CN113598093B - Artificial breeding method of barbel grahami - Google Patents

Artificial breeding method of barbel grahami Download PDF

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CN113598093B
CN113598093B CN202110872817.8A CN202110872817A CN113598093B CN 113598093 B CN113598093 B CN 113598093B CN 202110872817 A CN202110872817 A CN 202110872817A CN 113598093 B CN113598093 B CN 113598093B
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meal
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CN113598093A (en
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李文俊
韩崇
李陇旭
易祖盛
桂林
钟良明
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Guangzhou University
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    • A23K—FODDER
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23K20/158—Fatty acids; Fats; Products containing oils or fats
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/163—Sugars; Polysaccharides
    • A—HUMAN NECESSITIES
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    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
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    • A23K20/174—Vitamins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
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Abstract

The invention belongs to the technical field of freshwater fish breeding, and discloses an artificial breeding method of barbel grahami. The artificial breeding method comprises the following steps: parent fish is cultivated, artificial induction of labor, artificial insemination, embryonated egg hatching and seedling cultivation, the spiny barbel is bred artificially, the induction rate reaches more than 80%, the fertility rate reaches more than 90%, the fry survival rate reaches more than 90%, efficient artificial breeding of the spiny barbel is realized, the production efficiency is improved, and sustainable development and utilization of parent fish is guaranteed.

Description

Artificial breeding method of barbel grahami
Technical Field
The invention belongs to the technical field of freshwater fish breeding, and particularly relates to an artificial breeding method of barbel grahami.
Background
Light barb cheilbus hollandi (spinebabus hollandi), it is affiliated to Cypriniformes (Cypriniformes), cyprinidae (Cyprinidae), the cheilbus subfamily (Barbineae), the barb cheilus belongs to (spinebabus), each river system in south above the Yangtze river is main distribution in china, is a famous economic fish that has higher nutrition and medicinal value, receives market and culturist deeply. However, since the 80's of the 20 th century, the natural population resources of the barbel grahami have been rapidly declining due to various human factors, and many rivers in China have been difficult to trace. In order to protect the aquatic germplasm resource, more than 10 national-grade aquatic germplasm resource protection areas of the Sinocyclocheilus grahami are established in Guangdong province, guangxi province, jiangxi province, fujian province, hunan province, anhui province and other provinces by the Ministry of agriculture.
Due to the damage of the living environment of the origin, the population quantity and the quality of the spinibarbus grahami are seriously exhausted, and the risk of extinction of wild germplasm resources exists. Therefore, research works such as artificial breeding, cultivation and development need to be carried out on the hybrid plants so as to recover and continue germplasm resources and meet gradually-expanded market demands. At present, because of factors in aspects of reproduction biology, morphology and the like of the spiny barbell, diseases frequently occur under high-density cultivation, and the influence of drug residue, drug resistance enhancement, animal immunity reduction and the like caused by the large-scale use of antibacterial agent for prevention and treatment restricts the artificial reproduction and large-scale production of the spiny barbell; in addition, because the light barbed barbel has a small brooding amount which is only 1/15-1/5 of that of the Chinese carps, the conventional propagation technical means cannot make up the deficiency caused by the brooding amount and cannot meet the continuously expanded market demand.
Disclosure of Invention
The invention aims to provide a method for artificially breeding a barbel grahami.
The second aspect of the present invention is directed to provide the use of the artificial breeding method of the first aspect in fish breeding.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
the invention provides a first aspect of an artificial breeding method of Sinocyclocheilus grahami, which comprises the following steps:
s1, parent breeding:
a. and (3) daily cultivation: feeding parent fishes with the feed A, wherein the stocking density of the parent fishes is 100-300 tails/mu;
b. pre-partum cultivation: feeding the chicken with the feed B, and keeping the flowing water stimulation of the flushing water flow velocity of 5-20 cm/s;
c. pre-parturition ripening: injecting artificial hormone into parent fish to pre-induce spawning, and simultaneously stimulating by running water of 10-30 cm/s;
s2, artificial spawning induction: selectively mature parent fish, injecting artificial hormone twice, injecting female fish with the first needle and injecting female and male parent fish with the second needle, and keeping flowing water stimulation of 20-50 cm/s in the whole process;
s3, artificial fertilization;
s4, hatching fertilized eggs: artificially incubating the fertilized eggs obtained in the step S3 in a micro-flow water stimulation mode to obtain larval fish;
s5, seedling cultivation:
a. fry breeding: culturing the larval fish obtained in the step S4, and feeding initial baits after 3-5 days;
b. culturing and domesticating the young fishes: domesticating the food by using powdery feed;
the feed A comprises fish meal, wheat bran, soybean meal, rapeseed meal, flour, corn flour, soybean oil, beer yeast, betaine, choline chloride, taurine, potassium diformate, monocalcium phosphate, compound minerals and compound vitamins;
the feed B comprises feed C and animal fat;
the feed C comprises feed A, vitamin C, vitamin E, vitamin H, L-carnitine, mannan-oligosaccharide, beta-glucose and turmeric powder.
