CN102812913B - Cultivation method for hybridized all-female golden trout fries - Google Patents
Cultivation method for hybridized all-female golden trout fries Download PDFInfo
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- CN102812913B CN102812913B CN201210320756.5A CN201210320756A CN102812913B CN 102812913 B CN102812913 B CN 102812913B CN 201210320756 A CN201210320756 A CN 201210320756A CN 102812913 B CN102812913 B CN 102812913B
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Abstract
The invention provides a cultivation method for hybridized all-female golden trout fries. An androgen is utilized to induce golden trout parent fish in a mature period so as to obtain golden trout pseudo-milter, steelhead parent fish eggs are fertilized with golden trout pseudo-milter sperms, hatching is performed, and the hybridized all-female golden trout fries are obtained. The eyed rate of the fertilized eggs during a hatching period is over 81%, the fry hatching rate is over 92%, the female rate of the obtained fries is 100% by detection, the rate of golden trout (with golden yellow body color) of the fries is over 90%, the weight gaining rate is over 122% after the fries are cultivated for 30 days, and the survival rate is over 98%. Compared with fries generated by purebred golden trout mating, the cultivation method for the hybridized all-female golden trout fries maintains the golden trout rate to be unchanged, enables the golden trout fries having double advantages of being hybridized and all-female to be obtained, and has good economic benefit and market value.
Description
Technical field
The present invention relates to aquatic biological breeding technical field, relate to particularly the breeding method of hybridizing complete female golden trout seed.
Background technology
Salmon trout is one of fish that mainly cultivate in the world at present, under natural conditions, they grow in long-term water temperature more and are no more than 20 ℃, water quality and are oligotrophy, do not have, in the low thermal water or clear brook of chemical contamination and organic contamination, to be therefore referred to as again cold water fishes.Because salmon fishes has very high requirement to existence water quality, slightly pollute and cannot survive, so salmon trout soma is clean, without any pollution, be genuine pollution-free green food, taste relieved.In addition, salmon trout is the fish of a kind of high protein, higher fatty acid, low cholesterol (cholesterol level is very little).Compare with other fish, salmon trout contains the glycine that general fish lack, and the content of vitamin A, B is also higher than other fish, and other fish are can't be obtained especially for the content of trace elements iron.Salmon trout is Fresh & Tender in Texture, naivete taste, and without fishy smell, without scaling, people eats weekly twice generation that salmon trout just can prevent thrombotic disease.Italy's expert up-to-date result of study shows, eats the health products that contain salmon trout and can significantly reduce the risk of cardiac's die by visitation of God.Eat 1 gram of DHA refining with salmon trout fish oil every day, the risk of die by visitation of God will reduce heart attack time.Therefore, salmon trout is a kind of delicious meat and has the fish that health care is worth.Due to the quality of himself and the specific (special) requirements to living environment, determine the character of its green, environmental protection nutritive food, be therefore quite subject to consumer's favor.Salmon trout is China's main cold water cultured fishes at present, due to reasons such as long-term inbreeding, shortage systematic breedings, cause cultivating individuality and occurred that sex premature, individual miniaturization, disease the problem such as grow, can not meet the demand of consumer to large specification salmon trout, cause culture benefit to decline.
Research shows: hereditary conservative can be sharply shaken in hybridization, makes the inheritance of hybrid be imbued with motility, has larger plasticity, thereby can improve rapidly and significantly the vitality of hybrid, and then obtains hybrid vigour; Hybrid can be enriched genetic structure, by kinds different more than two hereditary basis or the above individual independent assortment of genes of kind, occurs new hereditary form by hybridization, and people can select good individuality then, cultivate into new varieties.Under normal circumstances, the sexual maturity of raun late 1 year than milter, declines because sexual maturity causes again damp production, meat, and for fear of the output and the quality that affect fish because of early sexual maturity, hybridizing gynoecy cultivation is an approach that effectively improves culture benefit.
Domestic research prepared by salmon trout hybrid seedlings mainly concentrates on the hybrid seedlings of hybrid seedlings preparation, rainbow trout and the golden trout of hybrid seedlings preparation, rainbow trout and the mountain female trout of rainbow trout and brook trout and prepares etc., research prepared by the complete female seed of salmon trout mainly concentrates on the preparation aspect of the purebred complete female seed of rainbow trout, its method is: with the complete female dliploid seed of Artificial gynogenesis technology acquisition, become pseudo-milter by the complete female dliploid seed of androgenic hormone induction of throwing something and feeding again, cultivate to sexual maturity and utilize former raun and false milter to carry out artificial propagation.The method operation is relatively loaded down with trivial details, need to first obtain complete female dliploid seed, again seed is carried out to androgen induction, cultivate to sexual maturity, after pseudo-milter gonadal maturation, carry out again follow-up fertilization work, nearly 3 years consuming time of whole process, therefore the method wastes time and energy, and owing to being pedigree breeding, therefore the seedling of cultivating lacks heterosis, hybrid vigor.
