CN101933492A - Method for artificially fertilizing giant salamander - Google Patents

Abstract

The invention relates to a method for artificially fertilizing giant salamander, which comprises the following steps of: after a female giant salamander begins to breed an ovum ribbon in a pond, artificially drawing the ovum ribbon of the giant salamander out; after placing the ovum ribbon into a smooth and anhydrous box, covering giant salamander semen which is artificially extruded and inspected to be better by a microscope on the ovum ribbon; respectively packaging into an incubating device after 30 minutes; placing into a flow pond for aerating and incubating; and incubating a fry by a giant salamander oosperm in the development process for 27 to 35 days under the condition of 18.5+/-0.5 DEG C. The invention has the characteristics of skillfully and accurately operating the artificial fertilization of the giant salamander under the condition of 18.5+/-0.5 DEG C and ensuring the average fertilization rate of the giant salamander reaches 80.56 percent.

Description

Giant salamander artificial insemination method

Technical field

The invention provides a kind of aquaculture technology, the method for giant salamander artificial insemination specifically.

Background technology

Giant salamander (Andriasdavidiansas) is commonly called as the giant salamander, it is the distinctive amphibian of China, it also is the ancient animal of bodily form maximum, longest-lived in the existing amphibian animal in the world, be in by the aquatic intermediary of tiding over of crossing to the Lu Sheng vertebrate, has important scientific research value for zoologizeing origin, phylogeny and geographical distribution etc., so the title of " living fossil " is arranged.Because giant salamander has high edible and medical value and ornamental value, therefore disorderly catching the phenomenon of hunting giant salamander excessively in recent years is becoming increasingly rampant, because the artificial destruction of habitat, gradually reduce even lose, cause wild giant salamander habitat fragmentation, limited giant salamander breeding distribution ability, reduced foraging areas, make the giant salamander hurried decline of quantity in recent years, the zone of most original giant salamander existence disappeares substantially.It is very few that the natural breeding spot of domestic existing giant salamander is openly reported.In order effectively to protect and save this precious living resources; the Ministry of Agriculture of country and fishery administration department have formulated multinomial protection and the policy and the measure of raising and train the breeding giant salamander carried out in encouragement; make domestic relevant area begin to pay attention to and be devoted to the activity of artificial domestication breeding giant salamander, success is being obtained aspect the giant salamander artificial propagation seedling in many provinces and regions.According to the PRELIMINARY RESULTS of our on-site inspection, in 17 provinces and regions of the former distribution of giant salamander, carry out giant salamander at present and protect mainly containing of work with moving: there are 2 tame giant salamander culture fields in Zhejiang Province, and about 1100 tails of parent's quantity are produced seedling tail surplus 50,000 per year; There are 5 families in Guangdong Province, and about 7000 tails of parent's quantity are produced tail surplus the seedling 20,000 per year; There are 13 families in the Hunan Province, and tail surplus parent's quantity 6000 is produced tail surplus the seedling 20,000 per year; There are 5 families in Guizhou Province, and tail surplus parent's quantity 1000 is produced tail surplus the seedling 15000 per year; There is 1 family in Yunnan Province, parent's quantity 200 tails; There are 4 families in Sichuan Province, parent's quantity 600 tails; There are 3 families the Shanxi Province, parent's quantity 400 tails; There is 1 family in Gansu Province, parent's quantity 150 tails; There are 3 families in the Hunan Province, parent's quantity 500 tails; There are 3 families in Jiangxi Province, parent's quantity 560 tails; Giant salamander is cultured to become salamander in ground such as the Fang County in Hubei Province, Shiyan; Have giant salamander culture family more than 3000 families in Shaanxi Province's Hanzhong City now, wherein obtained more than 40 giant salamander of fish-farming operation department approval foundation and raised and train in the breeding farm.

Domestic and international technology trends

Domestic original most of giant salamander existence area because of giant salamander quantity seldom, and artificial propagation techniques still is in and gropes the test initial stage, reproduction rate is lower, large-scale farming lacks seedling, thereby development speed is still slower in the short time.Giant salamander is oviparity, and is in vitro fertilization, lays eggs once in 1 year.Giant salamander hatches the back and grew 5 years in natural environment, and body reaches about 45～60cm, and body weight is at 3～6kg, and sexual gland is mature on the whole.

The growth of giant salamander egg cell, growth, ripe and degeneration, phase in the time of can being divided into 6 with its cytological characteristics (seeing Table 1), similar basically to fish, but yolk has its unique cytology characteristics.

