A method for artificial breeding of a species of the genus Onychostoma

Abstract

The application provides an artificial breeding method of Onychostoma sima, and belongs to the technical field of aquaculture. The method comprises the following steps: injecting the female parent of the breeding parent fish with a luteinizing hormone twice to artificially induce parturition, and obtaining fertilized eggs; using a microflow of 18-21 DEG C to artificially incubate the fertilized eggs, and controlling the water flow rate at different stages of incubation until the fry is hatched, and obtaining the fry of Onychostoma sima. The method of the application injects the luteinizing hormone in two injections, and strictly controls the injection dose of each injection, so that the well-developed eggs and sperm can be obtained, and the incubation conditions of the fertilized eggs are controlled, so that the parturition rate of Onychostoma sima is 100%, the fertilization rate is more than 89%, and the hatching rate is more than 86%, and the large-scale artificial breeding of Onychostoma sima is realized, which has important significance for protecting the endangered resource of Onychostoma sima.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of aquaculture, and particularly relates to an artificial breeding method of Garramirofronits. BACKGROUND

[0002] Garramirofronits belongs to Cypriniformes, Cyprinidae, Labeoninae and Garra. The front part of the body is cylindrical, the tail is flat, the back edge is arc-shaped, the chest and abdomen are flat, and the head is square-shaped. The unique feature is that there are two frontal protuberances in front of the nostrils of the head, each of which has a large and obvious conical keratinous protuberance at the front end. Garramirofronits is dark green in color, with dark gray fin strips and red tips. Garramirofronits lives in rivers with strong currents and many stones on the river bottom, and mainly feeds on algae, detritus and aquatic insect larvae. Garramirofronits is only distributed in the middle and lower reaches of the Lancang River and is a fish species unique to the Lancang River system. Garramirofronits grows fast, has tender and delicious meat, and is highly welcomed in the market. Garramirofronits has high economic value, development prospects and significance for popularization and cultivation.

[0003] With the development of the Lancang River cascade, six hydropower stations, namely Gongguoqiao, Xiaowan, Manwan, Dachaoshan, Nuozhadu and Jinghong, have been built and operated in the middle and lower reaches of the Lancang River, and have been in operation for more than 10 years. The construction and operation of the power stations have a significant impact on fish, especially Garramirofronits which produces drifting eggs. The dam blocks the migration of fish, changes the natural flow of the river, and turns the originally connected flowing river into a blocked slow-flowing habitat, which destroys the original spawning ground and habitat. The reduction of the river flow rate, the distance of the flowing water and the change of the water depth also affect the hatching of the drifting eggs. The change of the hydrological regime also affects the abundance of the bait organisms, which compresses the survival space of Garramirofronits and affects the population reproduction. In addition, the influence of human factors, the wild resources of Garramirofronits have decreased, and Garramirofronits needs to be protected.

[0004] In order to protect the population resources of Garramirofronits, it is necessary to provide an artificial breeding method of Garramirofronits, which provides guidance for artificial breeding, resource protection, large-scale production and development and utilization. SUMMARY

[0005] The present application relates to the technical field of aquaculture, and particularly relates to an artificial breeding method of Garramirofronits.

[0006] In order to achieve the above-mentioned purpose of the application, the present application provides the following technical scheme:

[0007] The present application provides an artificial breeding method of Garramirofronits, comprising the following steps:

[0008] S1. Collecting parents: 2-3 years old female and male parents with gonad development at the 5th stage are collected from June to August and then are put into temporary ponds for temporary cultivation;

[0009] S2. Artificially inducing spawning: 24-6 μg / kg of luteinizing hormone-releasing hormone A is injected into female parents, 11-13 h later, 27-9 μg / kg of luteinizing hormone-releasing hormone A and 5-7 mg / kg of diethylstilbestrol are injected into female parents; the water temperature is 20.5-21.5 ℃ after the second injection, and the water temperature is decreased by 0.4-0.6 ℃ per hour for 5-7 h, from 20.5-21.5 ℃ to 17.5-18.5 ℃, and then the water temperature is kept at 17.5-18.5 ℃, to stimulate the development of ovary of female parents;

[0010] At the same time when the second injection is performed on the female parents, 23.5-4.5 μg / kg of luteinizing hormone-releasing hormone A and 2.5-3.5 mg / kg of diethylstilbestrol are injected into male parents;

[0011] S3. Artificial insemination: 6-7 h after the second injection of the female parents, mature eggs are squeezed out by gently pressing the abdomen of the female fish, and then the sperm of the male fish is squeezed onto the eggs, 18-19 ℃ water is added to activate the sperm, and then the mucus is washed away after stirring for 110-130 s, to obtain the fertilized eggs;

[0012] S4. Hatching of the fertilized eggs: the fertilized eggs treated in S3 are placed in 18-21 ℃ water for hatching, the water flow rate is 0.4-0.6 m / s for 0-22 h, the water flow rate is 0.08-0.12 m / s for 22-26 h, the water flow rate is 0.4-0.6 m / s after 27 h, and the larvae are obtained after hatching for 23-25 h, and the larvae are further cultivated;

[0013] S4. After 23-25 h of hatching of the larvae, the larvae are transferred to a rearing tank for open-mouth fry, and open-mouth fry feed is fed; after the larvae open their mouths, the larvae are transferred to an outdoor cement pool for cultivation, to obtain the fish fry of the Oxyeleotris marmorata.

