Artificial breeding method of black head fish

Abstract

The invention provides an artificial breeding method of black head fish, and belongs to the technical field of aquaculture. The spawning induction method comprises the following steps: injecting spawning induction hormone into a female parent for breeding parent fish twice for artificial spawning induction to obtain fertilized eggs; during the incubation period of the fertilized eggs, micro-flow at 18-21 DEG C is adopted for artificial incubation, and the flow speed is controlled by water at different incubation stages until incubation and seedling emergence are achieved, so that the fry of the black head fish with the odd amount is obtained. According to the method, the oxytocic hormone is injected in a two-needle injection mode, the injection dosage of each needle is strictly controlled, ova and seminal fluid which are well developed can be obtained, meanwhile, the hatching conditions of the fertilized ova are controlled, the oxytocic rate of the black head fish is 100%, the fertilization rate is 89% or above, the hatching rate is 86% or above, large-scale artificial breeding of the black head fish is achieved, and the method is suitable for large-scale artificial breeding of the black head fish. The method has important significance for protecting the cyprinus carpio which is an endangered resource.

Description

Technical Field

[0001] The invention relates to the technical field of aquaculture, and in particular to an artificial breeding method for a garfish. Background Art

[0002] Garramirofronits belongs to the Cypriniformes, Cyprinidae, Labeoninae, genus Garra. The front of the body is cylindrical, the tail is flattened, the dorsal edge is arc-shaped, the chest and abdomen are flat, and the head is square. Its uniqueness lies in the frontal groove in front of the nostrils, forming two frontal processes, each of which has a large and obvious conical horny protrusion at the front. When alive, it is dark green, and each fin is dark gray with reddish ends. It lives in rivers with fast currents and rocky bottoms. It attaches to stones with suction cups, mainly scraping algae, and also eats debris and aquatic insect larvae. It is only distributed in the middle and lower reaches of the Lancang River in China, and is a fish unique to the Lancang River system. The strange-headed garramirofronits grows fast, has tender and delicious meat, is highly popular in the market, and has high economic value, development prospects and breeding promotion significance.

[0003] With the cascade development of the Lancang River, six hydropower stations, Gongguoqiao, Xiaowan, Manwan, Dachaoshan, Nuozadu and Jinghong, have been built and put into operation in the middle and lower reaches of the Lancang River. All of them have been put into operation for more than 10 years. The impact of the construction and operation of the power stations on fish has been very obvious, especially for fish that lay drifting eggs such as the strange-headed garfish. The dam blocked the migration of fish and changed the natural flow of the river. The originally connected flowing river section was transformed into a blocked slow-flow habitat, destroying the original spawning grounds and habitats; the reduction of river flow velocity, the reduction of water flow distance, and the change of water depth also affected the hatching of drifting eggs; the change of hydrological conditions also affected the abundance of bait organisms, compressed the living space of the strange-headed garfish, and also affected the reproduction of the population. In addition, due to the influence of human factors, the wild resources of the strange-headed garfish have declined and urgently need to be protected.

[0004] In order to protect the population resources of the odd-headed garfish, it is necessary to provide an artificial breeding method for the odd-headed garfish and provide guidance for artificial breeding, resource protection, large-scale production and development and utilization. Summary of the invention

[0005] The invention aims to provide an artificial breeding method for the odd-headed garfish.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The invention provides an artificial breeding method of a garfish, comprising the following steps:

[0008] S1. Parent fish collection: From June to August, female and male fish aged 2 to 3 years old with gonad development at stage V are collected and placed in temporary rearing ponds;

[0009] S2. Artificial induction of labor: Inject luteinizing hormone-releasing hormone A24-6μg / kg to the female parent, and 11-13 hours later inject luteinizing hormone-releasing hormone A27-9μg / kg + dioxin 5-7mg / kg to the female parent; the water temperature after the second injection is 20.5-21.5℃, and the water temperature is controlled to decrease by 0.4-0.6℃ every hour within 5-7 hours, from 20.5-21.5℃ to 17.5-18.5℃, and then continue to maintain 17.5-18.5℃ to stimulate the ovarian development of the female parent;

[0010] At the same time as the second injection of the female parent, the male parent is injected with luteinizing hormone-releasing hormone A2 3.5-4.5 μg / kg + dioconazole 2.5-3.5 mg / kg;

[0011] S3. Artificial insemination: 6-7 hours after the female parent completes the second injection of oxytocin, gently press the abdomen of the female fish to squeeze out the mature eggs, then squeeze the male fish sperm onto the eggs, add 18-19℃ water to activate the sperm, stir for 110-130s, and then wash off the mucus with 18-19℃ water to obtain fertilized eggs;

[0012] S4. Fertilized egg hatching: The fertilized eggs treated in S3 are placed in water at 18-21°C for hatching, with a water flow rate of 0.4-0.6 m / s for 0-22 hours, a water flow rate of 0.08-0.12 m / s for 22-26 hours, and a water flow rate of 0.4-0.6 m / s after 27 hours. After 23-25 ​​hours, the fry are obtained and cultured;

[0013] S4. 23 to 25 hours after the fry emerge from the membrane, transfer the fry to the fry culture tank and feed them with fry feed; after the fry start to hatch, transfer them to the outdoor cement pool for cultivation to obtain the odd-headed garfish fry.

[0014] Preferably, the weight of the female parent is ≥150 g, the weight of the male parent is ≥100 g, and semen flows out when the abdomen is squeezed.

