CN104041457B - A kind of Qinling Mountains fine-scaled graphite artificial fecundation method - Google Patents

A kind of Qinling Mountains fine-scaled graphite artificial fecundation method Download PDF

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CN104041457B
CN104041457B CN201410319444.1A CN201410319444A CN104041457B CN 104041457 B CN104041457 B CN 104041457B CN 201410319444 A CN201410319444 A CN 201410319444A CN 104041457 B CN104041457 B CN 104041457B
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qinling mountains
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邵俭
危起伟
吴金明
孙庆亮
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Yangtze River Fisheries Research Institute CAFS
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Abstract

The invention discloses a kind of Qinling Mountains fine-scaled graphite artificial fecundation method, step is: A, screening maturation, energetic Qinling Mountains fine-scaled graphite are as parent; B, parent's reinforced cultivating; C, artificial induced spawning: select by the end of March to mid-April, adopt two pin injections to hasten parturition; D, artificial semenpicking ovum: the parent after injection oxytocic drug divides and supports in spawning inducing pool, at spawning inducing pool one end erection water inlet pipe, select Qinling Mountains brachymystax lenok parent fish to adopt the extruding of the belly rhythm and pace of moving things to carry out live body semen collection ovum; E, artificial insemination: after adopting ovum, with fish physiological saline rinsing, squeeze on ovum by the seminal fluid of milter, and rear feather stirs, and carries out artificial insemination; F, artificial incubation: fertilized egg cleaned with pure water, have in the incubation tank of porous double-edged fine-toothed comb bottom pouring into and hatch, and swims out of now, shift out incubation equipment until fry punching, goes to fish basin and cultivate.Easy to implement the method, easy and simple to handle, its fertility rate and hatchability is high, and resulting bottle is healthy and strong, well-balanced for fish body, survival rate and bait utilization high, fast growth.

Description

A kind of Qinling Mountains fine-scaled graphite artificial fecundation method
Technical field
The invention belongs to endangered wildlife plasm resource protection technical field, be specially a kind of Qinling Mountains fine-scaled graphite artificial fecundation method.Its method is applicable to Qinling Mountains fine-scaled graphite Natural Population and recovers and artificial conservation aspect.
Background technology
Qinling Mountains fine-scaled graphite (Brachymystaxlenoktsinlingensis) is commonly called as colored fish, multicolored fish, plum blossom fish, is subordinate to salmon shape order (Saloniformes), land envelope type cold water fishes that salmon section (Salmonidae) is China endemic species, local distribution.Qinling Mountains fine-scaled graphite distribution is very narrow, habitat condition is strict, natural population is rare, only be distributed in the Heihe of Qinling Mountains Tai Baishan Mine Dong Lu, the stone river of northern foot, the Xushuihe Shui He of southern foot and too Baihe, thousand tributaries, river in Long County and Gansu the Upper Reaches of Wei River and tributary (Shaanxi Province animal institute etc., 1987).Due to the double influence of nature and human factor; Qinling Mountains fine-scaled graphite is perched and is increasingly worsened with living environment, and germ plasm resource is subject to heavy damage, has been put into wild (happy Pei fine jade and the Chen Yiyu of watching for animals of national secondary; 1998), very urgent to the protection of Qinling Mountains fine-scaled graphite.Along with artificial propagation, the reaching its maturity of seed rearing and various breeding way, China strengthens and payes attention to endangered species protection in addition, adopts artificial propagation techniques protection Qinling Mountains fine-scaled graphite resource to become the most effective means.But because Qinling Mountains fine-scaled graphite is individual miniaturized serious, sexual maturity stock number falls sharply, habitat conditions is harsh, in addition impatient hot-tempered, the kind trip of happiness jumping of Qinling Mountains fine-scaled graphite, skin mucus is few, and body is by thin squama, in body, fish-bone is relatively tiny, the features such as irritability is strong, its parent easily abrades onset lethal (Shi Deliang etc., 2012).In addition, Qinling Mountains fine-scaled graphite is carnivorous cold water fish, and the absolute fish brood amount of raun is little; In natural environment, fertilization rate, incubation rate and survival rate are low, and growth rate is slow, growth cycle long, artificial domestication is more difficult; Sexual maturity generally needs 3 ~ 4 years, and 500g sashimi (raw fish) long period is generally in 4 ~ 5 years, and this increases the difficulty that Qinling Mountains fine-scaled graphite germ plasm resource is recovered.In recent years, by practical exploration for many years, work out a kind of ripe, efficient Qinling Mountains brachymystax lenok parent fish and cultivate and artificial induced spawning technology, improve fertilization rate, incubation rate and weed survival rate.
