Breeding method of schizothorax grahami and application thereof

Abstract

The application relates to a breeding method of Schizothorax gongshanensis and application thereof, and belongs to the technical field of aquatic product culture. The application realizes large-scale artificial breeding of wild Schizothorax gongshanensis through steps of parent fish capturing, parent fish temporary breeding, artificial parturition induction, artificial insemination, hatching and fry breeding, the parturition induction rate, the fertilization rate, the hatching rate and the hatching rate are high, the whole artificial breeding process can be guaranteed, thereby technical support and theoretical basis are provided for protection and development and utilization of germplasm resources of Schizothorax gongshanensis.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of aquaculture, and in particular to a breeding method of Schizothorax gongshanensis and application thereof. BACKGROUND

[0002] Schizothorax gongshanensis, commonly known as Gongshan bowfish, belongs to Cypriniformes, Cyprinidae, Schizothoracinae and Schizothorax. The body is elongated, laterally compressed or slightly laterally compressed, with raised dorsal and ventral edges, and a round abdomen. The snout is blunt and round. The mouth is subventral or subinferior, horseshoe-shaped or arc-shaped, with the anterior mouth slit below the lower edge of the eye. The posterior end of the maxillary bone is below the intersection of the anterior edge of the eye and the nostril. The lower jaw has no horny part on the outside and a thin horny part on the inside. The lower lip is not developed, with two leaves, no papillae on the surface, and no intermediate leaf. The post-lip groove is interrupted. There are two pairs of well-developed barbels, which are about the same length, with the rostral barbels extending to the lower edge of the eye and the oral barbels extending to or almost to the opercular bone. It is mainly distributed in the main stream and tributaries of the Nujiang River above Fugong County in Nujiang Lisu Autonomous Prefecture, Yunnan Province, and prefers running water, tolerates low temperature, and has no long-distance migration habit. It feeds on benthic aquatic organisms.

[0003] Schizothorax gongshanensis is a protected fish in Nujiang Lisu Autonomous Prefecture, an endangered fish in the Red List of Chinese Biodiversity: Vertebrates Volume 5 Fishes (2021), and a Nujiang River endemic fish. It has tender and delicious meat and high nutritional value. Due to overfishing and invasion of alien species, the wild resources of Schizothorax gongshanensis have decreased dramatically, and it needs to be protected urgently. However, there is little research on Schizothorax gongshanensis, and there is no related report on artificial breeding of Schizothorax gongshanensis. SUMMARY

[0004] The present application aims to provide a breeding method of Schizothorax gongshanensis and application thereof, to solve the problem that the prior art has not disclosed a method of artificially breeding Schizothorax gongshanensis.

[0005] In order to achieve the above-mentioned application purpose, the present application provides the following technical solutions:

[0006] The present application provides a breeding method of Schizothorax gongshanensis, comprising the following steps:

[0007] (1) selecting male Schizothorax grahami with obvious rostral bead star, rough body surface, narrow abdomen, invaginated genital pore, and milky white semen flowing out after pressing abdomen as male parent fish, selecting female S. grahami with soft abdomen, obvious shaking, red and swollen genital pore, and granular feeling eggs as female parent fish B, and selecting female S. grahami with loose abdomen, obvious shaking, external genital pore, and eggs flowing out after pressing abdomen as female parent fish C; placing the male parent fish, female parent fish B and female parent fish C in a transport box, immersing in sodium chloride solution for 8-12 min, and then placing in water for temporary nursing to obtain temporary male parent fish, temporary female parent fish B and female parent fish C to be spawned respectively;

[0008] (2) after anesthetizing the temporary male parent fish and the temporary female parent fish B, injecting luteinizing hormone-releasing hormone LRH-A 25-6 μg / kg + diethylstilbestrol DOM 2.5-4 mg / kg + chorionic gonadotropin HCG 500-600 IU / kg to the temporary male parent fish, and injecting luteinizing hormone-releasing hormone LRH-A 10-12 μg / kg + diethylstilbestrol DOM 5-8 mg / kg + chorionic gonadotropin HCG 1000-1200 IU / kg to the temporary female parent fish B, and obtaining male parent fish to be spawned and female parent fish B to be spawned respectively after 10-12 h of injection;

[0009] (3) taking the eggs of the female parent fish B to be spawned or the female parent fish C to be spawned and the semen of the male parent fish to be spawned, mixing and standing the eggs and the semen to obtain fertilized eggs;

[0010] (4) obtaining initial hatched fry by breaking the membrane of the fertilized eggs after 110-148 h of incubation, and cultivating the initial hatched fry to obtain S. grahami fry.

