CN101884311B - Method for constructing tongue sole families and breeding superior families - Google Patents
Method for constructing tongue sole families and breeding superior families Download PDFInfo
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Abstract
The invention relates to a method for constructing tongue sole families and breeding good families, which comprises the technical content of breeding a tongue sole group, constructing the tongue sole families and screening a disease-resistant family and a fast growing family of tongue sole. The method comprises the following steps of: constructing the families through the group breeding and intraspecific hybridization technology; then, screening out the family resisting vibrio anguillarum disease from a large amount of family by an artificial infection approach, and screening out the fast growing family by growth comparison. The fries in the good families can be directly popularized and applied, or used for breeding a high-yield and disease-resistant good strain of tongue sole. The invention has the characteristics of strong pertinence, high breeding efficiency, obvious effect and the like. The established method for screening good tongue sole families provides a new technical approach for breeding the high-yield and disease-resistant good strain of tongue sole. The method is suitably popularized and applied to other cultured fishes, and has important significance and application value for breeding high-yield and disease-resistant strains of the cultured fishes.
Description
Technical field
The invention belongs to the fish fine-variety breeding method in the aquaculture field, is that a kind of family that can use in the seed selection of Cynoglossus semilaevis excellent strain makes up and selection.
Background technology
Cynoglossus semilaevis (
Cynoglossus semiliaevis) be the distinctive a kind of famous and precious economic seawater fish of China, belong to coastal waters warm water demersal fishes, China is coastal all distribution, is many with the Huanghai Sea, the Bohai Sea.Cynoglossus semilaevis is welcome by consumers in general deeply owing to its delicious flavour, fine and tender taste, nutritious, its market value is high, and the breed prospect is boundless.Cynoglossus semilaevis is one of principal item of present marine fish culture, and nearly ten thousand tons at present of its cultured outputs cost an arm and a leg, and market price reaches about 200 yuan/1000 grams, and the annual at present output value of cynoglossus semilaevis cultivation industry is about 2,000,000,000 yuans.
But along with the fast development of cynoglossus semilaevis cultivation industry, some problems occur in succession.Because long-term inbreeding causes the decline of seed growth rate, the resistance against diseases of Cynoglossus semilaevis artificial propagation to reduce, disease takes place frequent, causes cynoglossus semilaevis cultivation fry and fingerling mortality.What influence that the main disease of Cynoglossus semilaevis finds at present mainly comprises bacillary and viral disease, and is the bigger a kind of disease of harm Cynoglossus semilaevis by the bacteriosis that Vibrio anguillarum causes.Every year, due to illness evil caused cynoglossus semilaevis cultivation already to lose several hundred million yuans.Although take traditional prophylactic measures certain effect to be arranged as using medicines such as antibiotic class, but these medicines are easy to make pathogen to develop immunity to drugs, simultaneously breeding environment is also produced pollution, particularly at the fish cylinder accumulation, human beings'health is had potential hazard, thereby the application in actual production is restricted more and more.Therefore, cultivate fast growth, Cynoglossus semilaevis excellent strain that resistance against diseases is strong becomes the important topic of demanding urgently capturing in the cynoglossus semilaevis cultivation industry.
In view of the breed potentiality of Cynoglossus semilaevis and potential economic and social benefits, in recent years, China scientific worker has carried out big quantity research to Cynoglossus semilaevis genital regulating, sex controlling and cultural technique, comprise that mainly the method that adopts Artificial Control illumination and water temperature stimulates Cynoglossus semilaevis parent population maturation and natural spawning, this technology was succeedd in 2002, and had produced semi-smooth tongue sole offspring breed.In addition, studies show that, Cynoglossus semilaevis female individuals and male, have significant difference on growth rate, its female individuals is than the big 2-4 of male times of (Inst of Huanghai Sea Marine Products, Chinese Academy of Aquatic Product Science, Meng Tianxiang etc., 1988), after the Cynoglossus semilaevis fry was cultured through 1 year, female individuals can be grown the 500-750 gram, and male then has only the 50-150 gram.Because male was grown slow, influenced the quality of marketable fish, reduced the cultured output of Cynoglossus semilaevis, increased aquaculture cost, thereby the popularization of semi-smooth tongue sole offspring breed and the development of aquaculture industry have been had a strong impact on, Given this, in recent years, domestic scholars has been carried out system research to cynoglossus semilaevis gynogenesis and sex controlling technology.As: the research (Inst of Huanghai Sea Marine Products, Chinese Academy of Aquatic Product Science that filters out Cynoglossus semilaevis special numerator mark, Chen SL et al., 2007), set up molecular marker auxiliary sex control technology (Chen SL et al., 2008), with the female ratio of Cynoglossus semilaevis fry by bringing up to about 70% less than 50%; In addition, also set up Cynoglossus semilaevis artificial gynogenesis technology (Chen SL et al., 2009), thereby part has solved the male poky problem of Cynoglossus semilaevis.But be difficult to solve the problem that the Cynoglossus semilaevis germplasm is degenerated and disease takes place frequently by these technology, family selective breeding then is one of main means of carrying out disease-resistant high yield fine breeding.But up to the present, the research about tongue sole families foundation and quick growth and the strong superior families screening of premunition aspect does not appear in the newspapers as yet.