Summary of the invention
In view of the above problems, the object of the invention is to there are provided a kind of Qinling Mountains fine-scaled graphite artificial fecundation method, easy to implement the method, easy and simple to handle, parent population has stable egg laying amount and ejaculation volume, and its fertility rate and hatchability is high, resulting bottle is healthy and strong, well-balanced for fish body, survival rate and bait utilization high, there is stronger environmental resistance and premunition, fast growth.
In order to realize above-mentioned object, the present invention adopts following technical measures:
A kind of Qinling Mountains fine-scaled graphite artificial fecundation method, the steps include:
1) ripe, energetic Qinling Mountains fine-scaled graphite is screened as parent,
2) parent's reinforced cultivating:
A) be 1:2 ~ 3 by female-male proportion, stocking rate is 5 ~ 8 tails/m 3, the yellow mealworm or hang black light lamp night of throwing something and feeding, from starting by the end of February, increased water flow amount (0.05 ~ 0.1m 3/ s), promote the gonad development of Qinling Mountains brachymystax lenok parent fish.
B) at the beginning of 3 months, (2 ~ 8 days) start, every day monitors water temperature, water temperature reaches 6 ~ 7 DEG C, start to subtract food, after carried out povidone iodine water body disinfection (do not cut off the water, splash at water inlet, working concentration is 3 ~ 5mg/L) every 3 ~ 5 days, within before hastening parturition 5 ~ 7 days, stop eating, and stop water body disinfection.When water temperature is close to the bread worm (8 ~ 9 DEG C), male and female parent population carries out point pond and cultivates.
3) artificial induced spawning:
Select (28-31 day) to mid-April (13-17 day) by the end of March, adopt two pin injections to hasten parturition, first injects oxytocic drug DOM and LHRH-A for parent 2mixture, every 1000g Qinling Mountains fine-scaled graphite raun dosage is: DOM2 ~ 4mg+LHRH-A 27 ~ 10 μ g, Qinling Mountains fine-scaled graphite milter injected dose reduces by half.24 ~ 48h injects the second pin, and every 1000g Qinling Mountains fine-scaled graphite raun injected dose is DOM1 ~ 2mg+LHRH-A 23 ~ 5 μ g+ Cobastabs 124 μ g, Qinling Mountains fine-scaled graphite milter injected dose reduces by half or does not inject.
4) artificial semenpicking ovum:
Parent after injection oxytocic drug divides and supports in spawning inducing pool, and at spawning inducing pool one end erection water inlet pipe, from water surface elevation 0.5 ~ 1.0m, discharge controls at 0.1 ~ 0.15m 3/ s, surrounding 1.5m carry out corral, prevent parent population from jumping out spawning inducing pool, and after effector phase, select Qinling Mountains brachymystax lenok parent fish to adopt the extruding of the belly rhythm and pace of moving things to carry out live body semen collection ovum, female male parent ratio is 1:1 ~ 3.
5) artificial insemination:
After adopting ovum, with fish physiological saline rinsing 2 ~ 3 times, squeeze on ovum by the seminal fluid of milter rapidly, rear feather stirs 2 ~ 3 minutes gently, carries out artificial insemination.
6) artificial incubation:
Fertilized egg is carried out repeatedly (2 ~ 3 times) cleaning with pure water, is 30 seconds at every turn, has in the incubation tank of porous double-edged fine-toothed comb and hatch bottom slowly pouring into.Described hatching density is 80,000/m 3, adopt lower end water inlet, current stick as degree so that fish-egg is sticky, every day sooner or later water inlet pour into crude salt (Nacl content:>=90%) 2 ~ 3kg, until fry punching swim out of existing after, shift out incubation equipment, go to fish basin or net cage carries out seed rearing.
Screen body weight 0.25 ~ 0.4kg in described step 1, more than 3 ~ 4 ages sexually matured Qinling Mountains fine-scaled graphite is as parent.
