CN108967280B - Artificial propagation method of loaches climbing in lateral ditches - Google Patents
Artificial propagation method of loaches climbing in lateral ditches Download PDFInfo
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Classifications
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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
-
- 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)
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- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Fodder In General (AREA)
- Farming Of Fish And Shellfish (AREA)
Abstract
The invention discloses an artificial propagation method of lateral ditch mudskigurnus dabryanus, which specifically comprises the following steps: step S1: collecting and domesticating wild parent fishes; step S2: breeding parent fishes; step S3: selecting parent fish; step S4: artificial hasten parturition; the method specifically comprises the following steps: step S5: artificial insemination and incubation; the invention provides an artificial propagation technology of the loach with the lateral ditch and the rock climbing, which has high fertility rate and high hatchability rate and provides basic support for the recovery of the population of the loach with the lateral ditch and the rock climbing.
Description
Technical Field
The invention relates to the technical field of artificial propagation of lateral-furrow and cliff loaches, in particular to an artificial propagation method of the lateral-furrow and cliff loaches.
Background
Loach (Misgurni lateral wall)Beaufortialiui.Chang), belonging to the order Cyprinales, family Pinctada, subfamily aspidistrae, genus Trypanosoma. The front part of the body is flat, and the body width is obviously larger than the body height. The head is flat. The shape of the flat chisel. The anastomoses are subcutaneously enclosed to form an anastomotic fold. The herba seu radix Nothopanacis Delavayi is divided into 3 leaves, and the leaf space is provided with 2 pairs of small herba seu radix Nothopanacis Delavayi. The rear side of the kissing is provided with a lateral groove which is communicated with the head side. The front of the mouth is provided with an anastomosis groove. Lower mouth, very small mouth crack and arc shape. Lip flesh without mastoid process, and a pair of lips and lips are needed. The gill cleft was small and limited to the dorsal cranial side. The back fin from the starting point to the osculating end is slightly closer than the base of the tail fin, the back fin has no hard spine, and the outer edge is truncated. The even fin is flat. The pectoral fin begins at about the vertical line to the posterior edge of the eye and ends slightly beyond the ventral fin. The starting point of the ventral fin is slightly far from the starting point of the anal fin than the osculating end, and the rear edge is connected into a sucking disc shape, and the tail end is not close to the anus. The tail handle is short, and the length of the tail handle is smaller than the height of the tail handle. The anus is approximately equal to the distal end of the ventral fin and to the starting point of the anal fin. The body is fine-scaled. The head, chest and abdomen are naked without scales. Side line is finishedAnd (4) completing. The back and sides of the body were brown, with irregular brown patches on each fin. Mainly distributed in Minjiang and great river basins.
Due to the construction of a series of large hydraulic engineering in a large river crossing basin, the ecological environment of the area is influenced to a certain degree, and in addition, excessive fishing is carried out, the living environment of the loaches climbing from side ditches is worse and worse, and the wild resource amount is reduced sharply. So far, the research on the lateral channel perlitia anguillicaudati only has been reported about taxonomic research.
The artificial breeding work of the lateral ditch rock climbing loaches is carried out to recover and supplement the population quantity of the lateral ditch rock climbing loaches, and the method has important significance.
In the prior art, the artificial propagation technology of the mudskipper has not been reported.
Disclosure of Invention
The invention aims to provide an artificial propagation method of lateral ditch mudskilfurgus, and provides an artificial propagation technology of lateral ditch mudskilfurgus with high fertility rate and high hatching rate.
The invention is realized by the following technical scheme:
the artificial propagation method of the loach with the lateral ditch and the rock climbing specifically comprises the following steps:
step S1: collecting and domesticating wild parent fishes;
step S2: breeding parent fishes;
step S3: selecting parent fish;
step S4: artificial hasten parturition; the method specifically comprises the following steps:
step S41: selecting female fish; specifically, according to the determination data of the reproductive biology of the loaches with the lateral ditches and the crawl rocks, selecting female fishes according to the standard of 3.5g per fish;
step S42: artificially injecting an oxytocic; the oxytocic is a drug combination dose which is 1-3 mu g of LHRH-A2/g and 10-30 IU of HCG/g;
step S43: putting the parent fish injected with the oxytocic back into the circulating pool, keeping the water temperature at 15-17 ℃, and setting the water flow speed at 0.2-0.4 m/s;
step S5: artificial insemination and incubation; the method specifically comprises the following steps:
step S51: after the female fish is injected with the medicine for 16 hours, the female fish is fished out, and eggs on the abdomen are extruded to smoothly flow out of the cloaca, so that the eggs can be extruded;
step S52: sucking the redundant water on the surface of the female fish, and extruding the mature eggs into a dry culture dish;
step S53: obtaining sperms of the male fish; extruding the abdomen of the male fish to flow out of milky semen from the cloaca, and obtaining a solution with optimal sperm motility configuration by using the experimental result of sperm motility; flushing the eggs into a culture dish containing the eggs by using an injector, slightly shaking the culture dish by hands for 10-15 s, and standing for 30s to obtain fertilized eggs;
step S54: laying fertilized eggs in a beaker, placing the beaker at a water inlet and a water outlet of a temporary rearing basin, controlling the water temperature to be 15-17 ℃, and maintaining the microflow water flow rate to be 0.05-0.1 m/s for incubation; incubate in a beaker.
