CN112685684A - Zebra fish aerobic exercise capacity evaluation method - Google Patents
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Abstract
The invention discloses a zebra fish aerobic exercise capacity evaluation method, and belongs to the technical field of exercise capacity evaluation. It comprises the following steps: (1) measuring parameters of zebrafish followed by fasting; (2) transferring the zebra fish after fasting treatment to a lane of a respirator and carrying out adaptive training; (3) set up automatic three-stage mode of the lane of the respirometer: a flushing mode, a waiting mode and a measuring mode; (4) then increasing the flow rate of water until the zebra fish reaches the exhaustion state, recording real-time oxygen consumption data and the maximum swimming speed at the termination time, and obtaining the maximum oxygen consumption and the critical swimming speed of the zebra fish; (5) gradient movement intensity nodes are set according to the critical swimming speed of the zebra fish, blood lactic acid is detected by measuring blood slightly after each movement intensity node moves, and the blood lactic acid threshold of the zebra fish is obtained. The evaluation of the aerobic exercise capacity of the zebra fish is realized by measuring indexes such as the maximum oxygen consumption, the critical swimming speed, the blood lactic acid threshold and the like of the zebra fish.
Description
Technical Field
The invention belongs to the technical field of athletic ability evaluation, and particularly relates to an assessment method for zebra fish aerobic athletic ability.
Background
The aerobic exercise capacity is one of the aspects for evaluating the exercise capacity of the zebra fish, and the determination of the evaluation scheme is particularly important. At present, no article or patent about a method for evaluating the aerobic exercise capacity of zebra fish exists at home and abroad, and the zebra fish as a classical model organism is widely applied to a plurality of scientific research fields including the fields of sports science and sports medicine. Research on relevant aspects of zebra fish movement belongs to the frontier field at home, so that the establishment of a zebra fish aerobic movement capability assessment method system is of great importance. Through repeated experiments, a set of relatively perfect assessment method suitable for domestic zebra fish aerobic exercise capacity is found, and a reference can be provided for other scientific researchers to measure the zebra fish aerobic exercise capacity.
Disclosure of Invention
1. Problems to be solved
Aiming at the problems in the prior art, the invention provides a zebra fish aerobic exercise capacity assessment method, and particularly, the zebra fish aerobic exercise capacity assessment method is used for measuring the maximum oxygen consumption (MO) of zebra fish2max), critical swimming speed (U)crit) And blood lactic acid threshold and other indexes, and realizes the evaluation of the aerobic exercise capacity of the zebra fish.
2. Technical scheme
A zebra fish aerobic exercise capacity evaluation method comprises the following steps:
(1) preparing zebra fish, marking the zebra fish by numbers, measuring parameters of the zebra fish, and then performing fasting treatment;
(2) transferring the zebra fish after fasting treatment to a lane of a respirator, and carrying out adaptive training;
(3) set up automatic three-stage mode of the lane of the respirometer: a flushing mode, a waiting mode and a measuring mode;
(4) then gradually increasing the flow rate of water in a lane of the respirator until the zebra fish reaches an exhausted state, recording real-time oxygen consumption data and the maximum swimming speed at the termination time, and obtaining the maximum oxygen consumption and the critical swimming speed of the zebra fish;
(5) gradient movement intensity nodes are set according to the critical swimming speed of the zebra fish, blood lactic acid is detected by measuring blood slightly after each movement intensity node moves, and the blood lactic acid threshold of the zebra fish is obtained.
In the method for evaluating the aerobic exercise capacity of the zebra fish, the zebra fish in the step (1) is placed in an independent fish tank;
in the step (1), the parameters of the zebra fish comprise body length and body weight;
the time of fasting treatment in the step (1) is 24 h.
In the method for evaluating the aerobic exercise capacity of the zebra fish, the adaptive training in the step (2) is to adapt the zebra fish for 2 hours at the speed of 0.8 BL/s.
In the method for evaluating the aerobic exercise capacity of the zebra fish, the time of the flushing mode in the step (3) is 90 s;
the time for waiting for the mode in the step (3) is 30 s;
the time of the measurement mode in the step (3) is 5 min.
