The artificial breeding of giant salamanders has not been resolved due to many difficulties. It has not yet achieved large-scale production of seedlings, and its resources are difficult to be protected, proliferated, and utilized. For more than ten years, we have conducted in-depth research on the multidisciplinary knowledge of polypillary fish reproductive physiology, developmental biology, and reproductive biology, and discovered the constraints: First, male sexual maturation rate is low (mainly refers to poor semen quality); second is female and male. The development of gonads is not synchronized; the third is that it is very difficult to accurately determine the sexual maturity of the giant salamander with the naked eye. In response to these problems of developmental biology, we integrate the principles and techniques of reproductive physiology, reproductive biology and modern biotechnology, genetics, traditional Chinese medicine, pharmacology of traditional Chinese medicine, Chinese medicine engineering, wildlife nutrition and feed science, and other related disciplines. Innovated the artificial breeding technology of giant salamander. This innovative technology mainly includes the techniques of imitating the habitat of the crucian carp, artificially controlling the temperature, controlling the light to cultivate the brood, and feeding the traditional Chinese medicine “Shengjing Powder†and “the fodder for promoting follicle soup†to promote the development of female and male glands. This innovative technology has enabled the large-scale production of giant salamanders, providing a technological platform for the commercial breeding and protection, proliferation and utilization of giant salamanders. This article will provide a detailed introduction to the large-scale reproduction technology of giant salamanders.
1 Eco-environment fostered by broodstock Eco-environmental conditions bred by broodstock are important factors that affect the maturity of gonadal growth. Therefore, it is an important technique for fostering broodstocks to build imitated eco-environment and artificial temperature control, light control, water quality regulation, and feeding of artificial compound feeds.
1.1 Water temperature Water temperature is a determinant factor in the development of gonorrhoids. The main role of water temperature is to stimulate the hypothalamic secretion and release factors of the giant salamander fish to the anterior pituitary gland, and the pituitary gland produces two kinds of hormones (LH, FSH). These two hormones can regulate the occurrence of sperm and egg. Water temperature can also promote brood feeding to meet the nutrients needed for sperm cell development.
The water temperature is controlled at 16-24°C from March to September every year. The daily temperature of water should not exceed 1.5°C.
1.2 The Illuminati littoralis have the habit of backlighting, that is, fear of light. Therefore, during broodstock incubation, light is controlled at 300-5001x. In other words, brooding is performed under low light conditions
1.3 Feed The nutrient-rich feed is the material basis for the development of gonadal gonads. During broodstock incubation, the protein content in the feed is 42%-45.3%, sugar is 10%-18%, fat is 2%-3%, multidimensional and minerals is 1.5%, others are 7%-12 %, 2% to 3% of Chinese medicine spermatozoa and follicle-stimulating soup. The amino acid balance of protein in feed is extremely important and cannot be ignored. At the same time, according to different phases of gonad development in the broiler, it is necessary to add different amounts of traditional Chinese medicine Shengjing San and traditional Chinese medicine to promote follicle soup compound feed, which is a key technology for broodstock cultivation.
1.4 Water quality regulation During the incubation of the broodstock, a good water environment must be created, which is conducive to the healthy growth and development of the broodstock. The parental pool water quality requires a high degree of mineralization, hardness, and transparency. The pH is 6.5-7.5. In mountainous areas, mountain water and stream water can be used to cultivate broods, and the principle is to apply well water (after aeration) or tap water to cultivate brooders. Water quality regulation mainly involves changing water frequently, diligently cleaning and diligently disinfecting.
2 Brooding
2.1 Parental rearing The main body of the carp is 2 ways: one is female and male polyculture; the other is female and male alone.
2.2 Breeding From the research on the technique of broodfish reproduction in our 20 years, we found that females have a higher sexual maturation rate and a greater amount of cocking eggs; males have a low sexual maturation rate of only 26.2%, and this is 26.2% of males. The quality of semen is not all qualified. Artificial broodstock grown in artificial tunnels, basements, and other artificial projects have a higher rate of sexual maturation in males, but lower female sexual maturation rates. Therefore, the giant salamander has so far failed to achieve large-scale production of seedlings. Therefore, the cultivation of a large number of sexually mature broodstocks is a key technology for the large-scale production of giant salamanders.
The development of gonorrhoids has a regular pattern of seasonal cycles. It begins to develop from March to April each year and matures from July to October. According to the law of seasonal cycles, artificially cultivated broods mainly include winter cultivation, spring cultivation, prenatal cultivation and supplemented by physiological methods.
