CN1922991A - Seedling breeding method for dollar spot Scophthalmusmaximus - Google Patents
Seedling breeding method for dollar spot Scophthalmusmaximus Download PDFInfo
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Abstract
The invention relates to a method for breeding sole, which comprises: (1) cultivating parents that 1, adjusting water temperature that naturally decreasing 6-7Deg. C, and increasing to 12Deg. C; 2, adjusting light with lamp; 3, adjusting bait that uses shrimp, fish, etc, with vitamin and lecithin; 4, daily management that clearing and exchanging water; 5, using HCG and LHRH to accelerating growth; (2), picking up semen and eggs, and cultivating zygote, while the zygote is obtained by dry semination, and using sinke zygote and semi-sink zygote to cultivate the baby fish; (3), breeding seed that cultivating 3-5cm seeds in small and large pools, while the total survival rate can reach 28.0-30.0%, and the total survival rate at 8-9cm can reach 21%. The invention can improve the survival rate of seeds.
Description
Technical field
The invention belongs to the breed field of fish, relate to simultaneously that feeding of fish raised, container, heating or lighting device.
Background technology
Verasper variegate belongs to Pleuronectiformes, Pangasiidae, halibut genus, be large-scale left-eyed flounder, but quantity is few.Halibut has higher economic value and cultures value, is improved seeds that are worthy to be popularized.In the industry the basic research and the seedling growing process of the artificial breeding of verasper variegate a large amount of research work have been done in China's aquaculture: characteristic of division, habitat, the food of circle spot star dish are formed, grown and disease etc. has carried out studying (circle spot star dish biological characteristic research " Modern Fishery information " 2002,17[10] 25～27) as Chen Siqing etc.Wang Kaishun etc. observe (observation " Chinese aquatic science " 2003 that circle spot star dish embryo and prelarva grow, 10[6] .451～456) to its embryo and postembryonal development.Zhang Yongzhong etc. have studied the ultra microstructure (ultra microstructure and nuclear proparea special construction " animal journal " .2004 of circle spot star dish sperm, 50[4] .630～637) of round spot star dish sperm respectively with ESEM and perspective Electronic Speculum method.In addition, also just like the report of fish sperm and embryo's basic research such as superfreeze preservation, these all are provide the foundation technology and basic theory of the artificial breeding of circle spot star dish fish.Zheng Huidong has made Primary Study (artificial propagation and seedling growing process " Fujian aquatic products " .2003[3 of circle spot star dish] .15～17) to the artificial breeding of circle spot star dish.The achieving success though grow seedlings, however survival rate only has 18.3%.
Summary of the invention
At present prior art to the circle spot star dish existing defective of growing seedlings, the object of the present invention is to provide and a kind ofly can effectively improve fertilized egg hatching rate, fry quality and survival rate, and can realize batch production production, the verasper variegate artificial breeding technique of seed is provided for large-scale farming.
Technical scheme of the present invention comprises parent population accelerating, incubating oosperm, seed rearing three phases, and its concrete implementation step and main points are as follows:
One, parent population accelerating
Verasper variegate parent population male and female are grown asynchronous, the egg-laying time of female parent population is long, generally can continue about 2 months time, and male parent population gonad development is very fast, generally Zao than raun, duration is shorter, and in order to obtain ripe male and female parent population the same period, the present invention has adopted multiple accelerating method (ecological control method; Hormone accelerating method; The sperm freezing technology), guarantee that parent population is synchronous in the whole phase male and female of breeding:
1, ecological control method: control light, temperature control accelerating technology, many ponds light regulating technology.This technology has changed simply controls light, temperature control technique (only heat up, increase illumination) in the past.Water temperature and illumination are changed synchronously, make its gonad development change the process that a variation is arranged with natural environment, this is the just more approaching water environment of verasper variegate natural spawning is for laying eggs smoothly of parent population carried out the predecessor basis.Concrete grammar is as follows:
1.1 water temperature adjustment
Cooldown period: the parent fish rearing water temperature should reduce with natural temperature (12 ℃), and water temperature will arrive 6～7 ℃.
The constant temperature phase: natural temperature in the time of 6～7 ℃, constant temperature 7～10 days.
