CN111956783A - Artificial spawning induction mixture for micropterus salmoides and artificial breeding method - Google Patents
Artificial spawning induction mixture for micropterus salmoides and artificial breeding method Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
- A61K38/24—Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g. HCG; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
- A01K61/17—Hatching, e.g. incubators
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
- A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
- A61K31/454—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. pimozide, domperidone
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/08—Peptides having 5 to 11 amino acids
- A61K38/09—Luteinising hormone-releasing hormone [LHRH], i.e. Gonadotropin-releasing hormone [GnRH]; Related peptides
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
- A61P15/08—Drugs for genital or sexual disorders; Contraceptives for gonadal disorders or for enhancing fertility, e.g. inducers of ovulation or of spermatogenesis
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
Abstract
The invention discloses an artificial spawning induction mixture for micropterus salmoides and an artificial breeding method thereof, belonging to the technical field of aquatic animal breeding. The invention provides an artificial oxytocic mixture, which comprises: chorionic gonadotropin, luteinizing hormone releasing hormone analog No. 2, diutanone maleate. By adopting the artificial spawning induction mixture to carry out artificial breeding on the micropterus salmoides, the bred female fish has high brood amount, high ovum maturity and obviously improved fertilization rate and hatching rate, the breeding efficiency of the micropterus salmoides is effectively improved, and the time is saved.
Description
Technical Field
The invention relates to the technical field of aquatic animal breeding and reproduction, in particular to an artificial spawning induction mixing agent for parent micropterus salmoides and an artificial breeding method.
Background
The edible weever is beneficial to tonifying liver and kidney, benefiting spleen and stomach, reducing phlegm and relieving cough, and is also beneficial to wound healing and promoting the brain development of children. In recent years, with the gradual improvement of the quality requirements of consumers on aquatic products, weever has replaced four major domestic fishes, crucian carps, snakeheads and the like, becomes an important consumer product on dining tables of people, and the consumer market is continuously expanded. Because the parent fish and the offspring seed supply of the weever can not meet the market demand far, the price of the weever is always in the rising trend, and the weever has extremely high market value.
The weever is very wide in culture water area, can grow well in common fresh water ponds, fresh water factories, net cages and inland saline-alkali water areas, and has the advantages of excellent environmental adaptability, strong disease resistance, wide feed source and the like, so that the weever becomes an excellent artificial freshwater culture variety in China. The output of the prior fresh water cultured weever in Guangdong, Zhejiang and Jiangsu places is also considerable. According to data of 'Chinese fishery statistics yearbook 2019', the freshwater aquaculture yield of the weever in Guangdong province in 2018 reaches 25.84 ten thousand tons, and the weever is the first national place. While these favorable results are achieved, we notice that female weever generally matures in 1 year and belongs to fish with one pregnant egg and multiple ovulations. The amount of eggs in one pregnancy can reach 4-12 ten thousand grains, and the average amount of eggs in pregnancy is 6 ten thousand grains. Mature eggs can be discharged for 1-3 ten thousand times in each ovulation period, and can be discharged for 2-3 times in each reproductive cycle. The brooding amount, the egg discharging amount and the ovulation frequency of the parent fish are closely related to the nutrition, the peripheral environment and the management. In each spawning cycle of parent fish, 30-50% of ova may be locked or over-mature due to insufficient nutrition supply or failure of timely ovulation, thereby causing the problems of reduced quality of ova, low rate of fertilized ova and hatchability of ova, poor survival rate of offspring seeds and the like. Therefore, how to develop an artificial breeding method which can improve the single egg discharge amount of the micropterus salmoides, increase the fertilization rate and the hatching rate of the ova and improve the survival rate of the offspring seeds becomes a technical problem which needs to be solved urgently in the field.
Disclosure of Invention
In order to solve the problems in the prior art, the invention aims to provide an artificial spawning induction mixture which can obviously improve the egg discharging amount and the egg quality of micropterus salmoides and avoid the occurrence of egg occlusion.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides an artificial spawning induction mixture for micropterus salmoides, which comprises the following components: 1200-2000 IU/mL of chorionic gonadotropin, 150-220 mu g/mL of luteinizing hormone releasing hormone analogue No. 2 and 10-25 mg/mL of diutanone maleate.
Preferably, the chorionic gonadotropin in the artificial oxytocin mixture is 1500IU/mL, the luteinizing hormone releasing hormone analogue No. 2 is 200 mug/mL, and the diutanone maleate is 10 mg/mL.
The invention also aims to provide the application of the oxytocic mixture in artificial breeding of micropterus salmoides.