Further, in order to better implement the present invention, the step S2 specifically includes the following steps:
step S21: selecting male fish with body surface injury free and weight above 1.5g and female fish with body weight above 2.3 g; domesticated wild loaches climbing into lateral ditches are used as backup parent fishes for cultivation;
step S22: putting the reserved parent fish into an outdoor cement pond for cultivation, and culturing in the outdoor cement pond by using running water; the breeding density is 320 tails per cement pond, the water inflow of the cement pond is controlled to be 0.3m/s, the pH of the water for breeding is 6.9-7.7, the dissolved oxygen is 10.5-12 mg/L, and the water temperature is 15-20 ℃.
Step S23: and (5) feeding the parent fish in reserve.
Further, in order to better implement the present invention, the step S23 specifically includes the following steps:
step S231: feeding and cultivating, the concrete process is:
after the reserved parent fish is transferred into a cement pond, feeding once a day within 2 weeks, wherein the feeding amount is 1-2% of the weight of the parent fish every day, and feeding twice a day, wherein the bait is a tubificidae, chironomid larva and microparticle compound feed; the mass ratio of the tubificidae, the chironomus larvae and the microparticle compound feed is 2: 2: 1;
step S232: the feeding amount and the feeding times are reduced from the 3 rd week, because a large amount of algae grow in the pond to provide food for the parent fish at the moment, the bait put into the pond is observed in the 4 th week, and the parent fish can not be fed any more, so that the feeding of the parent fish can be stopped;
step S233: screening and intensive rearing are performed from the backup parent fish reared in step S232; the method specifically comprises the following steps: screening 60 mature parent fishes, and independently placing the mature parent fishes in another cement pond for reinforced cultivation, wherein the water flow speed is controlled to be 2.5-3.5 m/s; feeding the parent fish twice a day by 3-5% of the weight of the parent fish every day, wherein the bait is mixed feed of tubificidae, chironomid larvae and microparticles; the mass ratio of the tubificidae, the chironomus larvae and the microparticle compound feed is 2: 2: 1.
further, in order to better implement the present invention, the step S3 specifically includes: the female fish is selected from individuals with distended, soft and elastic abdomen, flowing feeling under light pressure and reddish genital pores, and the egg is squeezed to be uniform in size and light yellow; selecting an individual with an active individual, a slightly small body size and a long and sharp reproductive aperture from the male fish; the ratio of the female fish to the male fish is 1.2: 1.
further, in order to better implement the present invention, the step S42 specifically includes: and injecting an oxytocic drug from the base of the pectoral fin of the female fish at an angle of 45 degrees by adopting a needle injection method, wherein the needle insertion depth is 0.8-0.12 cm.
Further, in order to better implement the present invention, the wild parent fish domestication in the step S1 specifically includes a first stage of domestication and a second stage of domestication; the first stage domestication time is 35-40 days; the second stage domestication time is 120-150 days.
The first stage of domestication specifically means: adopting algae, moss, tubificidae and chironomid larvae growing on stone as initial feed for domestication for 35-40 days;
the second stage of domestication specifically comprises: domestication is carried out for 120-150 days by adopting tubificidae, chironomid larvae and microparticle compound feed as second-stage domestication bait; the tubificidae: chironomid larva: the mass ratio of the micro-particle compound feed is 1: 1: 2.
compared with the prior art, the invention has the following advantages and beneficial effects:
the invention provides an artificial propagation technology of the loach with the lateral ditch and the rock climbing, which has high fertility rate and high hatchability rate and provides basic support for the recovery of the population of the loach with the lateral ditch and the rock climbing.
Detailed Description
The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto.
Example 1:
the invention is realized by the following technical scheme:
the artificial propagation method of the loach with the lateral ditch and the rock climbing specifically comprises the following steps:
step S1: collecting and domesticating wild parent fishes;
when wild fish is collected, firstly, regional investigation is mainly used for investigating wild parent fish resources, a fixed sampling section is not arranged, and temporary points are arranged according to river reach conditions for investigating and collecting fish specimens; screening parent fish with strong physique and light injury, and soaking the parent fish for 1-3 minutes by adopting a mixed aqueous solution of penicillin and salt; the concentration and the proportion of the mixed aqueous solution specifically mean that 4.5 percent of salt water and 1kg of water are mixed with 0.5 ten thousand units of penicillin, and wild parent fishes are soaked for 1-3 minutes, so as to prevent and reduce saprolegniasis.
Selecting female fishes from the collected wild loaches with lateral furrows and climbing rocks, wherein the average weight of the female fishes is 3.5g, the weight of the male fishes is 2.0-2.5 g, the individuals are less sexually mature, analyzing anatomical conditions, collecting the gonads of the female fishes and the male fishes with the above weights to grow to IV, and the gonads of the female fishes and the male fishes reach the specifications of parent fishes and can be domesticated;
the first stage domestication: feeding and domesticating the collected wild parent fishes after the wild parent fishes are statically cultured for 25-30 days; in the first stage 21d, the parent fish feeds, wherein the bait is algae, moss, tubificidae and chironomid larvae; algae, moss, tubificidae and chironomid larvae growing on stone are used as initial feed;
the second stage domestication: the feed starts to be actively fed at 35-40 d, and the bait is compound feed of tubificidae, chironomid larvae and microparticles; the mass ratio of the compound feed is 1: 1: 2; performing the second stage of domestication.