Preferably, the feed A comprises 5-10% of fish meal, 20-27% of wheat bran, 20-27% of soybean meal, 15-20% of rapeseed meal, 5-9% of flour, 5-8% of corn flour, 2-5% of soybean oil, 2-5% of beer yeast, 1-3% of betaine, 0.2-0.5% of choline chloride, 0.05-0.15% of taurine, 1-5% of potassium diformate, 0.5-1% of monocalcium phosphate, 0.5-1.5% of compound minerals and 0.5-1.5% of compound vitamins by weight percentage.
More preferably, the feed A comprises 6-7% of fish meal, 25-26% of wheat bran, 25-26% of soybean meal, 18-19% of rapeseed meal, 5-6% of flour, 5-6% of corn flour, 3-5% of soybean oil, 3-5% of beer yeast, 1.6-2% of betaine, 0.3-0.5% of choline chloride, 0.1-0.15% of taurine, 2-5% of potassium diformate, 0.6-1% of monocalcium phosphate, 1-1.5% of compound minerals and 1-1.5% of compound vitamins by weight percentage.
Preferably, the feeding frequency of the feed A is 2-4 times/d, and the feeding rate is 1-3%.
Further preferably, the feeding frequency of the feed A is 2-3 times/d, and the feeding rate is 1-2%.
Preferably, the feed C comprises 92 to 96 percent of feed A, 0.01 to 0.07 percent of vitamin C, 0.01 to 0.07 percent of vitamin E, 0.01 to 0.07 percent of vitamin H, 0.5 to 1.5 percent of L-carnitine, 1 to 3 percent of mannan-oligosaccharide, 0.5 to 1.5 percent of beta-glucose and 0.5 to 1.5 percent of turmeric powder by weight percentage.
More preferably, the feed C comprises 94.85-96 wt% of feed A, 0.05-0.07 wt% of vitamin C, 0.01-0.05 wt% of vitamin E, 0.01-0.05 wt% of vitamin H, 1-1.5 wt% of L-carnitine, 2-3 wt% of mannan oligosaccharide, 1-1.5 wt% of beta-glucose and 0.5-1 wt% of turmeric powder.
Preferably, the feed B is formed by mixing the feed C and animal fat according to the mass ratio of (16-18): 1.
Preferably, the feeding frequency of the feed B in the prenatal cultivation in the step S1 is 2-4 times/d, and the feeding rate is 1-4%.
Further preferably, the feeding frequency of the feed B in the prenatal cultivation in the step S1 is 2-3 times/d, and the feeding rate is 1-2%.
Preferably, the animal fat is at least one of duck oil, chicken oil, lard, beef tallow and mutton tallow.
Preferably, the powdered feed in step S5 includes fish meal, shrimp meal, earthworm powder, rapeseed meal, starch, soybean meal, corn meal, brewer' S yeast, soybean oil, lactobacillus, mannooligosaccharide, beta-glucose, monocalcium phosphate, choline chloride, vitamin complex and mineral complex.
Further preferably, the powdered feed in step S5 comprises, by weight: 20 to 25 percent of fish meal, 5 to 10 percent of shrimp meal, 15 to 20 percent of earthworm powder, 10 to 15 percent of rapeseed meal, 3 to 7 percent of starch, 15 to 20 percent of soybean meal, 4 to 9 percent of corn flour, 3 to 8 percent of beer yeast, 3 to 8 percent of soybean oil, 0.5 to 1 percent of lactobacillus, 1 to 3 percent of mannan oligosaccharide, 0.5 to 1 percent of beta-glucose, 0.5 to 1 percent of calcium dihydrogen phosphate, 0.2 to 0.5 percent of choline chloride, 0.5 to 1.5 percent of compound vitamin and 0.5 to 1.5 percent of compound mineral.
Still more preferably, the powdered feed in the step S5 comprises, by weight: 22 to 25 percent of fish meal, 7 to 8 percent of shrimp meal, 16 to 18 percent of earthworm powder, 10 to 11 percent of rapeseed meal, 3 to 3.5 percent of starch, 15 to 18 percent of soybean meal, 5 to 9 percent of corn flour, 6.5 to 7.5 percent of beer yeast, 3 to 4 percent of soybean oil, 0.5 to 0.6 percent of lactobacillus, 1 to 2 percent of mannan oligosaccharide, 0.8 to 0.9 percent of beta-glucose, 0.8 to 1 percent of monocalcium phosphate, 0.4 to 0.5 percent of choline chloride, 1 to 1.5 percent of compound vitamin and 1 to 1.5 percent of compound mineral.
Preferably, the artificial hormone in step S1 is LRH-A2.
Preferably, the injection dosage of the artificial hormone in the step S1 is 1-4 mug/kg.
Further preferably, the injection dosage of the artificial hormone in the step S1 is 1 to 3 mug/kg.
Preferably, the parent fish in step S1 has an age of IV or more and a body weight of 1kg or more.
