Breeding method and application of schizothorax tribute

Abstract

The invention relates to a breeding method and application of schizothorax contribution and belongs to the technical field of aquaculture. According to the method, large-scale artificial breeding of the wild schizothorax tribute is achieved through the steps of parent fish capturing, parent fish temporary rearing, artificial spawning induction, artificial insemination, hatching, fry rearing and the like, the spawning induction rate, the fertilization rate, the hatching rate and the emergence rate are high, the whole artificial breeding process can be guaranteed, and the survival rate of the wild schizothorax tribute is increased. Therefore, technical support and theoretical basis are provided for germplasm resource protection, development and utilization of schizothorax tribute.

Description

Technical Field

[0001] The present invention relates to the technical field of aquaculture, and in particular to a breeding method of Gongshan schizothorax and application thereof. Background Art

[0002] The Gongshan schizothorax (Schizothorax gongshanensis), commonly known as the Gongshan bowfish, belongs to the order Cypriniformes, family Cyprinidae, subfamily Schizothorinae, genus Schizothorax; the body is elongated, laterally flattened or slightly laterally flattened, with raised dorsal and ventral margins and a rounded abdomen; the snout is blunt and rounded; the mouth is inferior or sub-inferior, horseshoe-shaped or arc-shaped, with the front end of the mouth below the horizontal line of the lower edge of the eye, and the rear end of the premaxilla below the angle between the anterior margin of the eye and the nostril; the outer part of the lower jaw has no keratin, but there is thin keratin on the inner side; the lower lip is underdeveloped, divided into left and right lobes, without mastoid processes on the surface, the anterior parts of the two lobes are not connected, and there is no middle lobe; the posterior lip groove is interrupted; there are two pairs of well-developed whiskers of approximately equal length, the snout whiskers extend to below the posterior margin of the eye, and the whiskers at the corners of the mouth extend to or almost to the gill cover. It is mainly distributed in the main and tributary rivers of Nujiang River above Fugong County, Nujiang Lisu Autonomous Prefecture, Yunnan Province. It likes flowing water, is resistant to low temperatures, has no long-distance migration habits, and feeds on benthic aquatic organisms.

[0003] The Gongshan schizothorax is a protected species in the Nujiang Lisu Autonomous Prefecture, listed as endangered in the "China Red List of Biodiversity: Vertebrates, Volume V, Freshwater Fishes (2021)," and endemic to the Nujiang River. Its meat is tender, delicious, and highly nutritious. Due to overfishing, illegal fishing, and invasive alien fish, the wild population of Gongshan schizothorax has declined sharply, making it an urgent conservation priority. However, research on Gongshan schizothorax is currently scarce, and there are no reports on its captive breeding. Summary of the Invention

[0004] The object of the present invention is to provide a breeding method for Gongshan schizothorax and application thereof, so as to solve the problem that the prior art has not disclosed a method for artificially breeding Gongshan schizothorax.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The present invention provides a breeding method for Gongshan schizothorax, comprising the following steps:

[0007] (1) Select male Gongshan schizothorax with obvious snout, rough body surface, narrow abdomen, inverted genital pore, and milky white semen flowing out when pressing the abdomen as male broodstock; select female Gongshan schizothorax with relatively soft abdomen, obvious shaking, red and swollen genital pore and protruding, and granular eggs as female broodstock B; select female Gongshan schizothorax with loose abdomen, obvious shaking, protruding genital pore and eggs flowing out when pressing the abdomen as female broodstock C; place the male broodstock, female broodstock B and female broodstock C in a transport box for transportation, soak them in sodium chloride solution for 8 to 12 minutes, and then place them in water for temporary culture to obtain temporary culture male broodstock, temporary culture female broodstock B and female broodstock C to be born respectively;