[0014] Preferably, the weight of the female parents is ≥150 g, and the weight of the male parents is ≥100 g, and the sperm is squeezed out by pressing the abdomen.

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

[0016] Preferably, when the sperm is activated, the water is added in an amount of 2-4 times the volume of the mixture of the fish eggs and the fish sperm.

[0017] Preferably, the hatching of the fertilized eggs is performed in a hatching ring, the water depth in the hatching ring is 58-62 cm, and the density of the fertilized eggs is 48-52 thousand eggs / m 3 .

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

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

[0020] Preferably, the parent fish are captured by placing a ground cage in a shallow place with stone bottom in the middle and lower reaches of the Lancang River in the evening.

[0021] Preferably, the captured parent fish are packed with bags, 3-4 L of water and 9-11 odd-nose catfish are added to each bag, and 0.4-0.6 g of anesthetic, 4-6 g of compound vitamin, and 0.8-1.2 mL of ginger paste are added at the same time, and then stirred, and after the air is exhausted, oxygen is added again, and the bag opening is tightly tied, and the size of the bag is 38-42 cm x 68-72 cm.

[0022] Preferably, the captured parent fish are soaked in 0.3%-0.5% salt water for 8-12 min and then placed in a temporary culture tank for temporary culture.

[0023] By adopting the above technical scheme, the present application has the following beneficial effects: the present application adopts the method of injecting the oxytocin by two injections, and strictly controls the dosage of each injection, so that the well-developed eggs and sperm can be obtained, and the hatching conditions of the fertilized eggs are controlled, so that the oxytocin production rate of the odd-nose catfish is 100%, the fertilization rate is more than 89%, and the hatching rate is more than 86%, and the scale artificial breeding of the Lhasa schizothoracn fish is realized, which has important significance for protecting the endangered resources of the odd-nose catfish. DETAILED DESCRIPTION

[0024] The present application provides an artificial breeding method of odd-nose catfish, which comprises the following steps:

[0025] S1. Parent fish capture: 2-3 year-old female and male fish with gonad development at the V stage are captured in June-August, and then are placed in a temporary culture tank for temporary culture;

[0026] S2. Artificial oxytocin induction: 24-6 μg / kg of luteinizing hormone-releasing hormone A is injected into the female parent, and 27-9 μg / kg of luteinizing hormone-releasing hormone A and 5-7 mg / kg of diethylstilbestrol are injected into the female parent after 11-13 h; 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℃ per hour for 5-7 h, from 20.5-21.5℃ to 17.5-18.5℃, and then the water temperature is maintained at 17.5-18.5℃, so as to stimulate the ovarian development of the female parent;

[0027] The male parent is injected with luteinizing hormone releasing hormone A 23.5-4.5 μg / kg + diethylstilbestrol 2.5-3.5 mg / kg at the same time when the female parent is injected with the second needle;

[0028] S3. Artificial insemination: 6-7 h after the female parent completes the injection of the second needle, the abdomen of the female fish is gently pressed to squeeze out mature eggs, and then the sperm of the male fish is squeezed onto the eggs, 18-19℃ water is added to activate the sperm, and after stirring for 110-130 s, the mucus is washed away with 18-19℃ water to obtain fertilized eggs;

[0029] S4. Fertilized egg hatching: the fertilized eggs treated in S3 are placed in 18-21℃ water for hatching, the water flow rate is 0.4-0.6 m / s for 0-22 h, the water flow rate is 0.08-0.12 m / s for 22-26 h, and the water flow rate is 0.4-0.6 m / s after 27 h, the membrane is out after 23-25 h, and the fry is obtained, and the culture is continued;

[0030] S4. After 23-25 h of fry membrane out, the fry is transferred to an open fry culture tank and fed with open fry feed; after the fry opens, it is transferred to an outdoor concrete pool for culture, and the odd-fin suckermouth catfish fry is obtained.