2.2.1 Winter Breeding Winter breeding refers to the post-natal physical fitness stage. The main purpose of broodstock cultivation at this stage is to provide a large amount of nutrients to the body of the giant salamander, so that it can quickly return to health after birth; secondly, it is necessary to accelerate the degradation and absorption of sperm and ovary so that the sperm and ovary can develop normally. The main tasks of winter cultivation are water temperature regulation and feeding. Water temperature regulation is divided into two phases: First, the water temperature is controlled at 18-23 °C, so that the broodstock ingest a large number of food, restore health, and then the water temperature is reduced to below 10 °C, so that the parental into hibernation, accelerate the absorption and degradation of the spermary ovary. The winter bait feed is based on compound feed, and 2% lysine and 1% multivitamins are added to the compound feed.
2.2.2 Spring Breeding In spring, the gonads begin to develop. The gonad development at this stage is an important development period from the second phase to the third phase. The main purpose of brood cultivation at this stage is to promote the development of gonad gonads, so that their gonads can be converted to phase III as soon as possible; secondly, high-quality diets should be fed to meet the nutrients required for gonad development. The main tasks of spring cultivation are regulating water temperature, feeding, adjusting light and regulating water quality. The regulation of water temperature means that the temperature of the water is controlled at 18-22°C, so that the parental body starts to feed early and the gonadal begins to develop early; feeding is added to the compound feed that promotes the gonorrhea development of Chinese traditional medicine.
2.3 Prenatal culturing This stage of development of gonads is the period from the end of stage III to the stage of initial transformation of stage V. It is an important stage of gonad development and maturation. Therefore, the aim of brood cultivation at this stage is to cultivate a large number of mature broods, which are mainly cultivated using ecological and physiological methods.
Ecological approach: Wild dolphins inhabit in streams and springs in deep mountain gorges. Therefore, when artificially fostering broodstock, it is necessary to build a cave that mimics the ecological environment (returned water, spring water). At the same time, adjust the light to 5001x. The water temperature is controlled at the optimal growth and development range of 18-23°C for the broodstock.
Physiological method: It is to inject the spermatozoa of the Chinese medicine that promote the development of gonads in the parent body into the body and promote the development and maturation of the gonads. The joints between the nervous system and the gonadal pituitary glands are important organs for the development and maturation of germ cells. The Chinese medicine Shengjing injection can promote the release of hormones from the pituitary of the giant salamander and increase the maturation rate of the salamander.
3 Artificial breeding new technology
3.1 The parental selection of sexual maturation features: male vents uplift circle prominent white spots, visible swelling around the inside. B-ultrasonic examination: The testis is 7-9cm long and 1-2cm wide; the testis is full, with no groove in the middle indicating maturity.
The female's abdomen is enlarged, soft and elastic. B-ultrasonic examination showed that the egg diameter was 6-7mm, and the ovary showed a single chain bead indicating maturity.
3.2 artificial production
3.2.1 oxytocin RES+LRH-A+DOM,LRH-A+HCG.
3.2.2 Effort dose (according to new standards)
Female: RES+LRH-A+DOM4-10μg/kg;
HCG l00-1 5001U/kg
LRH-A 0.2-150μg/kg
Male: The above dose is halved.
3.2.3 Method of Oxygen Production The parental oxytocin injection site is the intercostal space, and the needle is 0.3-0.5cm. The needle is injected with a human infusion soft plastic tube needle, which can prevent the body from swinging when the parent body is injected.
3.2.4 Fertility effect The time from parental ovulation injection to spawning is 4-11 days, and the best spawning time is 4-6 days. Therefore, from the 3rd day after urging, someone should be on duty to observe whether the female parent produces egg glue film.
3.2.5 Artificial Insemination The artificial insemination of salamander adopts the wet and dry method. The operation procedure is: When the female parent body produces the pectin film, it should be immediately carried by hand to put the egg in the enamel pot, and the other person will mature. A good male parent catches up, puts a person in a stretcher, and uses his hands to lightly push the abdomen of the male. When there is milky white semen, the semen can be directly squeezed into the egg, and the eggs can be lightly shaken to make the egg fertilize evenly. After 15 minutes, add purified water. After another 20 minutes, the eggs were changed 2 times. Finally, the egg is placed in a small plastic basket and suspended in the hatchery, which is the incubation phase.
4 artificial hatching
4.1 Incubation of ecological conditions
4.1.1 Water temperature and water source Fertilized egg hatching water temperature requirements are strictly controlled at 20-22 °C. The daily temperature of the water should not exceed 1.0°C. The water source can be tap water, spring water (spring water first to put a reservoir, later release hatch pool), well water (to be used after aeration). Spring water is easy to hatch, so it is necessary to increase the temperature of the spring when hatching with spring water.