[0008] (2) After the temporary male broodstock and the temporary female broodstock B are anesthetized, the temporary male broodstock are injected with luteinizing hormone-releasing hormone LRH-A2 5-6 μg / kg + dioxigenin DOM 2.5-4 mg / kg + human chorionic gonadotropin HCG 500-600 IU / kg, and the temporary female broodstock B is injected with luteinizing hormone-releasing hormone LRH-A2 10-12 μg / kg + dioxigenin DOM 5-8 mg / kg + human chorionic gonadotropin HCG 1000-1200 IU / kg. 10-12 hours after the injection, the male broodstock and the female broodstock B are obtained respectively;

[0009] (3) taking eggs from the expectant female broodstock B or the expectant female broodstock C and semen from the expectant male broodstock, mixing the eggs and semen, and allowing them to stand to obtain fertilized eggs;

[0010] (4) The fertilized eggs are incubated for 110 to 148 hours, and then the membranes are broken to obtain newly hatched fry, which are then cultivated to obtain Gongshan schizothorax fry.

[0011] Preferably, the transport box in step (1) contains MS-22220-50 mg / L and ginger paste 0.2-0.4 g / m 3 , multivitamins 0.3~0.5g / m 3 ;

[0012] The transport density is 12 to 16 tails / m 3 ;

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

[0014] Preferably, the anesthesia time in step (2) is 0.5 to 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 female broodstock to be born in step (3) to the number of male broodstock to be born is 2 to 4:1;

[0017] The female broodstock to be born include female broodstock to be born B and female broodstock to be born C;

[0018] The standing time is 2 to 3 minutes.

[0019] Preferably, the incubation flow rate in step (4) is 2-4 L / min, the incubation temperature is 17-19°C, and the incubation density is 0.5-20,000 grains / m 2 .

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

[0021] (a) No feeding was given to newly hatched larvae from 0 to 9 days after hatching;

[0022] (b) Feed rotifers once a day on the 10th to 15th day after hatching.

[0023] (c) Newly hatched larvae were fed powder feed twice a day from 16 to 35 days after hatching;

[0024] (d) The newly hatched fry were put into the pond on the 36th day after hatching, and the Gongshan schizothorax fry were obtained and fed with powder feed twice a day.

[0025] Preferably, the water temperature for the cultivation is 17-19°C, and the dissolved oxygen content of the cultivated water is ≥7 mg / L;

[0026] The feeding amount of the rotifer in step (b) is 4% to 6% of the body weight of the newly hatched larvae;

[0027] The particle size of the powder in 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;

[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 present invention also provides application of the breeding method in artificial breeding of Schizothorax gongshanensis.

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

[0031] The present invention realizes the large-scale artificial breeding of wild Gongshan schizothorax, which is the first of its kind both at home and abroad. The induced spawning rate reaches 100%, the fertilization rate and the hatching rate are both higher than 90%, the emergence rate is high, and the entire artificial breeding process can be guaranteed, thereby providing technical support and theoretical basis for the protection, development and utilization of the germplasm resources of Gongshan schizothorax. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1These are broodstock of Gongshan schizothoracine, of which (1) is female broodstock B and (2) is male broodstock;

[0033] Figure 2 The artificial insemination process of Gongshan schizothorax;

[0034] Figure 3 The embryonic development and newly hatched larvae of Gongshan schizothorax are shown in Figure 1, where (a) is the blastoderm stage, (b) is the two-cell stage, (c) is the four-cell stage, (d) is the multi-cell stage, (e) is the neurula stage, (f) is the muscle effect stage, (g) is the pre-embryo stage, (h) is the membrane rupture stage, (i) is a 3-day-old newly hatched larvae, and (j) is a 14-day-old newly hatched larvae.