[0031] The application collects Onychostoma sima in the breeding season, and obtains the selected Onychostoma sima after transportation and treatment. Specifically, in June-August every year, the ground cage is placed in the shallow place with stone bottom in the middle and lower reaches of Lancang River, and the Onychostoma sima is collected; the collected Onychostoma sima is packaged and transported by bag, the bag is preferably a straight plastic bag, the size of the straight plastic bag is 38-42 cm*68-72 cm, further preferably 39-41 cm*69-7 cm, more preferably 40 cm*70 cm, 3-4 L of water and 8-12 Onychostoma sima are added to each bag, the volume of the water is further preferably 3.2-3.8 L, more preferably 3.5 L, the number of Onychostoma sima in the straight plastic bag is 9-11, more preferably 10; 0.4-0.6 g of anesthetic, 4-6 g of compound vitamin and 0.8-1.2 mL of ginger extract are added at the same time, the anesthetic is preferably MS-222, the mass of the anesthetic is further preferably 0.45-0.55 g, more preferably 0.5 g, the mass of the compound vitamin is 4.5-5.5 g, further preferably 5 g, the volume of the ginger extract is further preferably 0.9-1.1 mL, more preferably 1 mL, then the air in the bag is exhausted, oxygen is filled, the bag opening is tightly tied, and the packaging bag is pressed with the arm to check whether there is water leakage or air leakage, if there is such phenomenon, it is replaced in time and repackaged. The method can be transported for 2-3 days; after arriving at the destination, the Onychostoma sima is warmed and watered, soaked in salt water and placed in a temporary breeding pool, the concentration of the salt water is 0.3-0.5%, further preferably 0.35-0.45%, more preferably 0.4%; the soaking time is 8-12 min, further preferably 9-11 min, more preferably 10 min. The temporary breeding pool is preferably a circular cement pool, the diameter is 1.8-2.2 m, further preferably 1.9-2.1 m, more preferably 2 m; the water outlet is in the middle of the temporary breeding pool, and the water inlet is located on one side of the pool wall.

[0032] After the Onychostoma sima is collected, the sexually mature female and male parents are selected from the selected Onychostoma sima. The sexually mature parents are 2-3 year-old female and male fish with the gonad development of the fifth stage, the male parent has a weight of more than 100 g, the pearl star is not obvious when the chest fin bar is touched, the abdomen is slightly rough, and the milky white semen flows out when the abdomen is lightly pressed, and the semen is scattered when entering the water; the female parent has a weight of more than 150 g, the chest fin bar is smooth when touched, and the abdomen is obviously swollen.

[0033] The present application selects parents, and then artificially induces parturition. The female and male parents are injected with an inducing agent to obtain parents to be induced. The ratio of female to male in the parents to be induced is 1.2-1.8:1, further preferably 1.3-1.7:1, and more preferably 1.5:1. The female parent is injected twice. The first injection is of luteinizing hormone-releasing hormone A2, and the injection dose is 4-6 μg / kg, further preferably 4.5-5.5 μg / kg, and more preferably 5 μg / kg. The second injection is of luteinizing hormone-releasing hormone A2+ diethylstilbestrol, and the injection dose of the luteinizing hormone-releasing hormone A2 is 7-9 μg / kg, further preferably 7.5-8.5 μg / kg, and more preferably 8 μg / kg. The injection dose of the diethylstilbestrol is 5-7 mg / kg, further preferably 5.5-6.5 mg / kg, and more preferably 6 mg / kg. The water temperature after the second injection is 20.5-21.5°C, further preferably 20.8-21.2°C, and more preferably 21°C. The water temperature is controlled to decrease by 0.4-0.6°C per hour, further preferably 0.5°C per hour, from 20.5-21.5°C to 17.5-18.5°C, further preferably 17.8-18.2°C, and more preferably 18°C, within 5-7 hours, further preferably 5.5-6.5 hours, and more preferably 6 hours, and then maintained at 17.5-18.5°C, further preferably 17.8-18.2°C, and more preferably 18°C, to stimulate the development of the ovary of the female parent. The male parent is injected with luteinizing hormone-releasing hormone A2+ diethylstilbestrol at the same time as the second injection of the female parent. The injection dose of the luteinizing hormone-releasing hormone A2 is 3.5-4.5 μg / kg, further preferably 3.8-4.2 μg / kg, and more preferably 4.0 μg / kg. The injection dose of the diethylstilbestrol is 2.5-3.5 mg / kg, further preferably 2.8-3.2 mg / kg, and more preferably 3.0 mg / kg.

[0034] The mature eggs are squeezed out by gently pressing the abdomen of the female fish 6-7 hours, further preferably 6.2-6.8 hours, and more preferably 6.5 hours, after the second injection of the female parent. The sperm of the male fish is then added to the eggs, and the sperm is activated by adding water. The mixture is stirred, and then the mucus is washed away to obtain the fertilized eggs. The water temperature is 18-19°C, further preferably 18.2-18.8°C, and more preferably 18.5°C. The amount of water added is 2-4 times, further preferably 2.5-3.5 times, and more preferably 3 times, the volume of the mixture of the fish eggs and the fish sperm. The stirring time is 110-130 seconds, further preferably 115-125 seconds, and more preferably 120 seconds. The water used to wash away the mucus has the same temperature as the water used to activate the sperm.