Temperature raising period: water temperature raises 1 ℃ weekly, reaches 12 ℃.
The constant temperature phase: when water temperature reaches 12 ℃, constant temperature is cultivated.
1.2 illumination adjustment
Lighting is adopted in gobo shading (dark layer); Intensity of illumination 500LX, light application time changes with the variation of water temperature, and concrete condition is as follows:
Shorten the phase sunshine: from cooldown period, light 1 hour (annotating: since the 11 hours light that contracts) contracts weekly.
The constant phase at sunshine: when illumination drops to 8 hours, permanent light is cultivated.
Raise the phase sunshine: according to the rising of water temperature, by increasing illumination in 1 hour weekly, reach 18 hours, permanent light is cultivated.
1.3 bait throwing in adjustment
Bait kind: shrimp 30%, clam worm 20%, yellow croaker 20%, clam 15%, oyster 10%, squid 5%.
Add vitamin: Vc, Ve, B B-complex weighs 1～2% with bait and adds, and throws something and feeds once every 10 days.
Promote the gonad development medicine: lecithin, add with fish body weight 0.5%.
1.4 daily management
Throw something and feed: once a day, throw capacity, different diet collocations are thrown something and fed.
Change water: change water every day once, dirty cleaning every day in the end once.
Wash fish: observe the parent population situation of ingesting, it is bad to ingest, dipping 100ppm formaldehyde, 1h..
Note observing: the parent population gonad development situation and the situation of ingesting.
2, hormone accelerating method:
Adopt hormone acceleratings such as HCG, LHRH, regulate the method for male and female rate of development.This technology is widely used for a long time, but consumption and method are also had nothing in common with each other owing to the kind difference of hastening parturition.Verasper variegate generally carries out hastening parturition for twice.HCG 400VI/KG for the first time; HCG 200VI/KG for the second time; LHRH-A320ug/kg.
3, sperm freezing technology
Grow " male earlier back is female " rule according to fish, be about to the male sperm freezing of fully-developed earlier, treat to use after the raun maturation.Though this technology is used on fish breed.But according to the kind difference of fish, the protection liquid of selection also has nothing in common with each other, and consumption and proportioning are also variant.Concrete grammar of the present invention is as follows: preserve container: liquid nitrogen container
Store method: V
TS-2: V
Sperm=7: 10
V
DMSO: V
Sperm=2: 10
V
Calf serum: V
Sperm=1: 10
Mix, put into test tube, liquid nitrogen container is put in sealing, on the liquid level 8～10 minutes, in the liquid level 5 minutes, puts into fully then.Holding time: 1 year.Preservation effect: survival rate 80% behind the sperm rewarming.
Two, obtain ovum and hatching
(1) obtains ovum
The present invention adopts: manually squeeze ovum, dry method insemination, the general raun egg-laying period is about 2 months, and the ovum of manually extruding can be divided into: overdone ovum, mature egg, underdone ovum, therefore, grasping and adopting ovum opportunity is one of test-tube emphasis.As adopting the labeling method in " molecular labeling assist family breeding " technology, determine that the best adopts ovum opportunity, raising fertilization rate and incubation rate.Labeling method: the parent population of selecting sexual gland to grow generally has two kinds of labeling methods (1) muscle of back marks (2) abdominal cavity mark.Kind different choice labeling method according to fish is also different, for verasper variegate, the present invention selects the muscle of back labeling method: squeeze into muscle of back (bullet must pass through sterilization) with mark small of the stock bullet, measure the coding of bullet then with inductor, write down data, just can determine egg-laying time and the egg laying amount of every parent population when laying eggs.
(2) hatching
Verasper variegate fertilized egg is special semibuoyant egg.Usually little inflation is hatched in the hatchery.But the fertilized egg that the present invention makes all is sunken at the bottom of the pond for heavy property and unfertilized egg in earlier stage, is difficult to separate, and changes conventional with novel hatching net cage (specification 1m * 1m * 1m) according to this kind situation; Make ovum come-up or suspension by strong inflation, carry out the hatching in early stage, the time is generally 96 hours (12 ± 0.5 ℃ of water temperatures).When development of fertilized ova arrives certain phase (body segment formation), fertilized egg shows as half buoyancy, and this moment, we collected the fertilized egg that suspends, and puts into the hatchery and carries out the later stage hatching, hatches prelarva after general 24 hours.The incubation rate of this moment is more than 90%, and 12 ± 0.5 ℃ of hatching water temperatures are changed water 1/2 every day, illumination 500LX, and early stage is inflation by force, and little inflation of later stage is hatched prelarva through 110-132 hour demoulding.