[0035] Figure 4 For Gongshan schizothorax fry;

[0036] Figure 5 Flowchart of the breeding method of the present invention. DETAILED DESCRIPTION

[0037] The present invention provides a breeding method for Gongshan schizothorax, comprising the following steps:

[0038] (1) Select male Gongshan schizothorax with obvious snout, rough body surface, narrow abdomen, inverted genital pore, and milky white semen flowing out when pressing the abdomen as male broodstock; select female Gongshan schizothorax with abdomen that is not soft enough, cannot or slightly shakes, and genital pore that is not red and swollen or slightly red and swollen but closed as female broodstock A; select female Gongshan schizothorax with abdomen that is relatively soft, can obviously shake, genital pore that is red and swollen and protruding, and granular eggs that can be felt as female broodstock B; select female Gongshan schizothorax with abdomen that is loose, can obviously shake, genital pore protruding, and eggs flowing out when pressing the abdomen as female broodstock C; release female broodstock A on site, place male broodstock, female broodstock B and female broodstock C in a transport box for transportation, soak them in sodium chloride solution for 8 to 12 minutes, preferably 8 minutes, and then place them in water at the source for temporary breeding, thereby obtaining temporary breeding male broodstock, temporary breeding female broodstock B and female broodstock C to be born respectively;

[0039] (2) After anesthetizing the temporarily cultured male broodstock and the temporarily cultured female broodstock B, the temporarily cultured male broodstock are injected with LRH-A2 5-6 μg / kg + DOM 2.5-4 mg / kg + HCG 500-600 IU / kg, preferably LRH-A2 5.5 μg / kg + DOM 3 mg / kg + HCG 550 IU / kg; at the same time, the temporarily cultured female broodstock B is injected with LRH-A2 10-12 μg / kg + DOM 5-8 mg / kg + HCG 1000-1200 IU / kg, preferably LRH-A2 11 μg / kg + DOM 6 mg / kg + HCG 1100 IU / kg; 10-12 hours after the injection, preferably 10 hours later, the male broodstock to be born and the female broodstock to be born are obtained respectively;

[0040] (3) taking eggs from the expectant female broodstock B or the expectant female broodstock C and semen from the expectant male broodstock, mixing the eggs and semen, allowing them to stand, and washing them to obtain fertilized eggs;

[0041] (4) The fertilized eggs are hatched for 110 to 148 hours, preferably 128 hours, and then the membranes are broken to obtain newly hatched fry, and the newly hatched fry are cultivated to obtain Gongshan schizothorax fry.

[0042] In the present invention, the transport box described in step (1) contains MS-22220-50 mg / L, preferably 30 mg / L; ginger paste 0.2-0.4 g / m 3 , preferably 0.2g / m 3 ; Multivitamins 0.3-0.5g / m 3 , preferably 0.3g / m 3 ; and oxygenate;

[0043] The transport density is 12 to 16 tails / m 3 , preferably 15 tails / m 3 ;

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

[0045] In the present invention, the anesthesia time in step (2) is 0.5 to 1 min, preferably 1 min; the anesthetic agent is MS-222;

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

[0047] In the present invention, the ratio of the number of the female broodstock to be born to the number of the male broodstock to be born in step (3) is 2 to 4:1, preferably 4:1;

[0048] The female broodstock to be born include female broodstock to be born B and female broodstock to be born C;

[0049] The standing time is 2 to 3 minutes, preferably 3 minutes;

[0050] The number of times of cleaning is 3 to 5 times, preferably 5 times.

[0051] In the present invention, the incubation flow rate of step (4) is 2 to 4 L / min, preferably 4 L / min; the incubation temperature is 17 to 19°C, preferably 18°C; the incubation density is 0.5 to 20,000 particles / m 2 , preferably 10,000 particles / m 2 .

[0052] In the present invention, the cultivation method in step (4) comprises the following steps:

[0053] (a) No feeding was given to newly hatched larvae from 0 to 9 days after hatching;

[0054] (b) Feed rotifers once a day on the 10th to 15th day after hatching.

[0055] (c) Newly hatched larvae were fed powder feed twice a day from 16 to 35 days after hatching;

[0056] (d) The newly hatched fry were fully fed and released into the pond on the 36th day after hatching, and the Gongshan schizothorax fry were obtained and fed with powder feed twice a day.