[0011] Preferably, the transport box in step (1) contains MS-222 20-50 mg / L, ginger extract 0.2-0.4 g / m 3 , and compound vitamin 0.3-0.5 g / m 3 .

[0012] The density of the transport is 12-16 tails / m 3 .

[0013] The initial concentration of the sodium chloride solution is 0.3%-0.5%.

[0014] Preferably, the anesthetizing time in step (2) is 0.5-1 min, and the anesthetic is MS-222.

[0015] The concentration of the anesthetic is 100-150 mg / L.

[0016] Preferably, the ratio of the number of the female parent fish to be spawned to the male parent fish to be spawned in step (3) is 2-4:1.

[0017] The to-be-labor female parent fish comprises to-be-labor female parent fish B and to-be-labor female parent fish C.

[0018] The standing time is 2-3 min.

[0019] Preferably, the flow rate of the hatching in step (4) is 2-4 L / min, the temperature of the hatching is 17-19 DEG C, and the density of the hatching is 0.5-20 thousand grains / m 2 .

[0020] Preferably, the method of the culturing in step (4) comprises the following steps:

[0021] (a) 0-9 d after the initial hatched fish is born, no feeding is performed;

[0022] (b) 10-15 d after the initial hatched fish is born, rotifers are fed once a day;

[0023] (c) 16-35 d after the initial hatched fish is born, powder is fed twice a day;

[0024] (d) 36 d after the initial hatched fish is born, the fish is put into a pond to obtain Gongshan schizothorax fry, and the powder is fed twice a day.

[0025] Preferably, the water temperature of the culturing is 17-19 DEG C, and the dissolved oxygen of the water body of the culturing is greater than or equal to 7 mg / L;

[0026] The feeding amount of the rotifers in step (b) is 4%-6% of the body weight of the initial hatched fish;

[0027] The particle size of the powder in step (c) is 50-70 meshes, and the feeding amount of the powder is 4%-6% of the body weight of the initial hatched fish;

[0028] The particle size of the powder in step (d) is 50-70 meshes, and the feeding amount of the powder is 4%-6% of the body weight of the Gongshan schizothorax fry.

[0029] The application further provides application of the breeding method in artificial breeding of Gongshan schizothorax.

[0030] The application has the following technical effects and advantages:

[0031] The application realizes large-scale artificial breeding of wild Gongshan schizothorax, which is first in China and abroad, the labor induction rate is 100%, the fertilization rate and the hatching rate are both higher than 90%, the hatching rate is high, and the whole artificial breeding process can be guaranteed, thereby providing technical support and theoretical basis for protection and development and utilization of Gongshan schizothorax germplasm resources. BRIEF DESCRIPTION OF DRAWINGS

[0032] Figure 1A Schizothorax gongshanensis artificial insemination process;

[0033] Figure 2 A Schizothorax gongshanensis artificial insemination process;

[0034] Figure 3 A Schizothorax gongshanensis artificial insemination process;

[0035] Figure 4 A Schizothorax gongshanensis artificial insemination process;

[0036] Figure 5 A Schizothorax gongshanensis artificial insemination process; DETAILED DESCRIPTION

[0037] The present application provides a Schizothorax gongshanensis breeding method, comprising the following steps:

[0038] (1) selecting a male Schizothorax gongshanensis with obvious snout beads, rough body surface, narrow abdomen, invaginated genital pore, and milky white semen flowing out when pressing the abdomen as a male parent fish, selecting a female Schizothorax gongshanensis with insufficiently soft abdomen, unable or slightly shaking, non-swollen or slightly swollen but closed genital pore as a female parent fish A, selecting a female Schizothorax gongshanensis with relatively soft abdomen, obvious shaking, swollen and protruding genital pore, and feeling granular eggs as a female parent fish B, and selecting a female Schizothorax gongshanensis with loose abdomen, obvious shaking, external genital pore, and egg flowing out when pressing the abdomen as a female parent fish C; releasing the female parent fish A on site, transporting the male parent fish, the female parent fish B and the female parent fish C in a transport box, soaking in a sodium chloride solution for 8-12 min, preferably 8 min, and then temporarily raising in the water of the source habitat to obtain a temporarily raised male parent fish, a temporarily raised female parent fish B and a female parent fish C to be born, respectively;

[0039] (2) after the anesthetizing the temporary male parent fish and the temporary female parent fish B, injecting LRH-A 25-6 μg / kg+DOM 2.5-4 mg / kg+HCG 500-600 IU / kg, preferably LRH-A 25.5 μg / kg+DOM 3 mg / kg+HCG 550 IU / kg to the temporary male parent fish; and injecting LRH-A 10-12 μg / kg+DOM 5-8 mg / kg+HCG 1000-1200 IU / kg, preferably LRH-A 11 μg / kg+DOM 6 mg / kg+HCG 1100 IU / kg to the temporary female parent fish B; obtaining the male parent fish to be spawned and the female parent fish B to be spawned respectively after 10-12 hours, preferably 10 hours of injection;

[0040] (3) taking the ovum of the female parent fish B to be spawned or the female parent fish C to be spawned and the semen of the male parent fish to be spawned, mixing, standing and washing the ovum and the semen to obtain the fertilized ovum;

[0041] (4) hatching the fertilized ovum for 110-148 hours, preferably 128 hours, and breaking the membrane to obtain the newly hatched fry, and culturing the newly hatched fry to obtain the Schizothorax grahami fry.

[0042] In the present application, the transport box in step (1) contains MS-222 20-50 mg / L, preferably 30 mg / L; ginger extract 0.2-0.4 g / m 3 , preferably 0.2 g / m 3 ; and compound vitamin 0.3-0.5 g / m 3 , preferably 0.3 g / m 3 ; and the transport is oxygenated.

[0043] The density of the transport is 12-16 tails / m 3 , preferably 15 tails / m 3 .

[0044] The initial concentration of the sodium chloride solution is 0.3%-0.5%, preferably 0.3%.

[0045] In the present application, the anesthetizing time in step (2) is 0.5-1 min, preferably 1 min; and the anesthetic is MS-222.

[0046] The concentration of the anesthetic is 100-150 mg / L, preferably 120 mg / L.

[0047] In the present application, the ratio of the number of the female parent fish to be spawned to the number of the male parent fish to be spawned in step (3) is 2-4:1, preferably 4:1.

[0048] The female parent fish to be spawned includes the female parent fish B to be spawned and the female parent fish C to be spawned.

[0049] The time for the standing is 2-3 minutes, preferably 3 minutes;

[0050] The number of times for the cleaning is 3-5 times, preferably 5 times.

[0051] In the present application, the flow rate for the hatching in step (4) is 2-4 L / min, preferably 4 L / min; the temperature for the hatching is 17-19℃, preferably 18℃; the density for the hatching is 0.5-20 thousand grains / m 2 , preferably 10 thousand grains / m 2 .

[0052] In the present application, the method for the culturing in step (4) comprises the following steps:

[0053] (a) from 0-9 days after the membrane shedding of the newly hatched fry, no feeding is performed;

[0054] (b) from 10-15 days after the membrane shedding of the newly hatched fry, rotifers are fed once a day;

[0055] (c) from 16-35 days after the membrane shedding of the newly hatched fry, powder is fed twice a day;

[0056] (d) on the 36th day after the membrane shedding of the newly hatched fry, the fish are fed to satiation and then put into a pond, and the powder is fed twice a day to obtain the Schizothorax grahami fry.

