CN106069897A - The method of colony/family that a kind of quick screening left-eyed flounder is high temperature resistant - Google Patents
The method of colony/family that a kind of quick screening left-eyed flounder is high temperature resistant Download PDFInfo
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- CN106069897A CN106069897A CN201610422052.7A CN201610422052A CN106069897A CN 106069897 A CN106069897 A CN 106069897A CN 201610422052 A CN201610422052 A CN 201610422052A CN 106069897 A CN106069897 A CN 106069897A
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- family
- colony
- high temperature
- ctm
- temperature resistant
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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
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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 present invention relates to marine fish culture and breeding technical field, specifically a kind of method based on the high temperature resistant colony/family of the lossless quick screening left-eyed flounder of dynamic method.Colony to be screened/family is individual, after fluorescence labeling, gradually rise cultivation temperature with the speed of 0.5 1 DEG C/h, observe the swimming state change of fish, record it and lose temperature when standing up ability, it is left-eyed flounder tolerable critical high temperature level (critical thermal maximum, CTM), the CTM value of each colony/family of statistical analysis.CTM height person is high temperature resistant colony/family；The horizontal no difference of science of statistics of CTM, is the colony/family of identical heat-resisting ability.Operating process of the present invention is simple, quick, and testing cost is low, effectively prevent the individual death during screening simultaneously, it is achieved that the lossless quick screening of colony/family that left-eyed flounder is high temperature resistant.
Description
Technical field
The present invention relates to marine fish culture and breeding technical field, specifically a kind of lossless quickly based on dynamic method
The method of the screening high temperature resistant colony/family of left-eyed flounder.
Background technology
The left-eyed flounder such as lefteye flounder and turbot is the important seawater economy cultivation fingerling of China, but it grew during summer
Speed substantially reduces.At present, breeding production utilizes deep sea water reduce water temperature as cold water source more, thus weaken the summer
The impact that left-eyed flounder is grown by Ji Gaowen, but which increase the production cost of breeding enterprise, and to the marine eco-environment
Cause potential impact.Therefore, need badly and carry out the seed selection of the high temperature resistant strain of left-eyed flounder.
Assessment to left-eyed flounder heat-resisting ability is the important foundation in high temperature resistant strain breeding, but presently relevant grinds
Study carefully and rely primarily on static method and carry out in production practices, i.e. more each colony/family death time under the high temperature conditions and dead
Colony/family that rate of dying differential screening is high temperature resistant.This traditional approach screening time cost is relatively big, and required time is short then several days, long
Then even several months several weeks, and for individual final most dead (death rate is higher than 50% more) of screening, not only cause one
Fixed culturing economic loss, and cause static method not to be suitable for even cannot be used for the temperature tolerance of negligible amounts colony/family
Shape is identified.In recent years, though microsatellite and AFLP equimolecular labelling technique obtain in the research of the high temperature resistant proterties of left-eyed flounder
Application, but the operation of this method is relatively complicated, and label screening efficiency is low, and the sample size of this method of what is more important detection is big, with 50
As a example by colony/family (each colony/family is chosen 20 tails and calculated), then need at least to analyze 1000 case samples, simultaneously its detection
Costly, the detection cycle is longer for expense, and this becomes the major reason of molecular marking technique few application in actual production.Cause
This, set up a kind of lossless fast appraisement method, it has also become screens high temperature resistant the urgent of colony/family of left-eyed flounder in production and is essential
Ask.Compared with traditional static method, dynamic method is a kind of method of Fast Evaluation species temperature resistant capability, can significantly reduce screening when
Between cost, this method has been used for the comparison of different fingerling or the high temperature resistant proterties of shell-fish at present, but utilize this method carry out with
There is not been reported in the screening of the one high temperature resistant colony/family of fingerling.Therefore, foundation carries out left-eyed flounder colony/family based on dynamic method
Being the evaluation method of heat-resisting ability, to promotion left-eyed flounder, high temperature resistant cultivation and breeding research and application have highly important
Effect.
Content of the invention
The shortcoming that present invention is generally directed to above-mentioned prior art, provides a kind of based on the lossless quick screening flounder sole of dynamic method
The method of colony/family that class is high temperature resistant.