The preferable second stage domestication can also adopt one or more of micro-particle S1, S3 and S5 feed mixed with tubificidae, micro-particle S1, S3 and S5 feed mixed with spirulina, diatom and eel powder mixed with tubificidae, the mixture of spirulina and diatom and Sanyou goldfish feed as the second stage domestication bait for domestication.
Step S2: breeding parent fishes;
selecting domesticated wild parent fish as a backup parent fish for breeding, wherein the backup parent fish is male fish with a body surface free of damage and a weight of more than 1.5g and female fish with a weight of more than 2.3 g;
putting the reserved parent fish into an outdoor cement pond for cultivation, and cultivating the parent fish in the cement pond by running water, wherein the cultivation density is 320 fish/pond, the water inflow of the cement pond is 0.3m/s, the pH value of cultivation water is 6.9-7.7, the dissolved oxygen is 10.5-12 mg/L, and the water temperature is 15-20 ℃;
in the first two weeks of cement pond cultivation, throw in bait twice every day respectively once every morning and evening, throw in bait for the first week and be tubificidae, chironomid larva, the A fodder that the microparticle cooperation formed, tubificidae in the A fodder: chironomid larva: the mass ratio of the microparticles is 1: 1: 2, feeding bait in the second week, namely feed B formed by matching tubificidae, chironomid larvae and micro-particles, wherein the tubificidae in the feed B: chironomid larva: the mass ratio of the micro-particles is 2: 1: 1.
in the third week of cement pond cultivation, the feeding amount and the feeding times are reduced; algae may provide a portion of the food since they have already grown in the cement pond beginning at week three; therefore, bait is fed once at 8 o' clock every night, the amount of the fed bait is 1-2% of the weight of the parent fish, the fed bait is C feed formed by mixing tubificidae and chironomus larvae, and the mass ratio of the tubificidae to the chironomus larvae in the C feed is 1: 1; then the feeding is gradually reduced, the proportion of tubificidae is reduced, and a large amount of algae growing in the pond can provide food for parent fish;
feeding baits in the 4 th week and observing, wherein when the parent fish is not eating the fed baits, the feeding of the baits can be stopped;
the reserve parent fish that were cultured for 4 weeks were screened and intensive cultured. The method specifically comprises the following steps: the method comprises the following steps of selecting female fish individuals to be complete and have no wound, slightly pressing the belly to be softer, enabling the female fish to see immature eggs through cloacal oosperm holes, selecting male fish individuals to be complete and have no wound, placing 60 mature parent fishes with good vitality in another cement pond independently, carrying out reinforced cultivation, increasing water flow, controlling the speed of the water flow to be 2.5-3.5 m/s, increasing feeding of baits, feeding the parents twice a day, wherein the feeding amount is 3-5% of the weight of the parent fishes every day, the baits are tubificidae larvae and corpuscle compound feeds, and the mass ratio of the baits is as follows: tubificidae: chironomid larva: the micro-particles are 2: 2: 1.
the operation has the effects that the flowing water is adopted to stimulate the fish body, the gonad of the fish body is more mature, and regular inspection of parent fish is facilitated before artificial propagation is carried out later; the water flushing strengthening cultivation is increased.
Step S3: selecting parent fish;
after intensive cultivation for two months, temporarily culturing 60 screened mature parent fishes in a temporary culture basin with the radius of 30cm, stimulating the parent fishes in the temporary culture basin by adopting flowing water circulation, controlling the water temperature to be 15-17 ℃, and keeping flowing water;
the female fish is selected from individuals with distended, soft and elastic abdomen, flowing feeling under light pressure and reddish genital pores, and the egg is squeezed to be uniform in size and light yellow; selecting an individual with an active individual, a slightly small body size and a long and sharp reproductive aperture from the male fish; the ratio of the male ratio to the female ratio is 1.2: 1.
step S4: artificial hasten parturition; the method specifically comprises the following steps:
step S41: selecting female fish; specifically, according to the determination data of the reproductive biology of the loaches with the lateral ditches and the crawl rocks, selecting female fishes according to the standard of 3.5g per fish;
step S42: artificially injecting an oxytocic; the oxytocic is a drug with the combined dose of 1-3 mu g LHRH-A2(iv)/g and 10-30 IU HCG/g;
step S43: putting the parent fish injected with the oxytocic back into the circulating pool, keeping the water temperature at 15-17 ℃, and setting the water flow speed at 0.2-0.4 m/s;
step S5: artificial insemination and incubation; the method specifically comprises the following steps:
step S51: after the female fish is injected with the medicine for 16 hours, the female fish is fished out, and eggs on the abdomen are extruded to smoothly flow out of the cloaca, so that the eggs can be extruded;
step S52: sucking the redundant water on the surface of the female fish, and extruding the mature eggs into a dry culture dish;
because the female fish is not anesthetized, the water on the surface of the female fish needs to be wiped off, the female fish is fixed by one hand, the female fish extrudes eggs by the other hand, and the extruded eggs are washed into a culture dish by physiological saline by an assistant due to the small and slightly viscous eggs; after squeezing, the male fish is caught again, namely water on the surface of the male fish is wiped off by using a roll paper, the female fish is fixed by using one hand, and the sperm is squeezed by using the other hand.
