CN110876343B - Method for improving artificial breeding efficiency of Leptobotia elongata - Google Patents

Method for improving artificial breeding efficiency of Leptobotia elongata Download PDF

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CN110876343B
CN110876343B CN201910921418.9A CN201910921418A CN110876343B CN 110876343 B CN110876343 B CN 110876343B CN 201910921418 A CN201910921418 A CN 201910921418A CN 110876343 B CN110876343 B CN 110876343B
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fish
parent
female
injection
male
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CN110876343A (en
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陈春娜
黄颖颖
杜军
龙治海
林珏
周波
龚全
李正一
戴绍祥
卢华
徐飞
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Institute Of Aquaculture Sichuan Academy Of Agricultural Sciences
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; 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
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; 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
    • A01K61/13—Prevention or treatment of fish diseases
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; 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
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K61/00—Culture of aquatic animals
    • A01K61/60—Floating cultivation devices, e.g. rafts or floating fish-farms
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K61/00—Culture of aquatic animals
    • A01K61/60—Floating cultivation devices, e.g. rafts or floating fish-farms
    • A01K61/65—Connecting or mooring devices therefor
    • 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

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  • Life Sciences & Earth Sciences (AREA)
  • Environmental Sciences (AREA)
  • Marine Sciences & Fisheries (AREA)
  • Zoology (AREA)
  • Animal Husbandry (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Farming Of Fish And Shellfish (AREA)

Abstract

The invention relates to the technical field of artificial fish breeding, in particular to a method for improving the artificial breeding efficiency of Leptobotia elongata. The method comprises the following specific steps of firstly carefully screening parent fishes, temporarily breeding the parent fishes in a polyvinyl chloride net cage, classifying the parent fishes, selecting different induced spawning needle pitch time and drug metering according to different development degrees, facilitating the protection of the parent fishes and the synchronous maturation of oocytes, reducing the proportion of immature oocytes in the induced spawning oocytes from 10% to 3%, then carrying out induced spawning treatment to uniformly mix sperms and eggs, and finally nursing the parent fishes, so that the injury of the spawning parent fishes and the death of the postpartum parent fishes are reduced to the maximum extent, limited loach resources are protected, the mortality rate of the postpartum parent fishes in the traditional method is 70% -100%, and the mortality rate under the technical condition is 10%.

Description

Method for improving artificial breeding efficiency of Leptobotia elongata
Technical Field
The invention relates to the technical field of artificial fish breeding, in particular to a method for improving the artificial breeding efficiency of Leptobotia elongata.
Background
Leptobotia elongata (Bleeker) belongs to the family of Misgurnidae, the genus Bosgurni, the largest fish of the family of Misgurnidae, is distributed in the middle and upstream and in tributaries of Yangtze river in China, is a unique species in China, and occupies an extremely important position in the whole ecological system of the living water area of the species. The loach has higher economic value, high protein content, rich nutritional value and quite reasonable amino acid variety quantity and proportion; meanwhile, the long and thin loaches also have a place in the field of ornamental fishes, and have won a gold prize in the national ornamental fish evaluation of Singapore in 1989. Due to the mutation of the environment, the population quantity and the distribution area of the wild loaches are obviously reduced, and the wild loaches are listed in Chinese endangered animal red leather books. Liu Jun carries out quantitative analysis on the threat degree of the special fishes in Yangtze river, and considers that the Liu Jun carries out three-level urgent protection on the rare species of Changyuan loaches. In recent years, with the increasing emphasis of the country on environmental protection and ecological restoration work, the proliferation and releasing tasks of the long and thin loaches are also developed continuously, the long and thin loach fries are obtained by breeding wild parent fishes caught in rivers at present, the technical requirement of artificial breeding is high, and in artificial domestication and breeding, the fact that the male and female parent fishes of the long and thin loaches are not synchronous in development, male fish sperm is difficult to obtain, the amount of the sperm is small, the quality is low, and therefore when artificial insemination is conducted, the fertilization rate of fish eggs is low, meanwhile, the stress resistance of the long and thin loaches is poor, the parent fishes are extremely easy to die in the artificial breeding process, and the continuous fry obtaining is not facilitated.
