CN102812913A - 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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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 the aquatic biological breeding technical field, relate to the breeding method of hybridizing complete female golden trout seed particularly.
Background technology
The salmon trout is one of fish of mainly culturing in the world at present; 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 under the natural conditions, therefore are referred to as cold water fishes again.Because salmon fishes has very high requirement to existence water quality, slightly pollute and promptly can't survive, so salmon trout soma is clean, there is not any pollution, be genuine pollution-free green food, it is relieved to taste.In addition, the 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, the salmon trout contains the glycine that general fish lack, and the content of vitamin A, B also is higher than other fish, and other fish are can't be obtained especially for the content of trace elements iron.The salmon trout is Fresh & Tender in Texture, the naivete flavor, and no fishy smell need not to scale, and the people eats twice generation that the salmon trout just can prevent thrombotic disease weekly.Up-to-date result of study of Italy expert shows, eats the risk that the health products that contain the salmon trout can reduce cardiac's die by visitation of God significantly.Eat 1 gram every day with the DHA that salmon trout fish oil refines, the risk of die by visitation of God in the time of will reducing heart attack.Therefore, the salmon trout is a kind of delicious meat and has the fish that health care is worth.Because himself quality and to the specific (special) requirements of living environment, determined the character of its green, environmental protection nutritive food, therefore quite receive consumer's favor.The salmon trout is China's main cold water property cultured fishes at present; Owing to reasons such as long-term inbreeding, shortage systematic breedings; Causing culturing individuality sex premature, individual miniaturization, disease have occurred and problem such as has grown; Can not satisfy the demand of consumer, cause culture benefit to descend big specification salmon trout.
Research shows: hybridization can be rapid shakes the conservative of heredity, makes the inheritance of hybrid be imbued with motility, has bigger plasticity, thereby can be rapidly and improve the vitality of hybrid significantly, and then obtains hybrid vigour; Hybrid can be enriched genetic structure, with two kind or above individual independent assortment of genes of kind that above hereditary basis is different, occurs new hereditary form through hybridization, and people can select good individuality then, cultivate into new varieties.Under the normal condition, the sexual maturity of raun late 1 year than milter descends because sexual maturity causes damp production, meat again, and for fear of the output and the quality that influence fish because of early sexual maturity, hybridizing the gynoecy breed is an approach that effectively improves culture benefit.
Domestic research to salmon trout hybrid seedlings preparation mainly concentrates on the hybrid seedlings preparation etc. of hybrid seedlings preparation, rainbow trout and golden trout of the women trout in hybrid seedlings preparation, rainbow trout and mountain of rainbow trout and brook trout; The research of the complete female seed preparation of salmon trout is mainly concentrated on the preparation aspect of the purebred complete female seed of rainbow trout; Its method is: obtain complete female dliploid seed with artificial induction's gynogenesis technology; Induce complete female dliploid seed to become pseudo-milter through the androgenic hormone of throwing something and feeding again, cultivate to sexual maturity and utilize former raun and false milter to carry out artificial propagation.This method operation is loaded down with trivial details relatively, needs to obtain earlier complete female dliploid seed, again seed is carried out androgen and induces; Cultivate to sexual maturity; Treat to carry out follow-up fertilization work again behind the pseudo-milter gonadal maturation, nearly 3 years consuming time of whole process, so this method wastes time and energy; And owing to be pedigree breeding, the seedling of cultivating lacks heterosis, hybrid vigor.
Gold trout (Oncorhynchus mykiss) body is golden yellow, is that the body colour separation of rainbow trout is planted, and is common cold water property cultured fishes.At present the research of the technology of preparing of hybridizing complete female golden trout seed is not seen that as yet report is arranged.
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 deficiency of prior art.
The breeding method of the complete female golden trout seed of hybridization provided by the invention may further comprise the steps:
1) obtains to have the pseudo-milter of golden trout of male sex function with the female golden trout parent population in androgenic hormone inductivity maturing stage;
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 step 1) said sexual maturing period is more than 3 ages, body weight >=2kg.