Summary of the invention
The purpose of this invention is to provide a kind of constructing tongue sole families and disease-resistant quick growth family method for screening, disease-resistant and high-yield variety is cultivated technical method is provided for Cynoglossus semilaevis, for fish culture already provides technological means disease-resistant and the high-yield variety seed selection.
The technology of the present invention thes contents are as follows: comprising: one, Cynoglossus semilaevis colony seed selection; Two, constructing tongue sole families; Three, the screening of the disease-resistant family of Cynoglossus semilaevis and the quick family of growing.
One, Cynoglossus semilaevis colony seed selection: comprising: the Cynoglossus semilaevis wild population, propagate the seed selection of colony artificially.
The seed selection of wild population: be to collect the Cynoglossus semilaevis fry or the fingerling of fishing for from Bohai Sea Gulf, select body surface not have scar, healthy energetic fry or fingerling are as wild population, put in a suitable place to breed in cement pit and culture, when fry grows into 12-18 centimetre, after identifying the sex of fry, female, milter seedling are separately cultured.After the 2-3 breed, from raun colony and milter colony, respectively select individual big 40-50 tail fish respectively as female, male parent population according to 1% ratio again, be used for family and make up.
Propagate the seed selection of colony artificially: propagate the fry that colony is the artificial propagation of cynoglossus semilaevis cultivation field artificially, filtering out the fast individuality of growth from the fry that cultivate cynoglossus semilaevis cultivation field, Shandong cultivates, selection percentage is 0.5-1%, promptly from the female adult fish of 10000 tail Cynoglossus semilaevis of propagating 2-3 artificially, select maximum individual 50 tails as female parent population, from propagate the male adult fish of 2500-5000 tail Cynoglossus semilaevis in 2 years artificially, select individual big 25-50 tail as male parent population, through behind the reinforced cultivating, use parent population as breeding, be used for family and make up.
Two, constructing tongue sole families:
From the Cynoglossus semilaevis wild population with propagate artificially and choose sexually matured healthy parent population the colony, annual 6-9 month by temperature control control photoinduction parent population gonad development and maturation.When the raun belly obviously swells, sexual gland length reaches the long about half of body, female milter is carried out artificial induced spawning, obtain seminal fluid of ripe milter and the ovum of ripe raun.Be used to carry out the dry method insemination from the seminal fluid of 1 tail milter with from the ovum of 1 tail raun, the fertilized egg that mating is produced is placed in the net cage with miniflow water and little inflation hatches, and a net cage is only put the fertilized egg of a family.When fry is about to incubation of membrane, it is transferred in the cylinder of growing seedlings of 2-3 cubic meter volume cultivate, only put the fry of a family in each cylinder, every cubic metre of water body is cultivated 5000-8000 tail prelarva.Should set up 20-40 family, each family should be cultivated 500-1000 tail fry and is used for disease-resistant and the screening high yield family at every turn.
Three, the screening of the disease-resistant family of Cynoglossus semilaevis and the quick family of growing.