[0015] Preferably, the ratio of the female parent to the male parent is 1.2-1.8:1.

[0016] Preferably, when activating sperm, the amount of water added is 2 to 4 times the volume of the mixture of fish eggs and fish semen.

[0017] Preferably, the fertilized eggs are incubated in an incubation ring, the water depth in the incubation ring is 58-62 cm, and the density of the fertilized eggs is 48,000-52,000 eggs / m 3 .

[0018] Preferably, the diameter of the open seedling cultivation tank is 0.8-1.2 m, the depth is 0.7-0.9 m, and the density of fry in the open seedling cultivation tank is 3800-4200 fry / m 3 , the water temperature is 20-22℃.

[0019] Preferably, the starter feed is S1 microparticle feed produced by Shandong Shengsuo Feed Co., Ltd., and the starter feed is fed once in the morning and evening every day, with each feeding accounting for 4.8-5.2% of the body weight of the farmed fish fry.

[0020] Preferably, the catching of broodstock is carried out by placing ground traps in the shallows with stone bottom in the middle and lower reaches of the Lancang River in the evening for catching.

[0021] Preferably, the captured broodstock are packed in bags, 3-4L of water and 9-11 odd-headed garfish are added to each bag, and 0.4-0.6g of anesthetic, 4-6g of multivitamins, and 0.8-1.2mL of ginger paste are added and stirred, the air is exhausted and then oxygenated, and the bag mouth is tied tightly, and the specifications of the bag are 38-42cm×68-72cm.

[0022] Preferably, the captured broodstock are soaked in 0.3% to 0.5% salt water for 8 to 12 minutes and then placed in a temporary holding pond for temporary holding.

[0023] By adopting the above technical scheme, the present invention has the following beneficial effects: the present invention adopts the method of injecting oxytocin in two injections, and strictly controls the injection dosage of each injection, so as to obtain well-developed eggs and semen, and at the same time control the hatching conditions of fertilized eggs, so that the induced labor rate of the odd-headed garfish is 100%, the fertilization rate is more than 89%, and the hatching rate is more than 86%, thereby realizing the large-scale artificial breeding of the Lhasa schizothorax, which is of great significance to the protection of the endangered resource of the odd-headed garfish. DETAILED DESCRIPTION

[0024] The invention provides an artificial breeding method of a garfish, comprising the following steps:

[0025] S1. Parent fish collection: From June to August, female and male fish aged 2 to 3 years old with gonad development at stage V are collected and placed in temporary rearing ponds;

[0026] S2. Artificial induction of labor: Inject luteinizing hormone-releasing hormone A24-6μg / kg to the female parent, and 11-13 hours later inject luteinizing hormone-releasing hormone A27-9μg / kg + dioxin 5-7mg / kg to the female parent; the water temperature after the second injection is 20.5-21.5℃, and the water temperature is controlled to decrease by 0.4-0.6℃ every hour within 5-7 hours, from 20.5-21.5℃ to 17.5-18.5℃, and then continue to maintain 17.5-18.5℃ to stimulate the ovarian development of the female parent;

[0027] At the same time as the second injection of the female parent, the male parent is injected with luteinizing hormone-releasing hormone A2 3.5-4.5 μg / kg + dioconazole 2.5-3.5 mg / kg;

[0028] S3. Artificial insemination: 6-7 hours after the female parent completes the second injection of oxytocin, gently press the abdomen of the female fish to squeeze out the mature eggs, then squeeze the male fish sperm onto the eggs, add 18-19℃ water to activate the sperm, stir for 110-130s, and then wash off the mucus with 18-19℃ water to obtain fertilized eggs;

[0029] S4. Fertilized egg hatching: The fertilized eggs treated in S3 are placed in water at 18-21°C for hatching, with a water flow rate of 0.4-0.6 m / s for 0-22 hours, a water flow rate of 0.08-0.12 m / s for 22-26 hours, and a water flow rate of 0.4-0.6 m / s after 27 hours. After 23-25 ​​hours, the fry are obtained and cultured;

[0030] S4. 23 to 25 hours after the fry emerge from the membrane, transfer the fry to the fry culture tank and feed them with fry feed; after the fry start to hatch, transfer them to the outdoor cement pool for cultivation to obtain the odd-headed garfish fry.

[0031] The present invention collects odd-headed garruloides in the breeding season, and obtains the alternative odd-headed garruloides after transportation and processing. Specifically, from June to August every year, ground traps are placed in the evening in the middle and lower reaches of the Lancang River, in the shallows with stone bottom, to catch odd-headed garruloides; the collected odd-headed garruloides are packed in bags for transportation, and the bags are preferably straight plastic bags, and the specifications of the straight plastic bags are 38-42cm×68-72cm, further preferably 39-41cm×69-7cm, and more preferably 40cm×70cm. 3-4L of water and 8-12 odd-headed garruloides are added to each bag, and the volume of the water is further preferably 3.2-3.8L, and more preferably 3.5L. The number of odd-headed garruloides in the straight plastic bag is 9-11, and more preferably 11. Preferably 10; add 0.4-0.6g anesthetic, 4-6g multivitamin, 0.8-1.2mL ginger paste at the same time, stir evenly, the anesthetic is preferably MS-222, the mass of the anesthetic is further preferably 0.45-0.55g, more preferably 0.5g, the mass of the multivitamin is 4.5-5.5g, more preferably 5g, the volume of the ginger paste is further preferably 0.9-1.1mL, more preferably 1mL, then exhaust the air in the bag, fill it with oxygen, tie the bag tightly, and press the bag lightly with your arm to see if there is any water or air leakage. If there is such a phenomenon, replace it in time and repackage it. This method can be transported for 2-3 days; after arriving at the destination, it is heated and watered, soaked in salt water and placed in a temporary holding pool for temporary holding, the concentration of the salt water is 0.3-0.5%, more preferably 0.35-0.45%, more preferably 0.4%; the soaking time is 8-12min, more preferably 9-11min, more preferably 10min. The temporary holding pool is preferably a circular cement pool with a diameter of 1.8 to 2.2 m, more preferably 1.9 to 2.1 m, and more preferably 2 m; the water outlet is in the middle of the temporary holding pool, and the water inlet is located on one side of the pool wall.