Preferably, the stocking density of the parent fish in the step S1 is 200-300 tails/mu.
Preferably, the dissolved oxygen of the water used for the daily cultivation in the step S1 is more than or equal to 5mg/L.
Preferably, the pH of the water used for the daily cultivation in step S1 is 7.0 to 9.0.
Preferably, the flowing water stimulation of 5-10 cm/S is kept in the prenatal cultivation process in the step S1.
Preferably, the flowing water stimulation of 20-30 cm/S is kept in the step S1 during the pre-parturition ripening.
Preferably, in the step S2, the artificial hormone injected by the first needle is LRH-A2, the injection amount is 1-3 μ g/kg, and the artificial hormone injected by the second needle is a mixed artificial hormone, wherein the mixed artificial hormone comprises HCG, DOM and LRH-A2.
Further preferably, the injection dosage of the artificial hormone injected by the first needle in the step S2 is 1 to 2 μ g/kg.
Further preferably, the injection dosage of the mixed artificial hormone in the step S2 is 400-600 IU/kg HCG, 3-6 mug/kg DOM and 3-5 mug/kg LRH-A2.
Still more preferably, the injection dosage of the mixed artificial hormone in the step S2 is 500-600 IU/kg HCG, 4-6 mug/kg HCG and 4-5 mug/kg LRH-A2.
Further preferably, when the female fish is injected by the second needle in the step S2, the injection dosage of the male fish is 1/3 to 1/2 of that of the female fish.
Further preferably, in the step S2, the interval between the injection of the artificial hormone by the first needle and the injection of the artificial hormone by the second needle is 12 to 14 hours.
Preferably, the step S2 is kept at 30-40 cm/S of running water stimulation all the time.
Preferably, the ratio of male to female of the parent fish in step S2 is (3 to 5): 1.
preferably, the artificial induced spawning effect time after the two-needle injection in step S2 is: 12 to 20 hours.
Preferably, the method for artificial fertilization in step S3 is dry fertilization.
Further preferably, the dry insemination method comprises the following specific steps: slightly pressing the abdomen of the female fish, extruding out the eggs, simultaneously diluting the semen extruded by the male fish with the random liquid, pouring the diluted semen into the eggs, and stirring and fertilizing with feathers.
Further preferably, the volume ratio of the semen to the anyhow liquid in the dry fertilization is (2-3): 1.
preferably, the dissolved oxygen of the water body in the artificial incubation of the fertilized eggs in the step S4 is more than or equal to 6mg/L, and the water temperature is 25-28 ℃.
Preferably, the initial bait in the step S5 is earthworm slurry and/or cooked egg yolk.
Preferably, the feeding frequency of the initial bait in the step S5 is 4-6 times/d, and the feeding rate is 10-40 g/ten thousand earthworm pulp and 1-3 cooked egg yolks/10 ten thousand earthworm pulp.
Further preferably, the feeding frequency of the initial bait in the step S5 is 4-5 times/d, and the feeding rate is 10-30 g/ten thousand earthworm pulp and 1-2 cooked egg yolks/10 ten thousand egg yolks.
Preferably, the compound minerals in the feed A and the powdered feed comprise NaCl and ZnSO 4 ·7H 2 O、FeSO 4 ·7H 2 O、MnSO 4 ·4H 2 O、CuSO 4 ·5H 2 O、CoCl 2 、Na 2 SeO 3 And KI.
Further preferably, the compound mineral in the feed A and the powder feed comprises 100-110 g/kg of NaCl and ZnSO 4 ·7H 2 O 23～25g/kg、FeSO 4 ·7H 2 O 22～25g/kg、MnSO 4 ·4H 2 O 8.1～9.4g/kg、CuSO 4 ·5H 2 O 2.2～3g/kg、CoCl 2 110～120mg/kg、Na 2 SeO 3 40-50 mg/kg and 30-32 mg/kg of KI.
Preferably, the compound vitamins in the feed A and the powdered feed comprise vitamin A and vitamin B 1 Vitamin B 2 Vitamin B 3 Vitamin B 5 Vitamin B 6 Vitamin B 12 Vitamin C, vitamin D, vitamin E, vitamin H and vitamin K 3 And vitamin M.
More preferably, the compound vitamins in the feed A and the powdered feed comprise 800-900 IU/g of vitamin A and vitamin B 1 0.3-0.5 IU/g and vitamin B 2 1.1-1.5 IU/g, vitamin B 3 3-4 IU/g and vitamin B 5 3-5 IU/g and vitamin B 6 0.5-0.7 IU/g and vitamin B 12 0.002-0.006 IU/g, vitamin C5.5-5.9 IU/g, vitamin D200-250 IU/g, vitamin E2-5 IU/g, vitamin H10-12 IU/g, vitamin K 3 2-6 IU/g and 0.2-0.5 IU/g of vitamin M.
Preferably, the artificial breeding method of the barbel grahami further comprises nursing of postpartum parent fishes.