1). the screening of the disease-resistant family of Cynoglossus semilaevis:
When each family fry of Cynoglossus semilaevis grew to 10-12cm, adopting soaking method carries out Vibrio anguillarum to each family fry to be infected, and estimated the sick ability of anti-vibrio anguillarum of each family fry, infected the cultivation of used Vibrio anguillarum bacterial strain and carried out according to conventional method.Infect experiment at first in advance and determine bacterium liquid half lethal concentration, the pre-fry that infects experiment usefulness is the population mixture from the equal number fry of 10 familys, and each family is got 50-100 tail fry and mixed.During pre-the infection, cultured bacterium liquid is diluted to the bacterium liquid that concentration is 2.0-3.0 OD with physiological saline, above-mentioned bacterium liquid is diluted to the (dilution 50 of different concentration with fresh seawater, 100,200,300,400,600 times), to mix fry and be divided into 6 groups at random, every group of 80-150 tail fry, every group of 80-150 tail mixed family fry is placed on soaks after 3-5 hour in the seawater of a concentration bacterium liquid, 6 groups of fries are placed on to soak in the seawater of 6 different bacterial concentrations and infect, water temperature is controlled at 21-23 ℃ during infection, after the infection fry is forwarded in the seawater of miniflow water and little inflation and raise, in subsequently 15-20 days, calculate the fry lethality of each concentration group, will cause that the bacterial concentration of 40-60% fry death is defined as half lethal concentration.
Carry out formal infection experiment according to half lethal concentration, the infection method is tested with pre-the infection.Different familys infect respectively, and each family infects 50-100 tail fry at every turn, repeated experiments 1 time.Infect with fish culture at 0.3-1m
3The fiberglass tank in.After the infection, carry out observed and recorded to state and the dead quantity of each family fry every day, measures dead individual total length and body weight.After infection 20-30 days, write down the fry survival quantity of all familys, measure total length and the body weight of survival fry.According to the quantity and the metainfective survival quantity of participating in the infection fry, calculate the fry survival rate of each family, estimate the disease-resistant performance of each family.Survival rate is defined as the strong family of premunition in the family more than 55%, survival rate is defined as the strong family of premunition in the family of 40-55%, survival rate is defined as the general family of premunition in the family of 20-40%, and survival rate is defined as the family of premunition difference in the family below 20%.
2), the Cynoglossus semilaevis screening of family of growing fast:
When each family fry of Cynoglossus semilaevis grows to total length and is 10-12cm, from the fry that each family does not infect, choose 200 tails, adopt the fry of the different familys of fluorescent pigment mark, the fry behind the mark is raised together with the m at 35-40
3Cement pit in.After culturing 8-15 month, collect each family fry and carry out total length and measured body weight.The fry sampling 60-100 tail of each family is measured, and calculates the mean of each family total length and body weight.Utilize daily gain rate (weight that increases during the breed/actual culturing time) to estimate the growth performance of different familys.The daily gain rate is big more, and its growth rate is fast more, growth performance is good more.The daily gain rate is defined as growing fast family in the family more than 0.25, and the family of daily gain rate between 0.2-0.25 is defined as the faster growing family; The daily gain rate is defined as the general family of growth rate in the family below 0.2.
The present invention and prior art contrast are characterized in:
The present invention adopts colony's seed selection and intraspecific hybridization technology to set up family, adopt artificial infection's approach from a large amount of familys, to filter out the family of anti-vibrio anguillarum disease then, filter out the fast family of growth by growth fraction, these superior families fries can directly be applicable, or are used for the cultivation of Cynoglossus semilaevis high-yield disease resisting improved seeds.
Pointed strong, the seed selection efficient height of the present invention, simple to operate, characteristics such as effect is obvious, the Cynoglossus semilaevis superior families screening technique of setting up is that new technological approaches has been opened up in the cultivation of Cynoglossus semilaevis high-yield disease resisting improved seeds, suit in other cultured fishes, to apply, to the significant and using value of cultured fishes high-yield disease resisting prevalent variety cultivation.
Embodiment:
Below by the structure of tongue sole families and the screening of quick growth and disease-resistant family, technology contents of the present invention is described in detail:
The technology of the present invention thes contents are as follows: comprising: one, Cynoglossus semilaevis colony seed selection; Two, constructing tongue sole families; Three, the screening of the disease-resistant family of Cynoglossus semilaevis and the quick family of growing.
One, Cynoglossus semilaevis colony seed selection: comprising: the Cynoglossus semilaevis wild population, propagate the seed selection of colony artificially.