Wherein, the method for the female golden trout parent population in said androgenic hormone inductivity maturing stage of step 1) does, the fish meal that will the contain androgenic hormone 40-100mg/kg golden trout parent population 70-100d that throws something and feeds; Afterwards, use the fish meal that does not contain androgenic hormone instead and threw something and fed 4-6 month, obtain to have the pseudo-milter of golden trout of male sex function.
Further, the method for the female golden trout parent population in said androgenic hormone inductivity maturing stage of step 1) does, the fish meal that will the contain androgenic hormone 40-50mg/kg golden trout parent population 85-90d that throws something and feeds; Afterwards, use the fish meal that does not contain androgenic hormone instead and threw something and fed 5 months, obtain to have the pseudo-milter of golden trout of male sex function.
Preferably, the throw something and feed feeding time of golden trout parent population of the above-mentioned fish meal that will contain androgenic hormone 40-50mg/kg is 90d.
Wherein, the androgenic hormone of step 1) is 17 α Methyltestosterones (17 α-methyltestosterone).
Further, before said 17 α Methyltestosterones are added into feed, dissolve with the alcohol of concentration >=60%, the mass volume ratio of 17 α Methyltestosterones and alcohol is 1mg:1～4ml.17 α Methyltestosterone solution after the dissolving evenly are sprayed on the parent population feed with after 10 times of the clear water dilutions, stir, make to mix, 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 said golden trout obtains through following method: the pseudo-milter spermary of Mechanical Crushing; After obtaining seminal fluid, activation of spermatozoa liquid and pseudo-milter seminal fluid are mixed into line activating according to volume ratio 20-30:1, said activation of spermatozoa liquid 1L prescription 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, regulating the 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, said 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 rainbow trout (Salmo gairdneri) raun ovum and activating is fertilized.
In the inventive method, the said incubation condition of step 3) is that fertilized egg is hatched 45-65d in 8-10 ℃ of breeding water.
In the inventive method, said 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 present invention uses the female golden trout parent population in androgenic hormone inductivity maturing stage to obtain to have the pseudo-milter of golden trout of 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; The complete female golden trout of hybridization of the inventive method preparation between the incubation period fertilized egg to send out a rate be more than 81%, the seed incubation rate reaches more than 92%, the fry that obtains is through detecting; Ratio of female is 100%; Seed gold trout rate (body colour is for golden yellow) reaches more than 90%, and it is more than 122% that seed is cultivated 30 days rates of body weight gain, and survival rate reaches more than 98%.The complete female golden trout offspring seed cultivation method of hybridization provided by the invention is compared with the seed that purebred golden trout mating produces; Both kept golden trout rate constant, and obtained to possess the golden trout seed of hybridization and complete female double dominant again, and the seedling time is short; Only about 9 months; Shorten the culture-cycle greatly, reduced aquaculture cost, had favorable economic benefit and market value.
Embodiment
Following examples further specify content of the present invention, but should not be construed as limitation of the present invention.Under the situation that does not deviate from the present invention's spirit and essence, modification or replacement to the inventive method, step or condition are done all belong to scope of the present invention.
If do not specialize the conventional means that used technological means is well known to those skilled in the art among the embodiment.Rainbow trout (Salmo gairdneri) parent population of the present invention and the golden trout parent population of blood-shot eye illness are eyed eggs that Beijing Fisheries Research Institute buys by Northern Europe agate bio tech ltd and cultivate and form.17 α Methyltestosterones are available from German SIGMA company, and purity is more than 98%.
The activation of spermatozoa liquid 1L prescription 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 hybridization complete female offspring seed cultivation method (1)
1, using purity is that 65% alcohol dissolves 17 α Methyltestosterones; The mass volume ratio of 17 α Methyltestosterones and alcohol is 1mg:2ml; After Methyltestosterone solution after the dissolving dilutes 10 times with clear water; Be sprayed on uniformly on the parent population feed with watering can, the content that makes 17 α Methyltestosterones in the feed is 60mg/kg.Stir repeatedly, it is mixed, it is subsequent use to dry in the shade in the ventilation.