Gold trout (Oncorhynchus mykiss) body is golden yellow, is a body colour separation kind for rainbow trout, is common cold water cultured fishes.At present the research of the technology of preparing of hybridizing complete female golden trout seed be there is not yet to report.
Summary of the invention
The object of the present invention is to provide the complete female golden trout offspring seed cultivation method of a kind of hybridization, to overcome the deficiencies in the prior art.
The breeding method of the complete female golden trout seed of hybridization provided by the invention, comprises the following steps:
1) obtain the pseudo-milter of golden trout with male sex function with the female golden trout parent population of androgenic hormone inducing maturation phase;
2) make the pseudo-milter sperm fertilization of rainbow trout (Salmo gairdneri) parent population ovum and golden trout;
3) incubating oosperm.
Wherein, the golden trout parent population in sexual maturing period is more than 3 ages described in step 1), body weight >=2kg.
Wherein, the method for the female golden trout parent population of androgenic hormone inducing maturation phase is described in step 1), by the golden trout parent population 70-100d that throws something and feeds containing the fish meal of androgenic hormone 40-100mg/kg; Afterwards, use the not fish meal containing androgenic hormone instead and throw something and feed 4-6 month, obtain the pseudo-milter of golden trout with male sex function.
Further, the method for the female golden trout parent population of androgenic hormone inducing maturation phase is described in step 1), by the golden trout parent population 85-90d that throws something and feeds containing the fish meal of androgenic hormone 40-50mg/kg; Afterwards, use the not fish meal containing androgenic hormone instead and throw something and feed 5 months, obtain the pseudo-milter of golden trout with male sex function.
Preferably, the above-mentioned feeding time by the golden trout parent population of throwing something and feeding containing the fish meal of androgenic hormone 40-50mg/kg is 90d.
Wherein, the androgenic hormone of step 1) is 17 α Methyltestosterones (17 α-methyltestosterone).
Further, described 17 α Methyltestosterones are added into before feed, dissolve with the alcohol of concentration >=60%, and the mass volume ratio of 17 α Methyltestosterones and alcohol is 1mg:1～4ml.10 times of 17 α Methyltestosterone solution clear water dilutions after dissolving are evenly sprayed on parent population feed afterwards, stir, make to mix, for subsequent use.
In the breeding method of the complete female golden trout seed of hybridization provided by the invention, step 2) the pseudo-milter sperm of described golden trout obtains by the following method: the pseudo-milter spermary of Mechanical Crushing, obtain after seminal fluid, activation of spermatozoa liquid and pseudo-milter seminal fluid are mixed into line activating according to volume ratio 20-30:1, described activation of spermatozoa liquid 1L formula is NaCl 7.60g, KCl 2.98g, CaCl
22H
2o 0.37g, MgCl
26H
2o 0.31g, NaHCO
30.21g, adding distil water is to 1000ml, and regulating pH value is 8.0-10.0; Activation condition is 2-6 ℃ and activates 3-4h.
Preferably, activation of spermatozoa liquid and pseudo-milter seminal fluid are mixed into line activating according to volume ratio 25:1, described activation of spermatozoa liquid pH value is 9.0; Activation condition is 4 ℃ and activates 3h.
Dry method fertilization method, the pseudo-milter seminal fluid of golden trout after getting the female fish-egg of rainbow trout (Salmo gairdneri) and activating is fertilized.
In the inventive method, incubation condition is that fertilized egg is hatched 45-65d in 8-10 ℃ of breeding water described in step 3).
In the inventive method, described golden trout is the golden trout of blood-shot eye illness.
The invention provides the application of said method in the complete female golden trout seed breeding work of hybridization.