Three, the cultivation of fry
(1) cultivates early stage
Nursery pond is behind cleaning and sterilizing, drop into prelarva after adding water to 1m, breeding density 5000～10000 tails/rice 3, prelarva are through 7 days initial feedings, 12～13 ℃ of water temperatures, throw something and feed behind prelarva opening wheel animalcule and add chlorella, 5/ml of wheel animalcule density, day throwing twice is changed water 2 times/day, each 1/2, bait throwing in is after changing water.Add " preventing sinking to the bottom anti-stream device " simultaneously and prevent that fry from sinking and dead, the cultivation time in early stage is 15 days.
" prevent sinking to the bottom anti-stream device ": be the equipment that a kind of recoil by current makes the sinking fry float again or suspend.
(2) later stage cultivates
Use specification to carry out the later stage cultivation, the prelarva of cultivating early stage is moved into for cultivating water body 10-15 times cement pit early stage.Later stage cultivates quantity of exchanged water and strengthens, and 2 times/day, each 2/3 to all, continued the wheel animalcule of throwing something and feeding on the 15th～35 day, throws something and feeds twice every day, and bait throwing in is after changing water, and water body wheel animalcule density keeps 5/ml; The fairy shrimp young of throwing something and feeding in the 20th～55 day day is thrown 1～2 time, and density remains on 1～2/ml; After the 45th day, be aided with large-scale copepoda and artifical compound feed every day, after the 20th day, postlarva ingests more vigorous, and ight soil is more, begins to inhale at the end, inhales the end every day once; Owing to there is the part postlarva to begin the attached end, inhale the necessary extreme care of end operation after the 40th day, the postlarva that sucks out puts together, and puts back to after the sterilization in the pond again, and the nurturing period, water pH in pond remained between 7.8～8.0, and dissolved oxygen is more than 4mg/L.
Adopt technology fertilized egg hatching rate of the present invention to reach 16～17%, cultivation back survival rate reached 40～50% in 15 days, and survival rate reached 70～80% in 40 days, and volt phase end survival rate reaches 80～90%, and the survival rate when growing into 3～5cm reaches 90～95%.The total survival rate of hatching when growing into 3～5cm reaches 28.0～30.0%.Total survival rate that fry is bred 8～9cm reaches more than 21%.
Outstanding advantage compared with prior art of the present invention is: the verasper variegate artificial breeding technique that provides is scientific and reasonable, practical, utilizes this technology can realize batch production production, for large-scale farming provides seed.Successfully solved female, the male parent population of verasper variegate and grown asynchronously, the anti-season operation makes the low guardian technique difficult problem that waits of survival rate of fish fry, makes the present invention grow seedlings to reach higher survival rate, and this is the prior art level that is beyond one's reach.
Below the invention will be further described with embodiment.
Embodiment
Embodiment
Verasper variegate parent population 800 tails were through 4 months domestication.
One. adopt following method to cultivate and accelerating:
1. water temperature adjustment
Cooldown period: the parent fish rearing water temperature should reduce with natural temperature (12 ℃), and water temperature will arrive 6～7 ℃.
The constant temperature phase: natural temperature in the time of 6～7 ℃, constant temperature 7～10 days.
Temperature raising period: water temperature raises 1 ℃ weekly, reaches 12 ℃.
The constant temperature phase: when water temperature reaches 12 ℃, constant temperature is cultivated.
2. illumination adjustment
Adopt lighting; Intensity of illumination 500LX, light application time changes with the variation of water temperature.
Shorten the phase sunshine: from cooldown period, and contract weekly light 1 hour of the light that contracted since 11 hours.
The constant phase at sunshine:, determine light application time according to the water temperature situation when illumination drops to 8 hours.
Sunshine temperature raising period: according to the rising of water temperature,, reach 18 hours by increasing illumination in 1 hour weekly.