[0032] After catching the odd-headed garrulidae, the present invention selects sexually mature male and female parents from the candidate odd-headed garrulidae. The sexually mature parents are 2-3 year old female and male fish with gonad development in stage V. The male parent weighs more than 100g, has inconspicuous pearl stars when touched along the direction of the pectoral fin rays, is slightly rough, and milky white semen flows out when the abdomen is lightly pressed and disperses when it enters the water; the female parent weighs more than 150g, is smooth when touched along the direction of the pectoral fin rays, and has an obviously swollen abdomen.

[0033] After selecting the parents, the present invention performs artificial induction of labor, injects an oxytocin into the male and female parents, and obtains the expectant parents, wherein the female-to-male ratio of the expectant parents is 1.2 to 1.8:1, more preferably 1.3 to 1.7:1, and even more preferably 1.5:1. The female parent is injected twice, the first injection of luteinizing hormone releasing hormone A2, the injection dose is 4-6 μg / kg, more preferably 4.5-5.5 μg / kg, more preferably 5 μg / kg; 11-13 hours later, preferably 11.5-12.5 hours later, more preferably 12 hours later, the female parent is injected with luteinizing hormone releasing hormone A2 + dioxigenin, the injection dose of luteinizing hormone releasing hormone A2 is 7-9 μg / kg, more preferably 7.5-8.5 μg / kg, more preferably 8 μg / kg; the injection dose of dioxigenin is 5-7 mg / kg, more preferably 5.5-6.5 mg / kg, more preferably 6 mg / kg. The water temperature after the second injection is 20.5-21.5°C, more preferably 20.8-21.2°C, more preferably 21°C, and is controlled within 5-7 hours, more preferably within 5.5-6.5 hours, more preferably within 6 hours, the water temperature is reduced by 0.4-0.6°C every hour, more preferably by 0.5°C every hour, from 20.5-21.5°C to 17.5-18.5°C, more preferably 17.8-18.2°C, more preferably 18°C, and then maintained at 17.5-18.5°C, and further The temperature is preferably 17.8-18.2°C, more preferably 18°C, to stimulate the ovarian development of the female parent; while the female parent is injected with the second injection, the male parent is injected with luteinizing hormone releasing hormone A2 + dioic acid, the injection amount of the luteinizing hormone releasing hormone A2 is 3.5-4.5 μg / kg, more preferably 3.8-4.2 μg / kg, more preferably 4.0 μg / kg; the injection amount of dioic acid is 2.5-3.5 mg / kg, more preferably 2.8-3.2 mg / kg, more preferably 3.0 mg / kg.

[0034] 6 to 7 hours, preferably 6.2 to 6.8 hours, and more preferably 6.5 hours after the female parent completes the second injection of the oxytocin, gently press the abdomen of the female fish to squeeze out the mature eggs, then squeeze the male fish sperm onto the eggs, then add water to activate the sperm, stir, and then wash off the mucus to obtain fertilized eggs. The temperature of the water is 18 to 19°C, preferably 18.2 to 18.8°C, and more preferably 18.5°C; the amount of water added is 2 to 4 times the volume of the mixed solution of fish eggs and fish sperm, preferably 2.5 to 3.5 times, and more preferably 3 times; the stirring time is 110 to 130 seconds, preferably 115 to 125 seconds, and more preferably 120 seconds; the temperature of the water for washing off the mucus is the same as the water temperature for activating the sperm.

[0035] The present invention hatches the fertilized eggs after obtaining them. The fertilized eggs of the odd-headed garruloides are drifting eggs. The fertilized eggs of the odd-headed garruloides are preferably placed in a hatching ring for micro-flowing water hatching. The water depth in the hatching ring is 58 to 62 cm, more preferably 59 to 61 cm, and more preferably 60 cm. The density of the fertilized eggs is 48,000 to 52,000 eggs / m 3 , more preferably 49,000 to 51,000 grains / m 3 , more preferably 50,000 particles / m 3 . Adjust the flow rate according to the hatching stage, maintain the flow rate at 0.4-0.6 m / s for 0-22 h, more preferably 0.45-0.55 m / s, more preferably 0.5 m / s; reduce the flow rate to 0.08-0.12 m / s for 22-26 h, more preferably 0.09-0.11 m / s, more preferably 0.1 m / s; restore the flow rate to 0.4-0.6 m / s after 27 h, more preferably 0.45-0.55 m / s, more preferably 0.5 m / s, control the hatching water temperature to 18-21 ° C, more preferably 19-20 ° C, more preferably 19.5 ° C, and after 23-25 ​​h, the fry are removed from the membrane to obtain the fry and continue to be cultured.