The pseudo-milter of golden trout that the present invention uses the female golden trout parent population acquisition of androgenic hormone inducing maturation phase to have male sex function; Make the pseudo-milter sperm fertilization of rainbow trout (Salmo gairdneri) parent population ovum and golden trout, hatching, obtains the complete female golden trout seed of hybridization.Experiment detects to be found, between the complete female golden trout incubation period of hybridization prepared by the inventive method fertilized egg to send out a rate be more than 81%, hatching rate reaches more than 92%, the fry obtaining after testing, ratio of female is 100%, seed gold trout rate (body colour is for golden yellow) reaches more than 90%, and 30 days rates of body weight gain of larval rearing are more than 122%, and survival rate reaches more than 98%.Compared with the seed that the complete female golden trout offspring seed cultivation method of hybridization provided by the invention produces with purebred golden trout mating, both kept golden trout rate constant, obtain again the golden trout seed that possesses hybridization and complete female double dominant, and the seedling time is short, only about 9 months, greatly shorten the culture-cycle, reduced aquaculture cost, there is good economic benefit and market value.
Embodiment
Following examples further illustrate content of the present invention, but should not be construed as limitation of the present invention.Without departing from the spirit and substance of the case in the present invention, the modification that the inventive method, step or condition are done or replacement, all belong to scope of the present invention.
If do not specialize, the conventional means that in embodiment, technological means used is well known to those skilled in the art.Rainbow trout (Salmo gairdneri) parent population of the present invention and the golden trout parent population of blood-shot eye illness are Beijing Fisheries Research Institute and are cultivated and formed by the eyed eggs of agate bio tech ltd, Northern Europe purchase.17 α Methyltestosterones are purchased from German SIGMA company, and purity is more than 98%.
The activation of spermatozoa liquid 1L formula that the present invention uses is NaCl 7.60g, KCl 2.98g, CaCl
22H
2o 0.37g, MgCl
26H
2o 0.31g, NaHCO
30.21g, adding distil water is to 1000ml.
Embodiment 1 is hybridized complete female offspring seed cultivation method (1)
1, dissolve 17 α Methyltestosterones with the alcohol that purity is 65%, the mass volume ratio of 17 α Methyltestosterones and alcohol is 1mg:2ml, Methyltestosterone solution after dissolving is with after 10 times of clear water dilutions, be sprayed on uniformly on parent population feed with watering can, the content that makes 17 α Methyltestosterones in feed is 60mg/kg.Repeatedly stir, it is mixed, dry in the shade in ventilation for subsequent use.
2, more than selecting for 3 ages, specification reaches 2kg, sexually matured female blood-shot eye illness gold trout parent population is put into independent culturing pool, the feed that contains 17 α Methyltestosterones having prepared in the step 1 of throwing something and feeding, feed 100d according to conventional raising method, change afterwards to throw normally and do not continue to feed 4 months containing the parent population feed of 17 α Methyltestosterones.In the female blood-shot eye illness gold trout of normally feeding again after 17 α Methyltestosterone inductions, select the upper jaw in form and be slightly longer than lower jaw, lower jaw front end is false milter (pseudo-milter) to the blood-shot eye illness gold trout parent population of the 17 α Methyltestosterone feeds of throwing something and feeding of upper process.Detect and find, the present invention induces after female blood-shot eye illness gold trout parent population by 17 α Methyltestosterones, and the sexual gland that obtains expection reverses milter, and the occurrence rate of pseudo-milter is 95%.
Take out the spermary of pseudo-milter, grind spermary with mortar, filtered through gauze, the activation of spermatozoa liquid dilution (V/V=1:20) that seminal fluid is 8.5 by pH value, deposits under 4 ℃ of conditions of refrigerator activation of spermatozoa, makes it have normal sperm function, and the activation of spermatozoa time is 3h.
Fish-egg adopts dry method fertilization.First adopt extrusion that fish-egg is clamp-oned in clean water basin fully-developed rainbow trout (Salmo gairdneri) raun, every 4 tail rauns are 1 group.Again the pseudo-milter seminal fluid of blood-shot eye illness gold trout through activating is poured in the basin of fish-egg, stirred immediately, by adding Spawning water with the volume ratio 150:1 that activates seminal fluid, stir gently, after 1min, water is poured out, 3 times so repeatedly, clean.
3, the hatching pail that can put into the Spawning water of 8 ℃ after fertilized egg is cleaned up is normally hatched.Go out fry through 45 days incubating oosperms.It is the complete female golden trout of flavous hybridization that the fry hatching is body colour.