[0057] In the present application, the water temperature for the culturing is 17-19℃, preferably 18℃; the dissolved oxygen content of the water for the culturing is ≥7 mg / L;

[0058] The rotifers in step (b) are taken from an outdoor pond, and the feeding amount of the rotifers is 4%-6% of the body weight of the newly hatched fry, preferably 5%;

[0059] The powder in steps (c)-(d) is preferably Tongwei 191 sinking powder, which is purchased from Kunming Tongwei Feed Co., Ltd.

[0060] The particle size of the powder in step (c) is 50-70 mesh, preferably 60 mesh; and the feeding amount of the powder is 4%-6% of the body weight of the newly hatched fry, preferably 5%;

[0061] The particle size of the powder in step (d) is 50-70 mesh, preferably 60 mesh; and the feeding amount of the powder is 4%-6% of the body weight of the Schizothorax grahami fry, preferably 5%.

[0062] The present application also provides the application of the breeding method in artificially breeding the Schizothorax grahami.

[0063] The technical solutions provided by the present application will be described in detail below in conjunction with the examples, but they should not be understood as limiting the scope of protection of the present application.

[0064] In the feed of this invention, rotifers are taken from outdoor ponds, and Tongwei 191 sinking feed is purchased from Kunming Tongwei Feed Co., Ltd.

[0065] In the reagents of this invention, the compound vitamins were purchased from Xinxiang Huachu Trade Co., Ltd.

[0066] In the device of this invention, the portable oxygen pump was purchased from Yuning Aquarium Equipment Co., Ltd., and the Yuxianke incubator was purchased from the Institute of Hydro-Engineering Ecology, Chinese Academy of Sciences, Ministry of Water Resources.

[0067] Example 1: Breeding of Gongshan Schizothorax

[0068] (1) Catching of broodstock: At 19:00 on September 5, 2024, a gillnet with a length of 5m, a depth of 1.5m, and a mesh diameter of 6cm was set up in the backwater bay of the main stream at 50m upstream of the confluence of the Ridongqu tributary in the Chawalong section of the Nujiang River. At 5:00 on September 6, 17 Gongshan schizothorax were caught, including 5 male Gongshan schizothorax with an average weight of 246g and 12 female Gongshan schizothorax with an average weight of 289g. Among them, those with obvious pearly spots on the snout, rough body surface, narrow and long abdomen, sunken genital opening, and milky white semen flowing out when the abdomen is pressed and the semen disperses when it comes into contact with water were selected. Four male Gongshan schizothorax were selected as male parent fish. Two female Gongshan schizothorax were selected as parent fish A, characterized by a less supple abdomen, inability to move or slight movement, and a closed genital opening with no or slight redness or swelling. These females were released on-site because they were unsuitable for artificial breeding. Six female Gongshan schizothorax were selected as parent fish B, characterized by a relatively supple abdomen, noticeable movement, a red and protruding genital opening, and the presence of granular eggs. Four female Gongshan schizothorax were selected as parent fish C, characterized by a relaxed abdomen, noticeable movement, a protruding genital opening, and the release of eggs upon abdominal pressure. Figure 1 As shown. Male broodstock, female broodstock B, and female broodstock C were transported in transport boxes. MS-2 2250 mg / L and ginger paste 0.4 g / m³ were added to the transport boxes. 3 0.5g / m² of multivitamins 3 A portable oxygen pump was used for oxygenation, and the transport density was set at 12 fish / m³. 3 ;

[0069] (2) Temporary rearing of broodstock: After being transported to the temporary rearing base, the male broodstock, female broodstock B and female broodstock C were conditioned and soaked in 0.3% sodium chloride solution for 8 minutes and then placed in a circular fiberglass temporary rearing tank with a diameter of 1m and a depth of 0.8m, with water entering from the side and draining from the center of the bottom. The temporary rearing tank contained 50% water from the source area, and temporary reared male broodstock, temporary reared female broodstock B and female broodstock C awaiting birth were obtained respectively.