Step S53: obtaining sperms of the male fish; extruding the abdomen of the male fish to flow out of milky semen from the cloaca, and obtaining a solution with optimal sperm motility configuration by using the experimental result of sperm motility; flushing the eggs into a culture dish containing the eggs by using an injector, slightly shaking the culture dish by hands for 10-15 s, and standing for 30s to obtain fertilized eggs;
the solution prepared by the optimal sperm motility obtained by the experimental result of the sperm motility specifically comprises the following components: by preparing NaCl solutions with different concentrations, the violent movement time, the quick movement time, the service life and the activation rate of the loach sperms with the lateral ditches and the rock climbing show a trend that the violent movement time, the quick movement time, the service life and the activation rate are increased and then decreased along with the increase of the concentration of the NaCl solution.
In 4% NaCl solution, the sperm has the longest violent movement time and life, and the activation rate is 100%;
in 3% NaCl solution, the sperm has the longest fast moving time and the activation rate is over 90%.
The concentration range of the most suitable NaCl for the sperm motility of the loach with the lateral ditch and the creeping rock is 3 to 4 percent; the solution with the most suitable sperm motility configuration is NaCl physiological saline with the concentration of 3 percent.
Step S54: laying fertilized eggs in a beaker, placing the beaker at a water inlet and a water outlet of a temporary culture basin, controlling the water temperature at 15-17 ℃, keeping micro-flow water, controlling the flow rate of the micro-flow water at 0.05-0.1 m/s, and incubating in the beaker.
And (3) incubating the larvae of the loaches with the lateral ditches at the temperature of 15-17 ℃ for 60h and 55min, and incubating the larvae at the temperature of 974.56 ℃ for h.
Other parts of this embodiment are the same as those of the above embodiment, and thus are not described again.
Example 2:
the invention is realized by the following technical scheme, and the artificial propagation method of the loach with lateral ditches and climbing rocks specifically comprises the following steps:
step S1: collecting and domesticating wild parent fishes;
when wild fish is collected, firstly, regional investigation is mainly used for investigating wild parent fish resources, a fixed sampling section is not arranged, and temporary points are arranged according to river reach conditions for investigating and collecting fish specimens; screening parent fish with strong physique and light injury, and soaking the parent fish in mixed water solution of penicillin and salt for 1 minute;
the concentration and proportion of the mixed aqueous solution specifically mean that 4.5 percent of salt water and 1kg of water are mixed with 0.5 ten thousand units of penicillin, and wild parent fish are soaked for 1 minute, so as to prevent and reduce saprolegniasis.
Selecting female fish with the average weight of 3.5g from the collected wild loaches with the lateral ditch and the rock climbing, selecting male fish with the weight of 2.5g for smaller sexual maturity, analyzing anatomical conditions, collecting the gonads of the female fish and the male fish with the weights to be developed to IV to reach the parent fish specification, and domesticating;
the first stage domestication: feeding and domesticating the collected wild parent fishes after the wild parent fishes are statically cultured for 30 d; in the first stage 21d the parent fish is fed; the first stage domesticating bait is algae, moss, tubificidae and chironomid larvae; algae, moss, tubificidae and chironomid larvae growing on stone are used as initial feed;
the second stage domestication: the feeding starts actively at 35d, and the domestication bait at the second stage is a compound feed of tubificidae, chironomid larvae and microparticles; the mass ratio of the domesticating bait in the second stage is 1: 1: 2; performing the second stage of domestication.
Step S2: breeding parent fishes;
selecting domesticated wild parent fish as a backup parent fish for breeding, wherein the backup parent fish is male fish with a body surface free of damage and a weight of more than 1.5g and female fish with a weight of more than 2.3 g;
putting the reserved parent fish into an outdoor cement pond for cultivation, and adopting running water cultivation in the cement pond, wherein the cultivation density is 320 fish/pond, the water inflow of the cement pond is 0.3m/s, the pH value of cultivation water is 7.0, the dissolved oxygen is 12mg/L, and the water temperature is 16 ℃;
in the first two weeks of cement pond cultivation, bait is thrown twice a day, once in the morning and at night, and the bait thrown in the first week is tubificidae: chironomid larva: the mass ratio of the micro-particle compound feed is 1: 1: 2, feeding bait for tubificidae in the second week: chironomid larva: the mass ratio of the micro-particle compound feed is 2: 1: 1.
in the third week of cement pond cultivation, the feeding amount and the feeding times are reduced; in the third week, algae grow, so that a part of food can be provided for the parent fishes, the parent fishes are fed once at 8 o' clock every day, the feeding amount is 1-2% of the weight of the parent fishes, and the fed baits are tubificidae: chironomid larvae is 1: 1; as a result of the increased time, the large amount of algae that grows in the pond will provide food for the parent fish; so the feeding and the proportion of tubificidae are gradually reduced.