The morphological feature of table 1 giant salamander egg cell development

It at the bottom of the 7-8 month peak period of giant salamander gonad maturity.Autumn in the first year, (10～November), ovary was in the I phase, winter, (11～January), ovary was in the II phase, and spring, (2～April), the ovary major part was in the III phase, and minority ontogeny is to the IV beginning, antenatal ovary (5～July) is then grown the phase to IV, and postpartum, ovary then degenerated to the VI phase rapidly.Winter, spermary was in II～III phase, was in III～IV beginning spring, and antenatal spermary is in the IV～V phase, and postpartum, spermary then was in the VI phase.The wild giant salamander ovum incubation period is 35～42d, brood ovum to hatch the date also different, can differ a couple of days.

In decades; the scientific research technical staff author of China carries out many-sided research to the propagation technique of giant salamander; make slow progress; though the scientific research personnel has taked a lot of measures respectively in recent years; and obtained good effect; but still be difficult to accomplish scale production, chief reason is that these measures all have certain limitation.

Find that in our research process because male and female are grown the rapid minimizing that asynchronous and interference of human activities has caused the giant salamander resource, when propagating artificially, the sperm viability of male generation is not enough, causes reproduction rate of giant salamanders extremely low in the natural propagation process for giant salamander.

The disclosed a kind of method that improves giant salamander rate of hatchability and survival rate of Chinese patent literature, application number: 99116491, it is characterized in that: (1) is built close salamander and cultured the pond: the high 1.2m in pond, the long 2m in pond, pond are shallow infundibulate at the bottom of the wide 1m pond, and trench digging 25cm is wide in the middle of the pond, the shop big 2cm in hole ²The 0.25 * 0.5m that slips through the net, the bottom of trench degree of depth is 5-50cm from the salamander nest to draining valve, ditch shape is the slide shape, has 1.5m * 1m=1.5m in the pond in vain ²Water surface space for activities, stay 0.5m * 1m=0.5m in the pond ²Build the salamander nest, in the salamander nest by the 0.2m * 0.5m=0.1m of 20c m eminence embedding at the bottom of the pond of bay wall on one side ²Slabstone, the high lid of 70cm nest top at the bottom of the pond, the nest door only stays the big hole for the giant salamander turnover of diameter 30c m, all the other all seal with slabstone; (2) build the pond of laying eggs, ejaculate: the high 1.5m on one side in pond, the high 1.2m of another side, wide 3m, long 3m stays the platform of 0.2m * 0.3m at the bottom of leaning on the pond, one side of the high 1.2m in pond, dig dark 0.5m downwards, the cistern of wide 3m at the bottom of remaining the pond; Be provided with in the pond and twine the ovum bolt; (3) keep good temperature, light and sanitary condition, the temperature of culturing in the pond is controlled at 16-20 ℃ winter, and spring 18-22 ℃, in 22-25 ℃ of summer, in 20-22 ℃ of autumn, the morning, saturating natural daylight intensity was 200Lux, noon 50Lux, afternoon 100-150Lux; Produce the seed pond and require to build the indoor of health in, indoor temperature is controlled at about 22 ℃, and natural daylight intensity is 100lux; In culturing the pond and in the product seed pond, definitely mosquito can not be arranged; With the woody mosquito repellent of fragrant flower yesterday; (4) the rational food of arranging in pairs or groups; In daily breeding process, feed a frog that had just eaten molluscan health for every other day close salamander; (5) stimulative ovulation ejaculation in good time: in the April～May in every year, feed the little fish of the injectivity hormone that a tail lives every other day according to the 15 days in advance ovulation prediction phase of close salamander September～October, short its produces seed ejaculation.

Summary of the invention

Purpose of the present invention just is to provide a kind of giant salamander artificial insemination method, its not only to giant salamander without any negative effect, repetitive manual fertilization and hatching successfully continuously, and can improve fertilization rate and incubation rate.

The object of the present invention is achieved like this:

Giant salamander artificial insemination method is: giant salamander artificial insemination method is: after treating that female giant salamander begins the output chalaza in the pond, manually the giant salamander chalaza is pulled out, be put in the smooth anhydrous box after,

With manually extrude and through microexamination preferably the giant salamander seminal fluid be covered on the chalaza, dilution slowly rotates basin slightly, makes the abundant combination of smart ovum, leave standstill after 10 minutes to add Chi Shui,

Treat to be sub-packed in the hatching utensil after 30 minutes, put into the oxygenation hatching of flowing water pond, under 18.5 ± 0.5 ℃ of conditions, giant salamander fertilized egg hatches seedling through 27～35 days growth courses,

Incubation period should be noted: (1) controls water temperature and illumination well; (2) anti-harness the river mouldy; (3) prevent embryo's " subsides shell ", regularly stir.

Characteristics of the present invention: the giant salamander artificial insemination is under 18.5 ± 0.5 ℃ of conditions, and skilled accurate the operation can make the average fertilization rate reached to 80.56% of giant salamander.