2, select more than 3 ages; Specification reaches 2kg; Sexually matured female blood-shot eye illness gold trout parent population is put into independent breed pond; The feed that contains 17 α Methyltestosterones that has prepared in the step 1 of throwing something and feeding is fed 100d according to the raising method of routine, changes the normal parent population feed that does not contain 17 α Methyltestosterones of throwing afterwards and continues to feed 4 months.In the female blood-shot eye illness gold trout of after inducing, normally feeding again through 17 α Methyltestosterones; Select on form the upper jaw and be longer than lower jaw slightly, the 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, after the present invention induced female blood-shot eye illness gold trout parent population through 17 α Methyltestosterones, the sexual gland that obtains expection reversed milter, and the occurrence rate of pseudo-milter is 95%.
Take out the spermary of pseudo-milter, grind spermary with mortar, filtered through gauze, it is that 8.5 activation of spermatozoa liquid dilutes (V/V=1:20) that seminal fluid uses pH value, deposits under 4 ℃ of conditions of refrigerator activation of spermatozoa, makes it have normal sperm function, the activation of spermatozoa time is 3h.
Fish-egg adopts the dry method fertilization.Adopt extrusion that fish-egg is clamp-oned in the clean water basin fully-developed rainbow trout (Salmo gairdneri) raun earlier, per 4 tail rauns are 1 group.To pour in the basin of fish-egg through the pseudo-milter seminal fluid of blood-shot eye illness gold trout that activates again, stir immediately,, stir gently, behind the 1min water poured out, 3 times so repeatedly, clean by adding the hatching water with the volume ratio 150:1 that activates seminal fluid.
3, the hatching pail that can put into 8 ℃ hatching water 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 that hatches is body colour.
The full raun fertilized egg of hybridization of adopting this method processing to obtain is sent out a rate and is reached 82%, and the seed incubation rate reaches 95%, and the female ratio of seed is 100%, and seed gold trout rate (being that body colour is golden yellow) reaches 93%.The inventive method explanation is compared with the seed gold trout rate that purebred golden trout mating produces, and the golden trout rate of the complete female golden trout seed of hybridization that the inventive method obtains is constant, hybridizes the double dominant that complete female golden trout seed has hybridization and complete female seed again simultaneously.
Embodiment 2 hybridization complete female offspring seed cultivation methods (2)
1, using purity is that 70% alcohol dissolves 17 α Methyltestosterones, and the mass volume ratio of 17 α Methyltestosterones and alcohol is 1mg:1ml.Methyltestosterone solution after the dissolving is sprayed on the parent population feed with watering can after diluting 10 times with clear water uniformly, and the content that makes 17 α Methyltestosterones in the feed is 20mg/kg.Stir repeatedly, it is mixed, it is subsequent use to dry in the shade in the ventilation.
2, select more than 3 ages; Specification reaches 2.1kg; Sexually matured female blood-shot eye illness gold trout parent population is put into independent breed pond; The feed that contains 17 α Methyltestosterones that has prepared in the step 1 of throwing something and feeding is fed 120d according to the raising method of routine, changes the normal parent population feed that does not contain 17 α Methyltestosterones of throwing afterwards and continues to feed 5 months.In the female blood-shot eye illness gold trout of after inducing, normally feeding again through 17 α Methyltestosterones; Select on form the upper jaw and be longer than lower jaw slightly, the 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, after the present invention induced female blood-shot eye illness gold trout parent population through 17 α Methyltestosterones, the sexual gland that obtains expection reversed milter, and the occurrence rate of pseudo-milter is 74%.
Take out the spermary of pseudo-milter, grind spermary with mortar, filtered through gauze, it is that 8.0 activation of spermatozoa liquid dilutes (V/V=1:20) that seminal fluid uses pH value, deposits under 4 ℃ of conditions of refrigerator activation of spermatozoa, makes it have normal sperm function, the activation of spermatozoa time is 3h.
Fish-egg adopts the dry method fertilization.Adopt extrusion that fish-egg is clamp-oned in the clean water basin fully-developed rainbow trout (Salmo gairdneri) raun earlier, per 4 tail rauns are 1 group.To pour in the same basin through the pseudo-milter seminal fluid of blood-shot eye illness gold trout that activates again, stir immediately,, stir gently, behind the 1min water poured out, 4 times so repeatedly, clean by adding the hatching water with the volume ratio 140:1 that activates seminal fluid.
3, the hatching pail that can put into 8 ℃ hatching water 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 that hatches is body colour.