[0057] In the present invention, the water temperature for cultivation is 17-19°C, preferably 18°C; the dissolved oxygen content of the cultivated water is ≥7mg / L;

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

[0059] The powdered material in steps (c) to (d) is preferably Tongwei 191 sediment, purchased from Kunming Tongwei Feed Co., Ltd.;

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

[0062] The present invention also provides application of the breeding method in artificial breeding of Schizothorax gongshanensis.

[0063] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0064] In the bait of the present invention, rotifers were taken from an outdoor pond, and Tongwei 191 sinking material was purchased from Kunming Tongwei Feed Co., Ltd.;

[0065] Among the reagents of the present invention, the multivitamins were purchased from Xinxiang Huaxu Trading Co., Ltd.

[0066] In the equipment of the present invention, the portable oxygen pump was purchased from Yuning Aquarium Equipment Co., Ltd., and the Yushchenko incubator was purchased from the Institute of Water Engineering Ecology of the Chinese Academy of Sciences under the Ministry of Water Resources.

[0067] Example 1: Breeding of Gongshan Schizothorax

[0068] (1) Capture of broodstock: At 19:00 on September 5, 2024, a gill net with a length of 5 m, a depth of 1.5 m and a mesh diameter of 6 cm was set in the backwater bay of the main stream 50 m upstream of the confluence of the Ridongqu tributary in the Chawalon section of the Nujiang River. At 5:00 on September 6, the net was closed and 17 Gongshan schizothorax were caught, including 5 male Gongshan schizothorax with an average weight of 246 g and 12 female Gongshan schizothorax with an average weight of 289 g. The broodstock with obvious snout stars, rough body surface, narrow abdomen, sunken genital pore, milky white semen flowing out when pressing the abdomen and semen that disperses when contacting water were selected. The male Gongshan schizothorax with strong fibrous tissue was selected as male broodstock, with a total of 4 tails; the female Gongshan schizothorax with an abdomen that was not soft enough, could not shake or was slightly shaken, and the genital pore was not red and swollen or was slightly red and swollen but closed was selected as female broodstock A, with a total of 2 tails, which could not be used for artificial breeding and were released on the spot; the female Gongshan schizothorax with a relatively soft abdomen that could shake obviously, a genital pore that was red and swollen and protruded, and granular eggs that could be felt was selected as female broodstock B, with a total of 6 tails; the female Gongshan schizothorax with a loose abdomen that could shake obviously, a protruding genital pore, and eggs flowing out when the abdomen was pressed was selected as female broodstock C, with a total of 4 tails, such as Figure 1 The male broodstock, female broodstock B and female broodstock C were placed in a transport box for transportation, and MS-22250mg / L and ginger paste 0.4g / m were added to the transport box. 3 , multivitamins 0.5g / m 3 , and use a portable oxygen pump for oxygenation, and set the transport density to 12 tails / m 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 are heated and watered, soaked in 0.3% sodium chloride solution for 8 minutes, and then placed in a circular fiberglass temporary rearing barrel with a diameter of 1 m and a depth of 0.8 m, with water entering from the side and draining from the center of the bottom. The temporary rearing barrel contains 50% water from the source of the source, and the temporary rearing male broodstock, temporary rearing female broodstock B and expectant female broodstock C are obtained respectively;

[0070] (3) Artificial induction of labor: After anesthetizing the temporarily cultured male broodstock and the temporarily cultured female broodstock B with 120 mg / LMS-222 for 1 minute, the temporarily cultured male broodstock were injected with LRH-A2 5 μg / kg + DOM 3 mg / kg + HCG 550 IU / kg at one time, and the temporarily cultured female broodstock B were injected with LRH-A2 10 μg / kg + DOM 6 mg / kg + HCG 1100 IU / kg at one time. Ten hours after the injection, the male broodstock and the female broodstock B were obtained.