[0035] The present application carries out hatching after obtaining the fertilized eggs, the fertilized eggs of the Chondrostoma ocellatum are the drifting eggs, preferably the fertilized eggs of the Chondrostoma ocellatum are placed in the hatching ring channel and hatched in the micro-flow water, the water depth in the hatching ring channel is 58-62 cm, further preferably 59-61 cm, more preferably 60 cm; the density of the fertilized eggs is 48-52 thousand pieces / m 3 , further preferably 49-51 thousand pieces / m 3 , more preferably 50 thousand pieces / m 3 . The flow rate is adjusted according to the hatching stage, the flow rate is kept at 0.4-0.6 m / s in 0-22 h, further preferably 0.45-0.55 m / s, more preferably 0.5 m / s; the flow rate is reduced to 0.08-0.12 m / s in 22-26 h, further preferably 0.09-0.11 m / s, more preferably 0.1 m / s; the flow rate is restored to 0.4-0.6 m / s after 27 h, further preferably 0.45-0.55 m / s, more preferably 0.5 m / s, the hatching water temperature is controlled at 18-21 DEG C, further preferably 19-20 DEG C, more preferably 19.5 DEG C, the membrane is out after 23-25 h, the fry is obtained, and the culture is continued.

[0036] The present application further preferably 23.5-24.5 h, more preferably 24 h after the fry membrane is out, the fry is transferred to the opening fry culture tank for culture, the diameter of the opening fry culture tank is 0.8-1.2 m, further preferably 0.9-1.1 m, more preferably 1 m; the depth of the opening fry culture tank is 0.7-0.9 m, further preferably 0.75-0.85 m, more preferably 0.8 m; the opening fry culture tank has independent water inlet and outlet, the water outlet is in the middle of the bottom, a water pipe with a diameter of 0.6 m is inserted into the water outlet, and holes are drilled on the side of the pipe. The density of the fry in the opening fry culture tank is 3800-4200 tails / m 2 , further preferably 3900-4100 tails / m 2 , more preferably 4000 tails / m 2 ; the water temperature is 20-22 DEG C, further preferably 20.5-21.5 DEG C, more preferably 21 DEG C. The conventional opening fry feed (micro-capsule feed) is fed 4 times a day, the opening fry feed is preferably S1 micro-particle feed produced by Shandong Shengsuo Feed Co., Ltd., and the weight of the cultured fry is 4.8-5.2% per time, further preferably 4.9-5.1%, more preferably 5%. The Chondrostoma ocellatum is transferred to the outdoor cement pool after opening (about 10 d after the membrane is out), and the cement pool is disinfected with quicklime 5-7 d before the transfer, the amount of quicklime used is 18-22 kg / mu, further preferably 19-21 kg / mu, more preferably 20 kg / mu. The Chondrostoma ocellatum is put into the pond in the morning after being fed to satiation, and the feed is fed 2 times a day after being put into the pond, and the water is replaced by 1 / 3 every week, and only daily management is needed to obtain the Chondrostoma ocellatum fry.

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

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

[0039] Example 1

[0040] On July 25, 2024, in the evening, the ground cage was placed at the shallow place with stone bottom in the flowing water section of the middle and lower reaches of the Lancang River, and the oddhead fish would scrape the algae growing on the stones into the ground cage; the collected oddhead fish was packed in a 40 cm x 70 cm straight plastic bag, 4 L of water and 10 oddhead fish were added to each bag, 0.5 g of MS-222, 5 g of compound vitamin, and 1 mL of ginger paste were added and stirred evenly, the air in the bag was discharged first, then oxygen was filled, the bag opening was tightly tied with a rubber ring, and the packaging bag was lightly pressed with the arm to check whether there was water leakage or air leakage, if there was such phenomenon, it should be replaced and repacked; after arriving at the destination, the temperature and water were adjusted, and after being soaked in 0.3% saline for 10 min, it was placed in a 2 m diameter circular cement pool for temporary cultivation, the water was discharged in the middle of the cement pool, and the water inlet was located on one side of the pool wall.

[0041] On July 27, 2024, among the selected oddhead fish collected on July 25, 2 two-year-old male oddhead fish with an average weight of 105 g were selected, which had no obvious beads along the fin ray direction of the pectoral fin, were slightly rough, had whiteish yellow seminal fluid flowing out when the abdomen was lightly pressed, and scattered immediately after entering the water; 3 two-year-old female oddhead fish with an average weight of 158 g were selected, which had smooth fin rays along the fin ray direction of the pectoral fin, and the abdomen was obviously swollen; the ratio of males to females was 3:2.