Cultivate pond in described step 2 in a step and be designed to rivers and canals shape cement pit, form drop, gravel and stone is laid at the bottom of pond, inlet end arranges sparge pipe or the facility that pumps up water (single-phase submersible pump QD6-32/3-1.1, bore 40mm), below water inlet, the depth of water strengthens, and form artificial pool (1.2 ~ 1.5m), each channel volume is about 16 ~ 20m 3, mean depth of water is 0.5 ~ 0.8m, and draining is convenient, and pond water quality transparency is 0.5 ~ 0.7m, and dissolved oxygen is higher than 8mg/L.
When water temperature reaches 8 ~ 12 DEG C when hastening parturition in described step 3, carry out artificial induced spawning.
Described step 3 adopts two pin injections, DOM and LHRH-A 2intermixture, every tail parent injection volume is less than or equal to 2ml mixing dosage, and first injects oxytocic drug DOM and LHRH-A for parent 2intermixture, every 1000g Qinling Mountains fine-scaled graphite raun dosage is: DOM3mg+LHRH-A 28 ~ 9 μ g, 24 ~ 48h injects the second pin, and every 1000g Qinling Mountains fine-scaled graphite raun injected dose is DOM1.5mg+LHRH-A 24 μ g+ Cobastabs 124 μ g.
Spawning inducing pool mean depth of water 0.5m in described step 4, erection water inlet pipe mouth of pipe height dried up 0.5m, surrounding fine-structure mesh carries out corral, prevents fish body from jumping out and abrading.
First with fish physiological saline rinsing after described step 5 ovum artificial collection, rear being squeezed by seminal fluid is rapidly stirred with feather on ovum.
After described step 6 artificial insemination, eluriate 2 ~ 3 times with pure water rapidly, be 30 seconds at every turn, and have in the incubation tank of porous double-edged fine-toothed comb bottom slowly pouring into and hatch, hatching density is 80,000/m 3, adopt lower end water inlet, current stick as degree so that fish-egg is not sticky.
Described step 6 morning and evening every day pours crude salt 2 ~ 3kg at water inlet and carries out water body filtration sterilization.
When described step 6 water temperature is 8 ~ 9 DEG C, the incubation of membrane time is 450 ~ 480h.
When described step 6 water temperature is 10 ~ 12 DEG C, the incubation of membrane time is 408 ~ 445h.
The present invention compared with prior art, has the following advantages and effect:
State a kind of Qinling Mountains fine-scaled graphite artificial fecundation method, by artificial regulatory envirment factor, make male and female fish gonad development basic synchronization; ensure that artificial induced spawning carries out smoothly; method is simple, and workable, practical application is good; its fertilization rate is higher than 90.00%; incubation rate is higher than 83.00%, and survival rate is higher than 90.00%, the same with the breeding of all the other salmon trout cold water fishes; be applicable to the large-scale production of seed completely, thus recover for the protection of Qinling Mountains fine-scaled graphite species resource and Natural Population thereof.
Embodiment
Below in conjunction with concrete embodiment, the present invention is further described.
Embodiment 1:
A kind of Qinling Mountains fine-scaled graphite artificial fecundation method, the steps include:
1. Juvenile stage:
Screen body focuses on ripe, the energetic Qinling Mountains fine-scaled graphite of 0.25 ~ 0.4kg as parent.
2. parent's reinforced cultivating:
Under natural conditions, Qinling Mountains fine-scaled graphite life habit is special, and habitat condition is harsh, and its milter gonad development is early than raun.Therefore, in order to impel parent to adapt to Artificial facilities, and ensureing that male and female fish gonad development is synchronous, carrying out artificial propagation smoothly, need to carry out Ecology simulation reinforced cultivating to parent, and the gonad development of the female male parent of artificial regulatory.