Embodiment

The present invention will be further described below in conjunction with embodiment,

This invention is being carried out giant salamander in the artificial propagation production test, and at first strict technical standard by preferred formulation selects to culture demonstration place, base and construction ecological culture pond.Selected place has the natural water source condition, and its water quality, water temperature satisfy the physiological requirements that giant salamander produces growth and gonad maturity, and wherein pH value is between 6.5-7.3, and dissolved oxygen is more than 5.6ml/L; Water temperature 10-25 ℃, optimum water temperature 18-22 ℃.Institute builds cultures the pond and meets fully that giant salamander grows and the needed environment of life habit, and promptly bionical attitude environment is for cultivating the healthy material base that salamander provides most critical of planting.Strengthen simultaneously the cultivation of male close salamander and parent are matched selection,, keep perennial draingage, regulate measures such as discharge and water temperature, promote male salamander gonad development, improve its sperm viability by the abundant bait of desired nutritional that different development stage is thrown something and fed; It is ripe synchronously that the giant salamander parent reaches physiology and modal sperm and ovum; Select the female salamander of parent more than 8 ages, the required important germplasm condition of artificial efficient breeding giant salamander has been satisfied in the healthy individual pairing of male salamander more than 5 years.

The glued membrane whether female parent has ovum in the output note to be observed in the back of hastening parturition, and laid eggs in 3～7 days son, row of the giant salamander after hastening parturition is smart, and 3 days after hastening parturition begin to check whether male giant salamander produces essence and sperm quality simultaneously.

The method of checking the giant salamander sperm quality is: squeeze and get 1～2 of giant salamander seminal fluid, drip on slide, add one of spermatozoa diluent, place microscopically to observe, sperm bunchy or agglomerating distribution or individualism, but sperm only rotates or does not have then fertility difference or at all can not do in fertilization of any activity at all, and the sperm that has only individualism and do gun is only the measured sperm of matter, just can be used for artificial insemination.So male giant salamander is performed mark, be used for artificial insemination when the time comes.

Giant salamander artificial insemination way is: after treating that female giant salamander begins the output chalaza in the pond, manually the giant salamander chalaza is pulled out, after being put in the smooth anhydrous box, with manually extrude and through microexamination preferably the giant salamander seminal fluid be covered on the chalaza, dilution slightly, slowly rotate basin, make the abundant combination of smart ovum, leave standstill after 10 minutes and add Chi Shui, treat to be sub-packed in the hatching utensil after 30 minutes, put into the oxygenation hatching of flowing water pond, under 18.5 ± 0.5 ℃ of conditions, giant salamander fertilized egg hatches seedling through 27～35 days growth courses.Incubation period should be noted: (1) controls water temperature and illumination well; (2) anti-harness the river mouldy; (3) prevent embryo's " subsides shell ", regularly stir.

Giant salamander artificial insemination result (seeing Table 3).

Table 3 giant salamander artificial spawning, fertilization result

The result draws by experiment, and the giant salamander artificial insemination is under 18.5 ± 0.5 ℃ of conditions, and skilled accurate the operation can make the average fertilization rate reached to 80.56% of giant salamander.

Giant salamander artificial incubation result (seeing Table 4).

The fertilization of table 4 giant salamander, hatch results

The result draws by experiment, and giant salamander artificial incubation is accurately controlled incubation condition under 18.5 ± 0.5 ℃ of conditions, can make the average incubation rate of giant salamander reach 83.51%.

Behind the giant salamander spawning and fertilization, sometimes the chalaza is held out outside the hole, can be adopted the artificial incubation measure.The concrete grammar of artificial incubation is: set nylon bolting silk framework is put into the hatchery, and framework about about 30cm, evenly is placed in fertilized egg in the framework at the bottom of the pond then; Adopt miniflow water between the incubation period, cause embryonic death, on the hatchery, hide with black plastic cloth for fear of the light irradiation.Running check embryonic development situation is in time rejected unfertilized egg between the incubation period, to avoid water quality deterioration.

Claims (1)

1. giant salamander artificial insemination method is characterized in that:

Giant salamander artificial insemination method is: after treating that female giant salamander begins the output chalaza in the pond, manually the giant salamander chalaza is pulled out, be put in the smooth anhydrous box after,

With manually extrude and through microexamination preferably the giant salamander seminal fluid be covered on the chalaza, dilution slowly rotates basin slightly, makes the abundant combination of smart ovum, leave standstill after 10 minutes to add Chi Shui,

Treat to be sub-packed in the hatching utensil after 30 minutes, put into the oxygenation hatching of flowing water pond, under 18.5 ± 0.5 ℃ of conditions, giant salamander fertilized egg hatches seedling through 27～35 days growth courses,

Incubation period should be noted: (1) controls water temperature and illumination well; (2) anti-harness the river mouldy; (3) prevent embryo's " subsides shell ", regularly stir.