In the method for evaluating the aerobic exercise capacity of the zebra fish, the flow rate increase mode of water in the lane of the respirator in the step (4) is increased by 2.7BL/s every 14 min;
the standard for judging the exhaustion state of the zebra fish in the step (4) is that the zebra fish stops at the tail of the lane for more than 20 s;
the calculation formula of the critical swimming speed in the step (4) is
Ucrit＝U+(t/Δt)*ΔU；
Wherein U is the maximum swimming speed (cm/s) reached by the experimental fish in the test; wherein Δ U is the speed increment (2.7 BL/s); wherein Δ t is the time interval (14 min); where t is the time (min) of adherence at maximum speed.
In the method for evaluating the aerobic exercise capacity of the zebra fish, the treatment method before measuring the body length and the body weight in the step (1) is as follows, the zebra fish is placed into anesthetic for anesthesia, the zebra fish is taken out and placed on a wet towel, the body length is measured by a vernier caliper, the measurement part is from the head of the zebra fish to the tail, the zebra fish is wiped dry, then the zebra fish is placed into an electronic balance for weighing the body weight, and the numerical value is recorded.
In the method for evaluating the aerobic exercise capacity of the zebra fish, the mass concentration of the anesthetic is 40mg/L, and the method comprises the steps of adding 0.04g of tricaine into 1000ml of water and uniformly mixing the three.
In the method for evaluating the aerobic exercise capacity of the zebra fish, the gradient exercise intensity nodes in the step (5) are 0%, 20%, 40%, 60%, 80% and 100% of the critical swimming speed;
and (5) moving each movement intensity node for 5 min.
In the method for evaluating the aerobic exercise capacity of the zebra fish, the method for taking the blood in a trace amount in the step (5) is to take the blood from the dorsal artery or the posterior main vein of the zebra fish by using a trace blood taking needle.
In the method for evaluating the aerobic exercise capacity of the zebra fish, the blood lactic acid threshold of the zebra fish in the step (5) is obtained by drawing an increasing curve of the blood lactic acid changing along with exercise intensity.
3. Advantageous effects
Compared with the prior art, the invention has the beneficial effects that:
in conclusion, the maximum oxygen consumption, the critical swimming speed and the blood lactic acid threshold are used as the detection indexes of the aerobic exercise capacity of the zebra fish for the first time, a uniform measurement standard is provided for measuring the aerobic exercise capacity of the zebra fish, and the detection method is scientific and reasonable. The model organism zebra fish selected by the invention has the advantage of verification of aerobic movement of living animals, and is beneficial to improving the experimental efficiency and reducing the experimental cost.
Drawings
FIG. 1 is a blood vessel anatomical diagram of a zebra fish; the left side of the figure is a full-view diagram of the zebra fish, and the right side of the figure is a cross section of the zebra fish;
FIG. 2 is a graph of the hemospastic behavior of zebra fish;
FIG. 3 is a diagram showing the blood collection of zebra fish and the detection of the zebra fish by using a trace blood lactic acid detector of EKF company of Germany;
FIG. 4 is a graph showing the oxygen consumption MO of zebra fish measured in example 12A drawing;
FIG. 5 is a graph of the measured zebra fish lactic acid threshold in example 1;
FIG. 6 is a graph showing the relationship between blood lactate concentration and relative exercise intensity in human in example 1; wherein FIG. 6 is derived from the following literature (Wang Yuan, Suzhou Job. sports physiology [ M ]. people sports Press: Beijing, 2012: 321);
FIG. 7 shows the oxygen consumption MO of zebra fish measured by the improved scheme 12A drawing;
FIG. 8 is a graph of oxygen consumption rate of fish and swimming speed in modification 1; wherein FIG. 8 is derived from the following documents (herquan super, royal lead, Liuhuijie, etc.. swimming characteristics study of juvenile chubs under different sport modes [ J ]. freshwater fishery, 2018,48(2): 3-9.);
FIG. 9 shows the oxygen consumption MO of zebra fish of 3 months old measured in the modified scheme 22A drawing;
FIG. 10 shows the oxygen consumption MO of 23-month-old zebra fish measured in the modified scheme 22A drawing;
FIG. 11 shows the oxygen consumption MO of zebra fish of 3 and 23 months old measured in modified scheme 22A drawing;
FIG. 12 is a graph showing the relationship between the maximum oxygen consumption of zebra fish at 3 months and 23 months in the modified embodiment 2;
FIG. 13 is a graph showing the relative critical swimming speed of zebra fish of 3 months old and 23 months old according to modification 2.