The complete female fish fertilized egg of hybridization that adopts the method processing to obtain is sent out a rate and is reached 82%, and hatching rate reaches 95%, and the female sex rate of seed is 100%, and seed gold trout rate (being that body colour is golden yellow) reaches 93%.Compared with the seed gold trout rate that the inventive method explanation produces with purebred golden trout mating, the golden trout rate of the complete female golden trout seed of hybridization that the inventive method obtains is constant, hybridizes complete female golden trout seed simultaneously and have again the double dominant of hybridization and complete female seed.
Embodiment 2 is hybridized complete female offspring seed cultivation method (2)
1, dissolve 17 α Methyltestosterones with the alcohol that purity is 70%, the mass volume ratio of 17 α Methyltestosterones and alcohol is 1mg:1ml.Methyltestosterone solution after dissolving is with after 10 times of clear water dilutions, is sprayed on uniformly on parent population feed with watering can, and the content that makes 17 α Methyltestosterones in feed is 20mg/kg.Repeatedly stir, it is mixed, dry in the shade in ventilation for subsequent use.
2, more than selecting for 3 ages, specification reaches 2.1kg, sexually matured female blood-shot eye illness gold trout parent population is put into independent culturing pool, the feed that contains 17 α Methyltestosterones having prepared in the step 1 of throwing something and feeding, feed 120d according to conventional raising method, change afterwards to throw normally and do not continue to feed 5 months containing the parent population feed of 17 α Methyltestosterones.In the female blood-shot eye illness gold trout of normally feeding again after 17 α Methyltestosterone inductions, select the upper jaw in form and be slightly longer than lower jaw, lower jaw front end is false milter (pseudo-milter) to the blood-shot eye illness gold trout parent population of the 17 α Methyltestosterone feeds of throwing something and feeding of upper process.Detect and find, the present invention induces after female blood-shot eye illness gold trout parent population by 17 α Methyltestosterones, and the sexual gland that obtains expection reverses milter, and the occurrence rate of pseudo-milter is 74%.
Take out the spermary of pseudo-milter, grind spermary with mortar, filtered through gauze, the activation of spermatozoa liquid dilution (V/V=1:20) that seminal fluid is 8.0 by pH value, deposits under 4 ℃ of conditions of refrigerator activation of spermatozoa, makes it have normal sperm function, and the activation of spermatozoa time is 3h.
Fish-egg adopts dry method fertilization.First adopt extrusion that fish-egg is clamp-oned in clean water basin fully-developed rainbow trout (Salmo gairdneri) raun, every 4 tail rauns are 1 group.Again the pseudo-milter seminal fluid of blood-shot eye illness gold trout through activating is poured in same basin, stirred immediately, by adding Spawning water with the volume ratio 140:1 that activates seminal fluid, stir gently, after 1min, water is poured out, 4 times so repeatedly, clean.
3, the hatching pail that can put into the Spawning water of 8 ℃ after fertilized egg is cleaned up is normally hatched.Go out fry through 55 days incubating oosperms.It is the complete female golden trout of flavous hybridization that the fry hatching is body colour.
The complete female fish fertilized egg of hybridization that adopts the method processing to obtain is sent out a rate and is reached 85%, and hatching rate reaches 94%, and the female sex rate of seed is 100%, and seed gold trout rate (being that body colour is golden yellow) reaches 94%.Compared with the seed gold trout rate that the inventive method explanation produces with purebred golden trout mating, the golden trout rate of the complete female golden trout seed of hybridization that the inventive method obtains is constant, hybridizes complete female golden trout seed simultaneously and have again the double dominant of hybridization and complete female seed.
Embodiment 3 is hybridized complete female offspring seed cultivation method (3)
1, dissolve 17 α Methyltestosterones with the alcohol that purity is 60%, the mass volume ratio of 17 α Methyltestosterones and alcohol is to be sprayed on uniformly on parent population feed with watering can after the Methyltestosterone solution clear water after 1mg:2ml dissolves dilutes 10 times, and the content that makes 17 α Methyltestosterones in feed is 40mg/kg.Repeatedly stir, it is mixed, dry in the shade in ventilation for subsequent use.
