CN113678760B - Cross breeding method for artificially breeding and breeding pelteobagrus fulvidraco with disease resistance - Google Patents
Cross breeding method for artificially breeding and breeding pelteobagrus fulvidraco with disease resistance Download PDFInfo
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Classifications
-
- 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
- A01K61/17—Hatching, e.g. incubators
-
- 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 discloses a hybridization breeding method for artificially breeding pelteobagrus fulvidraco with disease resistance, which comprises the following steps: s1 family cultivation: after the wild resource investigation of pelteobagrus fulvidraco and pelteobagrus vachelli is carried out, selecting healthy two-age pelteobagrus fulvidraco and two-age pelteobagrus vachelli without diseases and injuries, and carrying out selfing twice to obtain respective F1 generation and F2 generation; s2, disease resistance breeding: f1 and F2 generations are generated by respective selfing to perform aeromonas hydrophila resistance character breeding; providing more sufficient freshwater fish seedlings, enriching the variety of pond culture, being beneficial to the continuous proliferation of natural resource population and protecting the diversity of organisms. Meanwhile, the industrial association and the radiation degree are continuously improved, the industrial chain for producing, selling and selling is orderly extended, the development of industries such as living and fresh transportation, tool manufacturing for fishing, feed and bait is driven, the problem of the outlet of a large amount of local fishery labor force is solved, and the ecological and social benefits are remarkable.
Description
Technical Field
The invention relates to the technical field of breeding methods, in particular to a hybridization breeding method for artificially breeding pelteobagrus fulvidraco with disease resistance.
Background
Pelteobagrus fulvidraco (Pelteobagrus fulvidraco) belongs to the genus Pelteobagrus (pelteobarus) of the family catfish (Babridae) of the order catfish (Siluriformes), and is widely distributed in the rest of the water areas of China except western plateau and Xinjiang. Pelteobagrus fulvidraco is a benthic fish, and is an important economic fish because of delicious meat and high nutritive value, and is deeply favored by breeders and consumers. The pelteobagrus vachelli (Pelteobagrus vachelli) belongs to the same genus as pelteobagrus vachelli, is the relatively largest variety of individuals in the pelteobagrus vachelli, is mainly distributed in the Yangtze river and the tributaries thereof, has tender fish meat, delicious taste and rich nutrition, and is also an important economic fish. In recent years, although the artificial culture yield of pelteobagrus fulvidraco is increased, the market demand can not be met, and meanwhile, the demand for excellent pelteobagrus fulvidraco seedlings in production is continuously increased. Therefore, the hybrid pelteobagrus fulvidraco fries with excellent properties are cultivated and bred based on the hybridization breeding technology, the current industrial requirements can be met, the popularization and industrialization prospects are strong, and good economic benefits can be generated.
At present, the cross breeding of pelteobagrus fulvidraco (female) x pelteobagrus vachelli (female) can be bred into hybrid varieties. The growth speed and fertilization rate of the filial generation are higher than those of the parent, the heterosis is obvious, but the filial generation is often influenced by aeromonas hydrophila (Aeromonas hydrophila) in actual production, the disease is usually urgent and can not be cured by medicines after the disease, and the hybrid pelteobagrus fulvidraco fries with aeromonas hydrophila resistance can be cultivated by the implementation of the technology.
Disclosure of Invention
The invention aims to provide a cross breeding method for artificially breeding pelteobagrus fulvidraco with disease resistance, which solves the problems in the background technology.
In order to achieve the above purpose, the present invention provides the following technical solutions: a cross breeding method for artificially breeding and breeding pelteobagrus fulvidraco with disease resistance comprises the following steps:
s1, family cultivation: after carrying out wild resource investigation of pelteobagrus fulvidraco and pelteobagrus vachelli, selecting disease-free and injury-free healthy pelteobagrus vachelli and pelteobagrus vachelli; then generating F1 and F2 generation by selfing respectively;
s2, disease-resistant breeding: selecting diseased pelteobagrus fulvidraco and pelteobagrus vachelli individuals after S1 cultivation, carrying out 3 times of artificial infection tests after clinical anatomy observation and bacteria separation and identification, and selecting female pelteobagrus fulvidraco and male pelteobagrus vachelli of F1 generation and F2 generation groups with 3 times of infection with antibacterial advantages through the tests;
s3, crossbreeding of an antibacterial family: carrying out artificial spawning and artificial insemination on the antibacterial dominant object selected in the step S2;
s4, F1 generation anti-bacterial strain is obtained: repeating the steps of artificial spawning, artificial fertilization and artificial hatching in the step S3, and obtaining the pelteobagrus fulvidraco multiplied by pelteobagrus vachelli antibacterial fish fries through the hybridization breeding.