The invention also aims to provide an artificial breeding method of micropterus salmoides.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides an artificial breeding method of micropterus salmoides, which comprises the following steps:
(1) artificial hastening parturition: selecting parent fish, wherein the afternoon is 15: 00-16: injecting 00 female fish with the oxytocic mixture of claim 1 or 2; placing the female fish injected with the mixture and the male fish not induced to spawn in the same water area for temporary rearing, and separating the female fish and the male fish by using a net cage; the next morning 6: 00-7: 00 is male fish injected with the oxytocic mixture of claim 1 or 2; after the male fish hastening parturition for 3-6 hours, dismantling the net cage to ensure that the female fish is fully contacted with the male fish;
(2) collecting the ovum and sperm of parent fish, artificial fertilization, fertilized egg incubation and larva cultivation;
(3) and (3) continuously culturing the parent fish after ovulation for 8-12 d, then hastening parturition according to the step (1), and performing ovulation and fertilization for the second time.
Preferably, the injection amount of the oxytocic mixture in the step (1) is as follows: 0.8-1.2 mL of oxytocic mixture is injected into every 500g of female fish, and the injection amount of male fish is half of that of the current female fish.
Preferably, the temperature of the cultivation water before breeding of the parent fish is 22-24 ℃, and the water flow speed is 10-20L/min.
Preferably, the parent fish is fed with the feed twice a day during the breeding period, the protein level in the feed is 44-46%, and the grease level in the feed is 8-10%.
Preferably, every 2 days, 60-100 mg of the mixed solution of the vitamin E and the vitamin A is sprayed on each kilogram of the feed for feeding for 1 time.
Preferably, the mixture is prepared from vitamin E: the vitamin A is prepared according to the weight ratio of 1: 2.
Preferably, the incubation conditions in step (2) include: the density of the fertilized eggs is 1.5-2.5 ten thousand per liter, the water flow rate is 2-4L/min, and the water temperature is 22-24 ℃.
Preferably, after the weever is incubated to grow the membrane for 6-7 days, larva cultivation is carried out, wherein the larva cultivation conditions in the step (2) are as follows: the seedling density is 8000-10000 tails/m3The water flow rate is 5-8L/min, the water temperature is 22-24 ℃, and 60-80 mL/m of water is added into the water every 3 days3The EM bacterium of (1).
Compared with the prior art, the technical scheme of the invention has the following beneficial effects:
(1) according to the invention, by improving parent fish breeding and selection, the breeding efficiency of parent fish is effectively improved, the maturity of the selected parent fish is high, the brood amount of female fish is high, and the average brood amount is more than 8 ten thousand grains;
(2) the oxytocic and the oxytocic technology provided by the invention can effectively promote the gonad development of female fish and male fish, and improve the ovulation amount and the ovulation efficiency of the female fish, so that the egg discharging amount of the female fish is 4-6 ten thousand eggs each time, the ovulation amount can reach more than 90% of the total pregnant egg amount within 2 ovulation time, and the occurrence probability of egg locking and overmaturity is reduced;
(3) the invention provides the oxytocic and the oxytocic technology, which can effectively improve the quality of the ovum, the fertilization rate of the ovum is higher than 85%, the hatching rate is higher than 90%, and the survival rate of the offspring seeds is higher than 90%.
Detailed Description
The invention provides an artificial spawning induction mixture for micropterus salmoides, which comprises the following components: 1200-2000 IU/mL of chorionic gonadotropin, 150-220 mu g/mL of luteinizing hormone releasing hormone analogue No. 2 and 10-25 mg/mL of diutanone maleate. The invention obtains the oxytocic mixture by compounding the oxytocic substances, and can obviously improve the egg discharging amount of the largemouth bass and avoid the phenomena of ovum locking and overmaturity after being injected into the parent fish of the largemouth bass. The proportion is preferably 1500IU/mL of chorionic gonadotropin, 200 mu g/mL of luteinizing hormone releasing hormone analogue No. 2 and 10mg/mL of diutanone maleate. In the invention, chorionic gonadotropin, luteinizing hormone releasing hormone analogue No. 2 and diutanone maleate can be all commercially available products. In a specific embodiment of the invention, chorionic gonadotropin, luteinizing hormone releasing hormone analog No. 2, diutanone maleate were purchased from Ningbo second hormone plant.
The invention also provides the application of the induced spawning mixture in artificial breeding of micropterus salmoides.
The invention also provides an artificial breeding method of micropterus salmoides, which comprises the following steps:
(1) artificial hastening parturition: selecting parent fish, wherein the afternoon is 15: 00-16: injecting 00 female fish with the oxytocic mixture of claim 1 or 2; placing the female fish injected with the mixture and the male fish not induced to spawn in the same water area for temporary rearing, and separating the female fish and the male fish by using a net cage; the next morning 6: 00-7: 00 is male fish injected with the oxytocic mixture of claim 1 or 2; after the male fish hastening parturition for 3-6 hours, dismantling the net cage to ensure that the female fish is fully contacted with the male fish;
(2) collecting the ovum and sperm of parent fish, artificial fertilization, fertilized egg incubation and larva cultivation;
(3) and (3) continuously culturing the parent fish after ovulation for 8-12 d, then hastening parturition according to the step (1), and performing ovulation and fertilization for the second time.