Therefore, the artificial breeding efficiency of the long and thin loaches is improved, the continuous obtaining of batch seedlings is facilitated, and the method has great significance for germplasm resource protection of the long and thin loaches.
Disclosure of Invention
Aiming at the problems, the invention provides a method for improving the artificial propagation efficiency of the loaches to overcome the defects in the prior art. The stress reaction of parent fish of the loach is favorably slowed down, the synchronous maturation rate of spermocyte is improved, the movement time of sperms is prolonged, and the fertilization rate and the survival rate of the parent fish are greatly improved.
The technical scheme of the invention is as follows:
a method for improving the artificial propagation efficiency of Leptobotia elongata comprises the following specific steps:
a: temporarily culturing parent fishes: respectively putting the selected parent fish male and female individuals into a polyvinyl chloride net cage for temporary culture;
b: classifying parent fishes: slightly extruding along the opening direction of the parent fish cloaca, and classifying the male and female parent fishes according to the result;
female fish:
the first type: the discharged egg particles are round, even and white or milk white;
the second type: the sizes and the shapes of the discharged eggs are different and are yellowish;
in the third category: no egg is discharged from the cloaca;
male fish:
the first type: sperm microscopic examination of the sperm has motile sperm;
the second type: semen microscopic examination of no or few motile sperm;
in the third category: no semen flows out of the cloaca;
c: parent fish injection: injecting according to the classification result;
the parent fish injection method of the first type of examination result comprises the following steps: the female parent fish is injected by a two-needle method, the needle distance between the two needles is 8 hours, the male fish is injected by one time, the injection time is 4 hours before the induced spawning time, and the injection dosage is half of the second needle of the female fish;
the parent fish injection method of the second type of examination result comprises the following steps: injecting the female parent fish by a two-needle method, wherein the needle distance between the two needles is 12h, injecting the male fish by one time, wherein the injection time is performed simultaneously with the second needle of the female fish, and the injection dosage is half of that of the second needle of the female fish;
in order to protect the parent fish, the male and female parent fish with the third type of inspection result does not induce spawning;
d: spawning induction of parent fish: respectively placing the male and female fishes injected with the oxytocic needle in two polyvinyl chloride net cages, and hastening parturition after corresponding oxytocic time is reached;
fishing out the female fish, extruding the belly of the female fish to extrude mature eggs into a dry container;
fishing out the male fish, extruding the belly of the male fish, enabling semen to flow onto a cover glass sheet, putting the semen and the cover glass sheet into a container for accommodating eggs, adding a sperm activating solution, enabling the sperm and the eggs to be well submerged, slightly shaking to enable the sperm and the eggs to be uniformly mixed, and then moving the mixture to an incubator;
e: and (3) parent fish nursing: and (3) putting the parent fish after spawning induction back to the polyvinyl chloride net cage, putting the net cage close to the water inlet, keeping the water flow, feeding fresh water earthworms every day, controlling illumination, and transferring to a parent fish pond for cultivation after 10-15 days.
The selected parent fish is good in maturity, female fish requires that an individual is complete and has no wound, the abdomen is soft and slightly swollen, and the minimum individual is more than 200 g; selecting the individual with intact body surface, no wound and vitality.
The ratio of the amount of the parent fish to the amount of the parent fish is 1: 2-1: 3.
The polyvinyl chloride net cage adopted in the step A has the illumination intensity of 50-100 lx, the net cage needs to be placed near a water inlet, and the water flow is controlled to be 0.1m3And a/min period.
The parent fish injection method of the first type of inspection result specifically comprises the following steps: the female parent fish is injected by two-needle method, the first injection dosage is 0.5mgPG +0.5ug LHRH-A per kilogram of fish body2+1mlH2O, the second injection amount is 1mgPG +8ug LHRH-A per kilogram of fish body2+1mlH2O, when the water temperature is 23 +/-1 ℃, the needle distance between the two needles is 8 hours, and the spawning induction time is 6 hours after the second needle injection; the male fish adopts one-time injection, the injection time is 4 hours before the induced spawning time, and the injection dosage is half of that of the second needle of the female fish.