Preferably, the method for generating the F1 and F2 generations by selfing in the S1 is to divide the obtained wild pelteobagrus fulvidraco and pelteobagrus vachelli into pools for feeding; the pelteobagrus fulvidraco is sexually mature in two ages; in 5-7 months of the next year, artificial spawning is respectively carried out on female fish and male fish, and fertilization activities are carried out at night when weather changes from sunny to cloudy; artificial spawning is carried out to obtain larval fish, and the larval fish is cultivated until the larval fish can freely swim for 7-8 days; obtaining F1-generation pelteobagrus fulvidraco and F1-generation pelteobagrus vachelli generated by selfing, and continuing to carry out pond-dividing cultivation; the selfing breeding of the F1 generation is carried out in the next year, and the F2 generation is cultivated.
Preferably, the clinical anatomical observation in S2 is that the pelteobagrus fulvidraco with obvious disease symptoms is brought back to a laboratory for routine dissection examination, and the symptoms of the body surface, gill and viscera of the disease fish, the presence or absence of parasites and the presence or absence of parasites are observed and inspected by a method combining naked eyes observation and microscopic examination, and records are made.
Preferably, the method for separating and identifying the bacteria in the S2 is to sample heart, liver and brain tissues of the diseased fish under a sterile condition, streak and inoculate on a common solid culture medium; culturing in a constant temperature incubator at 30deg.C for 24 hr, picking dominant colony, inoculating into liquid culture medium, and purifying to obtain strain; placing the strain in a refrigerator at 4 ℃ for standby; taking purified isolated strains, and observing the morphology and staining characteristics of bacteria under a microscope oil microscope after gram staining; the strain is gram-negative bacteria; 1 drop of oxidase reagent is dropped on a plurality of single colonies of the fresh culture of the isolated strain, and the oxidase test results are positive; under the aseptic condition, the isolated strain after rejuvenation is picked up for 3 times and is blown and sucked uniformly in a test tube filled with sterile PBS liquid to prepare bacterial suspension, the bacterial culture liquid is used as a raw material, PCR amplification is carried out on the 16S sequence of the bacterial suspension, and the bacterial suspension is compared with sequence information in NCBI, and the species of the bacterial suspension is identified and determined.
Preferably, the method of the artificial infection test is to adjust the concentration of bacterial liquid to about 10cfu/mL by combining a Mitsubishi tube method and a coating counting method; carrying out 3 times of artificial infection tests, wherein the first artificial infection objects are F1 generations of pelteobagrus fulvidraco and pelteobagrus vachelli; taking individuals dying or just dying in the 1 st F1 generation test from the 2 nd artificial infection test, and then separating and purifying bacteria to infect F2 generation pelteobagrus fulvidraco and pelteobagrus vachelli; infection 3 all parent fish of the F1 and F2 generations were subjected to an infection test with 10cfu/mL of bacterial liquid in a secondary experiment.
Preferably, the weight of the female pelteobagrus fulvidraco in the antibacterial dominant object selected in S3 is more than 80g and the weight of the male pelteobagrus vachelli is more than 500 g.
Preferably, in the step S3, when the water temperature of the fish pond is stable to be more than 24 ℃, artificial spawning is implemented on the female pelteobagrus fulvidraco and the male pelteobagrus vachelli in the fish pond, and the method for artificial spawning comprises the steps of using oxytocin for every 1Kg of female pelteobagrus fulvidraco: chorionic gonadotrophin 2000-3000IU, luteinizing hormone releasing hormone 15-24 mug, and diburone 1.5-3.0mg; the female pelteobagrus fulvidraco adopts a 2-time injection method, the 1 st injection is 20-40% of the total oxytocin dosage of the female pelteobagrus fulvidraco, the 2 nd injection is 60-80% of the total oxytocin dosage of the female pelteobagrus fulvidraco, and the time interval between the two injections is 12-14h; 1 injection of male pelteobagrus vachelli, and the 2 nd injection time synchronization of female pelteobagrus vachelli, the injection amount is 40-60% of the total oxytocin dosage of female pelteobagrus vachelli; after oxytocin injection, female pelteobagrus fulvidraco and male pelteobagrus vachelli are temporarily raised in a fish pond, and the fish pond is oxygenated by an oxygen pump.