The full raun fertilized egg of hybridization of adopting this method processing to obtain is sent out a rate and is reached 85%, and the seed incubation rate reaches 94%, and the female ratio of seed is 100%, and seed gold trout rate (being that body colour is golden yellow) reaches 94%.The inventive method explanation is compared with the seed gold trout rate that purebred golden trout mating produces, and the golden trout rate of the complete female golden trout seed of hybridization that the inventive method obtains is constant, hybridizes the double dominant that complete female golden trout seed has hybridization and complete female seed again simultaneously.
Embodiment 3 hybridization complete female offspring seed cultivation methods (3)
1, using purity is that 60% alcohol dissolves 17 α Methyltestosterones; The mass volume ratio of 17 α Methyltestosterones and alcohol is to be sprayed on uniformly on the parent population feed with watering can after the Methyltestosterone solution after the 1mg:2ml dissolving dilutes 10 times with clear water, and the content that makes 17 α Methyltestosterones in the feed is 40mg/kg.Stir repeatedly, it is mixed, it is subsequent use to dry in the shade in the ventilation.
2, select more than 3 ages; Specification reaches 2.15kg; Sexually matured female blood-shot eye illness gold trout parent population is put into independent breed pond; The feed that contains 17 α Methyltestosterones that has prepared in the step 1 of throwing something and feeding is fed 93d according to the raising method of routine, changes the normal parent population feed that does not contain 17 α Methyltestosterones of throwing afterwards and continues to feed 5 months.In the female blood-shot eye illness gold trout of after inducing, normally feeding again through 17 α Methyltestosterones; Select on form the upper jaw and be longer than lower jaw slightly, the 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, after the present invention induced female blood-shot eye illness gold trout parent population through 17 α Methyltestosterones, the sexual gland that obtains expection reversed milter, and the occurrence rate of pseudo-milter is 99%.
Take out the spermary of pseudo-milter, grind spermary with mortar, filtered through gauze, it is that 9.0 activation of spermatozoa liquid dilutes (V/V=1:25) that seminal fluid uses pH value, deposits under 4 ℃ of conditions of refrigerator activation of spermatozoa, makes it have normal sperm function, the activation of spermatozoa time is 3.5h.
Fish-egg adopts the dry method fertilization.Adopt extrusion that fish-egg is clamp-oned in the clean water basin fully-developed rainbow trout (Salmo gairdneri) raun earlier, per 4 tail rauns are 1 group.To pour in the same basin through the pseudo-milter seminal fluid of blood-shot eye illness gold trout that activates again, stir immediately,, stir gently, behind the 1min water poured out, 4 times so repeatedly, clean by adding the hatching water with the volume ratio 145:1 that activates seminal fluid.
3, the hatching pail that can put into 9 ℃ hatching water 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 that hatches is body colour.
The full raun fertilized egg of hybridization of adopting this method processing to obtain is sent out a rate and is reached 90%, and the seed incubation rate reaches 96%, and the female ratio of seed is 100%, and seed gold trout rate (being that body colour is golden yellow) reaches 98%.The inventive method explanation is compared with the seed gold trout rate that purebred golden trout mating produces, and the golden trout rate of the complete female golden trout seed of hybridization that the inventive method obtains is constant, hybridizes the double dominant that complete female golden trout seed has hybridization and complete female seed again simultaneously.
Embodiment 4 hybridization complete female offspring seed cultivation methods (4)
1, using purity is that 70% alcohol dissolves 17 α Methyltestosterones, and the mass volume ratio of 17 α Methyltestosterones and alcohol is 1mg:3ml.Methyltestosterone solution after the dissolving is sprayed on the parent population feed with watering can after diluting 10 times with clear water uniformly, and the content that makes 17 α Methyltestosterones in the feed is 50mg/kg.Stir repeatedly, it is mixed, it is subsequent use to dry in the shade in the ventilation.