Further preferably, the postpartum care comprises the steps of: in order to disinfect parent fish in vitro, penicillin is injected at the same time, and feed C is fed;
further preferably, the in vitro sterilization of the parent fish is the in vitro sterilization of the parent fish by using a potassium permanganate solution.
Further preferably, the concentration of the potassium permanganate solution is 15-20 mg/L.
In a second aspect of the invention, there is provided the use of the artificial breeding method of the first aspect in fish breeding.
The invention has the beneficial effects that: 1. according to the invention, the artificial breeding of the barbel spinuloides is carried out by adopting the steps of parent breeding, artificial induced spawning, artificial fertilization, oosperm incubation, fry breeding and the like for the first time, the induced spawning rate reaches more than 80%, the fertilization rate reaches more than 90%, and the fry survival rate reaches more than 90%, so that the efficient artificial breeding of the barbel spinuloides is realized, the production efficiency is improved, and the sustainable development and utilization of parent fishes are ensured; 2. the artificial breeding method of the spinibarbus grahami is convenient to operate and control, and large-scale production and cultivation of the spinibarbus grahami can be realized; 3. the method can provide technical support and seedling guarantee for germ plasm resource protection of the spinulobus grahami and resource restoration of a natural population distribution water area of the spinulobus grahami, and lays a good foundation for large-scale seedling culture and artificial culture of the spinulobus grahami.
Detailed Description
The present invention will now be described in detail with reference to specific examples, but the scope of the present invention is not limited thereto.
The materials, reagents and the like used in the present examples are commercially available materials and reagents unless otherwise specified.
Example 1
An artificial breeding method of barbel grahami includes the following steps:
at the end of 5 months in 2019, the artificial propagation of the barbus glabra is carried out by applying the method disclosed by the invention to a fish breeding field of Jinyue aquatic product Limited company in Guangdong Shaoguan city.
(1) Parent breeding:
a. and (3) daily cultivation: at 1500m 2 Culturing parent of the spinibarbus grahami in a pond with the water depth of 1.3m, wherein the parent fish comes from the population of the water system of the northern river, the stocking density is 200 tails/mu, the parent age is more than IV, and the weight is more than 1 kg; feeding feed A (in weight percentage, comprising7% of fish meal, 25% of wheat bran, 26% of soybean meal, 18% of rapeseed meal, 5% of flour, 6% of corn flour, 3% of soybean oil, 3% of beer yeast, 1.6% of betaine, 0.3% of choline chloride, 0.1% of taurine, 2% of potassium diformate, 1% of monocalcium phosphate, 1% of compound mineral substances and 1% of compound vitamins), feeding frequency is 2 times/d, feeding rate is 2%, and an oxygenation facility is arranged to ensure that the dissolved oxygen content of the culture water body is more than or equal to 5mg/L and pH is 7.0-8.5.
b. Pre-partum cultivation: starting 2 months before spawning promotion, feeding the parent fish with the feed C (comprising 94.85% of feed A, 0.05% of vitamin C, 0.05% of vitamin E, 0.05% of vitamin H, 1% of L-carnitine, 2% of mannan-oligosaccharide, 1% of beta-glucose and 1% of turmeric powder in percentage by weight) coated with duck oil accounting for 6% of the weight of the feed C to stimulate the gonads of the parent fish to rapidly and synchronously develop, simultaneously improve the anti-stress capability and the immunity of the parent fish, feed the parent fish at a frequency of 2 times/d and a feed rate of 2%, and maintain running water stimulation with a running water speed of 5cm/s in the culture process.
c. Pre-parturition ripening: at 15d before induction, the basal part of the pectoral fin of the parent fish is injected with luteinizing hormone releasing hormone analogue A2 (LRHR-A2) in the abdominal cavity, the dosage is 1 mu g/kg, the angle between the needle and the fish body is 45 degrees, the depth of the needle is 1 cm-1.5 cm, and the dosages of male and female are the same. After injection, the culture is continued in the pond, meanwhile, the running water stimulation is enhanced (the flow rate is 20 cm/s), and an escape-proof net is arranged, wherein the net height is 2m.
(2) Artificial hasten parturition
The method comprises the following steps of selecting individuals with mature gonads from parent fishes which are intensively cultured and ripened prenatally, and carrying out artificial induced spawning at a water temperature of 23 ℃, wherein the male-female ratio is 3. The induced spawning process is divided into two injections with the interval of 12h, and 1 mu g/kg of LRH-A2 is injected into female fish for the 1 st time; the 2 nd male and female parent fish are injected with the medicine: 500IU/kg Chorionic Gonadotropin (HCG), 4 mug/kg deodourone maleate (Domperidone, DOM) and 5 mug/kg LRH-A2, diluted to 2mL with normal saline, halved male fish dose, and 12h effect time after two-needle injection; the whole course keeps 30cm/s running water stimulation, and the fish net is tightly covered above the pool to prevent the parent fish from jumping out of the pool, the covering net is preferably a loose-hole soft fishing net, and the injury of the fish in the jumping impact process is reduced.