[0042] The sex matured male and female parents were injected with two injections of the labor inducer, the first injection of the labor inducer was A24 μg / kg of luteinizing hormone releasing hormone for the female parent; 12 h later, the female parent was injected again with A27 μg / kg of luteinizing hormone releasing hormone + 5 mg / kg of diethylstilbestrol; at the same time of the second injection of the female parent, the male parent was injected with A23.5 μg / kg of luteinizing hormone releasing hormone + 2.5 mg / kg of diethylstilbestrol, and after the injection was completed, the female parent was allowed to give birth; at the same time, the water temperature of the parent to be born was controlled to be reduced by 0.5℃ every 1 h within 6 h after the second injection, from 20.5℃ to 17.5℃, and then maintained at 17.5℃, to stimulate the ovarian development of the female parent.

[0043] 7 h after the second injection of the labor inducer was completed, the male parent was observed to chase and hit the abdomen of the female parent with the head to perform dry artificial insemination, first, the mature eggs were squeezed out and placed in a clean and dry plastic basin by lightly pressing the abdomen of the female fish, then the sperm of the male fish was squeezed onto the eggs, 19℃ water was added to activate the sperm, and after stirring for 2 min, the mucus was washed away with 19℃ water to obtain the fertilized eggs.

[0044] The fertilized eggs were placed in a micro-flow incubation loop and incubated. The flow rate was adjusted according to the incubation stage: the flow rate was maintained at 0.5 m / s for 0 to 22 h, reduced to 0.1 m / s for 22 to 26 h, and restored to 0.5 m / s after 27 h. The incubation water temperature was controlled to be 18 to 21 ° C, and the eggs were hatched after 24 h. The artificial breeding method of the present embodiment had a 100% induced spawning rate, an 89.2% fertilization rate, and an 86.5% hatching rate, 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, which had 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) 4 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 well 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. Subsequent daily care was carried out to obtain about 75,000 fry of the strange-headed garfish.

[0046] Example 2

[0047] On the evening of August 5, 2024, traps were set up in the rocky shallows of the middle and lower reaches of the Lancang River. Twenty-eight garrulella were collected and packed into 40 cm × 70 cm straight plastic bags. Each bag contained 4 L of water and 10 garrulella. 0.5 g MS-222, 5 g of multivitamins, and 1 mL of ginger paste were added and mixed thoroughly. The air in the bag was first purged and then filled with oxygen. The bag was secured with a rubber band and gently pressed with the hand to check for leaks. If any were detected, the bag was replaced and repacked. After transport to the destination, the fish were heated and watered, then soaked in 0.5% saline for 10 minutes and placed in a 2 m diameter circular cement pool for temporary storage. The water outlet was located in the center of the pool, and the inlet was located on the side of the pool wall.

[0048] On August 8, we selected two male odd-headed garfish with an average weight of 111 g, which were slightly rough and had faint pearl stars when touched along the direction of the pectoral fin rays. When lightly pressed on the abdomen, milky white semen flowed out and dispersed when entering the water. We also selected three female odd-headed garfish 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 sex matured female and male parents are injected with two injections of the parturition stimulant. The first injection is 25 μg / kg of luteinizing hormone releasing hormone A for the female parent. 12 hours later, the female parent is injected again with 8 μg / kg of luteinizing hormone releasing hormone A + 6 mg / kg of deoxycorticosterone. At the same time the female parent is injected with the second injection, the male parent is injected with 4 μg / kg of luteinizing hormone releasing hormone A + 3 mg / kg of deoxycorticosterone. After the injection is completed, the parents are allowed to give birth. At the same time, within 6 hours after the second injection, the water temperature is decreased by 0.5°C every hour until it reaches 18°C. The temperature is then kept at 18°C to stimulate the development of the ovary of the female parent.

[0050] Six hours after the second injection of the parturition stimulant, the male parent chases the female parent and hits the abdomen of the female parent with the head to perform dry insemination. The mature eggs are squeezed out of the abdomen of the female parent and placed in a clean and dry plastic basin. The sperm of the male parent is then squeezed onto the eggs. Water at 18°C is added to activate the sperm. After 2 minutes of stirring, the mucus is washed away with water at 18°C to obtain the fertilized eggs. The fertilized eggs are placed in a hatching ring channel and hatched in a slow stream of water. The flow rate is adjusted according to the hatching stage. The flow rate is kept at 0.5 m / s for 0-22 hours, reduced to 0.1 m / s for 22-26 hours, and then restored to 0.5 m / s after 27 hours. The hatching water temperature is controlled at 18-21°C. The membrane is broken after 25 hours. The artificial breeding parturition rate is 100%, the fertilization rate is 90.3%, the hatching rate is 88.6%, and about 79,000 larvae are obtained.

[0051] Twenty-four hours after the membrane is broken (i.e., 49 hours after insemination), the larvae are transferred to a rearing tank with a diameter of 1 m, a depth of 0.8 m, and a separate water inlet and outlet. The water outlet is in the middle of the bottom and is inserted with a water pipe with a diameter of 0.6 m. The pipe is drilled with holes on the side to form an opening for the larvae. The larvae are fed with conventional opening feed (microcapsule feed) 4 times a day. Ten days after hatching, the larvae are transferred to an outdoor cement pool. Five days before the transfer, the pool is disinfected with 20 kg / mu of quicklime. The larvae are fed twice a day after being transferred to the pool. The water is changed by 1 / 3 every week. The subsequent daily care and maintenance are routine. About 77,000 larvae of the Miichthys miiuy are obtained.