Detailed Description
The invention is further described with reference to specific examples.
Experimental animals: adult wild zebra fish is adopted and purchased from national zebra fish resource center. The male and female zebra fish are separately raised under the standard conditions of 12h in darkness under the illumination of 12h and at 28 ℃, and the brine shrimp is fed regularly and quantitatively. When selecting experimental fish, taking normal sexually mature male zebra fish. Note that: BL is the body length of zebra fish. Note that: tricaine has Cas registry number 886-86-2, with the literal name 3-ethoxyanilide mesylate.
Before the actual implementation of example 1, the following tests were carried out:
(in the invention, the exploration of the aerobic exercise capacity of the zebra fish is mainly improved by means of a scheme for detecting the fish critical swimming speed reported in foreign documents, and a scheme suitable for evaluating the aerobic exercise capacity of the zebra fish in China is searched.)
Test protocol 1: the scheme measures the critical swimming speed and the maximum oxygen consumption, the original value of the maximum oxygen consumption should be increased, and the result shows that the maximum oxygen consumption is increased firstly and then reduced, so the scheme is not suitable. The method comprises the following specific steps: (1) after the body length and the body weight of the zebra fish are tested, the zebra fish is put in a lane at the speed of 5cm/s to adapt for 30min so as to eliminate the feeling of hypochondriac stress; (2) the swimming tunnel is automatically set to complete three stages within 10 min: the Flash time is 90s, the wait time is 30s and the measure time is 8 min; (3) the test is started by increasing 5cm/s every 10min, the water flow speed is continuously increased, the speed increment (delta U) is always 5cm/s, the time interval (delta t) is 10min each time until the experimental fish reaches the exhaustion state, and the judgment standard is that the experimental fish stops at the tail part of the lane for more than 20 s.
Test protocol 2: continuing searching for documents, finding a second scheme, firstly measuring the critical swimming speed, dividing the critical swimming speed into 25%, 50%, 75% and 100% movement speeds, measuring the oxygen consumption of 20min movement, and increasing the maximum oxygen consumption value of the zebra fish in the original scheme, but finding that the value is reduced through experimental measurement. The method comprises the following specific steps: (1) male wild zebrafish 3 months old were picked, anesthetized with tricaine (40mg/L) and measured for standard length (BL, cm) and body weight (BW, g) before introduction into the swimming tunnel; (2) placing a single zebra fish into the zebra fish swimming respiration measuring instrument, wherein the initial speed of the zebra fish is 5cms-1Increasing with 0.05ms every 10min-1Until the zebrafish are fatigued, which is determined as the time at which the fish can no longer maintain convection, and are swept towards the screen downstream of the tunnel and can not move by themselves within three seconds; visual observation ensured that no erratic swimming behavior or premature fatigue occurred during the swimming test; calculate U according to the following formulacrit：
Ucrit＝Ui+[Uii(Ti/Tii)]
Ui is the highest velocity maintained for the entire interval, Uii is the velocity increment (here 0.05 ms)-1) Ti is the time required from the beginning of the fatigue speed increment, Tii is the interval time (10 min is set here); absolute value (Ms)-1) Converted to standard body length per second (BLS)-1) Relative swimming speed in units; (3) slave UcritThe fish were treated for 0.05ms in the experiment-1Swim in circulating water for 2 h; the next morning a breath measurement is taken; at 5% UcritFor 20min at the minimum flow rate to measure the RMR, the conventional metabolic rate, i.e. the subsequent random measurements of 25%, 50%, 75% and 100% UcritOxygen consumption for 20min of exercise; after 20min at a constant speed, the respirator was reconnected to the continuous flow of water, which was filled with air, and the zebrafish was allowed to recover for 30 min; VO was then repeatedly measured at different swimming speeds2。