2, more than selecting for 3 ages, specification reaches 2.15kg, sexually matured female blood-shot eye illness gold trout parent population is put into independent culturing pool, the feed that contains 17 α Methyltestosterones having prepared in the step 1 of throwing something and feeding, feed 93d according to conventional raising method, change afterwards to throw normally and do not continue to feed 5 months containing the parent population feed of 17 α Methyltestosterones.In the female blood-shot eye illness gold trout of normally feeding again after 17 α Methyltestosterone inductions, select the upper jaw in form and be slightly longer than lower jaw, lower jaw front end is false milter (pseudo-milter) to the blood-shot eye illness gold trout parent population of the 17 α Methyltestosterone feeds of throwing something and feeding of upper process.Detect and find, the present invention induces after female blood-shot eye illness gold trout parent population by 17 α Methyltestosterones, and the sexual gland that obtains expection reverses milter, and the occurrence rate of pseudo-milter is 99%.
Take out the spermary of pseudo-milter, grind spermary with mortar, filtered through gauze, the activation of spermatozoa liquid dilution (V/V=1:25) that seminal fluid is 9.0 by pH value, deposits under 4 ℃ of conditions of refrigerator activation of spermatozoa, makes it have normal sperm function, and the activation of spermatozoa time is 3.5h.
Fish-egg adopts dry method fertilization.First adopt extrusion that fish-egg is clamp-oned in clean water basin fully-developed rainbow trout (Salmo gairdneri) raun, every 4 tail rauns are 1 group.Again the pseudo-milter seminal fluid of blood-shot eye illness gold trout through activating is poured in same basin, stirred immediately, by adding Spawning water with the volume ratio 145:1 that activates seminal fluid, stir gently, after 1min, water is poured out, 4 times so repeatedly, clean.
3, the hatching pail that can put into the Spawning water of 9 ℃ after fertilized egg is cleaned up is normally hatched.Go out fry through 53 days incubating oosperms.It is the complete female golden trout of flavous hybridization that the fry hatching is body colour.
The complete female fish fertilized egg of hybridization that adopts the method processing to obtain is sent out a rate and is reached 90%, and hatching rate reaches 96%, and the female sex rate of seed is 100%, and seed gold trout rate (being that body colour is golden yellow) reaches 98%.Compared with the seed gold trout rate that the inventive method explanation produces with purebred golden trout mating, the golden trout rate of the complete female golden trout seed of hybridization that the inventive method obtains is constant, hybridizes complete female golden trout seed simultaneously and have again the double dominant of hybridization and complete female seed.
Embodiment 4 is hybridized complete female offspring seed cultivation method (4)
1, dissolve 17 α Methyltestosterones with the alcohol that purity is 70%, the mass volume ratio of 17 α Methyltestosterones and alcohol is 1mg:3ml.After Methyltestosterone solution after dissolving dilutes 10 times with clear water, be sprayed on uniformly on parent population feed with watering can, the content that makes 17 α Methyltestosterones in feed is 50mg/kg.Repeatedly stir, it is mixed, dry in the shade in ventilation for subsequent use.
2, more than selecting for 3 ages, specification reaches 2.1kg, sexually matured female blood-shot eye illness gold trout parent population is put into independent culturing pool, the feed that contains 17 α Methyltestosterones having prepared in the step 1 of throwing something and feeding, feed 90d according to conventional raising method, change afterwards to throw normally and do not continue to feed 5 months containing the parent population feed of 17 α Methyltestosterones.In the female blood-shot eye illness gold trout of normally feeding again after 17 α Methyltestosterone inductions, select the upper jaw in form and be slightly longer than lower jaw, lower jaw front end is false milter (pseudo-milter) to the blood-shot eye illness gold trout parent population of the 17 α Methyltestosterone feeds of throwing something and feeding of upper process.Detect and find, the present invention induces after female blood-shot eye illness gold trout parent population by 17 α Methyltestosterones, and the sexual gland that obtains expection reverses milter, and the occurrence rate of pseudo-milter is 100%.
Take out the spermary of pseudo-milter, grind spermary with mortar, filtered through gauze, the activation of spermatozoa liquid dilution (V/V=1:25) that seminal fluid is 9.0 by pH value, deposits under 4 ℃ of conditions of refrigerator activation of spermatozoa, makes it have normal sperm function, and the activation of spermatozoa time is 3.5h.
Fish-egg adopts dry method fertilization.First adopt extrusion that fish-egg is clamp-oned in clean water basin fully-developed rainbow trout (Salmo gairdneri) raun, every 4 tail rauns are 1 group.Again the pseudo-milter seminal fluid of blood-shot eye illness gold trout through activating is poured in same basin, stirred immediately, by adding Spawning water with the volume ratio 150:1 that activates seminal fluid, stir gently, after 1min, water is poured out, 4 times so repeatedly, clean.