Feeding baits in the 4 th week and observing, wherein when the baits fed in the parent fishes are found out not to be damaged, the feeding of the baits can be stopped;
screening and strengthening cultivation of the backup parent fish after four weeks of cultivation; the method comprises the following steps of selecting female fish individuals to be complete and have no wound, slightly pressing the belly to be softer, enabling the female fish to see immature eggs through cloaking at the cloaca of the female fish, selecting male fish individuals to be complete and have no wound, placing 60 mature parent fishes with better vitality into another cement pond separately, carrying out reinforced cultivation, increasing water flow, controlling the speed of the water flow to be 2.5m/s, stimulating the parent fishes through running water, increasing the throwing of baits, feeding the parent fishes twice every day, wherein the feeding amount is 5% of the weight of the parent fishes, the baits are tubificidae, chironomid larvae and corpuscle matched feeds, and the proportion of the baits is as follows: tubificidae: chironomid larva: the mass ratio of the micro-particle compound feed is 2: 2: 1.
the screened 60 mature parent fishes are separately placed in another cement pond for reinforced cultivation, the function of the method is to stimulate the fish body by running water to ensure that the gonads of the parent fishes develop more mature, and the regular inspection of the parent fishes is convenient before the later artificial propagation; screening in 1 in the middle ten days of the month for increased water-flushing intensive cultivation, wherein the time for intensive cultivation is two months.
Step S3: selecting parent fish;
after intensive cultivation for two months, temporarily culturing the screened parent fishes subjected to intensive cultivation in a temporary culture basin with the radius of 30cm, stimulating the parent fishes in the temporary culture basin by adopting flowing water circulation, controlling the water temperature to be 16 ℃, and keeping flowing water;
the female fish is selected from individuals with distended, soft and elastic abdomen, flowing feeling under light pressure and reddish genital pores, and the egg is squeezed to be uniform in size and light yellow; selecting an individual with an active individual, a slightly small body size and a long and sharp reproductive aperture from the male fish; the ratio of the male ratio to the female ratio is 1.2: 1.
step S4: artificial hasten parturition; the method specifically comprises the following steps:
step S41: selecting female fish; specifically, according to the determination data of the reproductive biology of the loaches with the lateral ditches and the crawl rocks, selecting female fishes according to the standard of 3.5g per fish;
step S42: artificially injecting an oxytocic; the oxytocic is prepared from the following medicaments in part by weight: 2. mu.g LHRH-A2/g and 10IU HCG/g.
Step S43: putting the parent fish injected with the oxytocic back into the circulating pool, keeping the water temperature at 15-17 ℃, and setting the water flow speed at 0.2 m/s;
step S5: artificial insemination and incubation; the method specifically comprises the following steps:
step S51: after the female fish is injected with the medicine for 16 hours, the female fish is fished out, and eggs on the abdomen are extruded to smoothly flow out of the cloaca, so that the eggs can be extruded;
step S52: sucking the redundant water on the surface of the female fish, and extruding the mature eggs into a dry culture dish;
step S53: obtaining sperms of the male fish; squeezing the abdomen of the male fish to flow out of milky semen from the cloaca, flushing the normal saline with 3% concentration of NaCl with optimal sperm activity configuration obtained by the sperm activity experimental result into a culture dish containing the ovum by using an injector, slightly shaking the culture dish by hand for 10s, and standing for 30s to obtain a fertilized ovum;
step S54: laying fertilized eggs in a beaker, placing the beaker at a water inlet and a water outlet of a temporary culture basin, controlling the water temperature at 15 ℃, keeping micro-flowing water, and controlling the flow rate of the micro-flowing water to be 0.05m/s for incubation in the beaker. A water inlet pipe is arranged above the temporary rearing pot, and a water outlet pipe is arranged in the center of the pot.
And (3) after the embryo of the loach climbing the lateral ditch develops at 16 ℃ for 60h and 55min, starting hatching seedlings, and accumulating at 974.56 ℃ for h.
Other parts of this embodiment are the same as those of the above embodiment, and thus are not described again.
Example 3:
this example is the same procedure as example 2, except that the oxytocin dosage and composition are different; the adopted oxytocic drugs are as follows: 1. mu.g LHRH-A2/g and 30IU HCG/g.
Other parts of this embodiment are the same as those of the above embodiment, and thus are not described again.
Example 4:
this example is the same procedure as example 2, except that the oxytocin dosage and composition are different; the adopted oxytocic drugs are as follows: 2. mu.g LHRH-A2/g and 20IU HCG/g.
Other parts of this embodiment are the same as those of the above embodiment, and thus are not described again.
Example 5:
this example is the same procedure as example 2, except that the oxytocin dosage and composition are different; the adopted oxytocic drugs are as follows: 2. mu.g LHRH-A2/g and 30IU HCG/g.
Other parts of this embodiment are the same as those of the above embodiment, and thus are not described again.
Example 6:
this example is the same procedure as example 2, except that the oxytocin dosage and composition are different; the adopted oxytocic drugs are as follows: 1. mu.g LHRH-A2/g and 20IU HCG/g.
Other parts of this embodiment are the same as those of the above embodiment, and thus are not described again.
Example 7:
this example is the same procedure as example 2, except that the oxytocin dosage and composition are different; the adopted oxytocic drugs are as follows: 1. mu.g LHRH-A2/g and 10IU HCG/g.
Other parts of this embodiment are the same as those of the above embodiment, and thus are not described again.
Example 8:
the invention is realized by the following technical scheme, and the artificial propagation method of the loach with lateral ditches and climbing rocks specifically comprises the following steps:
step S1: collecting and domesticating wild parent fishes;
when wild fish is collected, firstly, regional investigation is mainly used for investigating wild parent fish resources, a fixed sampling section is not arranged, and temporary points are arranged according to river reach conditions for investigating and collecting fish specimens; screening parent fish with strong physique and light injury, and soaking in mixed water solution of penicillin and salt for 3 min;
the concentration and proportion of the mixed aqueous solution specifically mean that 4.5 percent of salt water and 1kg of water are mixed with 0.5 ten thousand units of penicillin, and wild parent fish are soaked for 3 minutes, so as to prevent and reduce saprolegniasis.