In the invention, the male and female parent fishes with normal body shape and color, strong body constitution and no disease and injury are selected for parent fish culture 1 month before parent fish breeding. As a preferable embodiment, the parent fish is placed in running water with the water temperature of 22-24 ℃ and the water flow speed of 10-20L/min for cultivation, the water temperature and the water flow speed are adjusted, the effect of stimulating the gonad development of the parent fish is achieved, and the reproductive capacity of the parent fish is improved. Wherein the water temperature is more preferably 23 ℃ and the water flow rate is more preferably 15L/min. The invention can adopt the conventional cultivation method for other environmental conditions of parent fish cultivation, such as illumination, ventilation and the like.
The invention feeds the parent fish every day, preferably 2 times a day during the breeding period. The protein level of the feed is preferably 44-46%, the oil level is preferably 8-10%, the gonad development level of the parent fish is promoted, and the egg laying capacity and the ovum level are improved. The content level of ash and crude fiber in the feed is not limited, and the feed can contain the ash and crude fiber in a conventional content. The feed of the invention further preferably has a protein level of 45% and a fat level of 9%. The feed of the invention can be prepared from fish oil: soybean oil: 50-60% of coconut oil: 20-30%: 20-30% of the total weight of the composition. The invention provides enough nutrition for the parent fish through reasonable proportioning of the nutrition level of the feed, and has the effects of promoting gonad development, improving ovum quality and promoting ovulation of the parent fish compared with the conventional special feed for the weever. The fish oil, the soybean oil and the coconut oil in the invention can be prepared from the conventional commercial products in the field.
In the period of feeding the parent fish with the feed, 60-100 mg of the mixed solution of the vitamin E and the vitamin A is sprayed on each kilogram of the feed every 2 days to feed the parent fish for 1 time, and more preferably 80mg of the mixed solution of the vitamin E and the vitamin A is sprayed on each kilogram of the feed. The vitamin mixed solution is prepared from vitamin E: the vitamin A is prepared according to the weight ratio of 1: 2. According to the invention, by adding the vitamin mixed liquor with a specific proportion, the quality of the ovum is obviously improved, and further the fertility rate, the hatching rate and the survival rate of the offspring seeds in the later period are improved. The vitamin E and the vitamin A in the invention are prepared from products which are commercially available and are conventional in the field.
The invention selects the pre-breeding female fish with thick and short body shape, expanded and soft abdomen, obvious ovary outline, uniform size of upper and lower abdomen, elasticity, slightly convex urogenital papilla, ruddy spawning period and 2 holes on the surface; selecting male fish which is thin and long in body shape, sunken and ruddy in urogenital papilla, has only 1 hole, has a small amount of milky semen flowing out of the abdomen and is scattered in water by slight pressure to artificially hasten parturition. The parent fish selection process is carried out by selecting parent fish with mature gonad according to the conventional method in the field.
In the artificial induced spawning stage, the induced spawning mixture is respectively injected into parent fishes, the injection positions are the inner sides of ventral fins of the fishes, needles are obliquely inserted, and the depth of the inserted needles is 1-1.2 cm, preferably 1.1 cm; the injection amount of the oxytocic mixture is as follows: 0.8-1.2 mL, more preferably 0.9-1 mL, is injected into every 500g female fish, and the injection amount of the male fish is half of that of the female fish.
The invention is implemented in the afternoon of 15: 00-16: and (5) injecting the oxytocic mixture into the female fish at 00 hours. Since the weever usually spawns in the evening when the light is weak, the single injection effect time of the spawning induction mixture is 24-25 h, so that the weever is selected from 15: 00-16: when the injection is injected at 00 hours, the female fish can be more favorably ovulated smoothly compared with other times, the single egg discharging amount can be further improved, and the quality of eggs cannot be influenced. The injection time of the male fish is 15-17 hours after the female fish is injected, and the injection time of the male fish is found to be 6: 00-7: 00 hours, the sperms of the male fish and the ova of the female fish can be ensured to be developed and matured synchronously, and the fertilization rate can be effectively improved in the artificial insemination process.
The female fish injected with oxytocin and the male fish not induced spawning are placed in the same water area for temporary rearing and separated by a net cage, and the female fish is temporarily reared in the net cage. The net cages are placed in the water storage tank, so that male fishes and female fishes can live in the same water body and are close to each other without influencing each other, and the male fishes can release certain sexual signals in the water body, so that sexual maturity of the female fishes is facilitated. And (3) dismantling the net cage 3-6 hours after the oxytocin is injected into the male fish, so that the female fish is fully contacted with the male fish, and the preferable time for dismantling the net cage is 4 hours after the oxytocin is injected into the male fish. Compared with a method for temporarily breeding parent fish alone, the method for temporarily breeding parent fish by using the net cages can obviously improve the single egg discharging amount of the female fish. The invention has no special requirements on the material of the net cage, and the mesh size only needs to ensure that parent fish does not pass through.