3, the hatching pail that can put into the Spawning water of 10 ℃ after fertilized egg is cleaned up is normally hatched.Go out fry through 55 days incubating oosperms.It is the complete female golden trout of flavous hybridization that the fry hatching is body colour.
The complete female fish fertilized egg of hybridization that adopts the method processing to obtain is sent out a rate and is reached 88%, and hatching rate reaches 95%, and the female sex rate of seed is 100%, and seed gold trout rate (being that body colour is golden yellow) reaches 97%.Compared with the seed gold trout rate that the inventive method explanation produces with purebred golden trout mating, the golden trout rate of the complete female golden trout seed of hybridization that the inventive method obtains is constant, hybridizes complete female golden trout seed simultaneously and have again the double dominant of hybridization and complete female seed.
Embodiment 5 is hybridized complete female offspring seed cultivation method (5)
1, dissolve 17 α Methyltestosterones with the alcohol that purity is 75%, the mass volume ratio of 17 α Methyltestosterones and alcohol is 1mg:4ml.After Methyltestosterone solution after dissolving dilutes 10 times with clear water, be sprayed on uniformly on parent population feed with watering can, the content that makes 17 α Methyltestosterones in feed is 100mg/kg.Repeatedly stir, it is mixed, dry in the shade in ventilation for subsequent use.
2, more than selecting for 3 ages, specification reaches 2kg, sexually matured female blood-shot eye illness gold trout parent population is put into independent culturing pool, the feed that contains 17 α Methyltestosterones having prepared in the step 1 of throwing something and feeding, feed 70d according to conventional raising method, change afterwards to throw normally and do not continue to feed 6 months containing the parent population feed of 17 α Methyltestosterones.In the blood-shot eye illness gold trout of normally feeding again after 17 α Methyltestosterone inductions, to select the upper jaw in form and be slightly longer than lower jaw, lower jaw front end is false milter (pseudo-milter) to the blood-shot eye illness gold trout parent population of the 17 α Methyltestosterone feeds of throwing something and feeding of upper process.Detect and find, the present invention induces after female blood-shot eye illness gold trout parent population by 17 α Methyltestosterones, and the sexual gland that obtains expection reverses milter, and the occurrence rate of pseudo-milter is 85%.
Take out the spermary of pseudo-milter, grind spermary with mortar, filtered through gauze, the activation of spermatozoa liquid dilution (V/V=1:25) that seminal fluid is 10.0 by pH value, deposits under 4 ℃ of conditions of refrigerator activation of spermatozoa, makes it have normal sperm function, and the activation of spermatozoa time is 4h.
Fish-egg adopts dry method fertilization.First adopt extrusion that fish-egg is clamp-oned in clean water basin fully-developed rainbow trout (Salmo gairdneri) raun, every 4 tail rauns are 1 group.Again the pseudo-milter seminal fluid of blood-shot eye illness gold trout through activating is poured in the basin of fish-egg, stirred immediately, by adding Spawning water with the volume ratio 130:1 that activates seminal fluid, stir gently, after 1min, water is poured out, 5 times so repeatedly, clean.
3, the hatching pail that can put into the Spawning water of 9 ℃ after fertilized egg is cleaned up is normally hatched.Go out fry through 52 days incubating oosperms.It is the complete female golden trout of flavous hybridization that the fry hatching is body colour.
The complete female fish fertilized egg of hybridization that adopts the method processing to obtain is sent out a rate and is reached 81%, and hatching rate reaches 92%, and the female sex rate of seed is 100%, and seed gold trout rate (being that body colour is golden yellow) reaches 90%.Compared with the seed gold trout rate that the inventive method explanation produces with purebred golden trout mating, the golden trout rate of the complete female golden trout seed of hybridization that the inventive method obtains is constant, hybridizes complete female golden trout seed simultaneously and have again the double dominant of hybridization and complete female seed.
Embodiment 6 is hybridized complete female offspring seed cultivation method (6)
1, dissolve 17 α Methyltestosterones with the alcohol that purity is 65%, the mass volume ratio of 17 α Methyltestosterones and alcohol is 1mg:2ml.After Methyltestosterone solution after dissolving dilutes 10 times with clear water, be sprayed on uniformly on parent population feed with watering can, the content that makes 17 α Methyltestosterones in feed is 80mg/kg.Repeatedly stir, it is mixed, dry in the shade in ventilation for subsequent use.