Parent fish injection method for second type inspection resultThe method comprises the following steps: the female parent fish is injected by two-needle method, the first injection dosage is 1mgPG +1ug LHRH-A per kilogram of fish body2+1mlH2O, the second injection amount is 1mgPG +10ug LHRH-A per kilogram of fish body2+1mlH2O, when the water temperature is 23 +/-1 ℃, the needle distance between the two needles is 12 hours, and the spawning induction time is 7 hours after the second needle injection; the male fish adopts one-time injection, the injection time and the second needle of the female fish are carried out simultaneously, and the injection dosage is half of that of the second needle of the female fish.
The polyvinyl chloride net cage in the step D is placed near the water inlet, and the water flow is controlled to be 0.3m3And/min. The sperm activating solution comprises the following raw materials in parts by weight: 140 parts of sodium chloride, 26 parts of potassium chloride, 43.3 parts of calcium chloride, 11.3 parts of magnesium chloride, and 0.05M Tris-HCl for adjusting the pH to 7.9, wherein the solvent for preparing the activation solution is distilled water.
The water flow in the step E is controlled to be 0.1m3And/min, feeding fresh and live tubificidae accounting for 15 percent of the weight of the parent fish into the net cage every day, and controlling the illumination intensity in the net cage to be 50 lx.
The main advantages of the invention are: 1. according to different development degrees of parent fish, different induced spawning needle distance time and drug metering are selected, which is beneficial to the protection of parent fish and the synchronous maturation of oocytes, and the proportion of immature oocytes in induced spawning oocytes is reduced from 10% to 3%.
2. The semen of Misgurni Anguillicaudati contains large amount of Na+、K+、Ca2+、Mg2+Plasma, which is the main component that contributes to the osmotic pressure of the seminal plasma. Ions contained in the sperm activating liquid play a certain role in the sperm motility of the leptotia longissima, the energy loss caused by osmotic pressure difference can be reduced, the sperm motility can be improved, the sperm activation ratio can be improved, the quick movement time of the sperm is improved from 14s to 23s, the sperm activation ratio is improved from 72% to 94%, the combination chance of the sperm and the eggs is greatly increased, and the fertilization rate is greatly improved;
under the combined action of the 2 influences, the artificial insemination rate is increased from 50% to more than 90%.
3. The injury of the spawning parent fish and the death of the postpartum parent fish are reduced to the maximum extent, the limited loach resources are protected, the mortality rate of the postpartum parent fish in the traditional method is up to 70-100%, and the mortality rate under the technical condition is 10%.
Detailed Description
The present invention is described in detail below by way of examples.
Example 1
1. Parent fish selection and temporary rearing
The parent fish is good in maturity, female fish requires that an individual is complete and has no wound, the abdomen is soft and slightly swollen, and the minimum individual is more than 200 g; the male fish is selected from individuals with complete body surface, no wound and good vitality. The ratio of male to female is 1: 2-1: 3. Temporarily breeding male and female individuals in a polyvinyl chloride net cage, wherein the specification of the net cage is 100cm multiplied by 80cm multiplied by 30cm, a piece of foam can be placed on the net cage to prevent the foam from escaping, the illumination intensity in the net cage is controlled to be 50-100 lx, the net cage is placed near a water inlet, and the water flow is controlled to be 0.1m3And a/min period.
2. Drug and duration of effect
Checking and classifying parent fish
Slightly extruding along the opening direction of the cloaca of the parent fish, and classifying the male and female parent fish into the following categories according to results:
female fish:
the first type: a small amount of eggs are discharged from the cloaca, and the eggs are round, smooth, uniform and white or milk white;
the second type: a small amount of eggs are discharged from the cloaca, and the eggs are different in size and shape and are yellowish;
in the third category: no eggs are discharged from the cloaca.
Male fish:
the first type: white semen flows out of the cloaca and is scattered when meeting water, and motile semen is detected by microscopic examination;
the second type: white semen flows out of the cloaca, is not scattered when meeting water, and has no or little motile sperm under microscopic examination;
in the third category: no semen flows out of the cloaca.