Test protocol 3: and measuring a third scheme through the newly searched documents, and finding that the numerical value does not reach an ideal value. Before measurement, experimental fish is subjected to fasting treatment for 24 hours; putting the fish into a swimming lane to adapt for 1-1.5h at the speed of 0.8BL/s so as to eliminate the stress influence of the transfer process and the new environment on the experimental fish; the test starts with increasing the water flow rate by 2.7BL/s every 20min, the speed increase (Δ U) is always 2.7BL/s, and the time interval (Δ t) for each time is 20 min. Until the experimental fish reaches the exhaustion state, the judgment standard is that the experimental fish stops at the tail of the lane for more than 20 s; wherein the Flash time is 4min, the wait time is 1min, and the measure time is 5 min; then, the fish-free empty measurement is carried out to detect the oxygen consumption (unit: mg O) of the microorganisms in the water2 kg-1h-1)；
The formula is as follows: SMR Sa-Sb
Wherein Sa is the oxygen consumption measured by zebrafish; sb is the microbial oxygen consumption of the zebra-free fish air test;
test protocol 4: on the basis of the test scheme 3, scheme improvement is carried out, the scheme carries out two times of measurement on the same batch of fish and carries out empty measurement, and the result shows that the maximum oxygen consumption value of the empty measurement is not greatly different from the first numerical value and the second numerical value, and the second time is reduced in exercise capacity compared with the first time. The method comprises the following specific steps: (1) the fish were fasted for 24h before testing, and placed in lane 0.8BL/s for 2h to eliminate the stress; (2) the swimming tunnel is automatically set to complete three stages within 10 min: the Flash time is 4min, the wait time is 1min, and the measure time is 5 min; (3) the test is started by increasing 2.7BL/s every 20min, the water flow speed is continuously increased, the speed increment (delta U) is always 2.7BL/s, each time interval (delta t) is 20min until the experimental fish reaches the exhaustion state, and the judgment standard is that the experimental fish stops at the tail part of a lane for more than 20 s; (4) recovering at 0.8BL/s, and performing second measurement after 60min rest; (5) and carrying out empty measurement after the second measurement. (data is required to be available, several times measurable).
Example 1
Through the results of the test scheme 4, although the overall results are deviated, the oxygen consumption of some experimental fishes is increased along with the increase of the exercise intensity, so that the improvement is carried out on the basis of the oxygen consumption, and the results of the five tests of the experimental fishes are expected, and finally the scheme is determined.
The zebra fish aerobic exercise capacity evaluation method comprises the following steps:
(1) preparing zebra fish and carrying out numbering marking (such as a fish tank with a number), measuring parameters of the zebra fish, and then carrying out fasting treatment;
(2) transferring the zebra fish after fasting treatment to a lane of a respirator, and carrying out adaptive training; wherein the respirator utilizes the respiratory System of the Danish logo System zebra fish lane;
(3) set up automatic three-stage mode of the lane of the respirometer: a flushing mode, a waiting mode and a measuring mode;
(4) then gradually increasing the flow rate of water in a lane of the respirator until the zebra fish reaches an exhausted state, recording real-time oxygen consumption data and the maximum swimming speed at the termination time, and obtaining the maximum oxygen consumption and the critical swimming speed of the zebra fish;
(5) as shown in fig. 1, 2 and 3, gradient exercise intensity nodes are set according to the critical swimming speed of the zebra fish, and after each exercise intensity node moves, blood lactate is detected by micro-measuring blood to obtain the blood lactate threshold of the zebra fish.
In the method for evaluating the aerobic exercise capacity of the zebra fish, the zebra fish in the step (1) is placed in an independent fish tank;
in the step (1), the parameters of the zebra fish comprise body length and body weight;
the time of fasting treatment in the step (1) is 24 h.
In the method for evaluating the aerobic exercise capacity of the zebra fish, the adaptive training in the step (2) is to adapt the zebra fish for 2 hours at the speed of 0.8 BL/s.
In the method for evaluating the aerobic exercise capacity of the zebra fish, the time of the flushing mode in the step (3) is 90 s;
the time for waiting for the mode in the step (3) is 30 s;
the time of the measurement mode in the step (3) is 5 min.