Preferably, the artificial fertilization method is that after spawning is induced for 6-8 hours, parent fish is found to chase, female pelteobagrus fulvidraco is taken out from a fish pond, egg grains are placed into a container after the abdomen of the female pelteobagrus fulvidraco is extruded, and then the body surface of the female pelteobagrus fulvidraco is wiped dry; simultaneously, wiping the body surface of the male pelteobagrus vachelli, cutting open the abdomen, taking out the testis, grinding the testis with a grinding bowl, adding physiological saline, stirring and pouring into the container, uniformly mixing with egg grains, adding yellow mud water for debonding, and finally cleaning fertilized eggs with clear water for 2/3 times.
Preferably, the artificial incubation is carried out by putting fertilized eggs into an incubation barrel for running water inflation incubation, wherein the quantity of fertilized eggs in the incubation barrel is 80 ten thousand-100 ten thousand grains/m < 3 >; the periphery of the hatching barrel is wrapped by 60 mesh fabrics.
Compared with the prior art, the invention has the beneficial effects that:
the method has the advantages that the large-scale production of pelteobagrus fulvidraco fries is realized, the application and popularization of key technology application and popularization demonstration bases of the fries for large-scale breeding are established, and the development of the traditional pelteobagrus fulvidraco fish farming industry and the development of new special freshwater aquaculture fish varieties are promoted; providing more sufficient freshwater fish seedlings, enriching the variety of pond culture, being beneficial to the continuous proliferation of natural resource population and protecting the diversity of organisms. Meanwhile, the industrial association and the radiation degree are continuously improved, the industrial chain for producing, selling and selling is orderly extended, the development of industries such as living and fresh transportation, tool manufacturing for fishing, feed and bait is driven, the problem of the outlet of a large amount of local fishery labor force is solved, and the ecological and social benefits are remarkable.
Drawings
FIG. 1 is a schematic representation of a cross-breeding method.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Referring to fig. 1, the present invention provides a technical solution: a cross breeding method for artificially breeding and breeding pelteobagrus fulvidraco with disease resistance is characterized by comprising the following steps:
s1, family cultivation: after carrying out wild resource investigation of pelteobagrus fulvidraco and pelteobagrus vachelli, selecting disease-free and injury-free healthy pelteobagrus vachelli and pelteobagrus vachelli; then generating F1 and F2 generation by selfing respectively;
s2, disease-resistant breeding: selecting diseased pelteobagrus fulvidraco and pelteobagrus vachelli individuals after S1 cultivation, carrying out 3 times of artificial infection tests after clinical anatomy observation and bacteria separation and identification, and selecting female pelteobagrus fulvidraco and male pelteobagrus vachelli of F1 generation and F2 generation groups infected for the 3 times with antibacterial advantages through the tests;
s3, crossbreeding of an antibacterial family: carrying out artificial spawning and artificial insemination on the antibacterial dominant object selected in the step S2;
s4, F1 generation anti-bacterial strain is obtained: repeating the steps of artificial spawning, artificial fertilization and artificial hatching in the step S3, and obtaining the pelteobagrus fulvidraco multiplied by pelteobagrus vachelli antibacterial fish fries through the hybridization breeding.
The method for generating F1 and F2 generations by selfing in the S1 comprises the steps of feeding the obtained wild pelteobagrus fulvidraco and pelteobagrus vachelli in separate pools; the pelteobagrus fulvidraco is sexually mature in two ages; in 5-7 months of the next year, artificial spawning is respectively carried out on female fish and male fish, and fertilization activities are carried out at night when weather changes from sunny to cloudy; artificial spawning is carried out to obtain larval fish, and the larval fish is cultivated until the larval fish can freely swim for 7-8 days; obtaining F1-generation pelteobagrus fulvidraco and F1-generation pelteobagrus vachelli generated by selfing, and continuing to carry out pond-dividing cultivation; and breeding the F1 generation in the next year, and breeding the F2 generation.