Selecting female fish with individual weight of 3.5g and male fish with individual weight of 2.0 to be less sexually mature from the collected wild loaches, analyzing anatomical conditions, collecting the gonads of the female fish and the male fish with the above weights to be developed to IV to reach the parent fish specification, and performing domestication;
the first stage domestication: feeding domestication of the collected wild parent fish is started after the wild parent fish is statically cultured 25; the parent fish feed around the first stage 21 d; the first stage domesticating bait is algae, moss, tubificidae and chironomid larvae; algae, moss, tubificidae and chironomid larvae growing on stone are used as initial feed;
the second stage domestication: the feeding starts actively at 30d, and the domestication bait at the second stage is a compound feed of tubificidae, chironomid larvae and microparticles; the mass ratio of the domesticating bait in the second stage is 1: 1: 2; performing the second stage of domestication.
Step S2: breeding parent fishes;
selecting domesticated wild parent fish as a backup parent fish for breeding, wherein the backup parent fish is male fish with a body surface free of damage and a weight of more than 1.5g and female fish with a weight of more than 2.3 g;
putting the reserved parent fish into an outdoor cement pond for cultivation, and adopting running water cultivation in the cement pond, wherein the cultivation density is 320 fish/pond, the water inflow of the cement pond is 0.3m/s, the pH value of cultivation water is 7.7, the dissolved oxygen is 12mg/L, and the water temperature is 15 ℃;
in the first two weeks of cement pond cultivation, bait is thrown twice a day, once in the morning and at night, and the bait thrown in the first week is tubificidae: chironomid larva: the mass ratio of the micro-particle compound feed is 1: 1: 2, feeding bait for tubificidae in the second week: chironomid larva: the mass ratio of the micro-particle compound feed is 2: 1: 1.
in the third week of cement pond cultivation, the feeding amount and the feeding times are reduced; in the third week, as algae grow, a part of food can be provided for the parent fish, the parent fish is fed once at 8 o' clock every day, the feeding amount is 2% of the weight of the parent fish, and the fed bait is tubificidae: chironomid larvae is 1: 1; as a result of the increased time, the large amount of algae that grows in the pond will provide food for the parent fish; so the feeding and the proportion of tubificidae are gradually reduced.
Feeding baits in the 4 th week and observing, wherein when the baits fed in the parent fishes are found out not to be damaged, the feeding of the baits can be stopped;
screening and strengthening cultivation of the backup parent fish after four weeks of cultivation; the method comprises the following steps of selecting female fish individuals to be complete and have no wound, slightly pressing the belly to be softer, enabling the female fish to see immature eggs through cloaking at the cloaca of the female fish, selecting male fish individuals to be complete and have no wound, placing 60 mature parent fishes with better vitality into another cement pond separately, carrying out reinforced cultivation, increasing water flow, controlling the speed of the water flow to be 3.5m/s, stimulating the parent fishes through running water, increasing the throwing of baits, feeding the parent fishes twice every day, wherein the feeding amount is 5% of the weight of the parent fishes, the baits are tubificidae, chironomid larvae and corpuscle matched feeds, and the proportion of the baits is as follows: tubificidae: chironomid larva: the mass ratio of the micro-particle compound feed is 2: 2: 1.
the screened 60 mature parent fishes are separately placed in another cement pond for reinforced cultivation, the function of the method is to stimulate the fish body by running water to ensure that the gonads of the parent fishes develop more mature, and the regular inspection of the parent fishes is convenient before the later artificial propagation; screening is carried out in 2 months for increasing the water flushing strengthening cultivation, and the strengthening cultivation time is two months.
Step S3: selecting parent fish;
after intensive cultivation for two months, temporarily culturing the screened parent fishes subjected to intensive cultivation in a temporary culture basin with the radius of 30cm, stimulating the parent fishes in the temporary culture basin by adopting flowing water circulation, controlling the water temperature to be 16 ℃, and keeping flowing water;
the female fish is selected from individuals with distended, soft and elastic abdomen, flowing feeling under light pressure and reddish genital pores, and the egg is squeezed to be uniform in size and light yellow; selecting an individual with an active individual, a slightly small body size and a long and sharp reproductive aperture from the male fish; the ratio of the male ratio to the female ratio is 1.2: 1.
step S4: artificial hasten parturition; the method specifically comprises the following steps:
step S41: selecting female fish; specifically, according to the determination data of the reproductive biology of the loaches with the lateral ditches and the crawl rocks, selecting female fishes according to the standard of 3.5g per fish;
step S42: artificially injecting an oxytocic; the oxytocic is prepared from the following medicaments in part by weight: 1. mu.g LHRH-A2/g and 30IU HCG/g.