(3) Artificial insemination
Inseminating by adopting a dry method inseminating mode, slightly pressing the abdomen of a female fish in an effector phase, extruding eggs into a clean and dry container, simultaneously diluting semen extruded by a male fish by using 0.5 time (v/v) of a fish-use Rev liquid, immediately pouring the diluted semen into the freshly extruded eggs, immediately and slightly stirring the diluted semen by using feathers for 2min for artificial insemination, standing the eggs for 1min, and then rinsing the fertilized eggs by using 0.5% sodium chloride solution for 1 time; the sperm, ovum and fertilized ovum should be protected from direct sunlight.
(4) Postpartum parent fish nursing
After the artificial spawning of parent fish, the parent fish is weak and often injured, so the postpartum care needs to be strengthened to improve the survival rate. The parent fish is disinfected in vitro by potassium permanganate solution (the concentration is 20 mg/L), meanwhile, penicillin (8 ten thousand units/kg) is injected into the parent fish, then the parent fish is placed into a pond with fresh water, and the feed C is continuously fed for a period of time to help the parent fish to recover as soon as possible.
(5) Fertilized egg hatching
Transferring the fertilized eggs after water absorption and expansion into an incubation frame, arranging a 30-mesh screen at the bottom, and putting the fertilized eggs with the density of about 30000 granules/m 2 Hatching by micro-fluid water jet; during incubation, the dissolved oxygen of the water body is ensured to be more than or equal to 6mg/L; incubating at 25 deg.C; and (5) the fry comes out of the film for 45-55 h.
(6) Fry rearing
a. Fry breeding:
transferring the fish fry after the fish fry emerges from the film to an area of 15m 2 Temporarily culturing in a cement seedling pond with water depth of 0.3m and stocking fish fry at 30000 tails/m 2 To avoid bringing in unhatched bad eggs; keeping the water quality of a water source fresh, keeping the dissolved oxygen of the water body over 6mg/L, keeping the temperature of the water body between 22 and 28 ℃, and avoiding direct irradiation of sunlight; the total length of the newly emerged larva fishes is 6-8 mm, the larva fishes do less movement, the colony stays at the water bottom, and a large yolk sac is coated outside the larva fishes to maintain life activities by internal nutrition; after 3-5 days of yolk sac absorption, the fry continuously floats to the water surface and swims to the bottom of the pondFeeding a small amount of initial bait (a small amount of earthworm slurry sieved by 80 meshes and cooked egg yolk filtered by rubbing by a 300-mesh sieve are dropped along the edge of the pond surface) 5 times a day, wherein the feeding rate is 20-30 g/ten thousand of earthworm slurry and 2/10 ten thousand of cooked egg yolk, so that the nutritional requirement of the egg yolk in the growth transition period is ensured, and simultaneously, the water is drained and changed 1 time by a siphon method every 2 days to keep the water body clean.
b. Culturing and domesticating the young fishes: continuously feeding earthworm slurry from 15-day-old young fish, and adding small amount of powdered feed (comprising fish meal 25%, shrimp meal 8%, earthworm powder 16%, rapeseed meal 10%, starch 3%, soybean meal 18%, corn flour 5%, beer yeast 6.5%, soybean oil 3% and lactobacillus 0.5% (10%) by weight 6 ～10 7 cfu/g), 1% of mannan-oligosaccharide, 0.8% of beta-glucose, 0.8% of monocalcium phosphate, 0.4% of choline chloride, 1% of compound vitamin and 1% of compound mineral substance, the feeding frequency of the powdery feed is 4 times/d, the feeding rate is 5%, in the stage, the seedling raising pool is filled with water to the water level of 50-60 cm, the pollution and pollution are absorbed and discharged every day, and a small amount of water is supplemented. The proportion of feeding the powdery feed is gradually increased from 25-day-old young fishes until the powdery feed is completely fed, the feeding frequency is 4 times/d, the feeding rate is 7%, the water level of the culture pond is increased to 70-80 cm at the stage, and the dissolved oxygen of the water body is kept to be more than 6 mg/L. The juvenile fish fries cultured for 40-50 days have the specification of 4-5 cm and strong survival ability, and can be transferred into a large pond water body to be matched with zooplankton for continuous culture;
wherein the compound minerals comprise salt 100g/kg and ZnSO 4 ·7H 2 O 23g/kg、FeSO 4 ·7H 2 O 22g/kg、MnSO 4 ·4H 2 O 8.1g/kg、CuSO 4 ·5H 2 O 2.2g/kg、CoCl 2 110mg/kg、Na 2 SeO 3 50mg/kg and KI 32mg/kg;
the compound vitamins include vitamin A800 IU/g, vitamin B 1 0.3IU/g, vitamin B 2 1.1IU/g, vitamin B 3 3IU/g and vitamin B 5 3IU/g and vitamin B 6 0.5IU/g, vitamin B 12 0.002IU/g, 5.5IU/g vitamin C, 200IU/g vitamin D, 5IU/g vitamin E, 10IU/g vitamin H, vitamin K 3 2IU/g and vitamin M0.2 IU/g.