2, select more than 3 ages; Specification reaches 2.1kg; Sexually matured female blood-shot eye illness gold trout parent population is put into independent breed pond; The feed that contains 17 α Methyltestosterones that has prepared in the step 1 of throwing something and feeding is fed 90d according to the raising method of routine, changes the normal parent population feed that does not contain 17 α Methyltestosterones of throwing afterwards and continues to feed 5 months.In the female blood-shot eye illness gold trout of after inducing, normally feeding again through 17 α Methyltestosterones; Select on form the upper jaw and be longer than lower jaw slightly, the 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, after the present invention induced female blood-shot eye illness gold trout parent population through 17 α Methyltestosterones, the sexual gland that obtains expection reversed milter, and the occurrence rate of pseudo-milter is 100%.
Take out the spermary of pseudo-milter, grind spermary with mortar, filtered through gauze, it is that 9.0 activation of spermatozoa liquid dilutes (V/V=1:25) that seminal fluid uses pH value, deposits under 4 ℃ of conditions of refrigerator activation of spermatozoa, makes it have normal sperm function, the activation of spermatozoa time is 3.5h.
Fish-egg adopts the dry method fertilization.Adopt extrusion that fish-egg is clamp-oned in the clean water basin fully-developed rainbow trout (Salmo gairdneri) raun earlier, per 4 tail rauns are 1 group.To pour in the same basin through the pseudo-milter seminal fluid of blood-shot eye illness gold trout that activates again, stir immediately,, stir gently, behind the 1min water poured out, 4 times so repeatedly, clean by adding the hatching water with the volume ratio 150:1 that activates seminal fluid.
3, the hatching pail that can put into 10 ℃ hatching water 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 that hatches is body colour.
The full raun fertilized egg of hybridization of adopting this method processing to obtain is sent out a rate and is reached 88%, and the seed incubation rate reaches 95%, and the female ratio of seed is 100%, and seed gold trout rate (being that body colour is golden yellow) reaches 97%.The inventive method explanation is compared with the seed gold trout rate that purebred golden trout mating produces, and the golden trout rate of the complete female golden trout seed of hybridization that the inventive method obtains is constant, hybridizes the double dominant that complete female golden trout seed has hybridization and complete female seed again simultaneously.
Embodiment 5 hybridization complete female offspring seed cultivation methods (5)
1, using purity is that 75% alcohol dissolves 17 α Methyltestosterones, and the mass volume ratio of 17 α Methyltestosterones and alcohol is 1mg:4ml.Methyltestosterone solution after the dissolving is sprayed on the parent population feed with watering can after diluting 10 times with clear water uniformly, and the content that makes 17 α Methyltestosterones in the feed is 100mg/kg.Stir repeatedly, it is mixed, it is subsequent use to dry in the shade in the ventilation.
2, select more than 3 ages; Specification reaches 2kg; Sexually matured female blood-shot eye illness gold trout parent population is put into independent breed pond; The feed that contains 17 α Methyltestosterones that has prepared in the step 1 of throwing something and feeding is fed 70d according to the raising method of routine, changes the normal parent population feed that does not contain 17 α Methyltestosterones of throwing afterwards and continues to feed 6 months.In the blood-shot eye illness of after inducing through 17 α Methyltestosterones, normally feeding the again gold trout, select on form the upper jaw and be longer than lower jaw slightly, the 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, after the present invention induced female blood-shot eye illness gold trout parent population through 17 α Methyltestosterones, the sexual gland that obtains expection reversed milter, and the occurrence rate of pseudo-milter is 85%.
Take out the spermary of pseudo-milter, grind spermary with mortar, filtered through gauze, it is that 10.0 activation of spermatozoa liquid dilutes (V/V=1:25) that seminal fluid uses pH value, deposits under 4 ℃ of conditions of refrigerator activation of spermatozoa, makes it have normal sperm function, the activation of spermatozoa time is 4h.
Fish-egg adopts the dry method fertilization.Adopt extrusion that fish-egg is clamp-oned in the clean water basin fully-developed rainbow trout (Salmo gairdneri) raun earlier, per 4 tail rauns are 1 group.To pour in the basin of fish-egg through the pseudo-milter seminal fluid of blood-shot eye illness gold trout that activates again, stir immediately,, stir gently, behind the 1min water poured out, 5 times so repeatedly, clean by adding the hatching water with the volume ratio 130:1 that activates seminal fluid.
3, the hatching pail that can put into 9 ℃ hatching water 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 that hatches is body colour.