The seed selection of wild population: be to collect the Cynoglossus semilaevis fry or the fingerling of fishing for from Bohai Sea Gulf, select body surface not have scar, healthy energetic fry or fingerling are as wild population, put in a suitable place to breed in cement pit and culture, when fry grows into 12-18 centimetre, after identifying the sex of fry, female, milter seedling are separately cultured.After the 2-3 breed, from raun colony and milter colony, respectively select individual big 40-50 tail fish respectively as female, male parent population according to 1% ratio again, be used for family and make up.
Propagate the seed selection of colony artificially: propagate the fry that colony is the artificial propagation of cynoglossus semilaevis cultivation field artificially, filtering out the fast individuality of growth from the fry that cultivate cynoglossus semilaevis cultivation field, Shandong cultivates, selection percentage is 0.5-1%, promptly from the female adult fish of 10000 tail Cynoglossus semilaevis of propagating 2-3 artificially, select maximum individual 50 tails as female parent population, from propagate the male adult fish of 2500-5000 tail Cynoglossus semilaevis in 2 years artificially, select individual big 25-50 tail as male parent population, through behind the reinforced cultivating, use parent population as breeding, be used for family and make up.
The present invention is since in October, 2003, the a large amount of Cynoglossus semilaevis fry or fingerlings of fishing for from Bohai Sea Gulf collected are cultured, collect and cultured the wild fry of tail more than 8000 altogether, when these fries grow into more than 13 centimetres, after identifying the sex of fry, female, milter seedling are separately cultured, through propagating artificially of 3-4, therefrom filtered out female adult fish tail more than 40 and male adult fish tail more than 40 of quick growth in 2007, put in a suitable place to breed in different cement pits and culture, be used for artificial induced spawning and family and make up.Propagate artificially the female fry of tail more than 10000 of colony for the artificial propagation of culturing and filter out the fastest female adult fish tail more than 50 of growth from cynoglossus semilaevis cultivation field, Shandong, with from the male fry of tail more than 5000, filter out male adult fish tail more than 50, be used for artificial induced spawning and family and make up.Through reaching sexual maturity behind the reinforced cultivating, select individual big, no wound, gonad development is good, the individuality with vigor is used fish as breeding.
Two, constructing tongue sole families:
The Cynoglossus semilaevis wild population that utilization filters out and propagate colony artificially as basic population adopts the method for intraspecific hybridization to set up tongue sole families.From the Cynoglossus semilaevis wild population with propagate artificially and choose sexually matured healthy parent population the colony, annual 6-9 month by temperature control control photoinduction parent population gonad development and maturation.When the raun belly obviously swells, sexual gland length reaches the long about half of body, female milter is carried out artificial induced spawning, obtain seminal fluid of ripe milter and the ovum of ripe raun.Be used to carry out the dry method insemination from the seminal fluid of 1 tail milter with from the ovum of 1 tail raun, to be placed on from the fertilized egg (i.e. family) that a tail milter and a tail raun mating produce in the net cage with miniflow water and little inflation and hatch, a net cage is only put the fertilized egg of a family.When fry is about to incubation of membrane, it is transferred in the cylinder of growing seedlings of 2-3 cubic meter volume cultivate, only put the fry of a family in each cylinder, every cubic metre of water body is cultivated 5000-8000 tail prelarva.Should set up 20-40 family, each family should be cultivated 500-1000 tail fry and is used for disease-resistant and the screening high yield family at every turn.
Tongue sole families among the present invention builds in September And October, 2008, mainly carries out family in Shandong Province Mingbo Aquatic Product Co., Ltd., Laizhou and makes up and culture.Adopt the fish that to extrude the seminal fluid and the ovum of hastening parturition out in wild population and 2 reproductive populations of cultured population as the parent who sets up family.The wild male Cynoglossus semilaevis parent population that is used to set up family is 27 tails; Male Cynoglossus semilaevis parent population 4 tails of cultured population; Female Cynoglossus semilaevis parent population is totally 35 tails.Artificial gather smart ovum dry method insemination, build together 35 of upright Cynoglossus semilaevis half sibs and family full-sibses.These family fries are cultured respectively, and its breed is carried out according to conventional method.