② injection
The parent fish injection method of the first type of examination result comprises the following steps: the female parent fish is injected by two-needle method, the first injection dosage is 0.5mgPG +0.5ug LH per kilogram of fish bodyRH-A2+1mlH2O, the second injection amount is 1mgPG +8ug LHRH-A per kilogram of fish body2+1mlH2O, when the water temperature is 23 +/-1 ℃, the needle distance between the two needles is 8 hours, and the spawning induction time is 6 hours after the second needle injection; the male fish adopts one-time injection, the injection time is 4 hours before the induced spawning time, and the injection dosage is half of that of the second needle of the female fish.
The parent fish injection method of the second type of examination result comprises the following steps: the female parent fish is injected by two-needle method, the first injection dosage is 1mgPG +1ug LHRH-A per kilogram of fish body2+1mlH2O, the second injection amount is 1mgPG +10ug LHRH-A per kilogram of fish body2+1mlH2O, when the water temperature is 23 +/-1 ℃, the needle distance between the two needles is 12 hours, and the spawning induction time is 7 hours after the second needle injection; the male fish adopts one-time injection, the injection time and the second needle of the female fish are carried out simultaneously, and the injection dosage is half of that of the second needle of the female fish.
In order to protect the parent fish, the male and female parent fish with the third kind of examination result is not induced to spawn.
The oxytocic is produced by Ningbo second hormone factory, and the selected specifications are as follows: LHRH-A2(10X 25 microgram), PG is carp pituitary gland.
3. Hasten parturition
Placing the male and female fishes injected with oxytocic needle in two polyvinyl chloride net cages respectively, placing the net cages near the water inlet, and controlling water flow at 0.3m3And a/min period. And manually hastening parturition after the parturition waiting time reaches the corresponding parturition hastening time.
Taking out the female fish, squeezing the gray eggs on the belly to flow out of the cloaca smoothly, squeezing the eggs, lightly putting the female fish into a parent fish fixer (patent No. ZL201720106816.1), and squeezing the mature eggs into a dry container.
Taking out the male fish, lightly putting the male fish into a parent fish fixing device, lightly putting a cover glass at a cloaca, extruding the abdomen of the male fish, enabling semen to flow onto the cover glass from the cloaca, putting the semen and the cover glass into a container for accommodating eggs, adding a sperm activating solution prepared in advance, wherein the dosage is suitable for just submerging egg grains, lightly shaking the container for 10s to uniformly mix the sperm and the eggs, standing for 1min, and finally moving the container into an incubator for incubation.
The sperm activating solution (taking 1L of activating solution as an example) used by the invention comprises the following raw materials: 140mg of sodium chloride, 26mg of potassium chloride, 43.3mg of calcium chloride, 11.3mg of magnesium chloride and 0.05M of Tris-HCl for adjusting the pH to 7.9, wherein the solvent for preparing the activating solution is distilled water, and the preparation method comprises the following steps:
solute mass calculation formula: mass of solute is solution mass concentration x volume of activating solution x solute molecular weight divided by 1000;
preparation step (taking 1L of activating solution as an example):
calculating the mass of sodium chloride required by the prepared activated liquid product according to a solute mass calculation formula, weighing 140mg of analytically pure sodium chloride with the mass, and putting the analytically pure sodium chloride into a beaker;
secondly, calculating the mass of potassium chloride required by the volume of the prepared activation liquid according to a solute mass calculation formula, weighing 26mg of analytically pure potassium chloride with the mass and the weighed sodium chloride, and putting the potassium chloride and the weighed sodium chloride into the same beaker;
thirdly, calculating the mass of calcium chloride required by the volume of the prepared activation liquid according to a solute mass calculation formula, weighing 43.3mg of analytically pure calcium chloride of the mass, and putting the calcium chloride into the same beaker;
fourthly, calculating the mass of the magnesium chloride required by the volume of the prepared activation liquid according to a solute mass calculation formula, weighing 11.3mg of analytically pure magnesium chloride with the mass, and putting the analytically pure magnesium chloride into the same beaker;
calculating the mass of Tris (hydroxymethyl) aminomethane (Tris-base) required by the volume of the prepared activation liquid according to a solute mass calculation formula, weighing 6.056g of analytically pure Tris-base with the mass, and putting the weighed Tris-base into the same beaker;
sixthly, adding distilled water with half volume of the activating solution into the beaker, stirring to fully dissolve the substances in the beaker, adding 320ml of 0.1M hydrochloric acid into the dissolved solution, mixing, transferring into a 1L volumetric flask, washing the beaker twice to three times by using a small amount of distilled water, pouring the washed solution into the volumetric flask together, finally using distilled water to fix the volume to the scale mark, and shaking uniformly.