After the female fish is injected with the oxytocic mixture for 24-26 hours, lightly pressing the abdomen of the female fish, and collecting ova; after injecting the oxytocic mixture into the male fish for 9-11 h, slightly wrapping the gonadal tissue by using a rubber head dropper in a sucking mode, and slightly sucking to take out semen; adding 2-3mL of 0.7% physiological saline into each 10000 of eggs to promote the opening of fertilization holes, adding 0.2-0.4 mL of semen, adding 3-5 mL of water to activate the fertilized eggs, closing the fertilization holes, stirring for 20-30 s, completing the fertilization process, and obtaining the fertilized eggs.
In the fertilized egg hatching process, the invention discovers that the fertilized egg density is adjusted to be 1.5-2.5 ten thousand grains per liter, the water flow speed is controlled to be 2-4L/min, the water temperature is 22-24 ℃, a good water body environment can be provided for the fertilized eggs, the hatching rate of the fertilized eggs is effectively improved, and the weever begins to hatch out of membranes after 70-72 hours. The invention further preferably adopts the water flow rate of 3L/min and the water temperature of 23-24 ℃. Other environmental conditions such as illumination, ventilation and the like in the incubation process of the fertilized eggs can adopt a conventional cultivation method, and the invention is not limited.
According to the method, after the weever is incubated out of the membrane for 6-7 days, the fries are fished out and put into a fry culture pond, and the density of the fries is 8000-10000 fries/m3Preferably 9000 tails/m3The fairy shrimp is used as initial feed; and continuously inflating during the cultivation period, controlling the water temperature to be 22-24 ℃, controlling the water flow speed to be 5-8L/min, changing water 1/3 every 3d, ensuring the temperature difference before and after water changing to be not more than 0.5 ℃, and ensuring the fresh water environment in a natural light period. The invention has no special requirements for the selection of the initial baits and the seedling culture pond, and can also be replaced by the conventional initial baits, culture barrels and the like.
Adding 60-80 mL/m of the culture medium into the seedling culture pond every 3 days3The EM strain (4) is preferably added at 70mL/m3The EM bacterium of (1). The addition of the EM bacteria can better improve the composition of beneficial bacteria in the water body, promote the intestinal development of the largemouth black bass and further improve the survival rate of fries.
According to the method, the ovulated parent fish is continuously cultivated for 8-12 days, preferably 10 days according to the parent fish cultivation method, then the spawning induction process is carried out, the ovaries of the female fish after two ovulations are basically emptied, the egg discharging amount of the two ovulations accounts for more than 90% of the total egg carrying amount of the female fish, the egg maturity is high, the fertility rate and the hatching rate are both remarkably improved, the breeding efficiency of the female fish is effectively improved, and the time is saved.
The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the specific embodiment of the invention, the parent micropterus salmoides is Youwei No. 2, which is purchased from the Zhujiang aquatic research institute of Chinese aquatic science institute, but is not a limitation on the technical scheme of the invention.
Example 1
A set of parent largemouth bass breeding and artificial breeding improvement technology comprises the following steps:
step 1, selecting male and female parent fishes which have normal body shape and body color, strong physique and no disease and injury for 1 month before breeding parent fishes. Putting the selected parent fish into flowing water with the water temperature of 24 ℃ and the water flow speed of 15L/min respectively. During the cultivation period, the feed with protein level of 44% and feed fat level of 10% (fish oil: soybean oil: coconut oil in proportion of 60%: 20%: 20%) was fed for 2 times per day. Every 2 days, 100mg of vitamin E and vitamin A mixed liquor (1: 2) is sprayed on each kilogram of feed to feed parent fish for 1 time, and gonad development and maturation are accelerated. Meanwhile, conventional cultured parent micropterus salmoides are selected as a control group, and special feed for micropterus salmoides produced by Zhejiang Xinxin Tianen aquatic feed limited company is fed, wherein the crude protein content is 45%, the fat content is 10%, the crude fiber content is 5%, and the crude ash content is 16%.
And 2, selecting 10 pre-bred female fish, wherein the female fish has a relatively thick and short body shape, an expanded and soft abdomen, an obvious ovary outline, uniform sizes of the upper and lower abdomens, elasticity, slightly convex urogenital papilla, a ruddy spawning period and 2 holes. The height of the female fish is as follows: the body width is 2: 1. the male fish 4 tails have a relatively thin and long body shape, the urogenital papilla is sunken and ruddy, only one hole is formed, and a small amount of milky semen flows out when the abdomen is lightly pressed and is dispersed in water.