3. bait throwing in adjustment
Bait kind: shrimp 30%, clam worm 20%, yellow croaker 20%, clam 15%, oyster 10%, squid 5%.
Add vitamin: Vc, Ve, B B-complex, consumption 1～2% was thrown something and fed once every 10 days.
Promote the gonad development medicine: lecithin, consumption 0.5%.
4. daily management
Throw something and feed: once a day, throw capacity, different diet collocations are thrown something and fed.
Change water: change water every day once, dirty cleaning every day in the end once.
Wash fish: observe the parent population situation of ingesting, it is bad to ingest, dipping 100ppm formaldehyde, 1h..
Note observing: the parent population gonad development situation and the situation of ingesting.
5. hormone accelerating method:
Adopt hormone acceleratings such as HCG, LHRH, regulate the method for male and female rate of development.Generally carry out hastening parturition for twice.HCG 400VI/KG for the first time; HCG 200VI/KG for the second time; LHRH-A3 20ug/kg.
Two. semen collection, adopt ovum and fertilized egg hatching: we select the muscle of back labeling method: squeeze into female parent population muscle of back with the mark small of the stock through the sterilization bullet, measure the coding of bullet then with inductor, write down data, just can determine egg-laying time and the egg laying amount of every parent population when laying eggs.After this batch parent population maturation, adopt kilogram surplus the ovum 30.Adopt simultaneously milliliter surplus the sperm 20, preserve with following liquid nitrogen method: preservation container: liquid nitrogen container.Sperm with the ratio of batching is:
V
TS-2: V
Sperm=7: 10
V
DMSO: V
Sperm=2: 10
V
Calf serum: V
Sperm=1: 10
With sperm and the batch mixes test tube of packing into after evenly, sealing.Put into liquid nitrogen container: on the liquid level 8～10 minutes, in the liquid level 5 minutes, put into fully then.Holding time can reach 1 year.Survival rate 80% behind the sperm rewarming.Successfully preserve milliliter surplus the sperm 20, but 4000 milliliters of mature eggs of normal process verasper variegate.
Adopt artificial crowded ovum, dry method insemination to obtain fertilized egg.Make 0.5KG fertilized egg altogether.
With specification is that 1m * 1m * 1m is hatched net cage, carries out the hatching in early stage with the demersal egg hatching m.Time is generally 96 hours (12 ± 0.5 ℃ of water temperatures), and when development of fertilized ova fertilized egg when body segment forms shows as half buoyancy, this moment, we collected the fertilized egg that suspends, and put into the hatchery and carried out the later stage hatching, hatched prelarva after general 24 hours.The incubation rate of this moment is more than 90%, and 12 ± 0.6 ℃ of hatching water temperatures are changed water 1/2 every day, illumination 500LX, and early stage is inflation by force, little inflation of later stage.
Three, the cultivation of fry
(1) cultivates early stage
Nursery pond adds water to and drops into prelarva 30000 tails, incubation rate 16.7%, breeding density 5000～10,000 tail/rice behind the 1m behind cleaning and sterilizing
3, prelarva is through 7 days initial feedings, 12～13 ℃ of water temperatures, and the wheel animalcule and add chlorella of throwing something and feeding behind the prelarva opening, 5/ml of wheel animalcule density day throws twice, changes water 2 times/day, and is each 1/2, and bait throwing in is after changing water.Add " preventing sinking to the bottom anti-stream device " simultaneously, prevent that fry from sinking and dead, the cultivation time in early stage is about 15 days.Survival rate 50%, " preventing sinking to the bottom anti-stream device " is the equipment that a kind of recoil by current makes the sinking fry float again or suspend.
(2) later stage cultivates
Use 15 meters
3The specification cement pit moves into the prelarva of cultivating early stage.Later stage cultivates quantity of exchanged water and strengthens, and 2 times/day, continued the wheel animalcule of throwing something and feeding in each 2/3, the 15～35 day, throw something and feed twice every day, bait throwing in is after changing water, and water body wheel animalcule density keeps 5/ml; The fairy shrimp young of throwing something and feeding in the 20th～55 day day is thrown 1-2 time, and density remains on this stage of 1～2/ml and improves constantly water temperature, heats up every day till 0.5 ℃ to 16 ℃, fry is grown fast, survival rate of fish fry 80%.