4.1.2 Dissolved Oxygen The amount of oxygen in the body of fertilized eggs should not be less than 5 mg/L.
4.1.3 Light Fertilized Eggs Lighting should be strictly controlled at about 300lx.
4.2 artificial incubation of fertilized eggs mainly using static water incubation and microfluidic water incubation. When water is hatched, change the water 1 time in the morning and in the evening.
Note 3 points during hatching: First, control the water temperature and light; Second, prevent and control water mold, that is, regular mildew net prevention during incubation, the concentration of 0.5-0.810-6, soaking the fertilized eggs for 10 minutes; third is to prevent Embryos "stick", that is regularly flipping fertilized eggs.
1 Eco-environment fostered by broodstock Eco-environmental conditions bred by broodstock are important factors that affect the maturity of gonadal growth. Therefore, it is an important technique for fostering broodstocks to build imitated eco-environment and artificial temperature control, light control, water quality regulation, and feeding of artificial compound feeds.
1.1 Water temperature Water temperature is a determinant factor in the development of gonorrhoids. The main role of water temperature is to stimulate the hypothalamic secretion and release factors of the giant salamander fish to the anterior pituitary gland, and the pituitary gland produces two kinds of hormones (LH, FSH). These two hormones can regulate the occurrence of sperm and egg. Water temperature can also promote brood feeding to meet the nutrients needed for sperm cell development.
The water temperature is controlled at 16-24°C from March to September every year. The daily temperature of water should not exceed 1.5°C.
1.2 The Illuminati littoralis have the habit of backlighting, that is, fear of light. Therefore, during broodstock incubation, light is controlled at 300-5001x. In other words, brooding is performed under low light conditions
1.3 Feed The nutrient-rich feed is the material basis for the development of gonadal gonads. During broodstock incubation, the protein content in the feed is 42%-45.3%, sugar is 10%-18%, fat is 2%-3%, multidimensional and minerals is 1.5%, others are 7%-12 %, 2% to 3% of Chinese medicine spermatozoa and follicle-stimulating soup. The amino acid balance of protein in feed is extremely important and cannot be ignored. At the same time, according to different phases of gonad development in the broiler, it is necessary to add different amounts of traditional Chinese medicine Shengjing San and traditional Chinese medicine to promote follicle soup compound feed, which is a key technology for broodstock cultivation.
1.4 Water quality regulation During the incubation of the broodstock, a good water environment must be created, which is conducive to the healthy growth and development of the broodstock. The parental pool water quality requires a high degree of mineralization, hardness, and transparency. The pH is 6.5-7.5. In mountainous areas, mountain water and stream water can be used to cultivate broods, and the principle is to apply well water (after aeration) or tap water to cultivate brooders. Water quality regulation mainly involves changing water frequently, diligently cleaning and diligently disinfecting.
2 Brooding
2.1 Parental rearing The main body of the carp is 2 ways: one is female and male polyculture; the other is female and male alone.
2.2 Breeding From the research on the technique of broodfish reproduction in our 20 years, we found that females have a higher sexual maturation rate and a greater amount of cocking eggs; males have a low sexual maturation rate of only 26.2%, and this is 26.2% of males. The quality of semen is not all qualified. Artificial broodstock grown in artificial tunnels, basements, and other artificial projects have a higher rate of sexual maturation in males, but lower female sexual maturation rates. Therefore, the giant salamander has so far failed to achieve large-scale production of seedlings. Therefore, the cultivation of a large number of sexually mature broodstocks is a key technology for the large-scale production of giant salamanders.
The development of gonorrhoids has a regular pattern of seasonal cycles. It begins to develop from March to April each year and matures from July to October. According to the law of seasonal cycles, artificially cultivated broods mainly include winter cultivation, spring cultivation, prenatal cultivation and supplemented by physiological methods.
2.2.1 Winter Breeding Winter breeding refers to the post-natal physical fitness stage. The main purpose of broodstock cultivation at this stage is to provide a large amount of nutrients to the body of the giant salamander, so that it can quickly return to health after birth; secondly, it is necessary to accelerate the degradation and absorption of sperm and ovary so that the sperm and ovary can develop normally. The main tasks of winter cultivation are water temperature regulation and feeding. Water temperature regulation is divided into two phases: First, the water temperature is controlled at 18-23 °C, so that the broodstock ingest a large number of food, restore health, and then the water temperature is reduced to below 10 °C, so that the parental into hibernation, accelerate the absorption and degradation of the spermary ovary. The winter bait feed is based on compound feed, and 2% lysine and 1% multivitamins are added to the compound feed.