Step 3, the experimental group adopts an artificial induced spawning mode to respectively prepare an induced spawning medicament of chorionic gonadotropin 1500IU/mL + luteinizing hormone releasing hormone analogue No. 2 mixture of 200 mug/mL + diutanone maleate 10 mg/mL. 0.8ml of the above-mentioned oxytocic mixture is injected into 500g of female fish. Injection time was 15 pm: 00, the injection part is the inner side of the ventral fin of the fish body, the needle is obliquely inserted, and the depth of the needle insertion is 1 cm. The male fish is 6: 00 the oxytocic mixture is injected in the same way as female fish, and the injection amount is reduced by half. And (3) adopting natural breeding in a control group, wherein the male-female ratio is 5: 2, 10 tails of female fish and 4 tails of male fish, placing the female fish and the male fish at 8m3The palm net sheet is placed on the bottom edge of the temporary rearing barrel.
Step 4, placing the female fish injected with oxytocin and the male fish not induced to spawn in the same round temporary rearing barrel with the volume of 8m3. The specification of the female fish temporarily cultured in the barrel is 2m3The net cage is isolated from the male fish. And (4) after the male fish is injected with oxytocin, dismantling the net cage to ensure that the female fish is fully contacted with the male fish to promote ovulation.
And step 5, after the female fish is injected for 24 hours, and after the male fish is injected for 9 hours, the effect time is reached. At this time, it can be obviously observed that the male fish surrounds the female fish, the belly and the genital orifice of the female fish are continuously contacted with the mouth, and the tail part of the male fish is also continuously flapped, which is a breeding signal that the male fish expects to be matched with the female fish. Gently squeezing the abdomen of the female fish, and collecting eggs with a stainless steel basin; the male fish adopts a sucking mode, and the sperm is taken out by light sucking after the neutral gland tissue is lightly wrapped by a rubber head dropper. Adding 0.2mL of semen into each 10000 ovum, adding 5mL of water for activation, and stirring for 30s to complete the fertilization process. After the fertilized eggs are debonded by yellow mud, the fertilized eggs are placed in hatching barrels, 20 ten thousand eggs are placed in each hatching barrel, the water flow speed is 3L/min, and the water temperature is 23 ℃. And hatching the fertilized eggs of the control group by adopting a conventional palm net piece in running water, collecting the fries after hatching for 7 days, and counting the number.
And 6, continuously culturing the female fish after ovulation according to the operation of the step 1 for 10 days, and then carrying out the spawning induction process. The control group is further cultivated for 10 days and then cultivated and bred.
TABLE 1 weight of parent fish in experimental group and control group
As can be seen from Table 1, after the parent fish breeding, the average weight of the experimental group 10 female fish is 846.33g, the average weight of the 4-tailed male fish is 682.48g, the average weight of the control group 10 female fish is 841.29g, and the average weight of the 4-tailed male fish is 713.52g, which indicates that the female fish and the male fish in the experimental group have no significant difference in weight (P >0.05) with the control group.
TABLE 2 comparison of the ovipositivity, fertilization rate and hatchability of the female fish between the experimental group and the control group
In the first ovulation period, 9 female fishes in the experimental group have smooth induced spawning, and the first and second egg discharge amount, fertility rate and hatchability of the 9 female fishes are respectively counted. In the control group, 10 female fish successively spawned within 3 days. As can be seen from Table 2, the experimental group showed higher twice egg laying amount, fertilization rate and hatching rate than the control group.
Example 2
A set of parent largemouth bass breeding and artificial breeding improvement technology comprises the following steps:
step 1, selecting male and female parent fishes which have normal body shape and body color, strong physique and no disease and injury for 1 month before breeding parent fishes. Putting the selected parent fish into flowing water with the water temperature of 22 ℃ and the water flow speed of 10L/min respectively. During the cultivation period, feed with protein level of 45% and feed oil level of 9% (fish oil: soybean oil: coconut oil in a ratio of 50%: 20%: 30%) is fed for 2 times every day. Every 2 days, 100mg of vitamin E and vitamin A mixed liquor (1: 2) is sprayed on each kilogram of feed to feed parent fish for 1 time, and gonad development and maturation are accelerated. Meanwhile, the parent micropterus salmoides cultured conventionally are selected as a control group. Meanwhile, conventional cultured parent micropterus salmoides are selected as a control group, and special feed for micropterus salmoides produced by Zhejiang Xinxin Tianen aquatic feed limited company is fed, wherein the crude protein content is 45%, the fat content is 10%, the crude fiber content is 5%, and the crude ash content is 16%.