[0036] The present invention transfers the fry to an open seedling cultivation tank for cultivation 23 to 25 hours after the fry emerge from the membrane, more preferably 23.5 to 24.5 hours after the fry emerge from the membrane, and more preferably 24 hours after the fry emerge from the membrane. The diameter of the open seedling cultivation tank is 0.8 to 1.2 m, more preferably 0.9 to 1.1 m, and more preferably 1 m; the depth of the open seedling cultivation tank is 0.7 to 0.9 m, more preferably 0.75 to 0.85 m, and more preferably 0.8 m; the open seedling cultivation tank has independent inlet and outlet ports, the outlet port is in the middle of the bottom, a 0.6 m diameter water pipe is inserted into the outlet port, and a hole is drilled on the side of the pipe. The density of fry in the open seedling cultivation tank is 3800 to 4200 tails / m 2 , more preferably 3900 to 4100 tails / m 2 , more preferably 4000 tails / m 2 ; The water temperature is 20-22°C, more preferably 20.5-21.5°C, and more preferably 21°C. Feed 4 times a day with conventional seedling feed (microcapsule feed), preferably S1 microparticle feed produced by Shandong Shengsuo Feed Co., Ltd., and feed 4.8-5.2% of the body weight of the farmed fry each time, more preferably 4.9-5.1%, and more preferably 5%. After the opening of the fish (about 10 days after the film is formed), it is transferred to an outdoor cement pool and disinfected with quicklime 5-7 days before the transfer. The amount of quicklime used is 18-22kg / mu, more preferably 19-21kg / mu, and more preferably 20kg / mu. The fish with the opening of the fish with the head of ...

[0037] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0038] The seedling feed in the embodiment of the present invention is S1 microparticle feed produced by Shandong Shengsuo Feed Co., Ltd.

[0039] Example 1

[0040] On the evening of July 25, 2024, ground traps were placed in the shallows with stone bottom in the middle and lower reaches of the Lancang River. The odd-headed grackles would scrape the algae attached to the stones and enter the ground traps; the collected odd-headed grackles were packed in 40cm×70cm straight plastic bags, 4L of water and 10 odd-headed grackles were added to each bag, and 0.5g MS-222, 5g of multivitamins, and 1mL of ginger paste were added and mixed evenly, the air in the bag was first discharged, and then filled with oxygen, the bag was tightened with a rubber band, and the packaging bag was lightly pressed with the arm to check for water or air leaks. If so, it was replaced and repackaged in time; after arriving at the destination, it was warmed and watered, soaked in 0.3% salt water for 10 minutes, and then placed in a circular cement pool with a diameter of 2m for temporary storage, with water outlet in the middle of the cement pool and the water inlet located on one side of the pool wall.

[0041] On July 27, 2024, among the alternative odd-fronted garfish collected on July 25, two 2-year-old male odd-fronted garfish were selected, which had unclear beads when touched along the direction of the pectoral fin rays, were slightly rough, and had milky white semen flowing out when the abdomen was lightly pressed, which dispersed as soon as it entered the water. The average weight was 105g; three 2-year-old female odd-fronted garfish were selected, which were smooth when touched along the direction of the pectoral fin rays, had an obviously swollen abdomen, and had an average weight of 158g; the male-to-female ratio was 3:2.

[0042] The sexually mature male and female parents are injected with oxytocin using a two-injection method. The first injection is of luteinizing hormone releasing hormone A24μg / kg for the female parent; 12 hours later, the female parent is injected with luteinizing hormone releasing hormone A27μg / kg + dioxicam 5mg / kg again; while the female parent is injected with the second injection, the male parent is injected with luteinizing hormone releasing hormone A23.5μg / kg + dioxicam 2.5mg / kg, and the mother is ready to give birth after the injection; at the same time, within 6 hours after the second injection, the water temperature of the expectant parent is controlled to decrease by 0.5℃ every 1 hour, from 20.5℃ to 17.5℃, and then maintained at 17.5℃ to stimulate the ovarian development of the female parent.

[0043] 7 hours after the female parent completed the second injection of oxytocin, the male parent could be observed chasing and hitting the female parent's abdomen with its head. Dry insemination was performed by first gently pressing the female fish's abdomen to squeeze out the mature eggs and place them in a clean and dry plastic basin. Then, the male fish's semen was squeezed onto the eggs, and 19°C water was added to activate the sperm. After stirring for 2 minutes, the mucus was washed off with 19°C water to obtain fertilized eggs.

[0044] The fertilized eggs were placed in a hatching loop for micro-flow incubation, and the flow rate was adjusted according to the hatching stage, the flow rate was maintained at 0.5 m / s for 0-22 hours, the flow rate was reduced to 0.1 m / s for 22-26 hours, and the flow rate was restored to 0.5 m / s after 27 hours, and the hatching water temperature was controlled to be 18-21° C., and the eggs were hatched after 24 hours. The artificial breeding induction rate of this embodiment was 100%, the fertilization rate was 89.2%, the hatching rate was 86.5%, and about 76,000 newly hatched fry were obtained.

[0045] 24 hours after the fry hatched (i.e. 48 hours after fertilization), the fry were transferred to an open fry cultivation tank with a diameter of 1m and a depth of 0.8m, with independent inlets and outlets. The outlet was in the middle of the bottom, and a 0.6m diameter water pipe was inserted into the outlet. There were holes drilled on the side of the pipe. They were fed with conventional open fry feed (microencapsulated feed) four times a day. They were transferred to an outdoor cement pool 10 days after hatching and disinfected with 20kg / mu of quicklime 5 days before being transferred to the outdoor pool. They were fed in the early morning and put into the pond. After being put into the pond, they were fed twice a day. 1 / 3 of the water was changed every week. Daily care was carried out afterwards, and about 75,000 fry of the strange-headed grayhead fish were obtained.