2.2.2 Spring Breeding In spring, the gonads begin to develop. The gonad development at this stage is an important development period from the second phase to the third phase. The main purpose of brood cultivation at this stage is to promote the development of gonad gonads, so that their gonads can be converted to phase III as soon as possible; secondly, high-quality diets should be fed to meet the nutrients required for gonad development. The main tasks of spring cultivation are regulating water temperature, feeding, adjusting light and regulating water quality. The regulation of water temperature means that the temperature of the water is controlled at 18-22°C, so that the parental body starts to feed early and the gonadal begins to develop early; feeding is added to the compound feed that promotes the gonorrhea development of Chinese traditional medicine.
2.3 Prenatal culturing This stage of development of gonads is the period from the end of stage III to the stage of initial transformation of stage V. It is an important stage of gonad development and maturation. Therefore, the aim of brood cultivation at this stage is to cultivate a large number of mature broods, which are mainly cultivated using ecological and physiological methods.
Ecological approach: Wild dolphins inhabit in streams and springs in deep mountain gorges. Therefore, when artificially fostering broodstock, it is necessary to build a cave that mimics the ecological environment (returned water, spring water). At the same time, adjust the light to 5001x. The water temperature is controlled at the optimal growth and development range of 18-23°C for the broodstock.
Physiological method: It is to inject the spermatozoa of the Chinese medicine that promote the development of gonads in the parent body into the body and promote the development and maturation of the gonads. The joints between the nervous system and the gonadal pituitary glands are important organs for the development and maturation of germ cells. The Chinese medicine Shengjing injection can promote the release of hormones from the pituitary of the giant salamander and increase the maturation rate of the salamander.
3 Artificial breeding new technology
3.1 The parental selection of sexual maturation features: male vents uplift circle prominent white spots, visible swelling around the inside. B-ultrasonic examination: The testis is 7-9cm long and 1-2cm wide; the testis is full, with no groove in the middle indicating maturity.
The female's abdomen is enlarged, soft and elastic. B-ultrasonic examination showed that the egg diameter was 6-7mm, and the ovary showed a single chain bead indicating maturity.
3.2 artificial production
3.2.1 oxytocin RES+LRH-A+DOM,LRH-A+HCG.
3.2.2 Effort dose (according to new standards)
Female: RES+LRH-A+DOM4-10μg/kg;
HCG l00-1 5001U/kg
LRH-A 0.2-150μg/kg
Male: The above dose is halved.
3.2.3 Method of Oxygen Production The parental oxytocin injection site is the intercostal space, and the needle is 0.3-0.5cm. The needle is injected with a human infusion soft plastic tube needle, which can prevent the body from swinging when the parent body is injected.
3.2.4 Fertility effect The time from parental ovulation injection to spawning is 4-11 days, and the best spawning time is 4-6 days. Therefore, from the 3rd day after urging, someone should be on duty to observe whether the female parent produces egg glue film.
3.2.5 Artificial Insemination The artificial insemination of salamander adopts the wet and dry method. The operation procedure is: When the female parent body produces the pectin film, it should be immediately carried by hand to put the egg in the enamel pot, and the other person will mature. A good male parent catches up, puts a person in a stretcher, and uses his hands to lightly push the abdomen of the male. When there is milky white semen, the semen can be directly squeezed into the egg, and the eggs can be lightly shaken to make the egg fertilize evenly. After 15 minutes, add purified water. After another 20 minutes, the eggs were changed 2 times. Finally, the egg is placed in a small plastic basket and suspended in the hatchery, which is the incubation phase.
4 artificial hatching
4.1 Incubation of ecological conditions
4.1.1 Water temperature and water source Fertilized egg hatching water temperature requirements are strictly controlled at 20-22 °C. The daily temperature of the water should not exceed 1.0°C. The water source can be tap water, spring water (spring water first to put a reservoir, later release hatch pool), well water (to be used after aeration). Spring water is easy to hatch, so it is necessary to increase the temperature of the spring when hatching with spring water.
4.1.2 Dissolved Oxygen The amount of oxygen in the body of fertilized eggs should not be less than 5 mg/L.
4.1.3 Light Fertilized Eggs Lighting should be strictly controlled at about 300lx.
4.2 artificial incubation of fertilized eggs mainly using static water incubation and microfluidic water incubation. When water is hatched, change the water 1 time in the morning and in the evening.
Note 3 points during hatching: First, control the water temperature and light; Second, prevent and control water mold, that is, regular mildew net prevention during incubation, the concentration of 0.5-0.810-6, soaking the fertilized eggs for 10 minutes; third is to prevent Embryos "stick", that is regularly flipping fertilized eggs.