[0052] Example 3

[0053] On July 30, 2024, at 5:00 pm, 28 tail of the fish were collected in the shallow waters of the Lancang River, with a bottom of stone blocks. The fish were packed in 40 cm x 70 cm straight plastic bags, with 4 L of water and 10 tail of the fish added to each bag. 0.5 g of MS-222, 5 g of compound vitamin, and 1 mL of ginger paste were also added and stirred evenly. The air in the bag was first exhausted, then oxygen was added. The bag was then tightly sealed with a rubber band, and the bag was lightly pressed with the arm to check for leaks. If there was a leak, the bag was replaced and re-packed. After transportation to the destination, the fish were soaked in 0.5% salt water for 10 minutes and then placed in a 2m diameter circular cement pool for temporary cultivation. The water was discharged from the middle of the pool, and the water inlet was located on one side of the pool wall.

[0054] On August 2, 2 tail of the fish were selected along the direction of the fin rays of the pectoral fin, with no obvious beads and slightly rough, and the average weight of the 2-year-old male fish was 108g. 3 tail of the fish were selected along the direction of the fin rays of the pectoral fin, with smooth and obvious abdominal swelling, and the average weight of the 2-year-old female fish was 157g. The ratio of male to female was 3:2.

[0055] The sex matured female and male parents were injected with the progestin, and the two needle method was used to inject the progestin. The first needle was injected with luteinizing hormone releasing hormone A 25 μg / kg for the female parent, and 12 hours later, the female parent was injected with luteinizing hormone releasing hormone A 28 μg / kg + diethylstilbestrol 6 mg / kg. At the same time, the male parent was injected with luteinizing hormone releasing hormone A 24 μg / kg + diethylstilbestrol 3 mg / kg, and the injection was completed after the second needle injection. The water temperature was controlled to decrease by 0.5℃ every 1h within 6h after the second needle injection, from 21.5℃ to 18.5℃, and then maintained at 18.5℃ to stimulate the development of the female parent's ovary.

[0056] 6 hours after the second injection of the female parent, the male parent was observed to chase and hit the female parent's abdomen with its head, and the dry insemination was performed. First, the mature eggs were squeezed out and placed in a clean and dry plastic basin, and then the male fish sperm was squeezed onto the eggs. 18℃ water was added to activate the sperm, and after 2 minutes of stirring, the mucus was washed away with water at 18℃. The fertilized eggs were placed in the hatching ring channel for micro-flow water hatching, and 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 then restored to 0.5m / s after 27h. The hatching water temperature was controlled at 18-21℃, and the membrane was out after 25h. The artificial breeding progestin rate was 100%, the fertilization rate was 89.8%, the hatching rate was 87.1%, and about 780,000 newly hatched fry were obtained.

[0057] The larvae are transferred to a 1 m diameter, 0.8 m deep, water outlet in the middle of the bottom, and a water inlet on one side of the wall, and a 0.6 m diameter water pipe is inserted into the water outlet, and the pipe is bored on the side to form an opening for the rearing tank. The larvae are fed with conventional starter feed (microcapsule feed) 4 times a day, and are transferred to an outdoor cement pool 10 days after hatching. The cement pool is disinfected with 20 kg / mu of quicklime 5 days before the transfer, and the larvae are fed 2 times a day after being released into the pool. The water is replaced 1 / 3 every week, and the subsequent daily management is carried out. About 760,000 fry of the odd-fin black carp are obtained.

[0058] Comparative Example 1

[0059] On July 24, 2024, in the evening, a cage was placed on a shallow shoal with a stone bottom in the middle and lower reaches of the Lancang River, and 21 odd-fin black carp were collected. The fish were packed in 40 cm x 70 cm straight plastic bags, 3-4 L of water and 10 odd-fin black carp were added to each bag, 0.5 g of MS-222, 5 g of compound vitamins, and 1 ml of ginger paste were added, and the mixture was stirred evenly. The air in the bag was first discharged, then oxygen was filled, and the bag opening was tightly tied with a rubber band. The bag was lightly pressed with the arm to check for leaks. If there is a leak, it must be replaced and repackaged. After transportation to the destination, the fish were soaked in 0.3% saline for 10 minutes and then placed in a 2 m diameter circular cement pool for temporary cultivation. The water outlet was in the middle of the cement pool, and the water inlet was on one side of the wall.