Preparing 0.1M hydrochloric acid: a volumetric flask with 8.36mL of hydrochloric acid added with water to reach 1000mL is used.
4. Postpartum parent fish nursing
Putting the parent fish back into the PVC net cage after artificial spawning induction, putting the net cage near the water inlet, keeping a certain water flow through the net cage, and controlling the water flow to be 0.1m3And/min. Fresh water earthworms which account for 15 percent of the weight of the parent fish are fed into the net cage every day, the illumination intensity in the net cage is controlled to be about 50lx, the parent fish is not disturbed to recover as much as possible except routine inspection every day, and the net cage is transferred into a parent fish pond for cultivation after 10 days.
Has the advantages that:
the method is characterized in that under the condition that the culture environment of parent loach is changed, the stress phenomenon of the parent fish is serious, and after the injection of an induced spawning needle, the parent fish can continuously swim along the wall of a pool, and the abdomen and the snout end can be scratched in a large area, so that the death rate of the parent fish after delivery is high. According to the invention, the male and female parent fishes are respectively placed in different polyvinyl chloride net cages to shield light and shade, so that the stress reaction of the parent fishes is favorably relieved, the parent fishes are prevented from being scratched, and the efficiency of artificial propagation is improved.
Secondly, in the prior art, the two-needle injection has large dosage, short needle distance interval time and unsatisfactory oxytocic effect, and the egg granules contain a large amount of immature egg granules. According to different growth degrees of the parent fish, the injection dosage is properly reduced, the interval time between two injection needles is prolonged, the drug effect of the first needle of the ripening needle can be fully exerted, the stress response of the parent fish to the oxytocic is reduced, the synchronous maturation of oocytes is facilitated, the proportion of immature oocytes is greatly reduced, and the insemination rate is effectively improved.
And thirdly, the number of sexually mature male loaches is small, the semen quantity is often small and the quality is uneven during artificial insemination, in the prior art, clear water is usually adopted to wash the semen near the reproductive hole of the male fish into a container for accommodating eggs to complete the insemination process, and the eggs are easy to absorb water and swell in advance before insemination, so that the insemination effect is influenced. The invention adopts the cover glass to collect the semen from the cloacae, the semen and the cover glass are put into the container for holding the ovum, and the special sperm activating solution is added, thereby effectively avoiding the problem of low fertilization rate caused by small amount of the semen and poor vitality.
Microscopic examination finds that the sperm of the long and thin loach has low activity and short rapid movement time which is only 14s and is obviously lower than that of other fishes, the sperm and the eggs have a certain distance before being combined in the artificial insemination process, and the sperm needs to move the distance as soon as possible after being discharged into a water body. Thus, the more viable sperm cells, the higher the fertilization rate in artificial reproduction. The ionic component added into the activating solution can reduce energy loss caused by osmotic pressure difference, play a role in prolonging the quick movement time of the sperms and obviously improve the activity and the activation ratio of the sperms.
The above description is only for the specific embodiments of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention, and the present invention shall be covered thereby. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.