[0071] (4) Artificial insemination: Gently squeeze the abdomen of the female broodstock B to be born several times to squeeze out the eggs completely into a dry and clean plastic basin, squeeze the semen of the male broodstock to be born onto the eggs and stir evenly to obtain a sperm-egg mixture, then add 4 times the volume of clean water of the sperm-egg mixture to activate the sperm, gently mix with a feather, let it stand for 3 minutes, and then rinse with clean water 5 times to wash away impurities, to obtain 22,000 fertilized eggs; artificially inseminate the female broodstock C according to the above method to obtain 15,000 fertilized eggs, as shown in the figure. Figure 2 As shown;

[0072] (5) Incubation and seedling cultivation: The fertilized eggs were placed in a Yushchenko incubator for incubation, with a controlled flow rate of 4 L / min, a temperature of 18°C, and a density of 10,000 seeds / m 2 , pick out dead eggs and disinfect them every morning and evening, and break the membrane after 128 hours of incubation to obtain 34,000 newly hatched fry. Figure 3 As shown; the newly hatched fry were placed in a round fiberglass culture barrel with a diameter of 1m, a depth of 0.8m, water inlet from the side and drainage at the bottom center for cultivation, the dissolved oxygen content of the water was set to ≥7mg / L, the water temperature was 18℃, and the water was changed regularly every day from 0 to 9 days after hatching without feeding; rotifers were fed once a day at 10:00 on the 10th to 15th day, and the feeding amount was 5% of the weight of the newly hatched fry; 60-mesh Tongwei 191 sinking material was fed once at 8:00 and 20:00 on the 16th to 35th day, and the sewage was sucked out once, and the feeding amount was 5% of the weight of the newly hatched fry; after being fully fed in the morning of the 36th day, 60-mesh Tongwei 191 sinking material was fed once at 8:00 and 20:00 every day, and the feeding amount was 5% of the weight of the Gongshan schizothorax fry; 30,000 Gongshan schizothorax fry were obtained on the 60th day, with an average total length of 4.5cm. Figures 4-5 and as shown in Table 1.

[0073] Example 2: Breeding of Gongshan Schizothorax

[0074] (1) Capture of broodstock: At 19:00 on September 19, 2024, a gill net with a length of 5 m, a depth of 1.5 m and a mesh diameter of 6 cm was set in the backwater bay of the main stream 50 m upstream of the confluence of the Ridongqu tributary in the Chawalon section of the Nujiang River. At 5:00 on September 20, the net was closed and 22 Gongshan schizothorax were caught, including 6 male Gongshan schizothorax with an average weight of 254 g and 16 female Gongshan schizothorax with an average weight of 301 g. Male Gongshan schizothorax with rough surface, narrow abdomen, sunken genital pore, milky white semen flowing out when pressing the abdomen and the semen dispersing when contacting water were selected as male broodstock, a total of 4 tails; female Gongshan schizothorax with soft abdomen, obvious shaking, red and swollen genital pore and protruding granular eggs were selected as female broodstock B, a total of 12 tails; female Gongshan schizothorax with loose abdomen, obvious shaking, protruding genital pore and eggs flowing out when pressing the abdomen were selected as female broodstock C, a total of 4 tails. The male broodstock, female broodstock B and female broodstock C were placed in a transport box for transportation, and MS-22230mg / L and ginger paste 0.2g / m were added to the transport box. 3 , multivitamins 0.3g / m 3 , and use a portable oxygen pump for oxygenation, and set the transport density to 12 tails / m 3 ;

[0075] (2) Temporary rearing of broodstock: After being transported to the temporary rearing base, the male broodstock, female broodstock B and female broodstock C are heated and watered, soaked in 0.5% sodium chloride solution for 12 minutes, and then placed in a circular fiberglass temporary rearing barrel with a diameter of 1 m and a depth of 0.8 m, with water entering from the side and draining from the center of the bottom. The temporary rearing barrel contains 40% source water, and the temporary rearing male broodstock, temporary rearing female broodstock B and expectant female broodstock C are obtained respectively;

[0076] (3) Artificial induction of labor: After anesthetizing the temporarily cultured male broodstock and the temporarily cultured female broodstock B with 120 mg / LMS-222 for 1 minute, the temporarily cultured male broodstock were injected with LRH-A2 6 μg / kg + DOM 4 mg / kg + HCG 600 IU / kg at one time, and the temporarily cultured female broodstock B were injected with LRH-A2 12 μg / kg + DOM 8 mg / kg + HCG 1200 IU / kg at one time. Twelve hours after the injection, the male broodstock and the female broodstock B were obtained.

