Artificial reproduction method of large salamander
Technical field the invention provides a kind of aquaculture technology, specifically, is a kind of artificial reproduction method of large salamander.
The background technology giant salamander common name giant salamander, its protection of resources, increment, utilization are the focuses of aquatic products scientific research personnel research.Though the report of the artificial propagation techniques aspect of giant salamander is more, really the scale reproductive success is less, and particularly artificial induced spawning and hatching repeat success seldom for years.Tracing it to its cause, is that parent does not reach sexual maturity or degeneration mostly, that is to say, fails to cultivate effectively the ripe and ovum fully-developed giant salamander parent of sperm development.Simultaneously, because the restriction of parent cultural method is female asynchronous with patrogenesis.Existing giant salamander parents culture and artificial induced spawning comparatively effective method are to inject hormone or throw something and feed to contain the special feed of Chinese herbal medicine composition, though can improve male giant salamander sexual mature rate and sperm viability, but because hormone and Chinese herbal medicine are to the influence of giant salamander physiological function, can not repeat continuously to succeed, be used for femalely, then can not lay eggs continuously.Therefore, this method is unfavorable for the breeding of giant salamander large-scale artificial.
Summary of the invention purpose of the present invention just is to provide a kind of artificial reproduction method of large salamander, it not only can improve female male giant salamander sexual mature rate, and can make female male gonad development synchronous, and to giant salamander without any negative effect, repetitive manual is hastened parturition and is hatched success continuously, and can improve fertilization rate and incubation rate.
For achieving the above object, embodiment of the present invention are: a kind of artificial reproduction method of large salamander is characterized in that its step comprises:
(1) the antenatal cultivation of parent: create the ecotope that adapts to the giant salamander gonad development, dark tunnel secret tunnel, spring, miniflow water, lay the gravel pebbles about 30cm at the bottom of the pond, the hole, cave that has cobble stone, boulder to build up in the pond, temperature control 20-22 ℃, light-operated 100lux, the special feed of giant salamander parents culture is the carp head, and its method is the season that the 3-4 month giant salamander parent since every year germinates, and gives male giant salamander every 1-2 day and throws something and feeds carp for the first time, give the female giant salamander carp secondary of throwing something and feeding weekly, feeding volume is the double centner giant salamander 2-3 kilogram carp head of throwing something and feeding;
(2) artificial induced spawning: the hormone of artificial induced spawning adopts carp brain hypophysis, LRH-A, HCG, and dosage is female 100-300 μ g/kg, male reducing by half, double injection; ,
(3) artificial insemination: during artificial insemination, use black wet cloth and muffle male giant salamander head, after sperm is extruded, add the dilution of 0.01% physiological saline, mix and add sperm nutrient solution with the chalaza, after 5-10 minute, add a small amount of distilled water, after 30 minutes, with the fertilization container water change twice;
(4) artificial incubation: spring is adopted in artificial incubation, miniflow water, and water temperature 18-20 ℃, each temperature difference is no more than 0.5 ℃, and water quality PH is 6.5-7.4;
It is that parent is cultivated feed that the present invention adopts the carp head, a because carp mesencephalohypophysial effect, can improve male parent sperm development maturing rate, and can make female gonad development synchronous, the carp head is a pure natural food, can not cause any health or physiological function damage, therefore can repeat continuously to hasten parturition the giant salamander parent.The artificial induced spawning that the present invention is specific and the ecological environmental condition of hatching can make fertilization rate and incubation rate reach respectively more than 80% with more than 85%, for the giant salamander scale artificial culture provides technical conditions.
Embodiment artificial reproduction method of large salamander provided by the invention is finished by following steps and method:
(1) the antenatal cultivation of parent: at first create the ecotope that is similar to the giant salamander gonad development under the natural environmental condition, dark tunnel secret tunnel, spring, miniflow water, lay the gravel pebbles about 30cm at the bottom of the pond, the hole, cave that has cobble stone, boulder to build up in the pond, temperature control 20-22 ℃, light-operated 100Lux, on this basis, from the season that annual 3-4 month giant salamander parent germinates, give throw something and feed carp 1 time of male giant salamander every day, give throw something and feed carp 2 times of female giant salamander weekly, feeding volume is 2kg/100kg;
(2) artificial induced spawning: oxytocic hormone adopts carp brain hypophysis, LPH-A, HCG, the female 200 μ g/kg of dosage, male 100 μ g/kg, double injection.
The present invention has done the experiment of different water temperatures to the effect time effects of laying eggs, and as can be seen, the back water temperature of hastening parturition is to the influence obvious (seeing Table 1) of the effect time of laying eggs.Facts have proved that the ripe parent back feritilization of ovum rate of output about 90-120 hour of hastening parturition is the highest, therefore, suitable water temperature of the present invention adopts 20-22 ℃;
Table 1 is hastened parturition the back water temperature to the lay eggs influence of effect time of giant salamander
Water temperature (℃) | The effect of the laying eggs time (h) | Sample size (tail) |
18 20 22 24 | 140 125 110 100 | 12 21 8 16 |
(3) artificial insemination: treat female in water behind the output egg jelly, immediately from the pond with male proposition, put into the cloth stretcher and muffle head with black wet cloth, a human hand upwards holds up female afterbody then, live chalaza one end with hand rest and slowly put into the enamel basin of disinfecting, after in the chalaza holder engagement, gently squeeze male umbilical region with have gentle hands immediately, extrude sperm, with 3 times of 0.01% physiological saline dilutions to seminal fluid, be poured into then in the chalaza, add several milliliters of sperm nutrient solutions, swing basin more gently, make its fertilization evenly.After 5-10 minute, add a small amount of distilled water, after 30 minutes, the water of basin is changed twice, enter the hatching stage;
(4) artificial incubation: adopt the miniflow mountain spring water, dark tunnel secret tunnel, dissolved oxygen amount 6mg/L with malachite solution prevention " water mold ", regularly stirs the embryo simultaneously.
Temperature is obvious to the influence of giant salamander embryonic development, can not be too high, can not be too low, and the present invention has done test (seeing Table 2) for this reason.Test shows that temperature is high more, and brooding time is short more, but temperature
Table 2 temperature is to the influence of giant salamander embryonic development
Water temperature (℃) | Brooding time (my god) | Embryo's (individual) | Incubation rate (%) |
12 16 18 20 22 | 47 45 39 28 26 | 120 80 450 960 520 | 80.0% 84.4% 95.7% 96.0% 81.2% |
Too high hatchability reduces, and therefore, preference temperature should be 18-20 ℃, and temperature difference per day is no more than 0.5 ℃.The acid-base value of water quality also has a significant effect to hatching, and pH value is higher than 8.2 or be lower than 6 embryos and can not grow, and by test, hatching water quality PH accommodation is between the 6.5-7.4;
(5) seedling is fed: at the growth of seedling initial stage after hatching, temperature is controlled at 16-20 ℃, feeds with aquatic black larva, and survival rate can reach more than 90%.