Claims (3)

1. The method for improving the artificial propagation efficiency of the long and thin loaches is characterized by comprising the following specific steps of:
a: temporarily culturing parent fishes: respectively putting the selected parent fish male and female individuals into a polyvinyl chloride net cage for temporary culture; the polyvinyl chloride net cage is characterized in that the illumination intensity in the net cage is 50-100 lx, the net cage needs to be placed near a water inlet, and the water flow rate is controlled to be about 0.1m for carrying out double-row cultivation/min;
b: classifying parent fishes: slightly extruding along the opening direction of the parent fish cloaca, and classifying the male and female parent fishes according to the result;
female fish:
the first type: the discharged egg particles are round, even and white or milk white;
the second type: the sizes and the shapes of the discharged eggs are different and are yellowish;
in the third category: no egg is discharged from the cloaca;
male fish:
the first type: sperm microscopic examination of the sperm has motile sperm;
the second type: semen microscopic examination of no or few motile sperm;
in the third category: no semen flows out of the cloaca;
c: parent fish injection: injecting according to the classification result;
the parent fish injection method of the first kind of inspection results of the female fish and the male fish specifically comprises the following steps: the female parent fish is injected by two-needle method, the first injection dosage is 0.5mgPG +0.5ug LHRH-A per kilogram of fish body2+1mlH2O, the second injection amount is 1mgPG +8ug LHRH-A per kilogram of fish body2+1mlH2O, when the water temperature is 23 +/-1 ℃, the needle distance between the two needles is 8 hours, and the spawning induction time is 6 hours after the second needle injection; the male fish is injected once, the injection time is 4 hours before the induced spawning time, and the injection dosage is half of that of the female fish through the second needle;
the parent fish injection method of the second-class inspection result of the female fish and the male fish specifically comprises the following steps: the female parent fish is injected by two-needle method, the first injection dosage is 1mgPG +1ug LHRH-A per kilogram of fish body2+1mlH2O, the second injection amount is 1mgPG +10ug LHRH-A per kilogram of fish body2+1mlH2O, when the water temperature is 23 +/-1 ℃, the needle distance between the two needles is 12 hours, and the spawning induction time is 7 hours after the second needle injection; the male fish adopts one-time injection, the injection time and the second needle of the female fish are carried out simultaneously, and the injection dosage is half of that of the second needle of the female fish;
in order to protect the parent fish, the male and female parent fish with the third type of inspection result does not induce spawning;
d: spawning induction of parent fish: respectively placing the male and female fishes injected with the oxytocic needle in two polyvinyl chloride net cages, and hastening parturition after corresponding oxytocic time is reached; placing the polyvinyl chloride net cage near the water inlet, and controlling the water flow rate to be 0.3 m/min;
fishing out the female fish, extruding the belly of the female fish to extrude mature eggs into a dry container;
fishing out the male fish, extruding the belly of the male fish, enabling semen to flow onto a cover glass sheet, putting the semen and the cover glass sheet into a container for accommodating eggs, adding a sperm activating solution, enabling the sperm and the eggs to be well submerged, slightly shaking to enable the sperm and the eggs to be uniformly mixed, and then moving the mixture to an incubator; the sperm activating solution comprises the following raw materials in parts by weight: 140 parts of sodium chloride, 26 parts of potassium chloride, 43.3 parts of calcium chloride, 11.3 parts of magnesium chloride, and adjusting the pH value to 7.9 by 0.05M Tris-HCl, wherein the solvent for preparing the activation solution is distilled water;
e: and (3) parent fish nursing: putting the parent fish after spawning induction back to the polyvinyl chloride net cage, putting the net cage near a water inlet, keeping the water flow, feeding fresh water earthworms every day, controlling illumination, and transferring to a parent fish pond for cultivation after 10-15 days; controlling water flow at 0.1 m/min, feeding fresh and live tubificidae 15% of the weight of the parent fish into the net cage every day, and controlling the illumination intensity in the net cage at 50 lx.
2. The method for improving the artificial propagation efficiency of the loaches according to claim 1, wherein the selected parent fishes are selected to have good maturity, female fishes require that individuals are intact without wounds, the abdomen is soft and slightly swollen, and the minimum individual weight is more than 200 g; selecting the individual with intact body surface, no wound and vitality.
3. The method for improving the artificial propagation efficiency of the loaches according to claim 1, wherein the ratio of the amount of the parent fishes to the amount of the parents fishes is 1: 2-1: 3.
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