Step 2, selecting 8 tails of female fish to be bred in advance, wherein the female fish has a relatively thick and short body shape, an expanded and soft abdomen, an obvious ovary outline, uniform upper and lower abdomen sizes, elasticity, slightly convex urogenital papilla, a ruddy spawning period and 2 holes, and is high in body height: the body width is preferably (3: 1). The male fish 4 tails have a relatively thin and long body shape, the urogenital papilla is sunken and ruddy, only one hole is formed, and a small amount of milky semen flows out when the abdomen is lightly pressed and is dispersed in water.
Step 3, respectively preparing oxytocin 1500IU/mL + luteinizing hormone releasing hormone analogue No. 2 200 mug/mL + diutanone maleate 10mg/mL mixture. 0.8mL of the above oxytocic mixture was injected per 500g of female fish. Injection time was 16 pm: 00, the injection part is the inner side of the ventral fin of the fish body, the needle is obliquely inserted, and the depth of the needle insertion is 1.1 cm. The male fish is 7: 00 the oxytocic mixture is injected in the same way as female fish, and the injection amount is reduced by half. And naturally breeding the female fish 8 tails and the male fish 4 tails in the control group.
Step 4, placing the female fish injected with oxytocin and the male fish not induced to spawn in the same round temporary rearing barrel with the volume of 4m3. Temporarily breeding female fish in barrel with specification of 1m3The net cage is isolated from the male fish. And (4) after the male fish is injected with oxytocin, dismantling the net cage to ensure that the female fish is fully contacted with the male fish to promote ovulation. The control group female fish and male fish are placed at 4m3The cultivation is carried out in a cultivation barrel, and a palm net piece is placed on the edge of the bottom of the barrel.
And step 5, after the female fish is injected for 26 hours, and after the male fish is injected for 11 hours, the effect time is reached. At this time, it can be obviously observed that the male fish surrounds the female fish, the belly and the genital orifice of the female fish are continuously contacted with the mouth, and the tail part of the male fish is also continuously flapped, which is a breeding signal that the male fish expects to be matched with the female fish. Gently squeezing the abdomen of the female fish, and collecting eggs with a stainless steel basin; the male fish adopts a sucking mode, and the sperm is taken out by light sucking after the neutral gland tissue is lightly wrapped by a rubber head dropper. Adding 0.2mL of semen into each 10000 ovum, adding 5mL of water for activation, and stirring for about 30 seconds to complete the fertilization process. After the fertilized eggs are debonded by yellow mud, the fertilized eggs are placed in hatching barrels, 20 ten thousand eggs are placed in each hatching barrel, the water flow speed is 3 liters/minute, and the water temperature is 24 ℃. And hatching the fertilized eggs of the control group by adopting a conventional palm net piece in running water, collecting the fries after hatching for 7 days, and counting the number.
And 6, after the bass is incubated for 72h, the fry starts to emerge from the membrane, and is in a flat swimming state after 7 days, and the fry is collected.
TABLE 3 weight of parent fish in experimental group and control group
As can be seen from Table 3, after the parent fish breeding, the average weight of the experimental group 8 female fish is 756.33g, the average weight of the 4-tailed male fish is 693.26g, the average weight of the control group 8 female fish is 765.23g, and the average weight of the 4-tailed male fish is 685.18g, which indicates that the female fish and the male fish in the experimental group have no significant difference in weight (P >0.05) with the control group.
TABLE 4 comparison of total ovulation amount and ovum quality of female fish in experimental group and control group
As can be seen from Table 4, the egg discharging amount of each female fish in the experimental group is 8.7-11.3 ten thousand, and the egg discharging amount of each female fish in the control group is 5.3-7.2 ten thousand, which indicates that the total egg laying amount of each female fish in the experimental group is significantly higher than that of the control group. Meanwhile, the blocking rate of the oocytes in the ova of the experimental group is 9.6-13.8%, and the blocking rate of the oocytes of the control group is 15.3-19.2%, which is obviously higher than that of the experimental group. The fertilization rate and the hatching rate of the two times in the experimental group are respectively 86.3-92.5% and 93.4-94.8%, and the fertilization rate and the hatching rate of the two times in the control group are respectively 67.2-72.8% and 74.1-75.5%, which are both obviously lower than those in the experimental group.
TABLE 5 Total emergence and survival rate of seedlings in experimental and control groups
As can be seen from Table 5, the seedling emergence amount of the seedlings bred according to the improved technology can reach 160 ten thousand, which is far higher than 76 ten thousand in the control group, and the survival rate of the seedlings can reach 90.3-91.2%, which is obviously higher than 82.5-84.5% of the control group. The improvement method is proved to greatly improve the utilization and the seedling breeding efficiency of parent fishes of micropterus salmoides, promote the high-quality seedling supply of micropterus salmoides and accelerate the industrial development.