The full raun fertilized egg of hybridization of adopting this method processing to obtain is sent out a rate and is reached 81%, and the seed incubation rate reaches 92%, and the female ratio of seed is 100%, and seed gold trout rate (being that body colour is golden yellow) reaches 90%.The inventive method explanation is compared with the seed gold trout rate that purebred golden trout mating produces, and the golden trout rate of the complete female golden trout seed of hybridization that the inventive method obtains is constant, hybridizes the double dominant that complete female golden trout seed has hybridization and complete female seed again simultaneously.
Embodiment 6 hybridization complete female offspring seed cultivation methods (6)
1, using purity is that 65% alcohol dissolves 17 α Methyltestosterones, and the mass volume ratio of 17 α Methyltestosterones and alcohol is 1mg:2ml.Methyltestosterone solution after the dissolving is sprayed on the parent population feed with watering can after diluting 10 times with clear water uniformly, and the content that makes 17 α Methyltestosterones in the feed is 80mg/kg.Stir repeatedly, it is mixed, it is subsequent use to dry in the shade in the ventilation.
2, select more than 3 ages; Specification reaches 2.2kg; Sexually matured female blood-shot eye illness gold trout parent population is put into independent breed pond; The feed that contains 17 α Methyltestosterones that has prepared in the step 1 of throwing something and feeding is fed 80d according to the raising method of routine, changes the normal parent population feed that does not contain 17 α Methyltestosterones of throwing afterwards and continues to feed 6 months.In the female blood-shot eye illness gold trout of after inducing, normally feeding again through 17 α Methyltestosterones; Select on form the upper jaw and be longer than lower jaw slightly, the 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, after the present invention induced female blood-shot eye illness gold trout parent population through 17 α Methyltestosterones, the sexual gland that obtains expection reversed milter, and the occurrence rate of pseudo-milter is 86%.
Take out the spermary of pseudo-milter, grind spermary with mortar, filtered through gauze, it is that 9.0 activation of spermatozoa liquid dilutes (V/V=1:30) that seminal fluid uses pH value, deposits under 4 ℃ of conditions of refrigerator activation of spermatozoa, makes it have normal sperm function, the activation of spermatozoa time is 4h.
Fish-egg adopts the dry method fertilization.Adopt extrusion that fish-egg is clamp-oned in the clean water basin fully-developed rainbow trout (Salmo gairdneri) raun earlier, per 4 tail rauns are 1 group.To pour in the basin of fish-egg through the pseudo-milter seminal fluid of blood-shot eye illness gold trout that activates again, stir immediately,, stir gently, behind the 1min water poured out, 3 times so repeatedly, clean by adding the hatching water with the volume ratio 130:1 that activates seminal fluid.
3, the hatching pail that can put into 8 ℃ hatching water 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 that hatches is body colour.
The full raun fertilized egg of hybridization of adopting this method processing to obtain is sent out a rate and is reached 85%, and the seed incubation rate reaches 95%, and the female ratio of seed is 100%, and seed gold trout rate (being that body colour is golden yellow) reaches 93%.The inventive method explanation is compared with the seed gold trout rate that purebred golden trout mating produces, and the golden trout rate of the complete female golden trout seed of hybridization that the inventive method obtains is constant, hybridizes the double dominant that complete female golden trout seed has hybridization and complete female seed again simultaneously.
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 hybridization that obtains, each embodiment gets 6000 tails.Each 6000 tail of purebred rainbow trout (Salmo gairdneri) and purebred golden trout.Average specification is 1.5～1.8g/ tail fry.
2, method
Under 8 ℃ of conditions of water temperature, above-mentioned fry is different batch different with preparation by kind, to put in a suitable place to breed respectively in 8 breed ponds, breeding density is 3000 tails/m
2The throw something and feed seed material of Beijing Hanye Technology Co., Ltd. is thrown something and fed 5 every day, and daily ration, feeding quantity is 5%, and water exchange capacity is 1 time/2h.Observe and write down body weight and the growth pattern of 30 days each fries.The result sees table 1.