[0077] (4) Artificial insemination: Gently squeeze the abdomen of the female broodstock B to be born several times to squeeze out the eggs completely into a dry and clean plastic basin, squeeze the semen of the male broodstock to be born onto the eggs and stir evenly to obtain a sperm-egg mixture, then add 3 times the volume of clean water of the sperm-egg mixture to activate the sperm, gently mix with a feather, let it stand for 3 minutes, and then rinse with clean water 3 times to wash away impurities, obtaining 44,000 fertilized eggs; artificially inseminate the female broodstock C according to the above method to obtain 15,000 fertilized eggs;

[0078] (5) Incubation and seedling cultivation: The fertilized eggs were placed in a Yushchenko incubator for incubation, with a flow rate of 2 L / min, a temperature of 17 °C, and a density of 10,000 seeds / m 2 , pick out dead eggs and disinfect them every morning and evening, and after incubation for 128 hours, break the membrane and obtain 56,000 newly hatched fry; place the newly hatched fry in a round fiberglass culture barrel with a diameter of 1m, a depth of 0.8m, and water inlet from the side and drainage from the bottom center for cultivation, set the dissolved oxygen content of the water to ≥7mg / L and the water temperature to 17℃, change the water regularly every day from 0th to 9th day after hatching, and do not feed; feed rotifers once a day at 10:00 on the 10th to 15th day and suck out the sewage, and the feeding amount is the newly hatched fry. 5% of the fish body weight; from the 16th to the 35th day, 60-mesh Tongwei 191 sinking feed was fed once at 8:00 and 20:00 every day and the sewage was sucked out, and the feeding amount was 5% of the weight of the newly hatched fry; after being fully fed in the early morning of the 36th day, 60-mesh Tongwei 191 sinking feed was fed once at 8:00 and 20:00 every day, and the feeding amount was 5% of the weight of the Gongshan schizothorax fry; on the 60th day, 52,000 Gongshan schizothorax fry were obtained, with an average total length of 4.4 cm. The results are shown in Table 1.

[0079] Example 3: Breeding of Gongshan Schizothorax

[0080] (1) Capture of broodstock: At 19:00 on September 12, 2024, a gill net with a length of 20 m, a depth of 2 m, and a mesh diameter of 8 cm was set in the backwater bay of the Chawalon section of the Nujiang River. At 5:00 on September 13, the net was closed and 24 Gongshan schizothorax were captured, including 6 male Gongshan schizothorax with an average weight of 254 g and 18 female Gongshan schizothorax with an average weight of 301 g. Male Gongshan schizothorax with obvious snout, rough body surface, narrow abdomen, sunken genital pore, and milky white semen flowing out when pressing the abdomen, which dispersed in water, were selected. The male broodstock were 5 in total; the female Gongshan schizothorax with an insufficiently soft abdomen, unable to shake or slightly shaken, and a genital pore that was not red and swollen or slightly red and swollen but closed were selected as female broodstock A, 3 in total, which were released on site because they could not be used for artificial breeding; the female Gongshan schizothorax with a relatively soft abdomen that could shake obviously, a genital pore that was red and swollen and protruding, and granular eggs that could be felt were selected as female broodstock B, 12 in total; the female Gongshan schizothorax with a loose abdomen that could shake obviously, a protruding genital pore, and eggs flowing out when the abdomen was pressed was selected as female broodstock C, 3 in total. The male broodstock, female broodstock B, and female broodstock C were placed in a transport box for transportation, and MS-22220mg / L and ginger paste 0.2g / m were added to the transport box. 3 , multivitamins 0.3g / m 3 , and use a portable oxygen pump for oxygenation, and set the transport density to 15 tails / m 3 ;

