the fish grabbing into the fishing net to the beginning of struggling from the water surface and the struggling times within 70 seconds, arranging the struggling times from short to long, taking 1 score of the first third flounder, 0 score of the middle third flounder and-1 score of the last third flounder; 1 point of the first third of flounder is taken according to the struggle times from high to low, 0 point of the middle third of flounder is taken, and-1 point of the last third of flounder is taken, two groups of flounder with the same data are placed in the same system, and fish schools after the first round of evaluation are recovered for 12-24 hours from stress;
s5, carrying out a second round of health evaluation on flatfish by using a full-automatic intermittent flow respiratory motion meter: before the full-automatic intermittent flow respiratory motion measuring instrument is used, disinfecting the instrument by using mild bleaching liquid every two days, washing the instrument for 3 times by using fresh water, soaking and washing the instrument clean, injecting fully aerated filtered natural seawater with the salinity of 25-35 per mill, and controlling the temperature by using TMP-REG temperature monitoring and controlling equipment;
monitoring a measurement environment to enable the measurement environment to reach a preset environment index, wherein the preset environment index is as follows: the water temperature in the monitoring system is 18 +/-0.1 ℃, the pH value is 7.5-8.5, the dissolved oxygen concentration is more than or equal to 6.8mg/L, the ammonia nitrogen is less than or equal to 0.1mg/L, the nitrite nitrogen is less than or equal to 0.1mg/L, and the sulfide is less than or equal to 0.1mg/L;
the size of the dissolved oxygen monitoring chamber is 48cm multiplied by 28cm multiplied by 4cm, the volume of the water body capable of being accommodated is 6.3L, the water body flow in the monitoring system is controlled by LDAQ-4, the dissolved oxygen is continuously monitored by using an electro-oxygen probe, the monitoring period is set to be 10 minutes as a cycle, wherein the 0 th to 4 th minutes are the process that an external water storage tank slowly adds new water into the monitoring chamber, so as to ensure that the dissolved oxygen of the water body in the monitoring chamber reaches 100 percent, the 5 th minute is the time for waiting for the data of a breathing chamber to be stable, the 6 th to 10 th minutes are the time for monitoring the change of the dissolved oxygen in the water body by using an AutoResp TM The software collects temperature and dissolved oxygen data, calculates corresponding respiration values, and performs at least 3 cycles without putting fishes in the system before starting to monitor the background oxygen consumption value caused by bacteria in the water body and controls the background oxygen consumption value to be within 5 percent of the maximum metabolic rate value of the adult fishes to be detected;
measuring the maximum metabolism rate, the standard metabolism rate and the absolute aerobic range of the flounder as the maximum aerobic respiration capacity index of the flounder: before 8 o' clock in the morning, flatfish is put into a rectangular temporary rearing chamber with preset environmental indexes from a rearing pond, fishing net is used for catching for 3 minutes continuously, and the fish tail part is touched by the fishing net to stimulate the fish tail part to continue explosive swimming when the fish to be detected decelerates or stops swimming; after 3 minutes, completely exhausting all individual physical strength, immediately flatly supporting the fish out of the water surface for 1 minute by using a fishing net, putting the fish into a monitoring chamber within 30 seconds, discharging and locking all gas in the monitoring chamber, covering the surface of the monitoring chamber by using a black baffle, minimizing external disturbance in a recovery stage, and carrying out 24 hours, wherein the maximum value of respiration is the maximum metabolic rate, the standard metabolic rate is obtained when the respiration in 1-2 cycles is stable, the absolute aerobic range is obtained by subtracting the standard newcastle metabolic rate from the maximum metabolic rate, and the maximum metabolic rate, the standard metabolic rate and the absolute aerobic range of each fish are recorded as respiratory metabolic indexes of the fish to be evaluated; respectively taking the first third of fishes to mark 1 minute, the middle third of fishes to mark 0 minute and the last third of fishes to mark-1 minute according to the arrangement of the maximum metabolism rate, the standard metabolism rate and the absolute aerobic range from high to low;
s6, in the third stage of the step S5, respiration monitoring under acute stimulation is carried out, and oxygen consumption change of the respiration monitoring is recorded and is used as an index for evaluating a respiration selection strategy of flounder under an acute stress condition: after 24 hours in the step S5, in a dissolved oxygen change monitoring period of 6-10 minutes of the system, carrying out oxygen consumption change evaluation for 5 minutes under acute stimulation to serve as a metabolism change index of flounder under the acute stress condition; removing a black baffle on the surface of a monitoring room before starting, simulating predators to interfere with the fish by using shadows generated by hands under light, wherein a cycle is set to 30 seconds, a tester continuously moves the hands above the monitoring room in 0-15 seconds, the resulting shadows simulate the predators under natural conditions, the interference is stopped in 16-30 seconds, and then the process is continued for 5 minutes, the change of dissolved oxygen in a water body is always monitored, the oxygen consumption value of flounder under interference is recorded, according to the change of oxygen consumption during interference, the obvious increase of oxygen consumption is recorded as 1 point, the unchanged oxygen consumption is recorded as 0 point, and the decrease of oxygen consumption is recorded as-1 point;