A) reinforced cultivating of Qinling Mountains brachymystax lenok parent fish: build Ecology analog ligand and cultivate pond, cultivate pond and be designed to rivers and canals shape cement pit, form drop, gravel and stone is laid at the bottom of pond, inlet end arranges sparge pipe or the facility that pumps up water, below water inlet, the depth of water strengthens, and form artificial pool (1.2 ~ 1.5m), each channel volume is about 16 or 17 or 18 or 19 or 20m 3, mean depth of water is 0.5 or 0.6 or 0.7 or 0.8m, and draining is convenient, and pond water quality transparency is 0.5 or 0.6 or 0.7m, and dissolved oxygen is higher than 8mg/L, and stocking rate is 20 or 25 or 35 or 40 tails/m 3, female-male proportion is 1:2, and yellow mealworm is parent population bait feeding, accompanies Enrofloxacin 1g/kg bait simultaneously, throws something and feeds continuously 3 days, use week about in one week.From by the end of February, increased water flow amount carries out stimulation by running water, promotes the gonad development of Qinling Mountains brachymystax lenok parent fish;
B) take out from mid-March and carry out artificial regulatory with well water, reduce pole temperature approach (8 ~ 12 DEG C), maintain day samming stable (10 ~ 11 DEG C), promote that Qinling Mountains fine-scaled graphite male and female parent population gonad development is synchronous; Carried out povidone iodine water body disinfection every 3 or 4 or 5 days, within before hastening parturition 5 or 6 or 7 days, stop throwing something and feeding, and stop water body disinfection.Water temperature divides pond to cultivate, to avoid parent population self-produced individually close to male and female parent population time the bread worm (8 ~ 9 DEG C).
3. artificial induced spawning:
(1) Sex judging and proportioning:
Qinling Mountains fine-scaled graphite raun touches hardness increase at breeding season pectoral fin, abdomeinal fin hand, and body colour is blue or green brown, and side is with orange red ribbon grain, and gonopore projection is brownish red; And milter is touched comparatively soft at breeding season pectoral fin, abdomeinal fin hand, body colour is pitchy, and side is with magneta colour striped, and gonopore taper is obviously in bronzing.
(2) selection of mature brood fish:
Qinling Mountains fine-scaled graphite raun: belly is and expands, softness, elasticity are strong, and touching stomach wall has plentiful and soft sense, is gonad development good.Qinling Mountains fine-scaled graphite milter: light pressure belly both sides have the seminal fluid of the sticky shape of milky to flow out and meet water scatters immediately.
(3) ocyodinic and dosage is injected:
Injecting method generally adopts intrathecal, and part is pectoral fin base portion or abdomeinal fin base portion, and adopt double injection, medicine is maleic acid DOM (DOM) and luteinizing hormone releasing hormone A 2(LHRH-A 2) mixture, the second injection amount reduces by half, add Cobastab 12mixing; Milter dosage all reduces by half use.
(4) drug use of artificial induced spawning:
Select late March to mid-April, respectively to two kinds of parents inject oxytocic drug DOM (maleic acid DOM, Ningbo, the three lives pharmaceutcal corporation, Ltd) and LHRH-A 2(luteinizing hormone releasing hormone A 2, Ningbo, the three lives pharmaceutcal corporation, Ltd) mixture, the dosage of every 1000g Qinling Mountains brachymystax lenok parent fish: DOM3mg+LHRH-A 28 ~ 9 μ g intermixtures; 24 or 28 or 32 or 36 or 40 or 44 or 48h inject the second pin, every 1000g Qinling Mountains brachymystax lenok parent fish injected dose is DOM1.5mg+LHRH-A 24 μ g+ Cobastabs 124 μ g intermixtures, above milter dosage reduces by half.Effect time is relevant to water temperature, water temperature 8 or 9 DEG C, and Qinling Mountains fine-scaled graphite raun Effect time is 48 or 55 or 62 or 70 or 82 or 88h, and milter Effect time is 36 or 39 or 44 or 46 or 48h; Water temperature 10 or 11 or 12 DEG C, Qinling Mountains fine-scaled graphite raun Effect time is 24 or 27 or 29 or 32 or 34 or 36h, and milter effect is 12 or 15 or 18 or 21 or 24h, and therefore raun will inject oxytocic drug early than 12h, the ecbolic effect timetable of Qinling Mountains fine-scaled graphite raun under different temperatures, in table 1.