[0081] (2) Temporary rearing of broodstock: After being transported to the temporary rearing base, the male broodstock, female broodstock B and female broodstock C are heated and watered, soaked in 0.3% sodium chloride solution for 8 minutes, and then placed in a circular fiberglass temporary rearing barrel with a diameter of 1 m and a depth of 0.8 m, with water entering from the side and draining from the center of the bottom. The temporary rearing barrel contains 30% source water, and the temporary rearing male broodstock, temporary rearing female broodstock B and expectant female broodstock C are obtained respectively;

[0082] (3) Artificial induction of labor: After anesthetizing the temporarily cultured male broodstock and the temporarily cultured female broodstock B with 120 mg / LMS-222 for 1 minute, the temporarily cultured male broodstock were injected with LRH-A2 5.5 μg / kg + DOM 3.5 mg / kg + HCG 500 IU / kg at one time, and the temporarily cultured female broodstock B were injected with LRH-A2 11 μg / kg + DOM 7 mg / kg + HCG 1000 IU / kg at one time. Ten hours after the injection, the male broodstock and the female broodstock B were obtained.

[0083] (4) Artificial insemination: Gently squeeze the abdomen of the female broodstock B to be born several times to squeeze out the eggs completely into a dry and clean plastic basin, squeeze the semen of the male broodstock to be born onto the eggs and stir evenly to obtain a sperm-egg mixture, then add 4 times the volume of clean water of the sperm-egg mixture to activate the sperm, gently mix with a feather, let it stand for 3 minutes, and then rinse with clean water 5 times to wash away impurities, obtaining 45,000 fertilized eggs; artificially inseminate the female broodstock C according to the above method to obtain 10,000 fertilized eggs;

[0084] (5) Incubation and seedling cultivation: The fertilized eggs were placed in a Yushchenko incubator for incubation, with a controlled flow rate of 4 L / min, a temperature of 18°C, and a density of 10,000 seeds / m 2 , pick out dead eggs and disinfect them every morning and evening, and after incubation for 128 hours, break the membrane and obtain 52,000 newly hatched fry; place the newly hatched fry in a round fiberglass culture barrel with a diameter of 1m, a depth of 0.8m, and water inlet from the side and drainage from the bottom center for cultivation, set the dissolved oxygen content of the water to ≥7mg / L and the water temperature to 18℃, change the water regularly every day from 0th to 9th day after hatching, and do not feed; feed rotifers once a day at 10:00 on the 10th to 15th day and suck out the sewage, and the feeding amount is the newly hatched fry. 5% of the fry body weight; from the 16th to the 35th day, 60-mesh Tongwei 191 sinking feed was fed once at 8:00 and 20:00 every day and the sewage was sucked out, and the feeding amount was 5% of the weight of the newly hatched fry; after being fully fed in the morning of the 36th day, 60-mesh Tongwei 191 sinking feed was fed once at 8:00 and 20:00 every day, and the feeding amount was 5% of the weight of the Gongshan schizothorax fry; on the 60th day, 50,000 Gongshan schizothorax fry were obtained, with an average total length of 4.5 cm. The results are shown in Table 1.

[0085] Table 1 Breeding results of Gongshan schizothorax

[0086]

[0087] The results show that the breeding method of the Gongshan schizothorax of the present invention can be used to carry out artificial breeding of wild Gongshan schizothorax, with an induced spawning rate of 100%, and a fertilization rate and a hatching rate both being above 90%.

[0088] As can be seen from the above examples, the present invention provides a breeding method and application of Gongshan schizothorax. The present invention achieves large-scale artificial breeding of wild Gongshan schizothorax, with high rates of induced fertilization, hatching, and emergence, ensuring a stable breeding process. This provides technical support and a theoretical basis for the conservation, development, and utilization of Gongshan schizothorax germplasm resources.