In the method for evaluating the aerobic exercise capacity of the zebra fish, the flow rate increase mode of water in the lane of the respirator in the step (4) is increased by 2.7BL/s every 14 min;
the standard for judging the exhaustion state of the zebra fish in the step (4) is that the zebra fish stops at the tail of the lane for more than 20 s;
the calculation formula of the critical swimming speed in the step (4) is
Ucrit＝U+(t/Δt)*ΔU；
Wherein U is the maximum swimming speed (cm/s) reached by the experimental fish in the test; wherein Δ U is the speed increment (2.7 BL/s); wherein Δ t is the time interval (14 min); where t is the time (min) of adherence at maximum speed.
In the method for evaluating the aerobic exercise capacity of the zebra fish, the treatment method before measuring the body length and the body weight in the step (1) is as follows, the zebra fish is placed into anesthetic for anesthesia, the zebra fish is taken out and placed on a wet towel, the body length is measured by a vernier caliper, the measurement part is from the head of the zebra fish to the tail, the zebra fish is wiped dry, then the zebra fish is placed into an electronic balance for weighing the body weight, and the numerical value is recorded.
In the method for evaluating the aerobic exercise capacity of the zebra fish, the mass concentration of the anesthetic is 40mg/L, and the method comprises the steps of adding 0.04g of tricaine into 1000ml of water and uniformly mixing the three.
In the method for evaluating the aerobic exercise capacity of the zebra fish, the gradient exercise intensity nodes in the step (5) are 0%, 20%, 40%, 60%, 80% and 100% of the critical swimming speed;
and (5) moving each movement intensity node for 5 min.
In the method for evaluating the aerobic exercise capacity of the zebra fish, the method for taking the blood in a trace amount in the step (5) is to take the blood from the dorsal artery or the posterior main vein of the zebra fish by using a trace blood taking needle. It should be noted that before micro blood sampling, the zebra fish is put into a rapid anesthetic solution with the mass concentration of 200mg/L (consistent with the preparation method of the anesthetic, the anesthetic mass is increased by 5 times), then the zebra fish is taken out and put on a wet towel, a self-made simple blood sampling needle is adopted, the puncturing part is a dorsal artery or a posterior main vein, and finally a micro blood lactic acid detector of German EKF company is adopted for detection.
In the method for evaluating the aerobic exercise capacity of the zebra fish, the blood lactic acid threshold of the zebra fish in the step (5) is obtained by drawing an increasing curve of the blood lactic acid changing along with exercise intensity.
Through retrieval, relevant articles and patents of a specific method for detecting the movement capacity of the zebra fish by using the blood lactic acid are not available at home and abroad. In the movement of a human body under increasing load, the blood lactate concentration increases along with the increase of the movement load, and when the movement intensity reaches a certain load, the starting point of the sharp rise of the blood lactate concentration is called a lactate threshold. A method for measuring blood lactic acid of fishes is not described at home and abroad, a large number of experiments are carried out on the method for measuring blood lactic acid of zebra fishes, a blood lactic acid curve of fishes is made, and the method has great significance on the research of the zebra fishes as model organisms applied to the field of sports medicine. The innovation of the blood sampling mode of zebra fish. The zebra fish has small volume, little blood volume and extremely difficult finding of blood taking parts, so that the zebra fish can continuously complete milk while avoiding irreversible trauma on the fish bodyThe detection process of the acid domain experimental scheme is a key problem of the detection technology. Firstly, in order to realize minimally invasive blood collection, a user uses a self-made capillary needle, a rubber hanging tube is used for covering the needle head, heparin sodium (5mg/ml) is used for infiltrating the capillary needle with pressure to achieve an anticoagulation effect, then the needle head is inserted into the posterior main vein of the anesthetized zebra fish, blood is sucked out with pressure, then the taken blood is placed on membrane paper, and a trace blood lactic acid detector of German EKF company is used for detection to obtain the blood lactic acid value of the zebra fish. Secondly, in order to realize the detection of each movement intensity node of the movement lactic acid threshold, a detection scheme of the movement detection lactic acid threshold is determined through a plurality of experiments. Firstly, measuring the