[0070] (3) Artificial spawning: Male broodstock and female broodstock B were anesthetized with 120mg / LMS-222 for 1 minute. The male broodstock was injected with LRH-A 25μg / kg + DOM 3mg / kg + HCG 550IU / kg. At the same time, the female broodstock B was injected with LRH-A 210μg / kg + DOM 6mg / kg + HCG 1100IU / kg. Ten hours after the injection, male broodstock and female broodstock B were obtained respectively.

[0071] (4) Artificial insemination: Gently squeeze the abdomen of the spawning female parent fish B several times to completely squeeze the eggs into a dry, clean plastic basin. Squeeze the sperm of the spawning male parent fish onto the eggs and stir evenly to obtain a sperm-egg mixture. Add 4 times the volume of water to the sperm-egg mixture to activate the sperm. Gently mix with a feather, let stand for 3 minutes, and then rinse 5 times with clean water to remove impurities, obtaining 22,000 fertilized eggs. Artificial insemination was then performed on the female parent fish C using the same method, yielding 15,000 fertilized eggs. Figure 2 As shown;

[0072] (5) Hatching and Seedling Cultivation: Fertilized eggs were placed in a Yushchenko incubator for incubation, with the flow rate controlled at 4 L / min, the temperature at 18℃, and the density at 10,000 eggs / m³. 2 Dead eggs were removed and disinfected every morning and evening. After 128 hours of incubation, 34,000 newly hatched fry were obtained. Figure 3 As shown; newly hatched fry were placed in circular fiberglass rearing tanks with a diameter of 1m and a depth of 0.8m, with side water intake and central drainage. The dissolved oxygen level was set at ≥7mg / L and the water temperature at 18℃. From day 0 to 9 after hatching, the water was changed regularly daily without feeding. From day 10 to 15, rotifers were fed once daily at 10:00 AM, and waste was removed; the feeding amount was 5% of the fry's body weight. From day 16 to 35, 60-mesh Tongwei 191 sedimentary feed was fed twice daily at 8:00 AM and 8:00 PM, and waste was removed; the feeding amount was 5% of the fry's body weight. On day 36, after a full morning feeding, the fry were released into the pond and fed twice daily at 8:00 AM and 8:00 PM, with the feeding amount being 5% of the fry's body weight. On day 60, 30,000 Gongshan Schizothorax fry were obtained, with an average total length of 4.5cm. The results are as follows. Figures 4-5 As shown in Table 1.

[0073] Example 2: Breeding of Gongshan Schizothorax

[0074] (1) Brood fish capture: On September 19, 2024, at 19:00, a 5m long, 1.5m deep, 6cm mesh diameter gill net was arranged in the backwater bay 50m upstream of the junction of the Rizhao Chawalong section of the Nujiang River and the Huihekou upstream river section of the Rizhao Chawalong section of the Nujiang River. On September 20, 5:00, 22 Schizothorax gongshanensis were captured, including 6 male Schizothorax gongshanensis with an average weight of 254g and 16 female Schizothorax gongshanensis with an average weight of 301g. Four male brood fish with obvious beak stars, rough body surface, narrow abdomen, invaginated genital pore, and milky white semen flowing out when pressing the abdomen were selected as male brood fish. Twelve female brood fish with soft abdomen, obvious shaking, red and prominent genital pore, and granular feeling eggs were selected as female brood fish B. Four female brood fish with loose abdomen, obvious shaking, external genital pore, and egg flow when pressing the abdomen were selected as female brood fish C. The male brood fish, female brood fish B, and female brood fish C were transported in a transport box, 30mg / L MS-222, 0.2g / m 3 ginger paste, and 0.3g / m 3 compound vitamins were added to the transport box, and a portable oxygen pump was used for oxygenation. The transport density was set to 12 / m 3 ;

[0075] (2) Brood fish temporary cultivation: After transportation to the temporary cultivation base, the male brood fish, female brood fish B, and female brood fish C were warmed and watered, soaked in 0.5% sodium chloride solution for 12 minutes, and then placed in a 1m diameter, 0.8m deep circular glass steel temporary cultivation barrel with side water inlet and bottom central water outlet for temporary cultivation. The temporary cultivation barrel contained 40% of the original water, and temporary male brood fish, temporary female brood fish B, and female brood fish C were obtained.