For achieving the above object, the present invention uses the technical scheme to be:
The method of colony/family that a kind of quick screening left-eyed flounder is high temperature resistant, by individual for colony to be screened/family fluorescence mark
After note, gradually rise cultivation temperature with the speed of 0.5-1 DEG C/h, observe the swimming state change of fish, record its forfeiture and stand up energy
Temperature during power, is left-eyed flounder tolerable critical high temperature level (critical thermal maximum, CTM), system
The CTM value of each colony/family analyzed by meter.CTM height person is high temperature resistant colony/family；The horizontal no difference of science of statistics of CTM, is phase
Colony/family with heat-resisting ability.
Choose left-eyed flounder colony/family to be screened, from each family, randomly select the normal healthy individuals of body colour,
After family being marked in fluorescence labeling color mark part combination mode, all colony/familys are divided equally and raises together with, support temporarily
Domestication 3～5d, period holding water temperature is constant, and continuous oxygenation is stand-by.
After colony to be screened/family individual fluorescence labeling, it is cultivated water temperature and gradually rises with the speed of 0.5-1 DEG C/h,
During this, observe the swimming state change of fish, record it and lose temperature when standing up ability, be that left-eyed flounder is tolerable to be faced
Boundary's high temperature level CTM, record temperature levels (CTM) now, and fish is pulled out be transferred in normal temperature seawater to recover normal；Ratio
The CTM value of more each colony/family.CTM height person is high temperature resistant colony/family；The horizontal no difference of science of statistics of CTM, is identical resistance to
Colony/the family of high temperature capabilities.
The swimming state change of described observation fish takes gently to stir water body, when the significant reaction of fish is less, utilizes instrument careful
Being stood up, in 1min, fish body can not recover to normal body state as standard, and the initial stage observed frequency that heats up is low, and the later stage is gradually
Strengthen.
Advantages of the present invention and good effect are as follows:
1) application in biological temperature resistant capability is evaluated for the dynamic method is introduced the high temperature resistant colony/family of left-eyed flounder by the present invention
Screening.Based on the heat-resisting ability evaluation method of dynamic method, left-eyed flounder colony/family can be distinguished extremely simple, rapidly
Heat-resisting ability difference；
2) conventional high temperature resistant fish screening is to carry out in lethal temperature level, and not only cycle time is long, and tests
Animal is many can not survive, and limits the application in the case of coefficient amount of being in lacks this.The present invention carries out flounder based on dynamic method principle
The screening of colony/family that sole class is high temperature resistant, dramatically reduces screening time cost, and during confirming to be prevented effectively from screening
The death of fish, so that the colony/family of negligible amounts can be used for high temperature resistant screening.
Brief description
During 3 turbot families that the traditional static method that Figure 1A provides for the embodiment of the present invention measures survival at high temperature
Between figure.Wherein F1, F2 and F3 represent turbot family the 1st, family 2 and family 3 respectively, and on cylinder, different small English alphabet represent
Significant difference；
3 turbot families of the dynamic method mensuration that Figure 1B provides for embodiment of the present invention time-to-live at high temperature
Figure.Wherein F1, F2 and F3 represent turbot family the 1st, family 2 and family 3 respectively, and on cylinder, different small English alphabet represent poor
Different significantly.
Detailed description of the invention
The present invention, based on dynamic method principle, sets up the loseless method evaluating left-eyed flounder colony/family heat-resisting ability, keeps away
Exempt from the fish that cause of screening dead, greatly reduced screening time cost, be grinding of the degeneration-resistant proterties such as left-eyed flounder is high temperature resistant
Study carefully and provide strong technical support.
Specifically a kind of principle based on the biological temperature resistant capability of Judgment of dynamic method, it is achieved left-eyed flounder is high temperature resistant group
The method of the lossless quick screening of body/family.Select different colony/familys, labeled, raise together with after, control water temperature is with less than 1
DEG C/speed of h gradually rises, until fish body loses ability of standing up, and records temperature levels now；Add up each colony/family
The critical high temperature level (critical thermal maximum, CTM) of tolerance, and each colony/family is ranked up, CTM
Value height represents that heat-resisting ability is strong, thus screens the acquisition high temperature resistant colony/family of left-eyed flounder.