critical swimming speed U according to the measuring scheme of the critical swimming speed of the zebra fishcritAnd dividing the increasing gradient of the zebra fish into U of 0%, 20%, 40%, 60%, 80%, 100%critAfter each exercise intensity node of six gradients moves for 5min, blood lactate concentration values of the exercise intensity nodes are obtained by measuring blood in a micro-scale manner and detecting the blood lactate by using a micro-scale blood lactate detector of Germany EKF company, as shown in FIG. 4. The zebra fish blood lactic acid threshold measuring method comprises the following steps: (1) firstly, measuring the body length and the body weight of the fish and measuring the critical swimming speed Ucrit(ii) a (2) Measuring the blood lactic acid value of a fish in a quiet state by adopting a micro blood sampling method, placing the fish into an anesthetic solution for anesthesia, taking the fish out, placing the fish on a wet towel, adopting a self-made simple blood sampling needle, pricking the fish into a dorsal artery or a posterior main vein, and detecting by adopting a micro blood lactic acid detector of German EKF company, wherein the zebra fish needs a certain time to repair a wound after the first blood sampling, and meanwhile, because the zebra fish is different from human beings, the blood volume contained in an individual is not enough to finish the blood sampling of six continuous detection points, the zebra fish is placed in E3 water containing methylene blue for recovery for 2 days after each blood sampling; (3) after the fish have recovered for 2 days, the fish are placed in lanes for 20% UcritMoving in water flow of the movement intensity node for 5min, taking out, detecting blood lactic acid by the micro blood sampling method, and recovering for 2 days; (4) then completing 40%, 60%, 80% and 100% U in sequencecritBlood lactate testing of exercise intensity nodesMeasuring; (5) the blood lactate threshold was obtained by plotting an increasing curve of blood lactate as a function of exercise intensity, as shown in fig. 5.
In addition, the blood lactic acid results of zebra fish measured by our test method are consistent with the trend of results measured by human (FIG. 6), and the lactic acid threshold of zebra fish detected in the test is 80% U of the relative exercise intensitycritLeft and right, meaning 80% U of the relative motion strengthcritLater, zebrafish will enter high-intensity anaerobic exercise. The results show that the method for detecting the lactic acid threshold of the zebra fish is scientific and effective, and the movement capability of the zebra fish is judged by judging the inflection points of the lactic acid thresholds of different zebra fish, so that a solid foundation is laid for the research on the related problems in the scientific field of movement by using the animal zebra fish in the classical model.
Other modification 1: (1) the fish were fasted for 24h before testing, and placed in lane 0.8BL/s for 2h to eliminate the stress; (2) the swimming tunnel is automatically set to complete three stages within 10 min: flushing (Flash) time is 4min, waiting (wait) time is 1min, and measuring (measure) time is 5 min; (3) the test is started by increasing 2.7BL/s every 20min, the water flow speed is continuously increased, the speed increment (delta U) is always 2.7BL/s, each time interval (delta t) is 20min until the experimental fish reaches the exhaustion state, and the judgment standard is that the experimental fish stops at the tail part of a lane for more than 20 s; (4) recovering at 0.8BL/s, and performing second measurement after 60min rest; (5) and carrying out empty measurement after the second measurement. The result shows that the tested oxygen consumption curve of the zebra fish (figure 7) does not accord with the trend of the oxygen consumption curve of the common fish (figure 8) reported by the literature, and the oxygen consumption curves of a plurality of zebra fish basically increase and then decrease, so that the situation shows that the scheme is not suitable for domestic oxygen consumption measurement of the zebra fish; a large number of literatures are consulted to find that the body length and the body weight of domestic zebra fish are smaller than those of foreign zebra fish due to environmental factors although the strains are the same. Therefore, in order to explore a scheme suitable for evaluating the aerobic exercise capacity of domestic zebra fish, the increasing time of 20min of the originally tested zebra fish is gradually reduced to 14min through repeated tests. Meanwhile, in order to reduce the influence of water flow of the zebra fish with smaller body size in the flushing and oxygen changing process, the flushing and oxygen changing time in the test scheme 1 is shortened from 4min to 90s, and the waiting time is shortened from 1min to 30 s.