Example 3
In order to explore the efficacy of the oxytocic mixture prepared by compounding the oxytocic, the following single-factor experiment is carried out:
according to the parent fish breeding mode in the experimental group of the embodiment 1 of the invention, parent fish breeding and selection are carried out, female fish 32 tails and male fish 16 tails with the same specification and the same breeding time are selected, and are randomly divided into four groups, namely an experimental group 1, an experimental group 2, an experimental group 3 and a control group, wherein each group comprises 8 female fishes and 4 male fishes, and artificial induced spawning is carried out on the experimental groups 1-3: the experimental group 1 selects oxytocin as a mixture of chorionic gonadotropin 1500IU/mL, luteinizing hormone releasing hormone analogue No. 2 200 mug/mL and diutanone maleate 10 mg/mL; the oxytocic selected by the experimental group 2 is chorionic gonadotropin 1500 IU/mL; the oxytocin selected in experimental group 3 is the luteinizing hormone releasing hormone analogue No. 2 200. mu.g/mL. The injection modes of the oxytocic or the oxytocic mixture are as follows: 0.8mL of the oxytocic mixture is injected into every 500g of female fish, and the injection time is 16 in the afternoon: 00, the injection part is the inner side of the ventral fin of the fish body, the needle is obliquely inserted, and the depth of the needle insertion is 1.1 cm. Placing the female fish injected with the mixture and the male fish not induced to spawn in the same water area for temporary rearing, and separating the female fish and the male fish by using a net cage; the male fish is 7: 00 the oxytocic mixture is injected in the same way as female fish, and the injection amount is reduced by half. The 3 experiments only differ in the injected oxytocic and the other culture conditions are the same. The parent fish in the control group was not injected with oxytocic or oxytocic, but kept in the same manner as the experimental group. The weights of the female fish in each experimental group were not significantly different (see table 6 for details).
TABLE 6 weight of parent fish in experimental group and control group
The first egg laying amount and the second egg laying amount of four groups of female fishes are counted respectively (see table 7 in detail), and the following results are obtained: the first ovulation amount of the experimental group 1 can reach 3.8-4.2 ten thousand grains, the second ovulation amount can reach 4.2-4.8 ten thousand grains, and the single ovulation amount and the total ovulation amount in the experimental group 1 are obviously higher than those of other experimental groups and control groups.
TABLE 7 amount of ova discharged twice in the experimental group and the control group
Example 4
In order to explore the influence of the feed on the ovulation time and oocyte development of the largemouth black bass during the parent fish breeding period, the following single-factor experiment is carried out:
selecting female fish 32 tails and male fish 16 tails with the same specification and similar development, randomly dividing the female fish and the male fish into four groups, namely an experimental group 1, an experimental group 2, an experimental group 3 and an experimental group 4, respectively, feeding the female fish and the male fish with 8 tails and 4 tails in the period of parent fish cultivation by respectively selecting different feeds: experiment group 1 feeds the feed with protein level 45% and feed fat level 9% (fish oil: soybean oil: coconut oil according to 50%: 20%: 30%) 2 times a day, and sprays vitamin E + vitamin A mixed solution (1: 2) of 80mg on each kilogram of feed to feed the parent fish 1 time every 2 days; the feed in the experimental group 2 is the same as that in the experimental group 1, the feeding frequency is 2 times per day, but every 2 days, 80mg of vitamin E is sprayed on each kilogram of feed to feed parent fish for 1 time; the experimental group 3 fed with the special feed (crude protein content 45%, fat 10%, crude fiber 5%, crude ash 16%) for micropterus salmoides produced by zhejiang euphorian aquatic feed limited company 2 times a day; the feed in experiment group 4 was the same as experiment group 3, and the feeding frequency was 2 times per day, but every 2 days, 80mg of vitamin E + vitamin A mixture (1: 2) was sprayed on each kilogram of feed to feed the parent fish 1 time. The experimental groups 1 to 4 are different in feed only fed during the period of parent fish cultivation, the rest cultivation conditions are the same, and the modes of later artificial induced spawning and the like are the same, and are the artificial breeding mode of the experimental group in the embodiment 1. Statistical analysis of ovulation time and oocyte development status of micropterus salmoides in four experiments (see table 8) revealed that: the first ovulation time of the largemouth bass in the experimental group 1 is 24-28 days and is shorter than that of the experimental group 2-3, the interval time between two ovulation in the experimental group 1 is 6-8 days and is obviously shorter than that of the experimental group 2-4, and the oocyte locking rate in the experimental group 1 is only 9.3-15.3 percent and is lower than that of the experimental group 2-4. Namely, after the feed formula and the feed additive in the experimental group 1 are combined for use, the advanced ovulation of the largemouth bass can be remarkably promoted, the interval period of two times of ovulation is shortened, and the oocyte blocking rate can be remarkably reduced.