2, more than selecting for 3 ages, specification reaches 2.2kg, sexually matured female blood-shot eye illness gold trout parent population is put into independent culturing pool, the feed that contains 17 α Methyltestosterones having prepared in the step 1 of throwing something and feeding, feed 80d according to conventional raising method, change afterwards to throw normally and do not continue to feed 6 months containing the parent population feed of 17 α Methyltestosterones.In the female blood-shot eye illness gold trout of normally feeding again after 17 α Methyltestosterone inductions, select the upper jaw in form and be slightly longer than lower jaw, lower jaw front end is false milter (pseudo-milter) to the blood-shot eye illness gold trout parent population of the 17 α Methyltestosterone feeds of throwing something and feeding of upper process.Detect and find, the present invention induces after female blood-shot eye illness gold trout parent population by 17 α Methyltestosterones, and the sexual gland that obtains expection reverses milter, and the occurrence rate of pseudo-milter is 86%.
Take out the spermary of pseudo-milter, grind spermary with mortar, filtered through gauze, the activation of spermatozoa liquid dilution (V/V=1:30) that seminal fluid is 9.0 by pH value, deposits under 4 ℃ of conditions of refrigerator activation of spermatozoa, makes it have normal sperm function, and the activation of spermatozoa time is 4h.
Fish-egg adopts dry method fertilization.First adopt extrusion that fish-egg is clamp-oned in clean water basin fully-developed rainbow trout (Salmo gairdneri) raun, every 4 tail rauns are 1 group.Again the pseudo-milter seminal fluid of blood-shot eye illness gold trout through activating is poured in the basin of fish-egg, stirred immediately, by adding Spawning water with the volume ratio 130:1 that activates seminal fluid, stir gently, after 1min, water is poured out, 3 times so repeatedly, clean.
3, the hatching pail that can put into the Spawning water of 8 ℃ after fertilized egg is cleaned up is normally hatched.Go out fry through 55 days incubating oosperms.It is the complete female golden trout of flavous hybridization that the fry hatching is body colour.
The complete female fish fertilized egg of hybridization that adopts the method processing to obtain is sent out a rate and is reached 85%, and hatching rate reaches 95%, and the female sex rate of seed is 100%, and seed gold trout rate (being that body colour is golden yellow) reaches 93%.Compared with the seed gold trout rate that the inventive method explanation produces with purebred golden trout mating, the golden trout rate of the complete female golden trout seed of hybridization that the inventive method obtains is constant, hybridizes complete female golden trout seed simultaneously and have again the double dominant of hybridization and complete female seed.
The complete female seed growth rate of hybridization and survival rate experiments experiment that embodiment 7 the present invention cultivate are used fish
Get embodiment 1-6 and cultivate the complete female golden trout seed of the hybridization obtaining, each embodiment gets 6000 tails.Purebred rainbow trout (Salmo gairdneri) and each 6000 tails of purebred golden trout.Average specification is 1.5～1.8g/ tail fry.
2, method
Under 8 ℃ of conditions of water temperature, above-mentioned fry, by kind difference and preparation batch difference, to be put in a suitable place to breed respectively in 8 culturing pools, breeding density is 3000 tails/m
2.The throw something and feed seed material of Beijing Hanye Technology Co., Ltd., throws something and feeds 5 every day, and daily ration, feeding quantity is 5%, and water exchange capacity is 1 time/2h.Observe and record body weight and the growth pattern of 30 days each fries.The results are shown in Table 1.
The growth rate comparison (X ± SD) in 30 days of feeding of the different fries of table 1
| Kind | 0d | 30d | Rate of body weight gain % |
| The fry of embodiment 1 | 1.61±0.15 | 3.60±0.24 | 123.6 |
| The fry of embodiment 2 | 1.59±0.09 | 3.56±1.15 | 123.8 |
| The fry of embodiment 3 | 1.63±0.10 | 3.63±0.16 | 122.8 |
| The fry of embodiment 4 | 1.62±0.15 | 3.63±0.24 | 123.9 |
| The fry of embodiment 5 | 1.61±0.12 | 3.60±0.19 | 123.5 |
| The fry of embodiment 6 | 1.63±0.08 | 3.65±0.13 | 124.2 |
| The false milter of gold trout raun × rainbow trout (Salmo gairdneri) | 1.61±0.11 | 3.47±0.15 | 115.6 |
| Purebred rainbow trout (Salmo gairdneri) | 1.65±0.08 | 3.35±0.12 | 104.2 |
| Purebred golden trout | 1.63±0.13 | 3.17±0.18 | 94.6 |
Weightening finish experimental result finds, the complete female golden trout seed growth rate of hybridization that embodiment of the present invention 1-6 method is cultivated is obviously faster than the growth rate of purebred rainbow trout (Salmo gairdneri) and purebred golden trout.In the duration of test embodiment 1-6 fry phenomenon of not falling ill, survival rate is 98.8%, the purebred rainbow trout (Salmo gairdneri) survival rate of fish fry of check experiment group is 98.1%, and the survival rate of purebred golden trout is 94.2%, and the survival rate of the complete female golden trout seed of visible hybridization of the present invention is higher than purebred rainbow trout (Salmo gairdneri) and purebred golden trout.