Three, the screening of Cynoglossus semilaevis superior families
1). the screening of the disease-resistant family of Cynoglossus semilaevis:
When each family fry of Cynoglossus semilaevis grew to 10-12cm, adopting soaking method carries out Vibrio anguillarum to each family fry to be infected, and estimated the sick ability of anti-vibrio anguillarum of each family fry, infected the cultivation of used Vibrio anguillarum bacterial strain and carried out according to conventional method.Infect experiment at first in advance and determine bacterium liquid half lethal concentration, the pre-fry that infects experiment usefulness is the population mixture from the equal number fry of 10 familys, and each family is got 50-100 tail fry and mixed.During pre-the infection, cultured bacterium liquid is diluted to the bacterium liquid that concentration is 2.0-3.0 OD with physiological saline, above-mentioned bacterium liquid is diluted to the (dilution 50 of different concentration with fresh seawater, 100,200,300,400,600 times), to mix fry and be divided into 6 groups at random, every group of 80-150 tail fry, every group of 80-150 tail mixed family fry is placed on soaks after 3-5 hour in the seawater of a concentration bacterium liquid, 6 groups of fries are placed on to soak in the seawater of 6 different bacterial concentrations and infect, water temperature is controlled at 21-23 ℃ during infection, after the infection fry is forwarded in the seawater of miniflow water and little inflation and raise, in subsequently 15-20 days, calculate the fry lethality of each concentration group, will cause that the dead bacterial concentration of fry of 40-60% is defined as half lethal concentration.
Carry out formal infection experiment according to half lethal concentration, the infection method is tested with pre-the infection.Different familys infect respectively, and each family infects 50-100 tail fry at every turn, repeated experiments 1 time.Infect with fish culture at 0.3-1m
3The fiberglass tank in.After the infection, carry out observed and recorded to state and the dead quantity of each family fry every day, measures dead individual total length and body weight.After infection 20-30 days, write down the fry survival quantity of all familys, measure total length and the body weight of survival fry.According to the quantity and the metainfective survival quantity of participating in the infection fry, calculate the fry survival rate of each family, estimate the disease-resistant performance of each family.Survival rate is defined as the strong family of premunition in the family more than 55%, survival rate is defined as the strong family of premunition in the family of 40-55%, survival rate is defined as the general family of premunition in the family of 20-40%, and survival rate is defined as the family of premunition difference in the family below 20%.
The present invention has carried out infection experiment in the tongue sole families to foundation in 2008 in 2009.Coinfection 18 familys.According to above-mentioned standard, we filter out 2 of the strong familys of premunition (family No. 2 and No. 12) from 18 familys, and it infects the average survival rate in back and is respectively 79.25% and 56.85%, is higher than other family.From 18 familys, also obtain 4 of the more intense familys of premunition (No. 3, No. 10, No. 14 and No. 19), 5 of the familys (No. 6, family, No. 7, No. 15, No. 16 and No. 30) that premunition is general, and 1 of the family (table 1) of premunition difference.
Table 1: the sick ability of the different family fry of Cynoglossus semilaevis anti-vibrio anguillarum relatively
2). grow the fast screening of family of Cynoglossus semilaevis:
When each family fry of Cynoglossus semilaevis grows to total length and is 10-12cm, from each family, choose 200 tail fries, adopt the fry of the different familys of fluorescent pigment mark, the fry behind the mark is raised together with the m at 35-40
3Cement pit in.After culturing 8-15 month, collect each family fry and carry out total length and measured body weight.The fry sampling 60-100 tail of each family is measured, and calculates the mean of each family total length and body weight.Utilize daily gain rate (weight that increases during the breed/actual culturing time) to estimate the growth performance of different familys.The daily gain rate is big more, and its growth rate is fast more, growth performance is good more.The daily gain rate is defined as growing fast family in the family more than 0.25, and the family of daily gain rate between 0.2-0.25 is defined as the faster growing family; The daily gain rate is defined as the general family of growth rate in the family below 0.2.Just can filter out the family of quick growth by this method.