Step S43: putting the parent fish injected with the oxytocic back into the circulating pool, keeping the water temperature at 17 ℃, and setting the water flow speed at 0.4 m/s;
step S5: artificial insemination and incubation; the method specifically comprises the following steps:
step S51: after the female fish is injected with the medicine for 16 hours, the female fish is fished out, and eggs on the abdomen are extruded to smoothly flow out of the cloaca, so that the eggs can be extruded;
step S52: sucking the excess water on the surface of the female fish, and extruding the mature eggs into a dry culture dish;
step S53: obtaining sperms of the male fish; extruding the abdomen of the male fish to flow out of milky semen from the cloaca, flushing normal saline with 3% concentration of NaCl with optimal sperm activity obtained by the sperm activity experimental result into a culture dish containing the ovum by using an injector, slightly shaking the culture dish by hand for 10-15 s, and standing for 30s to obtain a fertilized ovum;
step S54: laying fertilized eggs in a beaker, placing the beaker at a water inlet and a water outlet of a temporary culture basin, controlling the water temperature at 16 ℃, keeping micro-flow water, and controlling the flow rate of the micro-flow water to be 0.05m/s for incubation in the beaker.
A water inlet pipe is arranged above the temporary rearing pot, and a water outlet pipe is arranged in the center of the pot.
And (3) after the embryo of the loach climbing the lateral ditch develops at 16 ℃ for 60h and 55min, starting hatching seedlings, and accumulating at 974.56 ℃ for h.
Example 9:
the invention also sets a comparative example verification experiment:
comparative example 1:
this comparative example 1 differs from example 2 in the dosage and composition of the oxytocic; the oxytocic drugs adopted are LHRH-A2, HCG and DOM, and the dosages of the oxytocic drugs are LHRH-A23-5 mug/g, HCG 30-50 IU/g and DOM 2-3 mug/g;
comparative example 2:
this comparative example 2 differs from example 2 in the dosage and composition of the oxytocic; the oxytocic drugs adopted are LHRH-A2 and DOM, and the dosages are LHRH-A23-5 mu g/g and DOM 2-3 mu g/g;
comparative example 3:
this comparative example 3 differs from example 2 in the dosage and composition of the oxytocic; the adopted oxytocic drugs are LHRH-A2 and HCG, and the dosages of the oxytocic drugs are LHRH-A23-5 mu g/g and HCG 30-50 IU/g;
comparative example 4:
this comparative example 4 differs from example 2 in the dosage and composition of the oxytocic; the adopted oxytocic drugs are LHRH-A2 and HCG, and the dosages of the oxytocic drugs are LHRH-A21-3 mu g/g and HCG 10-30 IU/g;
comparative example 5:
this comparative example 5 differs from example 2 in the dosage and composition of the oxytocic; the adopted oxytocic drugs are LHRH-A2 and HCG, and the dosages of the oxytocic drugs are LHRH-A20.1-1 mu g/g and HCG 1-10 IU/g;
the artificial propagation method of the embodiment 3 and the comparative examples 1-5 is adopted for propagation, and the experimental data structure is shown in the following table 1:
TABLE 1
The doses of LHRH-A2 and DOM in Table 1 are in μ g/g; the dosage unit of HCG is IU/g;
as can be seen from Table 1: compared with the proportion of 1-5, the loaches with the lateral ditches and the rock creepers are matched through all the steps under the same breeding condition, so that the loaches with the lateral ditches and the rock creepers are high in fertility rate and hatching rate.
Comparative example 6:
this comparative example 6 differs from example 2 in the dosage and composition of the oxytocin; the difference is the water flow rate in step S2, the water flow rate in step S43, and the water flow rate in step S54; the water flow rate in step S2 was 2.5m/S, the water flow rate in step S43 was 0.2m/S and the water flow rate in step S54 was 0.1 m/S.
Comparative example 7:
this comparative example 7 differs from example 2 in the dosage and composition of the oxytocic; the difference is the water flow rate in step S2, the water flow rate in step S43, and the water flow rate in step S54; the water flow rate in step S2 was 3.5m/S, the water flow rate in step S43 was 0.4m/S and the water flow rate in step S54 was 0.1 m/S.
Comparative example 8:
this comparative example 8 differs from example 8 in the dosage and composition of the oxytocin; the difference is the water flow rate in step S2, the water flow rate in step S43, and the water flow rate in step S54; the water flow rate in step S2 was 2m/S, the water flow rate in step S43 was 0.3m/S, and the water flow rate in step S54 was 0.075 m/S.
Comparative example 9:
this comparative example 9 differs from example 2 in the dosage and composition of the oxytocin; the difference is the water flow rate in step S2, the water flow rate in step S43, and the water flow rate in step S54; the water flow rate in step S2 was 2.5m/S, the water flow rate in step S43 was 0.3m/S, and the water flow rate in step S54 was 0.1 m/S.
Comparative example 10:
this comparative example 10 differs from example 2 in the dosage and composition of the oxytocin; the difference is the water flow rate in step S2, the water flow rate in step S43, and the water flow rate in step S54; the water flow rate in step S2 was 3.5m/S, the water flow rate in step S43 was 0.3m/S and the water flow rate in step S54 was 0.05 m/S.
Comparative example 11:
this comparative example 10 differs from example 2 in the dosage and composition of the oxytocin; the difference is the water flow rate in step S2, the water flow rate in step S43, and the water flow rate in step S54; the water flow rate in step S2 was 0.8m/S, the water flow rate in step S43 was 0.1m/S, and the water flow rate in step S54 was 0.01 m/S.
The artificial propagation method of the embodiment 8 and the comparative examples 6-11 is adopted for propagation, and the experimental data structure is shown in the following table 2:
TABLE 2
From table 2, it can be seen that the loaches climbing the lateral ditches are matched with each other through each step in comparison with the comparative proportion of 6-11 under the same breeding conditions, and the fertility rate and the hatchability of the loaches climbing the lateral ditches can be effectively increased within the range of the water flow speed required by the invention.