[0046] Example 2

[0047] On the evening of August 5, 2024, in the middle and lower reaches of the Lancang River, traps were placed in the shallows with stones on the bottom, and 28 strange-headed murrelets were collected and packed in 40cm×70cm straight plastic bags. 4L of water and 10 strange-headed murrelets were added to each bag. At the same time, 0.5g MS-222, 5g multivitamins, and 1mL ginger paste were added and stirred evenly. The air in the bag was first exhausted, and then filled with oxygen. The bag was tied with a rubber band, and the bag was gently pressed with the arm to see if there was any water or air leakage. If there was any leakage, it should be replaced and repackaged in time. After being transported to the destination, it was heated and watered, soaked in 0.5% salt water for 10 minutes, and then placed in a circular cement pool with a diameter of 2m for temporary storage. The water was discharged in the middle of the cement pool, and the water inlet was located on one side of the pool wall.

[0048] On August 8, we selected two male odd-headed grackles with an average weight of 111 g, which had faint beads when touched along the direction of the pectoral fin rays. They were slightly rough, and had milky white semen flowing out when the abdomen was lightly pressed, and dispersed when they entered the water. We also selected three female odd-headed grackles with an average weight of 159 g, which were smooth when touched along the direction of the pectoral fin rays, and had an obviously swollen abdomen. The male-to-female ratio was 3:2.

[0049] The sexually mature male and female parents are injected with oxytocin using a two-injection method. The first injection is of luteinizing hormone releasing hormone A25μg / kg for the female parent; 12 hours later, the female parent is injected with luteinizing hormone releasing hormone A28μg / kg + dioxicam 6mg / kg again; while the female parent is injected with the second injection, the male parent is injected with luteinizing hormone releasing hormone A24μg / kg + dioxicam 3mg / kg, and the mother is prepared for delivery after the injection; at the same time, within 6 hours after the second injection, the water temperature of the expectant parent is controlled to decrease by 0.5℃ every 1h, from 21℃ to 18℃, and then maintained at 18℃ to stimulate the ovarian development of the female parent.

[0050] 6 hours after the female parent completed the second injection of oxytocin, the male parent could be observed chasing and hitting the female parent's abdomen with his head, and dry insemination was performed. First, the female fish's abdomen was gently pressed to squeeze out the mature eggs and place them in a clean and dry plastic basin. Then, the male fish's semen was squeezed onto the eggs, and 18°C ​​water was added to activate the sperm. After stirring for 2 minutes, the mucus was washed off with 18°C ​​water to obtain fertilized eggs. The fertilized eggs were placed in a hatching ring for micro-flow water incubation. The flow rate was adjusted according to the hatching stage. The flow rate was maintained at 0.5m / s for 0-22h, reduced to 0.1m / s for 22-26h, and restored to 0.5m / s after 27h. The hatching water temperature was controlled at 18-21°C, and the membranes emerged after 25h. The artificial breeding rate was 100%, the fertilization rate was 90.3%, and the hatching rate was 88.6%. About 79,000 newly hatched fry were obtained.

[0051] 24 hours after the fry hatched (i.e. 49 hours after fertilization), the fry were transferred to an open fry cultivation tank with a diameter of 1m and a depth of 0.8m, with independent inlets and outlets. The outlet was in the middle of the bottom, and a 0.6m diameter water pipe was inserted into the outlet. There were holes drilled on the side of the pipe. They were fed with conventional open fry feed (microencapsulated feed) four times a day. They were transferred to an outdoor cement pool 10 days after hatching and disinfected with 20kg / mu of quicklime 5 days before being transferred to the outdoor pool. They were put into the pond after being fully fed in the early morning. After being put into the pond, they were fed twice a day. 1 / 3 of the water was changed every week. Daily care was performed afterwards, and about 77,000 fry of the odd-headed grayling were obtained.

[0052] Example 3

[0053] On the evening of July 30, 2024, in the middle and lower reaches of the Lancang River, traps were placed in the shallows with stones on the bottom, and 28 strange-headed murrelets were collected and packed in 40cm×70cm straight plastic bags. 4L of water and 10 strange-headed murrelets were added to each bag. At the same time, 0.5g MS-222, 5g multivitamins, and 1mL ginger paste were added and stirred evenly. The air in the bag was first exhausted, and then filled with oxygen. The bag was tied with a rubber band, and the bag was gently pressed with the arm to see if there was any water or air leakage. If there was any leakage, it should be replaced and repackaged in time. After being transported to the destination, it was heated and watered, soaked in 0.5% salt water for 10 minutes, and then placed in a circular cement pool with a diameter of 2m for temporary storage. The water was discharged in the middle of the cement pool, and the water inlet was located on one side of the pool wall.

[0054] On August 2, we selected two male odd-headed grackles, with an average weight of 108 g, and they had faint beads when touched along the direction of the pectoral fin rays. They were slightly rough, and milky white semen flowed out when the abdomen was lightly pressed, and dispersed when they entered the water. We also selected three female odd-headed grackles, with an average weight of 157 g, and they were smooth when touched along the direction of the pectoral fin rays. The abdomen was obviously swollen, and the male-female ratio was 3:2.