Specifically,
1) colony/family mark is raised together with
Select left-eyed flounder colony/family to be assessed, from each family, randomly select healthy normally of body colour
Body, after being marked family in fluorescence labeling color mark part combination mode, divides equally all colony/familys to 4
In experiment cylinder；
2) dynamic method screening
The family having marked carrying out 3～5d adapt to, period keeps water temperature constant, continuous oxygenation.After experiment starts, with control
Temperature device control water temperature at the uniform velocity raises with the speed less than 1 DEG C/h；
3) fish body status monitoring
In temperature-rise period, take the observation plan of " loose at first but afterwards tight " (the initial stage observed frequency that i.e. heats up is low, and the later stage is gradually increased)
Slightly.When gently stirring water body, when the significant reaction of fish is less, the instruments such as fishnet are utilized carefully to be stood up, can not with fish body in 1min
Recover to normal body state be standard, record temperature levels (CTM) now, and fish is pulled out be transferred in normal temperature seawater with
Recover normal；The CTM value of each colony/family of statistical analysis, sequence determines each family temperature resistant capability difference (CTM value and colony/family
Be temperature resistant capability be inverse ratio).
Below in conjunction with the embodiment of the present invention, the technical scheme in the embodiment of the present invention is clearly and completely described.
Described embodiment is a part of embodiment of the present invention, but is not all, of embodiment.Based on the embodiment in the present invention, this
Field those of ordinary skill is obtained every other embodiment under inscribing before not making creative work, broadly fall into the present invention and protect
The scope protected.
Embodiment:
1) families selecting
The background informations such as the monthly age and the body size (such as body weight and total length) that confirm each family, choose 3 turbot families
(being respectively labeled as F1, F2, F3) carries out the differentiation of dynamic method heat-resisting ability；
2) family mark
From each family, observe and carefully fish for body colour normal healthy individuals 32 tail, being placed in amino between 60mg/L
Ethyl benzoate mesylate (MS-222) solution is anaesthetized 10min, then puts down gently on flat board, take the fluorescence labeling preparing
Entry needle, carries out embeddability fluorescence labeling to each family individuality, 3 familys carry out that afterbody is red respectively, yellow and blueness glimmering
Signal.
3) family is raised together with
Dividing equally the family after mark at random to 4 experiment cylinders, every cylinder repeats as an experiment, it is ensured that every cylinder each
Family has 8 tails.Then, all familys carry out the environment adaptation of at least 3～5d under constant temperature, and period keeps flowing water (often little
When quantity of exchanged water be 0.5m3), continuous oxygenation (> 6mg/L), water temperature (T0) temperature regulating device is remained stable, wherein T0Can be according to fingerling
Suitably adjust, the present embodiment is 17 DEG C；
4) dynamic method screening
Before experiment starts, again detect water temperature (T0).On this basis, temperature regulating device is utilized at the uniform velocity to rise with 1 DEG C/h speed
Height, and the state change of close observation fish.After being warming up to 30 DEG C, fishing gear is utilized gently to stir water body, if fish is without significant reaction, carefully
Stood up (flounder class: make on the right side of it health upward；Flounder class: make on the left of it health upward)；When fish cannot recover in 1min to
During normal body state, record temperature levels now, it is fish tolerable critical high temperature level (CTM).
5) Data acquisition and issuance
By data inputting Excel, calculate, with AVERAGE function, the CTM mean value that each experiment of each family repeats, and then
Calculate the CTM mean value that each family 4 experiment repeats, the final CTM value obtaining this family with AVERAGE function.Will be all
In family experiment repeats CTM average typing SPSS statistical software, select " analysis " " comparing average " " single factor test side
Difference analyses (one-way ANOVA) ", in " Multiple range test " option, select " Ducan ", thus analyze different family further
CTM difference.