The ecbolic effect time of Qinling Mountains fine-scaled graphite male and female parent population under table 1 different temperatures
4. artificial semenpicking ovum insemination:
Parent after injection oxytocic drug divides and supports in spawning inducing pool, strong stimulation by running water, and when parent population oestruses, pole happiness is jumped, run helter-skelter, and the male and female sometimes jump water surface reach 1m, and punching trip clearly against the current.Now, screening Qinling Mountains fine-scaled graphite parent is that ovum insemination is adopted in 1:1 ~ 2 by female-male proportion.
5. artificial incubation:
The fertilized egg of Qinling Mountains fine-scaled graphite is half adhesive egg, first with fish physiological saline 2 or 3 times carry out de-wash away glutinous after, adopt dry method insemination, seminal fluid is squeezed in ovum, stir 2 or 3 minutes, fully inseminate, insemination is carried out under dim lights, then the incubation tank putting into miniflow water is hatched, and hatching density is 80,000/m 3water body.Fertilized egg is orange-yellow, if pressing development of fertilized ova to neurula stage is fertilization Success criteria, its fertilized egg is 98%; If press development of fertilized ova to the orga-nogenesis later stage, its fertilization rate is 90%.Water temperature 8 or 9 DEG C, if be standard by 90% incubating oosperm membrane, about through 450 or 460 or 470 or 480h incubation of membrane; Water temperature 10 or 11 or 12 DEG C, if be standard by 90% incubating oosperm membrane, about through 408 or 415 or 422 or 431 or 438 or 441 or 445h incubation of membrane.Fry hatching, after 9 days, starts to occur schistostoma, after body colour deepens 50% fry appearance punching trip, carries out egg yolk and fairy shrimp opening; 90% fry shifts out incubation equipment after occurring that rushing parade is, moves to larvae cultivation net cage and carries out larval rearing.
Above-mentioned a kind of Qinling Mountains fine-scaled graphite artificial fecundation method, also has the following advantages:
1. there is stable egg laying amount: generally Qinling Mountains fine-scaled graphite gonad development milter comparatively raun early, interindividual variation is remarkable, and gonad development is asynchronous, and the absolute fish brood amount of raun is low, and fertilization rate, incubation rate and survival rate are all lower, scale breeding difficulty.The present invention is according to the biological property of Qinling Mountains fine-scaled graphite; in conjunction with various artificial regulatory means; carry out artificial induced spawning breeding; its fertilization rate reached 90%; incubation rate reaches 83.33%, survives and reach 92.9% after fingerling incubation of membrane, breeds the same with other salmon trout cold water fish; be applicable to the large-scale production of seed completely, thus recover for the protection of Qinling Mountains fine-scaled graphite species resource and Natural Population thereof.
2. there is stronger operability: the present invention is by artificial regulatory technology, and make the gonad development basic synchronization of milter, can ensure carrying out smoothly of artificial fingerling, its method is relatively simple and convenient, workable.
3. the Dominant Facies that has of filial generation obtained is to obviously: it is healthy and strong, well-balanced 1) to hatch fry fish body, and survival rate is high, and bait utilization is high; 2) environmental resistance is strong, changes greatly and has good tolerance period, have good adaptability in natural environment in flood period and water temperature.3) fry resistance against diseases is comparatively strong, and be easy to domestication, through the meticulous raising of 8 months, body length can reach 9 ~ 10cm, can carry out artificial releasing, was conducive to reconstruction and the recovery of germ plasm resource.