[0076] (3) Artificial induction of labor: After 1 minute of anesthesia of the temporary male brood fish and temporary female brood fish B with 120mg / L MS-222, the temporary male brood fish was injected with LRH-A 26μg / kg+DOM 4mg / kg+HCG 600IU / kg, and the temporary female brood fish B was injected with LRH-A 12μg / kg+DOM 8mg / kg+HCG 1200IU / kg. After 12 hours, the female brood fish C was obtained.

[0077] (4) Artificial insemination: The female brood fish B was gently squeezed several times, and the eggs were completely squeezed out into a dry and clean plastic basin. The semen of the male brood fish was squeezed onto the eggs and stirred evenly to obtain a sperm-egg mixture. Then, 3 times the volume of water was added to activate the sperm, and the mixture was gently mixed with a feather. After 3 minutes, the mixture was washed with clean water 3 times to remove impurities, and 44,000 fertilized eggs were obtained. The female brood fish C was artificially inseminated according to the above method, and 15,000 fertilized eggs were obtained.

[0078] (5) Incubation and fry cultivation: the fertilized eggs were placed in a Yuxian incubator for incubation, with a flow rate of 2 L / min, a temperature of 17°C, and a density of 100,000 eggs / m 2 , dead eggs were picked out and disinfected every morning and evening, and 128 h after incubation, 56,000 newly hatched fry were obtained by breaking the membrane; the newly hatched fry were placed in a circular glass steel cultivation barrel with a diameter of 1 m and a depth of 0.8 m, with water flowing in from the side and draining from the bottom center, and the dissolved oxygen in the water was set to ≥7 mg / L, and the water temperature was 17°C; after hatching, the water was changed at regular times every day from the 0th to the 9th day without feeding; from the 10th to the 15th day, rotifers were fed once a day at 10:00, and the feeding amount was 5% of the body weight of the newly hatched fry; from the 16th to the 35th day, 60-mesh Tongwei 191 sinking feed was fed twice a day at 8:00 and 20:00, and the feeding amount was 5% of the body weight of the newly hatched fry; on the 36th day, after the fish were fed to satiation in the morning, 60-mesh Tongwei 191 sinking feed was fed twice a day at 8:00 and 20:00, and the feeding amount was 5% of the body weight of the fry; on the 60th day, 52,000 fry of Schizothorax grahami were obtained, with an average total length of 4.4 cm, and the results are shown in Table 1.

[0079] Example 3: Breeding of Schizothorax grahami

[0080] (1) Parent fish capture: On September 12, 2024, at 19:00, a gill net with a length of 20 m, a depth of 2 m, and a mesh diameter of 8 cm was arranged in the backwater bay of the Chawalong section of the Nujiang River, and 24 Schizothorax grahami were captured by netting at 5:00 on September 13, including 6 male Schizothorax grahami with an average weight of 254 g and 18 female Schizothorax grahami with an average weight of 301 g; male parent fish with obvious snout beads, rough body surface, narrow abdomen, sunken genital pore, and milky white semen flowing out when pressing the abdomen and dispersing when the semen meets water were selected as male parent fish, a total of 5; female parent fish A with an abdomen that was not soft enough to shake or slightly shake, a genital pore that was not red or slightly red but closed, were selected for release into the wild, a total of 3; female parent fish B with a relatively soft abdomen, a clearly shaking abdomen, a red and prominent genital pore, and a granular feeling of eggs were selected as female parent fish B, a total of 12; female parent fish C with a relaxed abdomen, a clearly shaking abdomen, a protruding genital pore, and a flow of eggs when pressing the abdomen were selected as female parent fish C, a total of 3. The male parent fish, female parent fish B, and female parent fish C were placed in a transport box, MS-222 20 mg / L, ginger paste 0.2 g / m 3 , and compound vitamin 0.3 g / m 3 were added to the transport box, and a portable oxygen pump was used for oxygenation, and the transport density was set to 15 / m 3 ;