Carried out the growth comparative experiments of 18 familys among the present invention.Through cultivation more than a year, to set up in 2008 and mixed breed body weight and the total length of 18 family fries measure, obtained the mean value and the standard deviation (table 2) of the proterties such as total length, body weight of each family fry.Wherein, the average weight of No. 15 familys and average body are long the highest, and No. 28 family mean bodies are long and average weight is minimum.The result of multiple comparisons shows that No. 15 family body weight are obviously greater than other family.Consider the time order and function that family is set up, utilize the daily gain rate to estimate the growth performance of different familys.From the daily gain rate, the daily gain rate of No. 15 familys is the highest, is 0.260, according to above-mentioned standard, No. 15 familys is defined as growing fast family; And the daily gain rate of 4 familys such as No. 6, No. 7, No. 16 and No. 30 is respectively 0.217,0.229,0.246 and 0.208, should be for growing very fast family; And No. 1, No. 3, No. 4, No. 10, No. 12, No. 14 No. 19, No. 24, No. 27, the daily gain rate of No. 28, No. 33 and No. 34 familys is confirmed as the general family of growth rate all below 0.2.
Body length, weight average value, standard deviation and the daily gain rate of the different family fries of table 2 Cynoglossus semilaevis
Adopt said method, we have made up tongue sole families, and therefrom filter out the strong and family fast of growing of premunition, and these superior families fries can directly be applied, and also can be used for the high-yield disease resisting improved seeds and cultivate.
Claims (1)
1. constructing tongue sole families and breeding superior families method is characterized in that its technology contents comprises: (1), the seed selection of Cynoglossus semilaevis colony; (2), constructing tongue sole families; (3), the screening of the disease-resistant family of Cynoglossus semilaevis and the quick family of growing;
(1), Cynoglossus semilaevis colony seed selection: comprising: the Cynoglossus semilaevis wild population, propagate the seed selection of colony artificially;
The seed selection of wild population: collect the Cynoglossus semilaevis fry or the fingerling of fishing for from Bohai Sea Gulf, select body surface not have scar, healthy energetic fry or fingerling are as wild population, put in a suitable place to breed in cement pit and culture, when fry grows into 12-18 centimetre, after identifying the sex of fry, female, milter seedling are separately cultured; After the 2-3 breed, from raun colony and milter colony, respectively select individual big 40-50 tail fish respectively as female, male parent population according to 1% ratio again, be used for family and make up;
Propagate the seed selection of colony artificially: propagate the fry that colony is the artificial propagation of cynoglossus semilaevis cultivation field artificially, filtering out the fast individuality of growth from the fry that cultivate cynoglossus semilaevis cultivation field, Shandong cultivates, from the female adult fish of 10000 tail Cynoglossus semilaevis of propagating 2-3 artificially, select maximum individual 50 tails as female parent population again, from propagate the male adult fish of 2500-5000 tail Cynoglossus semilaevis in 2 years artificially, select individual big 25-50 tail as male parent population, through behind the reinforced cultivating, use parent population as breeding, be used for family and make up;
(2), constructing tongue sole families:
From the Cynoglossus semilaevis wild population with propagate artificially and choose sexually matured healthy parent population the colony, annual 6-9 month by temperature control control photoinduction parent population gonad development and maturation; When the raun belly obviously swells, sexual gland length reaches the long about half of body, female milter is carried out artificial induced spawning, obtain seminal fluid of ripe milter and the ovum of ripe raun; Be used to carry out the dry method insemination from the seminal fluid of 1 tail milter with from the ovum of 1 tail raun, the fertilized egg that mating is produced is placed in the net cage with miniflow water and little inflation hatches, and a net cage is only put the fertilized egg of a family; When fry is about to incubation of membrane, it is transferred in the cylinder of growing seedlings of 2-3 cubic meter volume cultivate, only put the fry of a family in each cylinder, every cubic metre of water body is cultivated 5000-8000 tail prelarva; Should set up 20-40 family, each family should be cultivated 500-1000 tail fry and is used for disease-resistant and the screening high yield family at every turn;
(3), the screening of the disease-resistant family of Cynoglossus semilaevis and the quick family of growing:
A. the screening of the disease-resistant family of Cynoglossus semilaevis:
When each family fry of Cynoglossus semilaevis grew to 10-12cm, adopting soaking method carries out Vibrio anguillarum to each family fry to be infected, and estimated the sick ability of anti-vibrio anguillarum of each family fry, infected the cultivation of used Vibrio anguillarum bacterial strain and carried out according to conventional method; Infect experiment at first in advance and determine bacterium liquid half lethal concentration, the pre-fry that infects experiment usefulness is the population mixture from the equal number fry of 10 familys, and each family is got 50-100 tail fry and mixed; During pre-the infection, cultured bacterium liquid is diluted to the bacterium liquid that concentration is 2.0-3.0 OD with physiological saline, above-mentioned bacterium liquid is diluted to 50 with fresh seawater, 100,200,300,400,600 times of different concentration, to mix fry and be divided into 6 groups at random, every group of 80-150 tail fry, every group of 80-150 tail mixed family fry is placed on soaks after 3-5 hour in the seawater of a concentration bacterium liquid, 6 groups of fries are placed on to soak in the seawater of 6 different bacterial concentrations and infect, water temperature is controlled at 21-23 ℃ during infection, after the infection fry is forwarded in the seawater of miniflow water and little inflation and raise, in subsequently 15-20 days, calculate the fry lethality of each concentration group, will cause that the bacterial concentration of 40-60% fry death is defined as half lethal concentration;
Adopt half lethal concentration to carry out formal infection experiment, the infection method is tested with pre-the infection; Different familys infect respectively, and each family infects 50-100 tail fry at every turn, repeated experiments 1 time; Infect with fish culture at 0.3-1m
3The fiberglass tank in; After the infection, carry out observed and recorded to state and the dead quantity of each family fry every day, measures dead individual total length and body weight; After infection 20-30 days, write down the fry survival quantity of all familys, measure total length and the body weight of survival fry; According to the quantity and the metainfective survival quantity of participating in the infection fry, calculate the fry survival rate of each family, estimate the disease-resistant performance of each family; Survival rate is defined as the strong family of premunition in the family more than 55%, survival rate is defined as the strong family of premunition in the family of 40-55%, survival rate is defined as the general family of premunition in the family of 20-40%, and survival rate is defined as the family of premunition difference in the family below 20%;
Grow the fast screening of family of B, Cynoglossus semilaevis:
When each family fry of Cynoglossus semilaevis grows to total length and is 10-12cm, from the fry that each family does not infect, choose 200 tails, adopt the fry of the different familys of fluorescent pigment mark, the fry behind the mark is raised together with the m at 35-40
3Cement pit in; After culturing 8-15 month, collect each family fry and carry out total length and measured body weight; The fry sampling 60-100 tail of each family is measured, and calculates the mean of each family total length and body weight; Utilize the daily gain rate, that is: the weight that increases during the breed/actual culturing time is estimated the growth performance of different familys; The daily gain rate is big more, and its growth rate is fast more, growth performance is good more; The daily gain rate is defined as growing fast family in the family more than 0.25, and the family of daily gain rate between 0.2-0.25 is defined as the faster growing family, and the daily gain rate is defined as the general family of growth rate in the family below 0.2.
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| CN102630616B (en) * | 2012-04-23 | 2013-01-23 | 中国水产科学研究院黄海水产研究所 | Method for improving ratio of female fish in cynoglossus semilaevis cultured fingerlings |
| CN103392632B (en) * | 2013-06-24 | 2015-04-29 | 浙江省海洋水产研究所 | Half-smooth tonguefish fluorescence labeling method |
| CN103947607B (en) * | 2014-05-08 | 2015-09-23 | 上海海洋大学 | A kind of construction method of grass carp anti-bacterial sepsis strain |
| CN107287323B (en) * | 2017-07-14 | 2020-10-27 | 海阳市黄海水产有限公司 | Molecular breeding method of new species cynoglossus semilaevis |
| CN107581105B (en) * | 2017-09-13 | 2020-06-23 | 唐山市维卓水产养殖有限公司 | Method for improving fry proportion of female cynoglossus semilaevis |
| CN109479760A (en) * | 2018-10-29 | 2019-03-19 | 南宁学院 | A kind of breeding method of mudskipper fry |
| CN115553234B (en) * | 2022-03-17 | 2023-09-29 | 南通龙洋水产有限公司 | Breeding method of broad-spectrum antibacterial tilapia fine variety |
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| CN100370896C (en) * | 2005-01-13 | 2008-02-27 | 莱州明波水产有限公司 | Breeding method of broodstock of half-smooth tongue sole |
| CN100337603C (en) * | 2005-08-08 | 2007-09-19 | 中国水产科学研究院黄海水产研究所 | Technological method for temperature and light control of cynoglossus semilaevis Gunther parent fish spawning |
| CN101699996A (en) * | 2009-11-20 | 2010-05-05 | 石斌 | Method for rapidly breeding Cynoglossus Semilaevis Gunther in soil pond |
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