[0060] On July 26, 2 two-year-old male odd-fin black carp with an average weight of 110 g were selected for the selection of odd-fin black carp, with no obvious pearl stars along the fin bar direction of the pectoral fin, slightly rough, and white semen flowing out of the abdomen when pressed lightly, and the fish dispersed immediately after entering the water. Two two-year-old female odd-fin black carp with an average weight of 155 g were selected for the selection of odd-fin black carp, with smooth fin bar direction along the pectoral fin, and the abdomen was significantly enlarged. The ratio of females to males was 3:2.

[0061] The female parent was injected with 212 μg / kg of luteinizing hormone releasing hormone A and 6 mg / kg of diethylstilbestrol, and the male parent was injected with half the dose of the female parent. After the injection was completed, the fish were allowed to spawn, and the water temperature was 26°C.

[0062] The development of the female parent was checked 6 hours after the injection was completed, and the eggs could not be successfully discharged. The eggs were 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 2 minutes of stirring, the mucus was washed away with water at 26°C. The fertilized eggs were placed in a hatching ring channel and hatched in a small stream of water. After about 6 hours, all the eggs died. The artificial breeding induction rate was 33.33%, the fertilized egg rate was 0%, and the hatching rate was 0%.

[0063] Comparative Example 2

[0064] On July 28, 2024, 2 two-year-old male Pterophyllum scalare with an average weight of 112 g were selected from the alternative odd-numbered head fish caught on July 24 (the same batch as Comparative Example 1), which had no obvious beads along the fin bar direction of the pectoral fin, were slightly rough, and had a light white discharge when the abdomen was lightly pressed. The discharge dispersed immediately after entering the water. Three two-year-old female Pterophyllum scalare with an average weight of 163 g were selected, which were smooth along the fin bar direction of the pectoral fin, and the abdomen was obviously swollen. The ratio of females to males was 3:2.

[0065] The two-needle method was used for injection of oxytocin. The first needle was injected with luteinizing hormone-releasing hormone A 25 μg / kg for the female parent. Twelve hours later, the female parent was injected again with luteinizing hormone-releasing hormone A 28 μg / kg + diethylstilbestrol 6 mg / kg. At the same time as the second injection of the female parent, the male parent was injected with an oxytocin agent at half the dose of the second injection of the female parent. After the injection was completed, the water temperature was 26°C.

[0066] The development was checked 6 hours after the injection of the female parent. The egg granules could be squeezed out, but the ovulation was not smooth, and most of the egg granules were not separated. Then the eggs were squeezed out and placed in a clean and dry plastic basin. The male fish sperm 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 away with water at 26°C, and the fertilized eggs were obtained.

[0067] The fertilized eggs were placed in a hatching ring channel with a flow rate of 0.5 m / s and a water temperature of 26°C. The membrane was broken after 21 hours, and the fry was obtained. The artificial insemination induction rate was 66.67%, the fertilization rate was 3.2%, the hatching rate was 10%, and about 0.02 million fry were obtained.

[0068] After 24 hours of fry membrane rupture (i.e. 49 hours after insemination), the fry was transferred to a diameter of 1 m, a depth of 0.8 m, with independent water inlet and outlet, the outlet was in the middle of the bottom, and a diameter of 0.6 m water pipe was inserted into the outlet, the pipe side was drilled with an opening fry breeding tank. Feed the fry 4 times a day with conventional starter feed (microcapsule feed). Ten days after hatching, the fry was transferred to an outdoor cement pool. Five days before transfer to the outdoor pool, the pool was disinfected with 20 kg of quicklime per mu. The fry was fed twice a day after being fed in the morning. The water was changed by 1 / 3 every week, and the subsequent daily maintenance was carried out. About 0.01 million fry of Pterophyllum scalare were obtained.

[0069] Comparative Example 3

[0070] On the evening of August 4, 2024, a ground trap was placed in the shallows with rocky bottoms in the middle and lower reaches of the Lancang River. 28 odd-headed garfish were collected and packed in 40cm×70cm straight plastic bags. 4L of water and 10 odd-headed garfish were added to each bag. 0.5g MS-222, 5g multivitamins, and 1ml ginger paste were added and mixed evenly. The air in the bag was first exhausted, then filled with oxygen. The bag was tightened with a rubber band and the bag was gently pressed with the arm to check for water or air leaks. If so, it was replaced and repackaged in time. After transportation to the destination, the fish were heated and watered, soaked in 0.5% salt water for 10 minutes, and then placed in a temporary holding pond with a diameter of 2m in a circular cement pool with water outlet in the middle and the water inlet located on the side of the pool wall.

[0071] On August 7, we selected two male garrulilla fish, weighing an average of 109g, each aged 2 years. The fish had a slightly rough surface when touched along the pectoral fin rays, and milky white semen flowed out when the fish entered the water. The fish also had a smooth surface when touched along the pectoral fin rays, and a noticeably enlarged abdomen. The fish weighed an average of 157g, each aged 2 years. The male-to-female ratio was 3:2.