[0089] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A method for breeding Gongshan schizothorax, characterized in that, The following steps are involved: (1) Select male Gongshan schizothorax with obvious snout, rough body surface, narrow abdomen, inverted genital pore, and milky white semen flowing out when pressing the abdomen as male broodstock; select female Gongshan schizothorax with relatively soft abdomen, obvious shaking, red and swollen genital pore and protruding, and granular eggs as female broodstock B; select female Gongshan schizothorax with loose abdomen, obvious shaking, protruding genital pore and eggs flowing out when pressing the abdomen as female broodstock C; place the male broodstock, female broodstock B and female broodstock C in a transport box for transportation, soak them in sodium chloride solution for 8 to 12 minutes, and then place them in water for temporary culture to obtain temporary culture male broodstock, temporary culture female broodstock B and female broodstock C to be born respectively; (2) After the temporary male broodstock and the temporary female broodstock B are anesthetized, the temporary male broodstock are injected with luteinizing hormone-releasing hormone LRH-A2 5-6 μg / kg + dioxigenin DOM 2.5-4 mg / kg + human chorionic gonadotropin HCG 500-600 IU / kg, and the temporary female broodstock B is injected with luteinizing hormone-releasing hormone LRH-A2 10-12 μg / kg + dioxigenin DOM 5-8 mg / kg + human chorionic gonadotropin HCG 1000-1200 IU / kg. 10-12 hours after the injection, the male broodstock and the female broodstock B are obtained respectively; (3) taking eggs from the expectant female broodstock B or the expectant female broodstock C and semen from the expectant male broodstock, mixing the eggs and semen, and allowing them to stand to obtain fertilized eggs; (4) The fertilized eggs are incubated for 110 to 148 hours, and then the membranes are broken to obtain newly hatched fry, which are then cultivated to obtain Gongshan schizothorax fry.

2. The breeding method according to claim 1, characterized in that The transport box described in step (1) contains MS-22220~50mg / L and ginger paste 0.2~0.4g / m 3 , multivitamins 0.3~0.5g / m 3 ; The transport density is 12 to 16 tails / m 3 ; The initial concentration of the sodium chloride solution is 0.3% to 0.5%.

3. The breeding method according to claim 2, characterized in that The anesthesia time in step (2) is 0.5 to 1 min, and the anesthetic used is MS-222; The concentration of the anesthetic is 100-150 mg / L.

4. The breeding method according to claim 3, characterized in that The ratio of the number of the female broodstock to be born in step (3) to the number of the male broodstock to be born is 2 to 4:1; The female broodstock to be born include female broodstock to be born B and female broodstock to be born C; The standing time is 2 to 3 minutes.

5. The breeding method according to claim 4, characterized in that The incubation flow rate of step (4) is 2-4 L / min, the incubation temperature is 17-19°C, and the incubation density is 0.5-20,000 grains / m 2 .

6. The breeding method according to claim 5, characterized in that The cultivation method in step (4) comprises the following steps: (a) No feeding was given to newly hatched larvae from 0 to 9 days after hatching; (b) Feed rotifers once a day on the 10th to 15th day after hatching. (c) Newly hatched larvae were fed powder feed twice a day from 16 to 35 days after hatching; (d) The newly hatched fry were put into the pond on the 36th day after hatching, and the Gongshan schizothorax fry were obtained and fed with powder feed twice a day.

7. The breeding method according to claim 6, characterized in that The water temperature for the cultivation is 17-19°C, and the dissolved oxygen content of the water body for the cultivation is ≥7mg / L; The feeding amount of the rotifer in step (b) is 4% to 6% of the body weight of the newly hatched larvae; The particle size of the powder in 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 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.

8. Use of the breeding method according to any one of claims 1 to 7 in artificial breeding of Schizothorax gongshanensis.

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