[0081] (2) Parent fish temporary breeding: After being transported to the temporary breeding base, the male parent fish, female parent fish B and female parent fish C were over-temperature and over-water, soaked in 0.3% sodium chloride solution for 8 min, and then placed in a circular glass steel temporary breeding barrel with a diameter of 1 m and a depth of 0.8 m, with water entering from the side and water being discharged from the bottom center, containing 30% of the source water. Temporary breeding male parent fish, temporary breeding female parent fish B and female parent fish C were obtained.

[0082] (3) Artificial induced production: After 1 min of anesthesia of the temporary breeding male parent fish and the temporary breeding female parent fish B with 120 mg / L MS-222, the temporary breeding male parent fish was injected with LRH-A 25.5 μg / kg + DOM 3.5 mg / kg + HCG 500 IU / kg at one time, and the temporary breeding female parent fish B was injected with LRH-A 211 μg / kg + DOM 7 mg / kg + HCG 1000 IU / kg at one time. After 10 h, the female parent fish C was obtained.

[0083] (4) Artificial insemination: The abdomen of the female parent fish B was gently squeezed several times to completely squeeze the eggs into a dry and clean plastic basin. The semen of the male parent fish was squeezed onto the eggs and stirred evenly to obtain a sperm-egg mixture. Then, 4 times the volume of clean water was added to activate the sperm, and the mixture was gently mixed with a feather. After 3 min, the mixture was washed with clean water for 5 times to remove impurities, and 45,000 fertilized eggs were obtained. The female parent fish C was artificially inseminated according to the above method, and 10,000 fertilized eggs were obtained.

[0084] (5) Incubation and fry cultivation: The fertilized eggs were placed in a Yuxenko incubator for incubation, with a flow rate of 4 L / min, a temperature of 18°C, and a density of 10,000 eggs / m 2 . Dead eggs were picked out daily and disinfected. After 128 h of incubation, the membrane was broken to obtain 52,000 newly hatched fry. The newly hatched fry were placed in a circular glass steel breeding barrel with a diameter of 1 m and a depth of 0.8 m, with water entering from the side and water being discharged from the bottom center. The dissolved oxygen content of the water was set to be greater than or equal to 7 mg / L, and the water temperature was set to be 18°C. After the membrane was broken, the water was changed every day from the 0th to the 9th day without feeding. From the 10th to the 15th day, rotifers were fed once a day at 10:00, and the water was sucked. The feeding amount was 5% of the body weight of the newly hatched fry. From the 16th to the 35th day, 60-mesh Tongwei 191 sinking material was fed twice a day at 8:00 and 20:00, and the water was sucked. The feeding amount was 5% of the body weight of the newly hatched fry. On the 36th day, the fry was fed with 60-mesh Tongwei 191 sinking material twice a day at 8:00 and 20:00 after being fed to satiation in the morning. The feeding amount was 5% of the body weight of the fry. On the 60th day, 50,000 fry of Schizothorax grahami were obtained, with an average total length of 4.5 cm. The results are shown in Table 1.

[0085] Table 1 Breeding results of Schizothorax grahami

[0086]

[0087] The results show that the breeding method of Schizothorax gongshanensis can carry out artificial breeding of wild Schizothorax gongshanensis, the rate of inducing parturition is 100%, and the fertilization rate and the hatching rate are all above 90%.

[0088] From the above examples, the application provides a breeding method of Schizothorax gongshanensis and application thereof. The application realizes large-scale artificial breeding of wild Schizothorax gongshanensis, and the rate of inducing parturition, the fertilization rate, the hatching rate and the emergence rate are high, the whole artificial breeding process can be guaranteed, thereby providing technical support and theoretical basis for the protection and development and utilization of germplasm resources of Schizothorax gongshanensis.