Other modification 2: (1) the fish were fasted for 24h before testing, and placed in lane 0.8BL/s for 2h to eliminate the stress; (2) the swimming tunnel is automatically set to complete three stages within 7 min: flushing (Flash) time is 90s, waiting (wait) time is 30s, and measuring (measure) time is 5 min; (3) the test starts by increasing 2.7BL/s every 14min, the water flow speed is continuously increased, the speed increment (delta U) is always 2.7BL/s, each time interval (delta t) is 14min until the experimental fish reaches the exhaustion state, and the judgment standard is that the experimental fish stops at the tail part of the lane for more than 20 s. As shown in fig. 9, 10, 11, 12, and 13, the results show that: the oxygen consumption curve of the zebra fish at the age of 3 months and the oxygen consumption curve of the zebra fish at the age of 23 months conform to the trend of the oxygen consumption curves of the fishes, the maximum oxygen consumption of the zebra fish at the age of 3 months is very significantly different from the maximum oxygen consumption of the zebra fish at the age of 23 months, and the maximum oxygen consumption of the zebra fish at the age of 3 months is higher than the maximum oxygen consumption at the age of 23 months. The relative critical swimming speed of the zebra fish of 3 months old is higher than that of the zebra fish of 23 months old, and the statistical analysis has obvious difference. The maximum oxygen consumption of the zebra fish is gradually reduced along with the increase of the age, the relative critical swimming speed is gradually reduced, and the aerobic exercise capacity is gradually reduced. The result accords with the development rule that the aerobic exercise capacity is reduced along with the increase of age, thereby further proving the scientificity and feasibility of the scheme for evaluating the aerobic exercise capacity of the zebra fish.
In conclusion, the maximum oxygen consumption, the critical swimming speed and the blood lactic acid threshold are used as the detection indexes of the aerobic exercise capacity of the zebra fish for the first time, a uniform measurement standard is provided for measuring the aerobic exercise capacity of the zebra fish, and the detection method is scientific and reasonable. The model organism zebra fish selected by the invention has the advantage of verification of aerobic movement of living animals, and is beneficial to improving the experimental efficiency and reducing the experimental cost.
Other descriptions:
1. the curve of the maximum oxygen consumption test result of the zebra fish is different from the curve of the maximum oxygen uptake test result of human beings, and a maximum oxygen consumption platform does not appear, so that the maximum oxygen consumption of the fish is judged only by taking the oxygen amount measured when the exercise capacity is increased gradually as the maximum oxygen consumption, but in the latest research on testing the maximum oxygen uptake of the human beings, the oxygen uptake measured when the load capacity is increased gradually is determined as the maximum oxygen uptake and is not accurate. At the present stage, the most advanced test for the maximum oxygen consumption of the zebra fish is basically the respiratory system of the zebra fish lane from Denmark, and only the consumption of oxygen in water in one period in the exercise process is measured, and the generation amount of carbon dioxide in the water cannot be measured to determine the respiratory quotient. Due to the technical limitation, a secondary index lactic acid threshold for judging the maximum oxygen uptake needs to be adopted, the index of the lactic acid threshold reflects the value of the maximum oxygen uptake, and finally the index of the aerobic exercise capacity of the zebra fish is accurately judged.
2. The assessment of the oxygen content and the motion ability of the zebra fish is to perform a corresponding index test on the zebra fish which is not subjected to motion training, but the zebra fish is normal under the condition that the individual difference of the zebra fish is poor due to the problem of the individual difference, in the test for assessing the oxygen content and the motion ability of the person, no test scheme is suitable for all persons, the zebra fish is also one, the zebra fish does not have an accurate reference value for the oxygen motion intensity at the present stage, and the problem of the individual difference of the age exists, so that the mark for assessing the oxygen motion ability of the zebra fish only has a relative value and does not have an absolute value, at the present stage, the maximum oxygen uptake is reflected by the index of the lactic acid threshold of the secondary index, further exploration is needed, but the lactic acid threshold value tested by the motion scheme is not problematic, and the lactic acid threshold value of the zebra fish in the resting state is equivalent to the lactic, the threshold lactate outcome curves tested also agree with the threshold lactate outcome curves for humans, so a exercise regimen for testing lactate threshold may be used.
3. The indexes for measuring aerobic exercise ability are divided into critical swimming speed, maximum oxygen consumption and blood lactic acid, so that the blood lactic acid and the maximum oxygen content are in parallel relation, and the aerobic exercise ability is also evaluated.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Furthermore, it should be understood that although the present specification describes embodiments, not every embodiment contains only a single technical patent, and such description of the specification is for clarity only, and those skilled in the art should be able to make the specification as a whole, and the technical patents in the embodiments may be appropriately combined to form other embodiments understood by those skilled in the art.