[0055] The sexually mature male and female parents are injected with oxytocin using a two-injection method. The first injection is of luteinizing hormone releasing hormone A25μg / kg for the female parent; 12 hours later, the female parent is injected with luteinizing hormone releasing hormone A28μg / kg + dioxicam 6mg / kg again; while the female parent is injected with the second injection, the male parent is injected with luteinizing hormone releasing hormone A24μg / kg + dioxicam 3mg / kg, and the mother is prepared for delivery after the injection; at the same time, within 6 hours after the second injection, the water temperature of the expectant parent is controlled to decrease by 0.5℃ every 1 hour, from 21.5℃ to 18.5℃, and then maintained at 18.5℃ to stimulate the ovarian development of the female parent.

[0056] 6 hours after the female parent completed the second injection of oxytocin, the male parent could be observed chasing and hitting the female parent's abdomen with his head, and dry insemination was performed. First, the female fish's abdomen was gently pressed to squeeze out the mature eggs and place them in a clean and dry plastic basin. Then, the male fish's semen was squeezed onto the eggs, and 18°C ​​water was added to activate the sperm. After stirring for 2 minutes, the mucus was washed off with 18°C ​​water to obtain fertilized eggs. The fertilized eggs were placed in the incubation ring for micro-flow water incubation. The flow rate was adjusted according to the incubation stage. The flow rate was maintained at 0.5m / s for 0-22h, and the flow rate was reduced to 0.1m / s for 22-26h. After 27h, the flow rate was restored to 0.5m / s. The incubation water temperature was controlled at 18-21°C, and the membranes emerged after 25h. The artificial breeding induction rate was 100%, the fertilization rate was 89.8%, and the hatching rate was 87.1%. About 78,000 newly hatched fry were obtained.

[0057] 24 hours after the fry hatched (i.e. 49 hours after fertilization), the fry were transferred to an open fry cultivation tank with a diameter of 1m and a depth of 0.8m, with independent inlets and outlets. The outlet was in the middle of the bottom, and a 0.6m diameter water pipe was inserted into the outlet. The side of the pipe had holes drilled in it. They were fed with conventional open fry feed (microencapsulated feed) four times a day. They were transferred to an outdoor cement pool 10 days after hatching and disinfected with 20kg / mu of quicklime 5 days before being transferred to the outdoor pool. They were fed in the early morning and put into the pond. After being put into the pond, they were fed twice a day. 1 / 3 of the water was changed every week. Daily care was performed afterwards, and about 76,000 fry of the odd-headed grayling were obtained.

[0058] Comparative Example 1

[0059] On July 24, 2024, in the evening, in the middle and lower reaches of the Lancang River, a trap was placed in the shallows with stones on the bottom, and 21 strange-headed murrelets were collected and packed in 40cm×70cm straight plastic bags. 3-4L of water and 10 strange-headed murrelets were added to each bag. At the same time, 0.5gMS-222, 5g of multivitamins, and 1ml of ginger paste were added and stirred evenly. The air in the bag was first exhausted, and then filled with oxygen. The bag was tightened with a rubber band, and the bag was gently pressed with the arm to see if there was any water or air leakage. If there was any leakage, it should be replaced and repackaged in time. After being transported to the destination, it was heated and watered, soaked in 0.3% salt water for 10 minutes, and then placed in a circular cement pool with a diameter of 2m for temporary storage. The water was discharged in the middle of the cement pool, and the water inlet was located on one side of the pool wall.

[0060] On July 26, we selected two male odd-fronted garruloides from the candidate fish. They had faint beads when touched along the direction of the pectoral fin rays, were slightly rough, and milky white semen flowed out when the abdomen was lightly pressed, which dispersed when they entered the water. They were 110g, aged two years old. We also selected three female odd-fronted garruloides, which had smooth texture when touched along the direction of the pectoral fin rays, had obviously swollen abdomen, and were 155g, aged two years old. The male-female ratio was 3:2.

[0061] The single-injection method was used to induce labor. The female parent was given a one-time injection of luteinizing hormone-releasing hormone A2 12 μg / kg + dioconazole 6 mg / kg. At the same time, the male parent was given a one-time injection of half the dose of the female parent's injection of oxytocin. After the injection, the mother was allowed to wait for labor. The water temperature was 26°C.

[0062] The development of the eggs was checked 6 hours after the injection of the female parent, but they could not be discharged smoothly. The eggs could not be forced out until the 18th hour, but they did not separate. The eggs were then squeezed out and placed in a clean and dry plastic basin. The male fish semen was squeezed onto the eggs, and water at 26°C was added to activate the sperm. After stirring for 2 minutes, the mucus was washed off with water at 26°C, and the fertilized eggs were placed in an incubation ring for micro-flow water incubation. After about 6 hours, all of them died. The artificial breeding rate was 33.33%, the fertilized eggs were 0%, and the hatching rate was 0%.