Breeding parents of the Sinocycheilus grahami Beijiang population at this time, hastening parturition for 143 tails of female fishes, hastening parturition for 121 tails successfully, and ensuring that the spawning hastening rate is 84.62 percent and no parent loss exists; obtaining 24.5 ten thousand eggs and 22.5 ten thousand fertilized eggs, wherein the fertilization rate is 91.84%; hatching 20.7 ten thousand young fries, wherein the hatching rate is 92.00 percent; after 40-50 days of cultivation, 19.8 thousands of barbus barbel seedlings with the average size of 4-5 cm are obtained, the survival rate of seedling cultivation is 95.65%, and high-efficiency artificial breeding is realized.
Example 2
An artificial breeding method of Sinocyclocheilus grahami comprises the following steps:
in the middle ten days of the 5 th month of 2020, in the fish fingerling field of jinyue aquatic products limited company, guangdong Shaoguan city, the artificial propagation of the spiny barbel can be carried out by applying the method of the invention.
(1) Parent breeding:
a. and (3) daily cultivation: at 1500m 2 Culturing parent of the spinibarbus grahami in a pond with the water depth of 1.3m, wherein the parent fish comes from the population of the water system of the northern river, the stocking density is 200 tails/mu, the parent age is more than IV, and the weight is more than 1 kg; the feed A (by weight percentage, including 6% of fish meal, 26% of wheat bran, 25% of soybean meal, 19% of rapeseed meal, 6% of flour, 5% of corn flour, 3% of soybean oil, 3% of beer yeast, 2% of betaine, 0.3% of choline chloride, 0.1% of taurine, 2% of potassium diformate, 0.6% of monocalcium phosphate, 1% of compound mineral and 1% of compound vitamin) is fed at a feeding frequency of 2 times/d and a feeding rate of 2%, and an oxygenation facility is provided to ensure that the dissolved oxygen content of the culture water body is not less than 5mg/L and the pH value is 7.0-8.5.
b. Pre-partum cultivation: starting 2 months before spawning induction, feeding the feed C (comprising 94.85% of feed A, 0.05% of vitamin C, 0.05% of vitamin E, 0.05% of vitamin H, 1% of L-carnitine, 2% of mannan-oligosaccharide, 1% of beta-glucose and 1% of turmeric powder in percentage by weight) coated with duck oil accounting for 6% of the weight of the feed C to stimulate the gonads of the parent fishes to rapidly and synchronously develop, simultaneously improve the anti-stress capability and the immunity capability of the parent fishes, feed frequency is 2 times/d, feed rate is 2%, and water flow stimulation with water flow velocity of 5cm/s is maintained in the culture process.
c. Pre-parturition ripening: at 15d before induction, the basal part of the pectoral fin of the parent fish is injected with luteinizing hormone releasing hormone analogue A2 (LRHR-A2) in the abdominal cavity, the dosage is 1 mu g/kg, the angle between the needle and the fish body is 45 degrees, the depth of the needle is 1 cm-1.5 cm, and the dosages of male and female are the same. After injection, the culture is continued in the pond, meanwhile, the running water stimulation is enhanced (the flow rate is 20 cm/s), and an escape-proof net is arranged, wherein the net height is 2m.
(2) Artificial hasten parturition
The method comprises the following steps of selecting individuals with mature gonads from parent fishes which are intensively cultured and ripened prenatally, and carrying out artificial induced spawning at a water temperature of 23 ℃, wherein the male-female ratio is 3. The induced spawning process is divided into two injections with the interval of 12h, and 1 mu g/kg of LRH-A2 is injected into female fish for the 1 st time; the 2 nd male and female parent fish are injected with the medicine: 500IU/kg Chorionic Gonadotropin (HCG), 4 mug/kg deodourone maleate (Domperidone, DOM) and 5 mug/kg LRH-A2, diluted to 2mL with normal saline, halved male fish dose, and 12h effect time after two-needle injection; the whole course keeps 30cm/s running water stimulation, and the fish net is tightly covered above the pool to prevent the parent fish from jumping out of the pool, the covering net is preferably a loose-hole soft fishing net, and the injury of the fish in the jumping impact process is reduced.
(3) Artificial insemination
Inseminating by adopting a dry method inseminating mode, slightly pressing the abdomen of a female fish in an effector phase, extruding eggs into a clean and dry container, simultaneously diluting semen extruded by a male fish by using 0.5 time (v/v) of a fish-use Rev liquid, immediately pouring the diluted semen into the freshly extruded eggs, immediately and slightly stirring the diluted semen by using feathers for 2min for artificial insemination, standing the eggs for 1min, and then rinsing the fertilized eggs by using 0.5% sodium chloride solution for 1 time; the sperm, ovum and fertilized egg should be protected from direct sunlight.
(4) Postpartum parent fish nursing
After the artificial spawning of parent fish, the parent fish is weak and often injured, so the postpartum care needs to be strengthened to improve the survival rate. The parent fish is disinfected in vitro by potassium permanganate solution (the concentration is 20 mg/L), meanwhile, penicillin (8 ten thousand units/kg) is injected into the parent fish, then the parent fish is placed into a pond with fresh water, and the feed C is continuously fed for a period of time to help the parent fish to recover as soon as possible.