Claims (5)
1. a breeding method for the complete female golden trout seed of hybridization, is characterized in that, comprises the following steps:
1) obtain the pseudo-milter of golden trout with male sex function with the female golden trout parent population of androgenic hormone inducing maturation phase; The golden trout parent population in described sexual maturing period is more than 3 ages, body weight >=2kg; The method of the female golden trout parent population of described androgenic hormone inducing maturation phase is, by the golden trout parent population 70-100d that throws something and feeds containing the fish meal of androgenic hormone 40-100mg/kg; Afterwards, use the not fish meal containing androgenic hormone instead and throw something and feed 4-6 month, obtain the pseudo-milter of golden trout with male sex function;
2) make the pseudo-milter sperm fertilization of the female fish parent fish ovum of rainbow trout (Salmo gairdneri) and golden trout; Described golden trout is the golden trout of blood-shot eye illness; The pseudo-milter sperm of described golden trout obtains by the following method: the pseudo-milter spermary of Mechanical Crushing, obtain after seminal fluid, activation of spermatozoa liquid and pseudo-milter seminal fluid are activated according to volume ratio 20-30:1, described activation of spermatozoa liquid 1L formula is NaCl7.60g, KCl2.98g, CaCl
22H
2o0.37g, MgCl
26H
2o0.31g, NaHCO
30.21g adding distil water is to 1000ml, and regulating pH value is 8.0-10.0; Activation condition is 2-6 ℃ and activates 3-4h;
3) incubating oosperm, incubation condition is that fertilized egg is hatched 45-65d in 8-10 ℃ of breeding water.
2. a kind of breeding method of hybridizing complete female golden trout seed as claimed in claim 1, it is characterized in that, described in step 1), the method for the female golden trout parent population of androgenic hormone inducing maturation phase is, by the golden trout parent population 85-95d that throws something and feeds containing the fish meal of androgenic hormone 40-50mg/kg; Afterwards, use the not fish meal containing androgenic hormone instead and throw something and feed 5 months, obtain the pseudo-milter of golden trout with male sex function.
3. a kind of breeding method of hybridizing complete female golden trout seed as claimed in claim 1, it is characterized in that, the androgenic hormone of step 1) is 17 α Methyltestosterones, before described 17 α Methyltestosterones are added into feed, dissolve 17 α Methyltestosterones with the alcohol that purity is 65%, the mass volume ratio of 17 α Methyltestosterones and alcohol is 1mg:2ml, Methyltestosterone solution after dissolving is with after 10 times of clear water dilutions, be sprayed on uniformly on parent population feed with watering can, the content that makes 17 α Methyltestosterones in feed is 60mg/kg; Repeatedly stir, it is mixed, dry in the shade in ventilation for subsequent use.
4. a kind of breeding method of hybridizing complete female golden trout seed as claimed in claim 1, is characterized in that, activation of spermatozoa liquid and pseudo-milter seminal fluid are activated according to volume ratio 25:1, and described activation of spermatozoa liquid pH value is 9.0; Activation condition is 4 ℃ and activates 3h.
5. the application of the method for the complete female golden trout seed of the arbitrary described hybridization of claim 1-4 in the complete female golden trout seed breeding work of hybridization.
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| CN103583427B (en) * | 2013-11-06 | 2015-09-23 | 广西壮族自治区水产科学研究院 | Pseudo-male and complete female bolti producing method for seed |
| CN106417256B (en) * | 2016-10-12 | 2020-01-31 | 中国水产科学研究院黑龙江水产研究所 | method for improving sperm utilization rate of rainbow trout pseudo-male fish |
| CN114532259B (en) * | 2022-03-01 | 2022-10-18 | 中国水产科学研究院黑龙江水产研究所 | Large-scale rainbow trout hologynic triploid seed production method |
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