After the 45th day, be aided with large-scale copepoda and artifical compound feed every day, after the 20th day, postlarva ingests more vigorous, and ight soil is more, begins to inhale at the end, inhales the end every day once; After the 40th day owing to there is the part postlarva to begin the attached end, inhale the necessary extreme care of end operation, the postlarva that sucks out puts together, with putting back in the pond again after the 5ppm furans immersion sterilization in 20 minutes, nurturing period, water PH in pond remained between 7.8～8, dissolved oxygen is more than 4mg/L, and general 5～7 days of the phase end of volt, this moment, bait was with mixed bait S
3With fairy shrimp be main, 16 ℃ of water temperatures are changed water 3 times, full dose, this stage survival rate 80%, specification 2cm, enter volt end state after, fry is basicly stable, mortality reduces, just when bait is changed, the part fry is dead because of ingesting, and a small amount of death occurs, and should change flowing water into, 16 ℃ of water temperatures this moment.This stage survival rate 90%, quantity are 8640 tails, specification 3～5cm.Just can normally culture the stage this moment.Our this batch fry specification 7～8cm of present stage, quantity 6400 tails, well-grown, situation is stable.
Claims (4)
1. the seedling-cultivating method of dollar spot Scophthalmusmaximus is characterized in that processing step is:
(1) parent fish rearing and accelerating:
1) water temperature adjustment
Cooldown period: 12 ℃ of water temperatures of parent fish rearing are reduced to 6～7 ℃ with natural temperature;
The constant temperature phase: natural temperature in the time of 6～7 ℃, constant temperature 7～10 days;
Temperature raising period: water temperature raises 1 ℃ weekly, reaches 12 ℃;
The constant temperature phase: when water temperature reaches 12 ℃, constant temperature is cultivated;
2) illumination adjustment
Adopt lighting; Intensity of illumination 500LX, light application time changes with the variation of water temperature;
Shorten the phase sunshine: from cooldown period, since the 11 hours light that contracts, light 1 hour contracted weekly;
The constant phase at sunshine:, determine light application time according to the water temperature situation when illumination drops to 8 hours;
Sunshine temperature raising period: according to the rising of water temperature,, reach 18 hours by increasing illumination in 1 hour weekly;
3) bait throwing in adjustment
Bait kind: shrimp 30%, clam worm 20%, yellow croaker 20%, clam 15%, oyster 10%, squid 5%;
Add vitamin: Vc, Ve, B B-complex weighs 1～2% with bait and adds, and throws something and feeds once in per 10 days;
Promote the gonad development medicine: lecithin, add with fish body weight 0.5%;
4) daily management
Throw something and feed: once a day, throw capacity, different diet collocations are thrown something and fed;
Change water: change water every day once, dirty cleaning every day in the end once;
Wash fish: observe the parent population situation of ingesting, it is bad to ingest, dipping 100ppm formaldehyde, 1h.;
5) hormone accelerating method:
Adopt HCG, LHRH hormone accelerating, generally carry out hastening parturition for twice: HCG 400VI/KG for the first time; HCG 200VI/KG for the second time; LHRH-A3 20ug/kg;
(2) semen collection, adopt ovum and fertilized egg hatching: carry out liquid nitrogen frozen after the artificial semen collection of going ahead of the rest and preserve, treat the after ripening raun adopt artificial squeeze ovum, sperm rewarming after the dry method insemination obtain fertilized egg; With specification is that 1m * 1m * 1m is hatched net cage, carries out early stage hatching with the demersal egg hatching m: 12 ± 0.5 ℃ of water temperatures 96 hours; When development of fertilized ova fertilized egg when body segment forms shows as half buoyancy, this moment, we collected the fertilized egg that suspends, and put into the hatchery and carried out 24 hour later stage 12 ± 0.6 ℃ of water temperatures and hatch prelarva; Change water 1/2 every day, illumination 500LX, early stage is inflation by force, little inflation of later stage;
(3) cultivation of fry
1) cultivates early stage
Nursery pond drops into prelarva, breeding density 5000～10,000 tail/rice after adding water to 1m behind cleaning and sterilizing