(5) Fertilized egg hatching
Transferring the fertilized eggs after water absorption and expansion into an incubation square frame, arranging a 30-mesh screen at the bottom of the square frame, and putting the fertilized eggs with the density of about 30000 eggs/m 2 Hatching by micro-fluid water jet; during incubation, the dissolved oxygen of the water body is ensured to be more than or equal to 6mg/L; incubating at 25 deg.C; and (5) the fry comes out of the film for 45-55 h.
(6) Fry rearing
a. Fry breeding:
transferring the fish fry after the fish fry emerges from the film to an area of 15m 2 Temporarily culturing in a cement seedling pond with water depth of 0.3m and stocking fish fry at 30000 tails/m 2 To avoid bringing in unhatched bad eggs; keeping the water quality of the water source fresh, keeping the dissolved oxygen of the water body over 6mg/L, keeping the temperature of the water body between 22 and 28 ℃, and avoiding direct irradiation of sunlight. The total length of the newly emerged larva fishes is 6-8 mm, the larva fishes do less movement, the colony stays at the water bottom, and a large yolk sac is coated outside the larva fishes to maintain life activities by internal nutrition; after the yolk sacs are absorbed by half, the larvae continuously float up to the water surface and swim to the bottom of the pool to start foraging, a small amount of initial feed (a small amount of 80-mesh sieved earthworm slurry and 300-mesh filtered cooked egg yolk are dripped along the edge of the pool surface) is fed at the time, the feeding frequency is 5 times/d, the feeding rate is 20-30 g/ten thousand tails of the earthworm slurry and 2/10 ten thousand tails of the cooked egg yolk, the nutritional requirement of the larvae in the growth transition period is ensured, and water is drained and changed 1 time every 2 days by a siphon method to keep the water body clean.
b. Culturing and domesticating the young fishes: continuously feeding the earthworm slurry from the young fish of 15 days old, and mixing with a small amount of powdered feed (by weight percentage, including fish meal 22%, shrimp meal 7%, earthworm powder 18%, rapeseed meal 11%, starch 3.5%, soybean meal 15%, corn flour 5%, beer yeast 7.5%, soybean oil 4%, lactobacillus 0.6% (10%) 6 ～10 7 cfu/g), 2 percent of mannan-oligosaccharide, 0.9 percent of beta-glucose, 1 percent of monocalcium phosphate, 0.5 percent of choline chloride, 1 percent of compound vitamin and 1 percent of compound mineral substance), the feeding frequency of the powdery feed is 4 times/d, the feeding rate is 5 percent, and the stage is to grow seedlingsThe pond is filled with water to reach the water level of 50-60 cm, sewage suction and discharge are carried out every day, and a small amount of water is supplemented. The proportion of feeding the powdery feed is gradually increased from 25-day-old young fishes until the powdery feed is completely fed, the feeding frequency is 4 times/d, the feeding rate is 7%, the water level of the culture pond is increased to 70-80 cm at the stage, and the dissolved oxygen of the water body is kept to be more than 6 mg/L. The juvenile fish fries cultured for 40-50 days have the specification of 4-5 cm and strong survival ability, and can be transferred into a large pond water body to be matched with zooplankton for continuous culture.
Wherein the compound minerals comprise salt 100g/kg and ZnSO 4 ·7H 2 O 23g/kg、FeSO 4 ·7H 2 O 22g/kg、MnSO 4 ·4H 2 O 8.1g/kg、CuSO 4 ·5H 2 O 2.2g/kg、CoCl 2 110mg/kg、Na 2 SeO 3 50mg/kg and KI 32mg/kg;
the compound vitamins include vitamin A800 IU/g, vitamin B 1 0.3IU/g, vitamin B 2 1.1IU/g, vitamin B 3 3IU/g, vitamin B 5 3IU/g, vitamin B 6 0.5IU/g, vitamin B 12 0.002IU/g, 5.5IU/g vitamin C, 200IU/g vitamin D, 5IU/g vitamin E, 10IU/g vitamin H, vitamin K 3 2IU/g and vitamin M0.2 IU/g.
Culturing parents of the Sinocyclocheilus grahami Beijiang population at this time, inducing spawning for 123 fish, successfully inducing spawning for 109 fish, and inducing spawning rate to be 88.62%, wherein no parent loss exists; obtaining 22.0 ten thousand eggs and 20.0 ten thousand fertilized eggs, wherein the fertilization rate is 90.91%; hatching 18.7 thousands of young fries, wherein the hatching rate is 93.50 percent; through 40-50 days of cultivation, 17.6 ten thousand of barbel seedlings with the average specification of 4-5 cm are obtained, the survival rate of the seedlings is 94.12%, and efficient artificial breeding is realized.