The different fries of the table 1 relatively (X ± SD) of 30 days growth rate that feeds
| 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 gold trout raun * false milter of 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 |
The weightening finish experimental result finds, the complete female golden trout seed growth rate of hybridization that embodiment of the 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 visible complete female golden trout survival rate of young of hybridization of the present invention is higher than purebred rainbow trout (Salmo gairdneri) and purebred golden trout.
Claims (10)
1. the breeding method of the complete female golden trout seed of hybridization is characterized in that, may further comprise the steps:
1) obtains to have the pseudo-milter of golden trout of male sex function with the female golden trout parent population in androgenic hormone inductivity maturing stage;
2) make the pseudo-milter sperm fertilization of rainbow trout (Salmo gairdneri) raun parent population ovum and golden trout;
3) incubating oosperm.
2. breeding method as claimed in claim 1 is characterized in that, the golden trout parent population in step 1) said sexual maturing period is more than 3 ages, body weight >=2kg.
3. breeding method as claimed in claim 1 is characterized in that, the method for the female golden trout parent population in said androgenic hormone inductivity maturing stage of step 1) does, the fish meal that will the contain androgenic hormone 40-100mg/kg golden trout parent population 70-100d that throws something and feeds; Afterwards, use the fish meal that does not contain androgenic hormone instead and threw something and fed 4-6 month, obtain to have the pseudo-milter of golden trout of male sex function.
4. breeding method as claimed in claim 3 is characterized in that, the method for the female golden trout parent population in said androgenic hormone inductivity maturing stage of step 1) does, the fish meal that will the contain androgenic hormone 40-50mg/kg golden trout parent population 85-95d that throws something and feeds; Afterwards, use the fish meal that does not contain androgenic hormone instead and threw something and fed 5 months, obtain to have the pseudo-milter of golden trout of male sex function.
5. breeding method as claimed in claim 1 is characterized in that, the androgenic hormone of step 1) is 17 α Methyltestosterones.
6. breeding method as claimed in claim 1; It is characterized in that step 2) the pseudo-milter sperm of said golden trout obtains through following method: the pseudo-milter spermary of Mechanical Crushing, behind the acquisition seminal fluid; Activation of spermatozoa liquid and pseudo-milter seminal fluid are activated according to volume ratio 20-30:1; Said activation of spermatozoa liquid 1L prescription 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, regulating the pH value is 8.0-10.0; Activation condition is 2-6 ℃ and activates 3-4h.
7. breeding method as claimed in claim 6 is characterized in that, activation of spermatozoa liquid and pseudo-milter seminal fluid are activated according to volume ratio 25:1, and said activation of spermatozoa liquid pH value is 9.0; Activation condition is 4 ℃ and activates 3h.
8. breeding method as claimed in claim 1 is characterized in that, the said incubation condition of step 3) is that fertilized egg is hatched 45-65d in 8-10 ℃ of breeding water.
9. like the arbitrary described breeding method of claim 1-8, it is characterized in that said golden trout is the golden trout of blood-shot eye illness.
10. the application of the arbitrary said method of claim 1-9 in the complete female golden trout seed breeding work of hybridization.
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| CN103583427A (en) * | 2013-11-06 | 2014-02-19 | 广西壮族自治区水产科学研究院 | Pseudo-male and full-female nile tilapia fingerling production method |
| CN106417256A (en) * | 2016-10-12 | 2017-02-22 | 中国水产科学研究院黑龙江水产研究所 | Method for increasing utilization rate of sperm of sex reversal milter of rainbow trout |
| CN114532259A (en) * | 2022-03-01 | 2022-05-27 | 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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Cited By (6)
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| CN103229742A (en) * | 2013-05-23 | 2013-08-07 | 北京市水产科学研究所 | Preparation method of oncorhynchus mykiss walbaum and oncorhynchus mykiss hybrid triploid offspring seed |
| CN103229742B (en) * | 2013-05-23 | 2014-12-10 | 北京市水产科学研究所 | Preparation method of oncorhynchus mykiss walbaum and oncorhynchus mykiss hybrid triploid offspring seed |
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| CN106417256A (en) * | 2016-10-12 | 2017-02-22 | 中国水产科学研究院黑龙江水产研究所 | Method for increasing utilization rate of sperm of sex reversal milter of rainbow trout |
| CN114532259A (en) * | 2022-03-01 | 2022-05-27 | 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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