312～13 ℃ of water temperatures, prelarva be through 7 days initial feedings, the wheel animalcule and add chlorella of throwing something and feeding behind the prelarva opening, 5/ml of wheel animalcule density, day throws twice, changes water 2 times/day, each 1/2, bait throwing in is after changing water, add " preventing sinking to the bottom anti-stream device " simultaneously, prevent that fry from sinking and dead, the cultivation time in early stage is about 15 days;
2) later stage cultivates
Use specification to carry out the later stage cultivation, the prelarva of cultivating early stage is moved into for cultivating water body 10-15 times cement pit early stage; Later stage cultivates quantity of exchanged water and strengthens, and 2 times/day, continued the wheel animalcule of throwing something and feeding in each 2/3, the 15～35 day, throw something and feed twice every day, bait throwing in is after changing water, and water body wheel animalcule density keeps 5/ml; The fairy shrimp young of throwing something and feeding in the 20th～55 day day is thrown 1-2 time, and density remains on 1～2/ml, and this stage improves constantly water temperature, heats up every day till 0.5 ℃ to 16 ℃, and fry is grown fast;
After the 45th day, be aided with large-scale copepoda and artifical compound feed every day, after the 20th day, postlarva ingests more vigorous, and ight soil is more, begins to inhale at the end, inhales the end every day once; After the 40th day owing to there is the part postlarva to begin the attached end, inhale the necessary extreme care of end operation, the postlarva that sucks out puts together, with putting back in the pond after the 5ppm furans immersion sterilization in 20 minutes, the nurturing period, water PH in pond remained between 7.8～8 again, and dissolved oxygen is more than 4mg/L, general 5～7 days of the phase end of volt, this moment, bait was based on mixed bait S3 and fairy shrimp, and 16 ℃ of water temperatures day are changed water 3 times, full dose, this stage specification 2cm, enter volt end state after, fry is basicly stable, just when bait is changed, should change flowing water into, 16 ℃ of water temperatures, this stage specification 3～5cm this moment; After this normally culture the stage.
2. according to the seedling-cultivating method of the described dollar spot Scophthalmusmaximus of claim 1, it is characterized in that described in the processing step that the artificial fish-egg that squeezes ovum is to take the labeling method of " molecular labeling assist family breeding " technology to determine that the best adopts ovum opportunity: the parent population that the selection sexual gland will be grown, squeeze into muscle of back with the mark small of the stock through the bullet of sterilization, measure the coding of bullet with inductor, write down data, just can determine egg-laying time and the egg laying amount of every parent population when laying eggs.
3. according to the seedling-cultivating method of the described dollar spot Scophthalmusmaximus of claim 1, it is characterized in that carrying out liquid nitrogen frozen after the artificial semen collection described in the processing step preserves, its sperm with the ratio of batching is:
V
TS-2: V
Sperm=7: 10
V
DMSO: V
Sperm=2: 10
V
Calf serum: V
Sperm=1: 10
With sperm and the batch mixes test tube of packing into after evenly, sealing is put into liquid nitrogen container: on the liquid level 8～10 minutes, in the liquid level 5 minutes, put into fully then; Holding time can reach 1 year.
4. according to the seedling-cultivating method of the described dollar spot Scophthalmusmaximus of claim 1, it is characterized in that in the processing step that fry cultivates in earlier stage that described anti-to sink to the bottom anti-stream device be the equipment that a kind of recoil by current makes the sinking fry float again or suspend.
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| CN101341856B (en) * | 2007-07-11 | 2010-09-15 | 中国科学院海洋研究所 | Artificial insemination method for improving germ cell fertility rate of verasper moseri germ cell |
| CN101428139B (en) * | 2007-11-07 | 2012-06-27 | 中国科学院海洋研究所 | Method for extending spermiation period of verasper moseri |
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| CN101341856B (en) * | 2007-07-11 | 2010-09-15 | 中国科学院海洋研究所 | Artificial insemination method for improving germ cell fertility rate of verasper moseri germ cell |
| CN101428139B (en) * | 2007-11-07 | 2012-06-27 | 中国科学院海洋研究所 | Method for extending spermiation period of verasper moseri |
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