[0063] Comparative Example 2

[0064] On July 28, 2024, among the alternative odd-headed garfish on July 24 (a batch caught on the same day as comparative example 1), two 2-year-old male odd-headed garfish were selected, which had inconspicuous pearl stars when touched along the direction of the pectoral fin rays, were slightly rough, had milky white semen flowing out when lightly pressed on the abdomen, and dispersed when entering the water, and had an average weight of 112g; three 2-year-old female odd-headed garfish were selected, which were smooth when touched along the direction of the pectoral fin rays, had an obviously swollen abdomen, and had an average weight of 163g; the male-female ratio was 3:2;

[0065] The two-injection method was used to induce labor. The first injection was luteinizing hormone-releasing hormone A25μg / kg for the female parent; 12 hours later, luteinizing hormone-releasing hormone A28μg / kg + dioconazole 6mg / kg were injected into the female parent again; at the same time as the second injection of the female parent, the male parent was injected with an oxytocin, and the injection dose was half of the second injection dose of the female parent. After the injection was completed, the mother was allowed to wait for delivery, and the water temperature was 26℃;

[0066] The development of the female parent was checked 6 hours after the injection. The eggs could be squeezed out, but ovulation was not smooth and most of the eggs did not separate. The eggs were then squeezed out and placed in a clean and dry plastic basin. The male fish semen was squeezed onto the eggs, and water at 26°C was added to activate the sperm. After stirring for 2 minutes, the mucus was washed off with water at 26°C to obtain fertilized eggs.

[0067] The fertilized eggs were placed in a hatching ring with micro-flow water for incubation, maintaining a flow rate of 0.5m / s and a water temperature of 26℃. The eggs emerged from the membranes after 21 hours and the fry were obtained. The artificial insemination had an induction rate of 66.67%, a fertilization rate of 3.2%, and a hatching rate of 10%. A total of about 200 newly hatched fry were obtained.

[0068] 24 hours after the fry hatched (i.e. 49 hours after fertilization), the fry were transferred to an open fry cultivation tank with a diameter of 1m and a depth of 0.8m, with independent inlets and outlets. The outlet was in the middle of the bottom, and a 0.6m diameter water pipe was inserted into the outlet. There were holes drilled on the side of the pipe. They were fed with conventional open fry feed (microencapsulated feed) four times a day. They were transferred to an outdoor cement pool 10 days after hatching and disinfected with 20kg / mu of quicklime 5 days before being transferred to the outdoor pool. They were fed in the pond in the early morning and fed twice a day after being put into the pond. 1 / 3 of the water was changed every week. Daily care was performed afterwards, and about 0.01 million fry of the odd-headed grayling were obtained.

[0069] Comparative Example 3

[0070] On the evening of August 4, 2024, in the middle and lower reaches of the Lancang River, traps were placed in the shallows with stones on the bottom, and 28 strange-headed murrelets were collected and packed in 40cm×70cm straight plastic bags. 4L of water and 10 strange-headed murrelets were added to each bag. At the same time, 0.5g MS-222, 5g multivitamins, and 1ml ginger paste were added and mixed evenly. The air in the bag was first exhausted, and then filled with oxygen. The bag was tightened with a rubber band, and the bag was gently pressed with the arm to see if there was any water or air leakage. If so, it should be replaced and repackaged in time. After being transported to the destination, it was heated and watered, soaked in 0.5% salt water for 10 minutes, and then placed in a temporary holding pool with a diameter of 2m, with water discharged in the middle and the water inlet located on the side of the pool wall 1;

[0071] On August 7, we selected 2 male garrulidae from the candidate fishes. They had inconspicuous pearl stars when touched along the direction of the pectoral fin rays, were slightly rough, had milky white semen flowing out when lightly pressed on the abdomen, and dispersed when entering the water. They had an average weight of 109g. We also selected 3 female garrulidae from the candidate fishes. They had smooth skin when touched along the direction of the pectoral fin rays, had obviously swollen abdomens, and had an average weight of 157g. The male-female ratio was 3:2.

[0072] Two injections of oxytocin are used for sexually mature male and female parents. The first injection is luteinizing hormone releasing hormone A25μg / kg for the female parent; 12 hours later, luteinizing hormone releasing hormone A28μg / kg + dioconazole 6mg / kg are injected into the female parent again; at the same time as the second injection of the female parent, the male parent is injected with oxytocin, and the injection dose is half of the second injection dose of the female parent. After the injection is completed, wait for delivery and keep the water temperature at 18-20℃;

[0073] 6 hours after the female parent completed the second injection of oxytocin, the male parent could be observed chasing and hitting the female parent's abdomen with his head, and then dry insemination was performed. First, the female fish's abdomen was gently pressed to squeeze out the mature eggs and put them in a clean and dry plastic basin. Then, the male fish's semen was squeezed onto the eggs, and water at a temperature of 19°C was added to activate the sperm. After stirring for 2 minutes, the mucus was washed off with water at a temperature of 19°C to obtain fertilized eggs.

[0074] The fertilized eggs were placed in a hatching ring with micro-flow water for incubation, the flow rate was maintained at 0.5m / s, and the hatching water temperature was controlled at 18-20℃. After 25 hours, the eggs emerged from the membranes and the fry were obtained. The artificial insemination had an induction rate of 100%, a fertilization rate of 56%, and a hatching rate of 62%. A total of about 32,000 newly hatched fry were obtained.

[0075] 24 hours after the fry hatched (i.e. 49 hours after fertilization), the fry were transferred to an open fry cultivation tank with a diameter of 1m and a depth of 0.8m, with independent inlets and outlets. The outlet was in the middle of the bottom, and a 0.6m diameter water pipe was inserted into the outlet. There were holes drilled on the side of the pipe. They were fed with conventional open fry feed (microencapsulated feed) four times a day. They were transferred to an outdoor cement pool 10 days after hatching and disinfected with 20kg / mu of quicklime 5 days before being transferred to the outdoor pool. They were fed in the pond in the early morning and fed twice a day after being put into the pond. 1 / 3 of the water was changed every week. Daily care was performed afterwards, and about 30,000 fry of the odd-headed grayling were obtained.