The clinical anatomy observation in the step S2 is that the pelteobagrus fulvidraco with obvious disease symptoms is carried back to a laboratory for routine section examination, and the symptoms of the body surface, gill and viscera of the disease fish, the existence of parasites and the like are observed and inspected by a method combining naked eyes observation and microscopic examination, and records are made.
The method for separating and identifying the bacteria in the S2 comprises sampling heart, liver and brain tissues of a diseased fish under a sterile condition, streaking and inoculating on a common solid culture medium; culturing in a constant temperature incubator at 30deg.C for 24 hr, picking dominant colony, inoculating into liquid culture medium, and purifying to obtain strain; placing the strain in a refrigerator at 4 ℃ for standby; taking purified isolated strains, and observing the morphology and staining characteristics of bacteria under a microscope oil microscope after gram staining; the strain is gram-negative bacteria; 1 drop of oxidase reagent is dropped on a plurality of single colonies of the fresh culture of the isolated strain, and the oxidase test results are positive; under the aseptic condition, the isolated strain after rejuvenation is picked up for 3 times and is blown and sucked uniformly in a test tube filled with sterile PBS liquid to prepare bacterial suspension, the bacterial culture liquid is used as a raw material, PCR amplification is carried out on the 16S sequence of the bacterial suspension, and the bacterial suspension is compared with sequence information in NCBI, and the species of the bacterial suspension is identified and determined.
Wherein, the method of the artificial infection test is to combine the Mitsubishi turbidimetry method and the coating counting method to adjust the concentration of bacterial liquid to about 10 cfu/mL; carrying out 3 times of artificial infection tests, wherein the first artificial infection objects are F1 generations of pelteobagrus fulvidraco and pelteobagrus vachelli; taking individuals dying or just dying in the 1 st F1 generation test from the 2 nd artificial infection test, and then separating and purifying bacteria to infect F2 generation pelteobagrus fulvidraco and pelteobagrus vachelli; infection 3 all parent fish of the F1 and F2 generations were subjected to an infection test with 10cfu/mL of bacterial liquid in a secondary experiment.
Wherein the weight of the female pelteobagrus fulvidraco in the antibacterial dominant object selected in the step S3 is more than 80g, and the weight of the male pelteobagrus vachelli is more than 500 g.
When the water temperature of the fish pond is stable and reaches more than 24 ℃, the artificial spawning is implemented for the female pelteobagrus fulvidraco and the male pelteobagrus vachelli in the fish pond, and the artificial spawning method comprises the steps of using oxytocin for every 1Kg of female pelteobagrus fulvidraco: chorionic gonadotrophin 2000-3000IU, luteinizing hormone releasing hormone 15-24 mug, and diburone 1.5-3.0mg; the female pelteobagrus fulvidraco adopts a 2-time injection method, the 1 st injection is 20-40% of the total oxytocin dosage of the female pelteobagrus fulvidraco, the 2 nd injection is 60-80% of the total oxytocin dosage of the female pelteobagrus fulvidraco, and the time interval between the two injections is 12-14h; 1 injection of male pelteobagrus vachelli, and the 2 nd injection time synchronization of female pelteobagrus vachelli, the injection amount is 40-60% of the total oxytocin dosage of female pelteobagrus vachelli; after oxytocin injection, female pelteobagrus fulvidraco and male pelteobagrus vachelli are temporarily raised in a fish pond, and the fish pond is oxygenated by an oxygen pump.
The artificial fertilization method comprises the steps of finding parent fish after spawning is induced for 6-8 hours, taking out female pelteobagrus fulvidraco from a fish pond, extruding the abdomen of the female pelteobagrus fulvidraco, placing egg particles into a container, and then wiping the body surface of the female pelteobagrus fulvidraco; simultaneously, wiping the body surface of the male pelteobagrus vachelli, cutting open the abdomen, taking out the testis, grinding the testis with a grinding bowl, adding physiological saline, stirring and pouring into the container, uniformly mixing with egg grains, adding yellow mud water for debonding, and finally cleaning fertilized eggs with clear water for 2/3 times.