[0072] The sexually mature male and female parents were injected with oxytocin using a two-injection method. The female parent was first injected with luteinizing hormone-releasing hormone A25μg / kg; 12 hours later, the female parent was injected with luteinizing hormone-releasing hormone A28μg / kg + dioxin 6mg / kg again; at the same time as the female parent was injected with the second injection, the male parent was injected with oxytocin, and the injection dose was half of the second injection dose of the female parent. After the injection, the mother was allowed to wait for delivery, and the water temperature was maintained at 18-20℃.

[0073] Six hours after the female parent completed the second injection of oxytocin, when the male parent was observed chasing and hitting the female parent's abdomen with his head, dry insemination was performed. First, the female fish's abdomen was gently pressed to squeeze out the mature eggs and placed 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 the fertilized eggs.

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

[0075] After 24 hours of the fry membrane (i.e. 49 hours after insemination), the fry is transferred to a diameter 1m, depth 0.8m, with independent water inlet and outlet, the outlet is in the middle of the bottom, the outlet is inserted with a diameter of 0.6m water pipe, the pipe side is drilled with an opening fry culture tank, the conventional opening fry feed (microcapsule material) is fed 4 times a day, and after 10 days of hatching, it is transferred to an outdoor cement pool, 5 days before being transferred to the outdoor pool, 20kg / mu of quicklime is used for disinfection, and the fish is fed in the morning, and after being fed in the pond, the feed is fed twice a day, and the water is changed 1 / 3 every week, and the subsequent is daily management, and about 300,000 fry of the odd-fin gourami is obtained.

[0076] Table 1. Statistics of artificial breeding of juvenile results

[0077]

[0078] From Table 1, it can be seen that in the comparative example 1, the female parent ovum cannot be discharged by one-time injection of labor induction; in the comparative example 2, the water temperature after labor induction is 26℃, which is higher than that of the examples, the female parent ovum can be discharged but the ovulation amount is low, the ovulation is difficult, and the separation degree is poor, while in the comparative example 3, the water temperature after injection is reduced and the water temperature for fertilization and hatching is reduced, which significantly improves the fertilized egg and hatching rate, but is still lower than that of the examples. While the examples 1 and 2 use the technology of the present application to carry out artificial breeding, through two-needle injection of labor induction, after the injection is completed, the temperature is controlled to stimulate, not only the artificial breeding of the odd-fin gourami is completed, but also the labor induction rate, the fertilization rate and the hatching rate are very high. Referring to the method, the artificial breeding, large-scale production and release of the odd-fin gourami can be realized, which has important significance for the protection of the odd-fin gourami.

[0079] From the above examples, it can be seen that the present application provides an artificial breeding method of odd-fin gourami, which can make the labor induction rate of the odd-fin gourami 100%, the fertilization rate more than 89%, and the hatching rate more than 86%.

[0080] The above is only the preferred embodiment of the present application, it should be pointed out that for the ordinary skilled in the art, without departing from the principles of the present application, a number of improvements and refinements can be made, which should be considered as the protection scope of the present application.

Claims

1. A method for artificial breeding of garfish, characterized in that: The following steps are involved: S1. Broodstock Collection: From June to August, 2-3 year old females and males at stage V of gonadal development are collected and placed in temporary holding ponds. S2. Artificial induction of labor: Inject the female parent with luteinizing hormone-releasing hormone A24-6 μg / kg. 11-13 hours later, inject the female parent with luteinizing hormone-releasing hormone A27-9 μg / kg and dioxinone 5-7 mg / kg. After the second injection, the water temperature is maintained at 20.5-21.5°C. Over the next 5-7 hours, the water temperature is lowered by 0.4-0.6°C every hour from 20.5-21.5°C to 17.5-18.5°C, and then maintained at 17.5-18.5°C continuously to stimulate ovarian development in 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 + dioxin 2.5-3.5 mg / kg; S3. Artificial insemination: 6-7 hours after the female parent receives the second injection of oxytocin, gently press the female's abdomen to squeeze out the mature eggs. Then, squeeze the male's semen onto the eggs. Add 18-19°C water to activate the sperm. Stir for 110-130 seconds, then wash away the mucus with 18-19°C water to obtain the fertilized eggs. S4. Fertilized egg incubation: The fertilized eggs treated in S3 were incubated in water at 18-21°C, 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 larvae were obtained and cultured; S4. 23-25 ​​hours after hatching, transfer the fry to a fry culture tank and feed them with fry feed. After hatching, transfer the fry to an outdoor cement pool for culture 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 a hatching ring with a water depth of 58 to 62 cm and a density of 48,000 to 52,000 eggs per cubic meter. 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 0.8-1.2m, the depth is 0.7-0.9m, and the density of fry in the open seedling cultivation tank is 3800-4200 fish / 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 broodstock are caught in the evening by placing ground traps in the shallows with stone bottom in the middle and lower reaches of the Lancang River.

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 garfish are added to each bag, 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 for temporary holding.

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