[0076] Table 1 Comparative Examples and Examples Artificial Breeding Results Statistics

[0077]

[0078] As can be seen from Table 1, Comparative Example 1 uses a one-time injection to induce labor, and the female parent's eggs cannot be discharged; Comparative Example 2 uses a two-needle injection to induce labor, but the water temperature after inducing labor is 26°C, which is higher than the embodiment. The female parent's eggs can be discharged but the ovulation volume is low, ovulation is difficult, and the separation is poor. Comparative Example 3 reduces the water temperature for delivery and the fertilization and incubation water temperature after injection, which significantly improves the fertilized eggs and hatching rate, but is still lower than the embodiment. While Examples 1 and 2 use the technology of the present invention to carry out artificial breeding work, and induce labor by two-needle injection. After the injection is completed, the temperature stimulation is controlled, which not only completes the artificial breeding of the odd-headed murrelet, but also the induction rate, fertilization rate and hatching rate are very high. Referring to this method, the artificial breeding, large-scale production and proliferation and release of the odd-headed murrelet can be realized, which is of great significance to the protection of the odd-headed murrelet.

[0079] It can be seen from the above embodiments that the present invention provides an artificial breeding method for the odd-headed garra, which can make the odd-headed garra's induced spawning rate 100%, the fertilization rate above 89%, and the hatching rate above 86%.

[0080] 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 principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. An artificial breeding method for the odd-headed garfish, characterized in that: The following steps are involved: S1. Parent fish collection: From June to August, female and male fish aged 2 to 3 years old with gonad development at stage V are collected and placed in temporary rearing ponds; S2. Artificial induction of labor: Inject luteinizing hormone-releasing hormone A24-6μg / kg to the female parent, and 11-13 hours later inject luteinizing hormone-releasing hormone A27-9μg / kg + dioxin 5-7mg / kg to the female parent; the water temperature after the second injection is 20.5-21.5℃, and the water temperature is controlled to decrease by 0.4-0.6℃ every hour within 5-7 hours, from 20.5-21.5℃ to 17.5-18.5℃, and then continue to maintain 17.5-18.5℃ to stimulate the ovarian development of the female parent; At the same time as the second injection of the female parent, the male parent is injected with luteinizing hormone-releasing hormone A2 3.5-4.5 μg / kg + dioconazole 2.5-3.5 mg / kg; S3. Artificial insemination: 6-7 hours after the female parent completes the second injection of oxytocin, gently press the abdomen of the female fish to squeeze out the mature eggs, then squeeze the male fish sperm onto the eggs, add 18-19℃ water to activate the sperm, stir for 110-130s, and then wash off the mucus with 18-19℃ water to obtain fertilized eggs; S4. Fertilized egg hatching: The fertilized eggs treated in S3 are placed in water at 18-21°C for hatching, with a water flow rate of 0.4-0.6 m / s for 0-22 hours, a water flow rate of 0.08-0.12 m / s for 22-26 hours, and a water flow rate of 0.4-0.6 m / s after 27 hours. After 23-25 ​​hours, the fry are obtained and cultured; S4. 23 to 25 hours after the fry emerge from the membrane, transfer the fry to the fry culture tank and feed them with fry feed; after the fry start to hatch, transfer them to the outdoor cement pool for cultivation to obtain the odd-headed garfish fry.

2. The artificial breeding method of the odd-headed garfish according to claim 1, characterized in that: The weight of the female parent is ≥150 g, the weight of the male parent is ≥100 g, and semen flows out when the abdomen is squeezed.

3. The artificial breeding method of the odd-headed garfish according to claim 2, characterized in that: The ratio of the female parent to the male parent is: 1.2～1.8:1。 4. The artificial breeding method of the odd-headed garfish according to claim 3, characterized in that: When activating sperm, the amount of water added is 2 to 4 times the volume of the mixture of fish eggs and fish semen.

5. The artificial breeding method of the odd-headed garfish according to claim 4, characterized in that: The fertilized eggs are incubated in the incubation ring, where the water depth is 58-62 cm and the density of the fertilized eggs is 48,000-52,000 eggs / m 3 .

6. The artificial breeding method of the odd-headed garfish according to claim 5, characterized in that: The diameter of the open seedling cultivation tank is 0.8-1.2 m, the depth is 0.7-0.9 m, and the density of fry in the open seedling cultivation tank is 3800-4200 fry / m 3 , the water temperature is 20-22℃.

7. The artificial breeding method of the odd-headed garfish according to claim 6, characterized in that: The starter fry feed is fed once in the morning and evening every day, and each feeding amount is 4.8-5.2% of the body weight of the cultured fry.

8. The artificial breeding method of the odd-headed garfish according to claim 1, characterized in that: The catching of broodstock is carried out by placing ground traps in the shallows with stone bottom in the middle and lower reaches of the Lancang River in the evening for catching.

9. The artificial breeding method of the odd-headed garfish according to claim 1, characterized in that: The captured broodstock are packed in bags, 3-4L of water and 9-11 odd-headed garfish are added to each bag, and 0.4-0.6g of anesthetic, 4-6g of multivitamins, and 0.8-1.2mL of ginger paste are added and stirred. The air is exhausted and then oxygenated, and the bag is tied tightly. The specifications of the bag are 38-42cm×68-72cm.

10. The artificial breeding method of the odd-headed garfish according to claim 8, characterized in that: The captured broodstock are soaked in 0.3% to 0.5% salt water for 8 to 12 minutes and then placed in a temporary holding pond.

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