Wherein, the artificial incubation is that fertilized eggs are put into an incubation barrel for running water aeration incubation, and the fertilized eggs in the incubation barrel are 80 ten thousand-100 ten thousand grains/m 3; the periphery of the hatching barrel is wrapped by 60 mesh fabrics.
Examples:
1. parent fish cultivation and breeding:
the test is carried out on a test base in a key laboratory for protecting fish resources at the upstream of the Yangtze river; after investigation and collection of wild resources of pelteobagrus fulvidraco and pelteobagrus vachelli stock of different geographical groups are carried out, selecting female parent 300 tails with the weight of more than 80g and male parent 5 tails with the weight of more than 500g of male pelteobagrus vachelli in 5 months, wherein the female parent is large in individual and the male parent is free from diseases and injuries; after the female fish is cultivated to sexual maturity, the female fish is strictly required to have round and full abdomen, the reproductive hole is light red, and the flow signs of the egg grains of the abdomen can be observed by inverting; the male fish is required to have larger reproduction protrusion and is peach red;
when the water temperature of the fish pond is stable and reaches more than 24 ℃, carrying out artificial spawning on female pelteobagrus fulvidraco and male pelteobagrus vachelli in the fish pond; the female pelteobagrus fulvidraco adopts a 2-time injection method, the 1 st injection is 20-40% of the total oxytocin dosage of the female pelteobagrus fulvidraco, the 2 nd injection is 60-80% of the total oxytocin dosage of the female pelteobagrus fulvidraco, and the time interval between the two injections is 12-14 hours; 1 injection of male pelteobagrus vachelli, and 2 nd injection time synchronization of female pelteobagrus vachelli, wherein the injection amount is 40-60% of total oxytocin consumption of female pelteobagrus vachelli, and pectoral fin base injection;
the oxytocin and the oxytocin for combined injection thereof are as follows: luteinizing hormone releasing hormone (Luteinizing hormone releasing hormone A, LHRH-A2), diesterone (DOM), and chorionic gonadotrophin (Human chorionicgonadotropin, HCG), which are mixtures of luteinizing hormone releasing hormone, diesterone, and chorionic gonadotrophin; 2500IU of chorionic gonadotrophin, 20 mug of luteinizing hormone releasing hormone and 10mg of diolone for each 1Kg female pelteobagrus fulvidraco; the oxytocin combination can obviously improve the propagation effect of the hybrid pelteobagrus fulvidraco, so that the effect time is reduced from 30 hours to 23 hours;
after oxytocin injection, temporarily culturing female pelteobagrus fulvidraco and male pelteobagrus vachelli in a fish pond, and oxygenation of the fish pond by an oxygen pump is carried out, and medium-strength running water is maintained; observing the activity condition of parent fish at regular time, after the parent fish has obvious oestrus symptoms, pulling a net timely for inspection so as to prevent the egg grains from overripening and degrading, taking out the female pelteobagrus fulvidraco from a pool, extruding the abdomen of the female pelteobagrus fulvidraco, placing the egg grains into a container, and then wiping the body surface of the female pelteobagrus fulvidraco; simultaneously, wiping the body surface of the male pelteobagrus vachelli, cutting open the abdomen, taking out the spermary, grinding the spermary with a grinding bowl, adding normal saline, stirring and pouring into the container, uniformly mixing with egg grains, adding yellow mud water for debonding, and finally cleaning fertilized eggs with clear water for 2-3 times; placing the hatching barrel into a hatching barrel for running water inflation hatching;
the fertilized eggs in the hatching barrel are 80 ten thousand to 100 ten thousand grains/m < 3 >; the upper half part of the artificial hatching barrel is a cylinder with the diameter of 0.5m, the lower half part is an inverted cone, microporous oxygenation equipment is erected in the barrel, and the bottom is provided with a water inlet pipe; wrapping the periphery of the barrel with 60 mesh cloth; the hatching water is treated river water, and the water inlet is filtered by a 80-mesh filter screen; keeping running water and inflating in the hatching process; forming pelteobagrus fulvidraco fries with excellent hybridization advantages, carrying out scale production, obtaining 500 groups of excellent parents, and establishing pelteobagrus fulvidraco X pelteobagrus vachelli disease-resistant family core groups;
2. carrying out F1 generation artificial breeding, pairing parent fishes one by one, and mating to reproduce F2 generation:
and selecting 300 female parents from 5 months after the third year, breeding the male parents again from 5 female parents to obtain F1 generation of pelteobagrus fulvidraco multiplied by pelteobagrus vachelli, then measuring the weight for 6 times in the growth period of 2 years, and breeding a variety with excellent growth characteristics as a family.