Claims (5)

1. a Qinling Mountains fine-scaled graphite artificial fecundation method, the steps include:
A, screening maturation, energetic Qinling Mountains fine-scaled graphite are as parent, and screen body weight 0.25 ~ 0.4kg, the Qinling Mountains fine-scaled graphite in 3 ~ 4 ages is as parent;
B, parent culture:
A) be 1:2 ~ 3 by female-male proportion, stocking rate is 5 ~ 8 tails/m 3, the yellow mealworm or hang black light lamp night of throwing something and feeding, from starting by the end of February, increased water flow amount 0.05 ~ 0.1m 3/ s;
B) started at the beginning of 3 months, every day monitors water temperature, water temperature reaches 6 ~ 7 DEG C, starts to subtract food, after carried out povidone iodine water body disinfection every 3 ~ 5 days: do not cut off the water, splash at water inlet, working concentration is 3 ~ 5mg/L, within before hastening parturition 5 ~ 7 days, stops eating, and stops water body disinfection, when water temperature is close to the bread worm, male and female parent population carries out point pond and cultivates;
C, artificial induced spawning: select by the end of March to mid-April, adopt two pin injections to hasten parturition, first injects oxytocic drug DOM and LHRH-A for parent 2mixture, every 1000g Qinling Mountains fine-scaled graphite raun dosage is: DOM2 ~ 4mg+LHRH-A 27 ~ 10 μ g, Qinling Mountains fine-scaled graphite milter injected dose reduces by half, and 24 ~ 48h injects the second pin, and every 1000g Qinling Mountains fine-scaled graphite raun injected dose is DOM1 ~ 2mg+LHRH-A 23 ~ 5 μ g+ Cobastabs 124 μ g, Qinling Mountains fine-scaled graphite milter injected dose reduces by half or does not inject;
D, artificial semenpicking ovum: the parent after injection oxytocic drug divides and supports in spawning inducing pool, at spawning inducing pool one end erection water inlet pipe, from water surface elevation 0.5 ~ 1.0m, discharge controls at 0.1 ~ 0.15m 3/ s, surrounding 1.5m carry out corral, and select Qinling Mountains brachymystax lenok parent fish to adopt the extruding of the belly rhythm and pace of moving things to carry out live body semen collection ovum, female male parent ratio is 1:1 ~ 3;
E, artificial insemination: after adopting ovum, with fish physiological saline rinsing 2 ~ 3 times, squeeze on ovum by the seminal fluid of milter, and rear feather stirs 2 ~ 3 minutes, carries out artificial insemination;
F, artificial incubation: fertilized egg is carried out 2 ~ 3 cleanings with pure water, and be 30 seconds at every turn, pour in incubation tank and hatch, described hatching density is 80,000/m 3, adopt lower end water inlet, current stick as degree so that fish-egg is sticky, and every day pours crude salt 2 ~ 3kg at water inlet sooner or later, until fry punching swim out of existing after, shift out incubation equipment, go to fish basin or net cage carries out seed rearing;
In described step B, a step is cultivated cultivation pond used and is designed to rivers and canals shape cement pit, forms drop, and lay gravel at the bottom of pond, inlet end arranges sparge pipe or the facility that pumps up water, and below water inlet, the depth of water strengthens, and form artificial pool, each channel volume is 16 ~ 20m 3, the depth of water is 0.5 ~ 0.8m, and draining is convenient, and pond water quality transparency is 0.5 ~ 0.7m, and dissolved oxygen is higher than 8mg/L; When water temperature reaches 8 ~ 12 DEG C when hastening parturition in described step C, carry out artificial induced spawning.
2. a kind of Qinling Mountains according to claim 1 fine-scaled graphite artificial fecundation method, is characterized in that: described step C adopts two pin injections, DOM and LHRH-A 2intermixture, every tail parent injection volume is less than or equal to 2ml mixing dosage, and first injects oxytocic drug DOM and LHRH-A for parent 2intermixture, every 1000g Qinling Mountains fine-scaled graphite raun dosage is: DOM3mg+LHRH-A 28 ~ 9 μ g, 24 ~ 48h injects the second pin, and every 1000g Qinling Mountains fine-scaled graphite raun injected dose is DOM1.5mg+LHRH-A 24 μ g+ Cobastabs 124 μ g.
3. a kind of Qinling Mountains according to claim 1 fine-scaled graphite artificial fecundation method, is characterized in that: spawning inducing pool mean depth of water 0.5m in described step D, and erection water inlet pipe mouth of pipe height dried up 0.5m, surrounding fine-structure mesh carries out corral.
4. a kind of Qinling Mountains according to claim 1 fine-scaled graphite artificial fecundation method, it is characterized in that: when described step F water temperature is 8 ~ 9 DEG C, the incubation of membrane time is 450 ~ 480h.
5. a kind of Qinling Mountains according to claim 1 fine-scaled graphite artificial fecundation method, it is characterized in that: when described step F water temperature is 10 ~ 12 DEG C, the incubation of membrane time is 408 ~ 445h.
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CN108605870A (en) * 2018-05-24 2018-10-02 陕西省水产研究所 A kind of Qinling Mountains fine-scaled graphite artificial acclimation method
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