In the present embodiment, the CTM value of 3 turbot families is respectively 32.5 DEG C (F1), 32.3 DEG C (F2) and 31.6 DEG C
(F3), owing to CTM value is proportional with family heat-resisting ability, tentatively judge that 3 turbot family heat-resisting ability orders are: F1
>F2>F3.SPSS 20.0 statistical analysis further confirms that, the CTM value difference of F1 and F2 is not different notable, and both is higher than F3.Combine
Conjunction is distinguished, the heat-resisting ability of F1 and F2 is close, and is above F3 (seeing Figure 1A).The present embodiment result and traditional static method
The selection result (seeing Figure 1B) is consistent.
The appraisement system based on dynamic method that the present invention sets up, not only operating process is extremely simple, and screening required time is relatively
Short (≤24h), and screening effect is good, it is achieved that the quick screening of colony/family that left-eyed flounder is high temperature resistant；And avoid screening
The death of period left-eyed flounder, it is achieved that the lossless screening of colony/family that left-eyed flounder is high temperature resistant.
Claims (4)
1. the method for the high temperature resistant colony/family of a quick screening left-eyed flounder, it is characterised in that: by colony/family to be screened
After individual fluorescence labeling, gradually rise cultivation temperature with the speed of 0.5-1 DEG C/h, observe the swimming state change of fish, record it
Lose temperature when standing up ability, be left-eyed flounder tolerable critical high temperature level (critical thermal
Maximum, CTM), the CTM value of each colony/family of statistical analysis, CTM height person is high temperature resistant colony/family；CTM level is without system
Difference learned by meter, is the colony/family of identical heat-resisting ability.
2. the quick method screening the high temperature resistant colony/family of left-eyed flounder as described in claim 1, it is characterised in that: choose
Left-eyed flounder colony/family to be screened, randomly selects the normal healthy individuals of body colour, with fluorescence labeling face from each family
After family is marked by color marker part combination mode, all colony/familys is divided equally and raises together with, domesticating 3～5d temporarily, the phase
Between keep water temperature constant, continuous oxygenation, stand-by.
3. the quick method screening the high temperature resistant colony/family of left-eyed flounder as described in claim 1 or 2, it is characterised in that: treat
After screening colony/family individual fluorescence labeling, it is cultivated water temperature and gradually rises with the speed of 0.5-1 DEG C/h, during this period, observes
The swimming state change of fish, records it and loses temperature when standing up ability, be left-eyed flounder tolerable critical high temperature level
CTM, record temperature levels (CTM) now, and fish is pulled out be transferred in normal temperature seawater to recover normal；Relatively more each colony/
The CTM value of family；CTM height person is high temperature resistant colony/family；The horizontal no difference of science of statistics of CTM, is identical heat-resisting ability
Colony/family.
4. the quick method screening the high temperature resistant colony/family of left-eyed flounder as described in claim 3, it is characterised in that: described
The swimming state change observing fish takes gently to stir water body, when the significant reaction of fish is less, utilizes instrument carefully to be stood up, with
In 1min, fish body can not recover to normal body state is standard, and the initial stage observed frequency that heats up is low, and the later stage is gradually increased.
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| CN201610422052.7A CN106069897A (en) | 2016-06-13 | 2016-06-13 | The method of colony/family that a kind of quick screening left-eyed flounder is high temperature resistant |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112931311A (en) * | 2021-02-09 | 2021-06-11 | 上海海洋大学 | Flounder health condition evaluation method |
| CN114931116A (en) * | 2022-05-17 | 2022-08-23 | 广东伟业罗非鱼良种有限公司 | Breeding method of tilapia strain with rapid growth and strong stress resistance under continuous high-temperature stress |
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| CN112931311A (en) * | 2021-02-09 | 2021-06-11 | 上海海洋大学 | Flounder health condition evaluation method |
| CN114931116A (en) * | 2022-05-17 | 2022-08-23 | 广东伟业罗非鱼良种有限公司 | Breeding method of tilapia strain with rapid growth and strong stress resistance under continuous high-temperature stress |
| CN114931116B (en) * | 2022-05-17 | 2023-09-08 | 广东伟业罗非鱼良种有限公司 | Breeding method of tilapia strain with rapid growth and strong stress resistance under continuous high temperature stress |
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