In the description of the present invention, the detailed description of the embodiments of the invention is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which may be made by those of ordinary skill in the art without the benefit of the present disclosure, are intended to be within the scope of the present disclosure wherein the terms "F1", "F2" are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated, whereby a feature defining "F1", "F2" may explicitly or implicitly include at least one such feature.
Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made therein without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (5)
1. A cross-breeding method for artificially breeding and selectively breeding pelteobagrus fulvidraco with disease resistance is characterized in that
The method comprises the following steps: s1, family cultivation: after carrying out wild resource investigation of pelteobagrus fulvidraco and pelteobagrus vachelli, selecting two-year pelteobagrus fulvidraco and two-year pelteobagrus vachelli which are free from diseases and injuries as parents; then generating F1 generation and F2 generation by selfing respectively;
s2, disease-resistant breeding: selecting diseased pelteobagrus fulvidraco and pelteobagrus vachelli individuals after S1 cultivation, carrying out 2 times of artificial infection tests after clinical anatomy observation and bacteria separation and identification, and selecting female pelteobagrus fulvidraco and male pelteobagrus vachelli of F2 generation group with antibacterial advantage through the tests;
s3, crossbreeding of an antibacterial family: carrying out artificial spawning and artificial insemination on the antibacterial dominant object selected in the step S2;
s4, obtaining a hybrid F1 generation: repeating the artificial spawning, artificial fertilization and artificial incubation steps in S2
Obtaining pelteobagrus fulvidraco multiplied by pelteobagrus vachelli antibacterial fish fries through the hybridization;
the method for generating F1 and F2 generations by respective selfing in S1 comprises the steps of feeding the obtained wild pelteobagrus fulvidraco and pelteobagrus vachelli in separate pools;
the pelteobagrus fulvidraco is sexually mature in two ages;
in 5-7 months of the next year, carrying out artificial induced spawning on female fish and male fish respectively;
artificial spawning is carried out to obtain larval fish, and the larval fish is cultivated until the larval fish can freely swim for 7-8 days;
obtaining F1-generation pelteobagrus fulvidraco and F1-generation pelteobagrus vachelli generated by selfing, and continuing to carry out pond-dividing cultivation; breeding F1 generation filial generation in the next year, and breeding F2 generation;
the clinical anatomical observation in the step S2 is that after the pelteobagrus fulvidraco with obvious disease symptoms is brought back to a laboratory, routine sectioning examination is carried out, and the symptoms of the body surface, gill and viscera of the disease fish, the existence of parasites and the existence of parasites are observed and checked by a method combining naked eyes observation and microscopic examination, and records are made;
the method for separating and identifying the bacteria in the S2 comprises the steps of sampling heart, liver and brain tissues of a diseased fish under a sterile condition, streaking and inoculating the heart, liver and brain tissues on a common solid culture medium; culturing in a 30 ℃ constant temperature incubator for 24 hours, picking dominant bacterial colonies, inoculating in a liquid culture medium, and purifying to obtain bacterial strains; placing the strain in a refrigerator at 4 ℃ for standby; taking purified isolated strains, carrying out gram staining on the isolated strains, and observing the morphology and staining characteristics of bacteria under a microscope oil microscope; the strain is gram-negative bacteria; 1 drop of oxidase reagent is dropped on a plurality of single colonies of the fresh culture of the isolated strain, and the oxidase test results are positive; under the aseptic condition, the isolated strain after rejuvenation is picked up for 3 times and is blown and sucked uniformly in a test tube filled with sterile PBS liquid to prepare bacterial suspension, the bacterial culture liquid is used as a raw material, PCR amplification is carried out on the 16S sequence of the bacterial suspension, and the bacterial suspension is compared with sequence information in NCBI, and the species of the bacterial suspension is identified and determined.