[0089] The above only describes the preferred embodiments of the application, and it should be noted that for those skilled in the art, without departing from the principles of the application, a number of improvements and refinements can be made, and these improvements and refinements should also be considered as the protection scope of the application.

Claims

1. A method for breeding Schizothorax grahami, characterized in that, It comprises the following steps: (1) selecting male Schizothorax grahami with obvious rostral bead, rough body surface, long and narrow abdomen, invaginated genital pore and milky white sperm flowing out after pressing abdomen as male parent fish, selecting female S. grahami with soft abdomen, obvious shaking, red and swollen genital pore and granular feeling as female parent fish B, and selecting female S. grahami with loose abdomen, obvious shaking, external genital pore and ovum flowing out after pressing abdomen as female parent fish C; placing the male parent fish, female parent fish B and female parent fish C in a transport box, immersing in sodium chloride solution for 8-12 min, and then placing in water for temporary cultivation to obtain temporary male parent fish, temporary female parent fish B and female parent fish C to be spawned; (2) after anesthetizing the temporary male parent fish and temporary female parent fish B, injecting luteinizing hormone-releasing hormone LRH-A2 5-6 μg / kg + diethylstilbestrol DOM 2.5-4 mg / kg + chorionic gonadotropin HCG 500-600 IU / kg to the temporary male parent fish, and injecting luteinizing hormone-releasing hormone LRH-A2 10-12 μg / kg + diethylstilbestrol DOM 5-8 mg / kg + chorionic gonadotropin HCG 1000-1200 IU / kg to the temporary female parent fish B, and then obtaining female parent fish to be spawned and female parent fish B to be spawned after 10-12 h; (3) taking ovum of the female parent fish B to be spawned or female parent fish C to be spawned and sperm of the male parent fish to be spawned, mixing and standing the ovum and sperm to obtain fertilized ovum; (4) breaking the membrane of the fertilized ovum after 110-148 h of incubation to obtain newly hatched fry, and cultivating the newly hatched fry to obtain S. grahami fry; The quantity ratio of the female parent fish to be spawned to the male parent fish to be spawned is 2-4:1; The female parent fish to be spawned comprises female parent fish B to be spawned and female parent fish C to be spawned; The standing time is 2-3 min; The hatching flow rate is 2-4 L / min, the hatching temperature is 17-19℃, and the hatching density is 0.5-20,000 grains / m 2 .

2. The breeding method according to claim 1, characterized by, The transport box in step (1) contains MS-222 20-50 mg / L, ginger paste 0.2-0.4 g / m 3 ; composite vitamin 0.3-0.5 g / m 3 ; The density of the transport is 12-16 fish / m 3 ; The initial concentration of the sodium chloride solution is 0.3%-0.5%.

3. The breeding method according to claim 2, characterized by, The anesthetizing time is 0.5-1 min, and the anesthetic is MS-222; The concentration of the anesthetic is 100-150 mg / L.

4. The breeding method according to claim 1, characterized by, The cultivating method of step (4) comprises the following steps: (a) not feeding from 0 to 9 d after the newly hatched fry breaks the membrane; (b) feeding rotifer once a day from 10 to 15 d after the newly hatched fry breaks the membrane; (c) feeding powder twice a day from 16 to 35 d after the newly hatched fry breaks the membrane; (d) releasing into a pond on the 36th day after the newly hatched fry breaks the membrane to obtain S. grahami fry, and feeding powder twice a day.

5. The breeding method according to claim 4, characterized by, The water temperature of the cultivation is 17-19 ℃, and the dissolved oxygen content of the water body is ≥7 mg / L; The feeding amount of the rotifer of step (b) is 4%-6% of the body weight of the newly hatched fry; The particle size of the powder of step (c) is 50-70 mesh, and the feeding amount of the powder is 4%-6% of the body weight of the newly hatched fry; The particle size of the powder of step (d) is 50-70 mesh, and the feeding amount of the powder is 4%-6% of the body weight of the S. grahami fry.

6. Application of the breeding method of any one of claims 1-5 in artificially breeding S. grahami.

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