putting flatfish into independent culture barrels one by one, covering half barrel openings of the culture barrels by covers, placing the flatfish in a semi-dark place, connecting each culture barrel with a water supply system of a culture pond to ensure that the water quality conditions are the same, and recovering the fish from the second round of evaluation stress in the culture barrels for 12-24 hours;
s7, carrying out stress solitary appetite recovery evaluation on flounder individuals one by bait fishing: after fasting, carrying out feeding reaction health evaluation on flatfish, feeding the flatfish on the uncovered side of a culture barrel or a fish pond with the illumination intensity of 40LX, preparing a fishing rod, replacing a fishhook at one end of a fishing line by a small wooden clamp, tying the other section of the fishing rod on the fishing rod, clamping a small fish to be fed on the wooden clamp, moving the fishing rod to drag the small fish to swim on the water surface, observing the reaction of the adult fish to be detected at the bottom of the pond, recording the adult fish which initiates feeding on a prey within 1,2 minutes as 0 when the adult fish which initiates feeding on the prey within 30 seconds, and recording the fish which has no interest in food or no feeding reaction within more than 2 minutes as-1;
s8, evaluating flounder by using sand burying behaviors: recording the time of beginning sand-burying behaviors of flounder, arranging the time of beginning sand-burying from short to long, taking 1 point of the first third of flounder, 0 point of the middle third of flounder and-1 point of the last third of flounder, then evaluating the sand coverage area, taking 1 point of the first third of flounder, 0 point of the middle third of flounder and-1 point of the last third of flounder;
s9, evaluating the circadian rhythm behavior of flounder by using a circadian rhythm behavior detector: disinfecting the equipment by using mild bleaching solution one day before the use, washing the equipment by using fresh water and soaking the equipment clean, wherein a light-color culture tank with the size of 70cm multiplied by 50cm multiplied by 30cm is used as a monitoring room; before 8 o' clock in the morning, respectively injecting 52L of fully aerated filtered natural seawater with salinity of 25-35 per thousand into each monitoring chamber, flatfish is gently placed into the monitoring chambers one by one, the photoperiod time is 12 hours, the illumination time is 12 hours, the camera tracks the swimming distance of each monitored flatfish, the swimming distance of each fish is reported once per minute, each fish is continuously monitored for 24 hours, and then the fish is fished out and placed back into the corresponding culture system to recover the fish for 12-24 hours in stress; taking whether obvious movement exists in the daytime and the total distance of the movement at night as scoring indexes, recording 1 score for no movement in the daytime, recording 0 score for no obvious movement and recording-1 score for obvious movement; the movement distance at night is arranged from high to low, the first third of flounder is counted for 1, the middle third of flounder is counted for 0, and the last third of flounder is counted for 1;
s10, according to the results of the capturing-reverse capturing behaviors, the metabolism indexes, the metabolism change after acute stress, the appetite recovery behaviors after stress orphism, the sand diving behaviors and the circadian rhythm behaviors in the steps S1 to S9, obtaining correlation distribution diagrams of different individuals and behavior test indexes by analyzing main components in XLSSTAT 2014, and obtaining a set of three different health conditions: the characteristic value is more than 2, the characteristic value is active fish, the characteristic value is-2 to 2, the characteristic value is intermediate fish, and the characteristic value is less than-2, the characteristic value is inactive fish; then the metabolism results are integrated to obtain the health conditions of different individuals, the fish with the integrated score from high to low is active fish with the integrated score of more than 9, the fish with the integrated score of-9 to 9 is intermediate fish, and the fish with the integrated score of less than-9 is inactive fish.
2. The method for evaluating the health status of flounder fish according to claim 1, wherein: in the step S1, the disinfectant is bleaching powder, the available residual chlorine in the disinfectant is more than 30ppm, the disinfectant is washed for 3 times by fresh water and soaked for 1 day, 0.02g/L yeast extract powder and 0.2ml/L photosynthetic bacteria are put into a circulating water system, the newly used circulating water system operates for not less than 30 days, and the system which is used continuously operates for one week.
3. The method for evaluating the health status of flounder fish according to claim 1, wherein: in step S3, the disinfection pond is filtered or diluted seawater containing 0.006 thousandth of povidone iodine, the salinity is 25 thousandth to 35 thousandth, the dissolved oxygen concentration is more than or equal to 6.8mg/L, and the throwing density of flounder seedlings introduced into the culture system is 5kg/m to 10kg/m 2 The feeding density of the flounder adult fish is 10-15kg/m 2 And aerating the sterilized water sufficiently by using an air pump, and soaking for 1h.
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