2. The method for artificially propagating and breeding pelteobagrus fulvidraco crossbreeding with disease resistance according to claim 1, which is characterized in that: and (3) the weight of the female pelteobagrus fulvidraco in the antibacterial dominant object selected in the step (S3) is more than 80g, and the weight of the male pelteobagrus vachelli is more than 500 g.
3. The method for artificially propagating and breeding pelteobagrus fulvidraco crossbreeding with disease resistance according to claim 1, which is characterized in that: in the step S3, when the water temperature of the fish pond is stable and reaches more than 24 ℃, artificial spawning is carried out on female pelteobagrus fulvidraco and male pelteobagrus vachelli in the fish pond, and the artificial spawning method comprises the steps of using spawning element for every 1Kg of female pelteobagrus fulvidraco: chorionic gonadotrophin 2000IU, luteinizing hormone releasing hormone 15 μg, diurone 5mg; the female pelteobagrus fulvidraco adopts a 2-time injection method, the 1 st injection is 40% of the total oxytocin consumption of the female pelteobagrus fulvidraco, the 2 nd injection is 60% of the total oxytocin consumption of the female pelteobagrus fulvidraco, and the time interval between the two injections is 12h; 1 injection of male pelteobagrus vachelli is synchronous with the 2 nd injection time of female pelteobagrus vachelli, and the injection amount is 50% of the total oxytocin consumption of the female pelteobagrus vachelli; after oxytocin injection, female pelteobagrus fulvidraco and male pelteobagrus vachelli are temporarily raised in a fish pond, and the fish pond is oxygenated by an oxygen pump.
4. The method for artificially breeding and breeding pelteobagrus fulvidraco crossbreeding with disease resistance according to claim 3, which is characterized in that: the artificial fertilization method comprises the steps of finding parent fish after 6.5 hours of spawning induction, taking out female pelteobagrus fulvidraco from a fish pond, extruding the abdomen of the female pelteobagrus fulvidraco, placing egg particles into a container, and then wiping the body surface of the female pelteobagrus fulvidraco; simultaneously, wiping the body surface of the male pelteobagrus vachelli, cutting open the abdomen, taking out the testis, grinding the testis with a grinding bowl, adding physiological saline, stirring and pouring into the container, uniformly mixing with egg grains, adding yellow mud water for debonding, and finally cleaning fertilized eggs with clear water for 3 times.
5. The method for artificially propagating and breeding pelteobagrus fulvidraco crossbreeding with disease resistance according to claim 4, which is characterized in that: the artificial incubation is to put fertilized eggs into an incubation barrel for running water inflation incubation, wherein the fertilized eggs in the incubation barrel are 80 ten thousand grains/m < 3 >; the periphery of the hatching barrel is wrapped by 60 mesh fabrics.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CA2371460A1 (en) * | 2001-12-13 | 2003-06-13 | Purdue Research Foundation | Cell culture system and methods of use |
| CN104480029A (en) * | 2014-11-20 | 2015-04-01 | 西北农林科技大学 | Wine yeast capable of low-yielding hydrogen sulfide and ethyl carbamate as well as screening method and application of wine yeast |
| CN106577388A (en) * | 2016-12-14 | 2017-04-26 | 华中农业大学 | Cross breeding method for Pelteobagrus fulvidraco variety namely Huangyou No. 1 with improved growth character |
| CN110463659A (en) * | 2019-09-11 | 2019-11-19 | 内江师范学院 | A kind of Kang Channel-catfish tarda hybridization Pelteobagrus fulvidraco excellent strain selection |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2371460A1 (en) * | 2001-12-13 | 2003-06-13 | Purdue Research Foundation | Cell culture system and methods of use |
| CN104480029A (en) * | 2014-11-20 | 2015-04-01 | 西北农林科技大学 | Wine yeast capable of low-yielding hydrogen sulfide and ethyl carbamate as well as screening method and application of wine yeast |
| CN106577388A (en) * | 2016-12-14 | 2017-04-26 | 华中农业大学 | Cross breeding method for Pelteobagrus fulvidraco variety namely Huangyou No. 1 with improved growth character |
| CN110463659A (en) * | 2019-09-11 | 2019-11-19 | 内江师范学院 | A kind of Kang Channel-catfish tarda hybridization Pelteobagrus fulvidraco excellent strain selection |
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