Claims (10)
1. A zebra fish aerobic exercise capacity evaluation method comprises the following steps:
(1) preparing zebra fish, marking the zebra fish by numbers, measuring parameters of the zebra fish, and then performing fasting treatment;
(2) transferring the zebra fish after fasting treatment to a lane of a respirator, and carrying out adaptive training;
(3) set up automatic three-stage mode of the lane of the respirometer: a flushing mode, a waiting mode and a measuring mode;
(4) then gradually increasing the flow rate of water in a lane of the respirator until the zebra fish reaches an exhausted state, recording real-time oxygen consumption data and the maximum swimming speed at the termination time, and obtaining the maximum oxygen consumption and the critical swimming speed of the zebra fish;
(5) gradient movement intensity nodes are set according to the critical swimming speed of the zebra fish, blood lactic acid is detected by measuring blood slightly after each movement intensity node moves, and the blood lactic acid threshold of the zebra fish is obtained.
2. The method for evaluating aerobic exercise capacity of zebra fish according to claim 1, wherein:
in the step (1), the zebra fish is placed in an independent fish tank;
in the step (1), the parameters of the zebra fish comprise body length and body weight;
the time of fasting treatment in the step (1) is 24 h.
3. The method for evaluating aerobic exercise capacity of zebra fish according to claim 1, wherein:
in the step (2), adaptive training is to adapt the zebra fish for 2h at the speed of 0.8 BL/s.
4. The method for evaluating aerobic exercise capacity of zebra fish according to claim 1, wherein:
the flushing mode in the step (3) lasts for 90 s;
the time for waiting for the mode in the step (3) is 30 s;
the time of the measurement mode in the step (3) is 5 min.
5. The method for evaluating aerobic exercise capacity of zebra fish according to claim 1, wherein:
in the step (4), the flow rate of water in the lane of the respirator is increased by 2.7BL/s every 14 min;
the standard for judging the exhaustion state of the zebra fish in the step (4) is that the zebra fish stops at the tail of the lane for more than 20 s;
the calculation formula of the critical swimming speed in the step (4) is
Ucrit＝U+(t/Δt)*ΔU；
Wherein U is the maximum swimming speed of the experimental fish in the test; wherein Δ U is the speed increment; wherein Δ t is a time interval; where t is the time of adherence at maximum speed.
6. The method for evaluating aerobic exercise capacity of zebra fish according to claim 2, wherein:
the processing method before measuring the body length and the body weight in the step (1) is that the zebra fish is placed into anesthetic for anesthesia, the zebra fish is taken out and placed on a wet towel, the body length is measured by a vernier caliper, the measuring part is from the head of the zebra fish to the tail of the zebra fish, the zebra fish is placed into an electronic balance after being wiped dry, the body weight is weighed, and the numerical value is recorded.
7. The method for evaluating aerobic exercise capacity of zebra fish according to claim 6, wherein:
the mass concentration of the anesthetic is 40mg/L, and the preparation method comprises the steps of adding 0.04g of tricaine into 1000ml of water and uniformly mixing the three.
8. The method for evaluating aerobic exercise capacity of zebra fish according to claim 1, wherein:
in the step (5), gradient exercise intensity nodes are 0%, 20%, 40%, 60%, 80% and 100% of the critical swimming speed;
and (5) moving each movement intensity node for 5 min.
9. The method for evaluating aerobic exercise capacity of zebra fish according to claim 1, wherein:
and (5) taking blood from the dorsal artery or the posterior main vein of the zebra fish by using a trace blood taking needle.
10. The method for evaluating aerobic exercise capacity of zebra fish according to claim 1, wherein:
and (5) obtaining the blood lactic acid threshold of the zebra fish in the step (5) by drawing an increasing curve of the blood lactic acid along with the change of the exercise intensity.
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| WO2019185965A1 (en) * | 2018-03-28 | 2019-10-03 | Consejo Superior De Investigaciones Científicas | Device and method for monitoring activity in fish |
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