TABLE 8 influence of different feed additive groups on ovulation time and oocyte development of Lateolabrax japonicus
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. An artificial spawning induction mixture for micropterus salmoides is characterized by comprising 1200-2000 IU/mL of chorionic gonadotropin, 150-220 [ mu ] g/mL of luteinizing hormone releasing hormone analogue 2 and 10-25 mg/mL of diutanone maleate.
2. The oxytocin mixture according to claim 1, characterized in that the oxytocin mixture comprises chorionic gonadotropin 1500IU/mL, luteinizing hormone releasing hormone analogue No. 2 200 μ g/mL and diutanone maleate 10 mg/mL.
3. The use of the oxytocic mixture as set forth in claim 1 in artificial breeding of micropterus salmoides.
4. An artificial breeding method of micropterus salmoides is characterized by comprising the following steps:
(1) artificial hastening parturition: selecting parent fish, wherein the afternoon is 15: 00-16: injecting 00 female fish with the oxytocic mixture of claim 1 or 2; placing the female fish injected with the mixture and the male fish not induced to spawn in the same water area for temporary rearing, and separating the female fish and the male fish by using a net cage; the next morning 6: 00-7: 00 is male fish injected with the oxytocic mixture of claim 1 or 2; after the male fish hastening parturition for 3-6 hours, dismantling the net cage to ensure that the female fish is fully contacted with the male fish;
(2) collecting the ovum and sperm of parent fish, artificial fertilization, fertilized egg incubation and larva cultivation;
(3) and (3) continuously culturing the parent fish after ovulation for 8-12 d, then hastening parturition according to the step (1), and performing ovulation and fertilization for the second time.
5. The method of claim 4, wherein the oxytocin mixture is injected in an amount of: 0.8-1.2 mL of oxytocic mixture is injected into every 500g of female fish, and the injection amount of male fish is half of that of the current female fish.
6. The method as claimed in claim 4, wherein the temperature of the water for cultivation before breeding of the parent fish is 22-24 ℃ and the water flow rate is 10-20L/min.
7. The method as claimed in claim 4, wherein the parent fish is fed with the feed twice a day during the rearing period, and the protein level in the feed is 44-46% and the fat level is 8-10%.
8. The method according to claim 7, wherein the mixed solution of vitamin E and vitamin A is sprayed to each kilogram of the feed for 1 time every 2 days, wherein the mixed solution is sprayed to each kilogram of the feed for 60-100 mg.
9. The method of claim 8, wherein the mixture is prepared from vitamin E: the vitamin A is prepared according to the weight ratio of 1: 2.
10. The method according to any one of claims 4 to 9, wherein the incubation conditions comprise: the fertilized egg density is 1.5-2.5 ten thousand grains per liter, the water flow rate is 2-4L/min, the water temperature is 22-24 ℃, and the larva cultivation is carried out after the weever is hatched out of the membrane for 6-7 days, wherein the cultivation conditions are as follows: the seedling density is 8000-10000 tails/m3The water flow rate is 5-8L/min, the water temperature is 22-24 ℃, and 60-80 mL/m of water is added into the water every 3 days3The EM bacterium of (1).
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| CN114051964A (en) * | 2021-11-15 | 2022-02-18 | 中国水产科学研究院淡水渔业研究中心 | Perch micropterus salmoides fry screening system and method |
| CN114569701A (en) * | 2022-03-21 | 2022-06-03 | 中国科学院水生生物研究所 | Artificial spawning induction method of finless eel based on gonadotropin releasing hormone analogue of finless eel |
| CN114885865A (en) * | 2022-03-29 | 2022-08-12 | 西双版纳云博水产养殖开发有限公司 | Spawning induction method suitable for female silurus riparia |
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| CN104798698A (en) * | 2014-01-23 | 2015-07-29 | 珠海市斗门区河口渔业研究所 | Artificial sea bass breeding method suitable for river mouth areas |
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| CN103749361A (en) * | 2014-01-13 | 2014-04-30 | 天津师范大学 | Method for incubating largemouth basses ahead of time |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112931312A (en) * | 2021-02-26 | 2021-06-11 | 海南晨海水产有限公司 | Artificial breeding method of seriolala quinqueradiata |
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| CN114569701A (en) * | 2022-03-21 | 2022-06-03 | 中国科学院水生生物研究所 | Artificial spawning induction method of finless eel based on gonadotropin releasing hormone analogue of finless eel |
| CN114569701B (en) * | 2022-03-21 | 2023-05-26 | 中国科学院水生生物研究所 | Artificial induced spawning method of finless eels based on finless eels gonadotrophin releasing hormone analogues |
| CN114885865A (en) * | 2022-03-29 | 2022-08-12 | 西双版纳云博水产养殖开发有限公司 | Spawning induction method suitable for female silurus riparia |
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