Other parts of this embodiment are the same as those of the above embodiment, and thus are not described again.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention in any way, and all simple modifications and equivalent variations of the above embodiments according to the technical spirit of the present invention are included in the scope of the present invention.
Claims (3)
1. The artificial propagation method of the loach with lateral ditches and climbing rocks is characterized by comprising the following steps: the method specifically comprises the following steps:
step S1: collecting and domesticating wild parent fishes;
the wild parent fish domestication in the step S1 specifically includes a first stage of domestication and a second stage of domestication; the first stage domestication time is 35-40 days; the second stage domestication time is 120-150 days;
the first stage of domestication specifically means: adopting algae, moss, tubificidae and chironomid larvae growing on stone as initial feed for domestication for 35-40 days;
the second stage of domestication specifically comprises: domestication is carried out for 120-150 days by adopting tubificidae, chironomid larvae and microparticle compound feed as second-stage domestication bait; the tubificidae: chironomid larva: the mass ratio of the micro-particle compound feed is 1: 1: 2;
step S2: breeding parent fishes;
step S21: selecting male fish with body surface injury free and weight above 1.5g and female fish with body weight above 2.3 g; domesticated wild loaches climbing into lateral ditches are used as backup parent fishes for cultivation;
step S22: putting the reserved parent fish into an outdoor cement pond for cultivation, and culturing in the outdoor cement pond by using running water; the breeding density is 320 tails per cement pond, the water inflow of the cement pond is controlled to be 0.3m/s, the pH of the water for breeding is 6.9-7.7, the dissolved oxygen is 10.5-12 mg/L, and the water temperature is 15-20 ℃;
step S23: feeding the reserved parent fish;
step S231: feeding and cultivating, which specifically comprises:
feeding baits once a day within 2 weeks after the backup parent fish is transferred into the cement pond, wherein the feeding amount is 1-2% of the weight of the parent fish every day, and the baits are tubificidae, chironomid larvae and microparticle compound feeds twice a day; the mass ratio of the tubificidae, the chironomus larvae and the microparticle compound feed is 2: 2: 1;
step S232: the feeding amount and the feeding times are reduced from the 3 rd week, and the feeding is stopped if the parent fish does not eat the fed baits in the 4 th week;
step S233: screening and intensive rearing are performed from the backup parent fish reared in step S232; the method specifically comprises the following steps: screening 60 mature parent fishes, and independently placing the mature parent fishes in another cement pond for reinforced cultivation;
the intensive cultivation specifically comprises the following steps: controlling the water flow speed to be 2.5-3.5 m/s; feeding the parent fish twice a day by 3-5% of the weight of the parent fish every day, wherein the fed baits are tubificidae, chironomidae larvae and microparticle compound feed; the mass ratio of the tubificidae, the chironomus larvae and the microparticle compound feed is 2: 2: 1;
step S3: selecting parent fish;
step S4: artificial hasten parturition; the method specifically comprises the following steps:
step S41: selecting female fish; according to the determination data of the breeding biology of the loaches climbing sideways, selecting female fish according to the standard of 3.5g per fish;
step S42: artificially injecting an oxytocic; the oxytocic is a pharmaceutical compound dose of 1-3 mu g LHRH-A2(iv)/g and 10-30 IU HCG/g;
step S43: putting the parent fish injected with the oxytocic back into the circulating pool, keeping the water temperature at 15-17 ℃, and setting the water flow speed at 0.2-0.4 m/s;
step S5: artificial insemination and incubation; the method specifically comprises the following steps:
step S51: after the female fish is injected with the medicine for 16 hours, the female fish is fished out, and eggs on the abdomen are extruded to smoothly flow out of the cloaca, so that the eggs can be extruded;
step S52: sucking the redundant water on the surface of the female fish, and extruding the mature eggs into a dry culture dish;
step S53: obtaining sperms of the male fish; extruding the abdomen of the male fish to flow out of milky semen from the cloaca, and obtaining a solution with optimal sperm motility configuration by using the experimental result of sperm motility; flushing the eggs into a culture dish containing the eggs by using an injector, slightly shaking the culture dish by hands for 10-15 s, and standing for 30s to obtain fertilized eggs;
step S54: laying fertilized eggs in a beaker, placing the beaker at a water inlet and a water outlet of a temporary rearing basin, controlling the water temperature to be 15-17 ℃, and maintaining the microflow water flow rate to be 0.05-0.1 m/s for incubation; incubate in a beaker.
2. The artificial propagation method of the lateral ditch mudskiguria according to claim 1, characterized in that: the step S3 specifically includes: the female fish is selected from individuals with distended, soft and elastic abdomen, flowing feeling under light pressure and reddish genital pores, and the egg is squeezed to be uniform in size and light yellow; selecting an individual with an active individual, a slightly small body size and a long and sharp reproductive aperture from the male fish; the ratio of the female fish to the male fish is 1.2: 1.
3. the artificial propagation method of the lateral ditch mudskiguria according to claim 1, characterized in that: the specific process of manually injecting the oxytocic in the step S42 is as follows: and injecting an oxytocic drug from the base of the pectoral fin of the female fish at an angle of 45 degrees by adopting a needle injection method, wherein the needle insertion depth is 0.8-0.12 cm.
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