While the embodiments of the present invention have been described in detail, the present invention is not limited to the embodiments, and various changes can be made without departing from the spirit of the present invention within the knowledge of those skilled in the art. Furthermore, the embodiments of the present invention and features of the embodiments may be combined with each other without conflict.

Claims (6)

1. An artificial breeding method of a spinibarbus hollandis is characterized by comprising the following steps:
s1, parent cultivation:
a. and (3) daily cultivation: feeding parent fishes with the feed A, wherein the stocking density of the parent fishes is 100 to 300 tails/mu;
b. pre-partum cultivation: feeding the chicken with feed B, and keeping the water flow velocity of 5 to 20cm/s for running water stimulation;
c. pre-parturition ripening: injecting artificial hormone into parent fish for pre-induced spawning, and simultaneously stimulating by running water at 10 to 30cm/s;
s2, artificial induced spawning: selectively injecting artificial hormones into mature parent fishes twice, injecting female fishes into a first needle and injecting male and female parent fishes into a second needle, and keeping running water stimulation of 20 to 50cm/s in the whole process;
s3, artificial fertilization;
s4, hatching fertilized eggs: artificially hatching the fertilized eggs obtained by fertilization in the step S3 in a micro-flow water stimulation mode to obtain larval fish;
s5, seedling cultivation:
a. fry breeding: culturing the fry obtained in the step S4, and feeding initial baits after 3 to 5 d;
b. culturing and domesticating the young fishes: domesticating the diet with powdery feed;
the feed A comprises fish meal, wheat bran, soybean meal, rapeseed meal, flour, corn flour, soybean oil, beer yeast, betaine, choline chloride, taurine, potassium diformate, monocalcium phosphate, compound minerals and compound vitamins;
the feed B comprises a feed C and animal grease;
the feed C comprises feed A, vitamin C, vitamin E, vitamin H, L-carnitine, mannan-oligosaccharide, beta-glucose and turmeric powder; the feed C comprises 92-96% of feed A, 0.01-0.07% of vitamin C, 0.01-0.07% of vitamin E, 0.01-0.07% of vitamin H, 0.5-1.5% of L-carnitine, 1-3% of mannan-oligosaccharide, 0.5-1.5% of beta-glucose and 0.5-1.5% of turmeric powder;
the female-male ratio of the parent fish in the step S2 is (3-5): 1;
when the female fish is injected by the second needle in the step S2, the injection dosage of the male fish is 1/3-1/2 of that of the female fish;
the powdered feed in the step S5 comprises: fish meal, shrimp meal, earthworm powder, rapeseed meal, starch, soybean meal, corn meal, beer yeast, soybean oil, lactobacillus, mannan oligosaccharide, beta-glucose, monocalcium phosphate, choline chloride, compound vitamins and compound mineral substances;
the initial bait in the step S5 is earthworm paste and/or cooked egg yolk; the feeding frequency of the opening bait is 4 to 6 times/d, the feeding rate is 10 to 40g/ten thousand tails of earthworm slurry, and 1 to 3/10 ten thousand tails of cooked egg yolk;
in the step S2, the artificial hormone injected by the first needle is LRH-A2, and the injection dosage is 1 to 3 micrograms per kg; the artificial hormone injected by the second needle is mixed artificial hormone; the mixed artificial hormone comprises HCG, DOM and LRH-A2.
2. The artificial breeding method according to claim 1, wherein the feed A comprises, by weight, 5% -10% of fish meal, 20% -27% of wheat bran, 20% -27% of soybean meal, 15% -20% of rapeseed meal, 5% -9% of flour, 5% -8% of corn flour, 2% -5% of soybean oil, 2% -5% of brewer's yeast, 1% -3% of betaine, 0.2% -0.5% of choline chloride, 0.05% -0.15% of taurine, 1% -5% of potassium diformate, 0.5% -1% of monocalcium phosphate, 0.5% -1.5% of composite mineral substances and 0.5% -1.5% of composite vitamins; the feeding frequency of the feed A is 2 to 4 times/d, and the feeding rate is 1 to 3 percent.
3. The artificial breeding method according to claim 1, wherein the animal fat is at least one of duck oil, chicken oil, lard, beef tallow and mutton tallow.
4. The artificial breeding method according to any one of claims 1 to 3, wherein the artificial hormone in the step S1 is LRH-A2; the injection dosage of the artificial hormone is 1 to 4 mu g/kg.
5. The artificial breeding method according to claim 1, wherein the complex minerals comprise NaCl and ZnSO 4 ·7H 2 O、FeSO 4 ·7H 2 O、MnSO 4 ·4H 2 O、CuSO 4 ·5H 2 O、CoCl 2 、Na 2 SeO 3 And KI; the compound vitamin comprises vitamin A and vitamin B 1 Vitamin B 2 Vitamin B 3 Vitamin B 5 Vitamin B 6 Vitamin B 12 Vitamin C, vitamin D, vitamin E, vitamin H, and vitamin K 3 And vitamin M.
6. Use of the artificial breeding method according to any one of claims 1 to 5 for fish breeding.
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