64 results about "Takifugu obscurus" patented technology

The invention provides an SNP locus related to rapid growth of takifugu obscurus and application. The SNP locus is located at position 724 of an MSTN gene with a nucleotide sequence shown as SEQ ID NO:1, and the base of the SNP locus is C or T. The SNP locus is used for breeding takifugu obscurus individuals with rapid growth potential. According to the method, by analyzing the correlation betweenthe locus genotype frequency and the growth traits of the takifugu obscurus, the SNP locus related to the growth traits is found at 724 base position of the MSTN gene of the takifugu obscurus, and the body weight, the body length and the body full length of individuals with TT homozygotic genotypes are obviously greater than phenotypic values of the growth traits of individuals with CC and CT genotypes (p smaller than 0.05). Thus, individuals with TT genotype at the locus can be preferentially selected as parents or for large-scale breeding in production.

The invention discloses a method for promoting synchronous development of sexual glands of takifugu obscurus male and female parent fishes, comprising the following steps of: screening parent fishes, rearing by adopting dim light in a parent fish rearing process, and promoting the development of the sexual gland of the takifugu obscurus by adopting means such as reducing illumination intensity, controlling water temperature, stimulating with running water and the like. Meanwhile, 1 per mill of Chinese herbal medicine, VE and edible salt are added into feed, animal feed such as lobster, musselmeat and the like are thrown and fed in the later stage of the rearing process to carry out strengthening rearing; pre-acceleration of maturation physiological regulating means is carried out twice, thus the problem that the development of the sexual glands of the takifugu obscurus male and female parent fishes is asynchronous, the situation that fry of the takifugu obscurus is in short supply can be alleviated, the takifugu obscurus can be artificially bred and released without consuming resource of wild parent fish, and a phenomenon that natural resource of the takifugu obscurus is sharply reduced owning to influence of economic activity of human can be avoided.

The invention discloses an SNP molecular marker related to the condition factor of Takifugu fasciatus and applications thereof. The gene sequence of SNP molecular marker source Takifugu fasciatus serotonin receptor 4 (HTR4) is as shown in SEQ ID NO:1, an SNP site exists at the 100th position of the sequence, and the base is C or G. The SNP locus disclosed by the invention is obviously related to the condition factor of the Takifugu fasciatus, and can be effectively used for molecular marker-assisted breeding of the Takifugu fasciatus, so that the breeding time of improved fish varieties can besaved. The method is accurate in operation, simple and practical.

The invention discloses primers and a method for quickly detecting Takifugu obscurus young fish sex difference single base mutation. According to the mononucleotide mutant sites on sex difference genes of Takifugu obscurus male and female individuals, a pair of external PCR (polymerase chain reaction) primer and internal PCR primer containing mispaired base for the mutant sites are designed to respectively amplify the sex difference gene sequences. By utilizing the primers, different sexes of DNA (deoxyribonucleic acid) samples of Takifugu obscurus can obtain different amplification results, thereby judging the sex difference of Takifugu obscurus. The method has the advantages of high speed and high accuracy, is simple to operate and suitable for popularization and application, can quickly and accurately identify the sex of the Takifugu obscurus young fish on the basis of extracting genome DNA (deoxyribonucleic acid) of Takifugu obscurus under 1 year, and has important application values in accelerating the Takifugu obscurus unisexual breeding progress and researching the quick PCR detection method in the field of molecular biology.

The invention discloses takifugu obscurus SNP molecular markers and application thereof in genetic breeding. The takifugu obscurus SNP molecular markers includes the SNP molecular marker TGF120 and the SNP molecular marker CIRP41, wherein the SNP molecular marker TGF120 is related to weight and standard head length, and the SNP molecular marker CIRP41 is related to cold resistance; amino acid sequences where the SNP molecular markers are located are shown in SEQ ID NO:1 and SEQ ID NO:2 respectively; the TGF120 is located at a 120th site, namely an m with a corresponding base of C or A, of thesequence shown in SEQ ID NO:1; and the CIRP41 is located at a 41th site, namely a y with a corresponding base of A or T, of the sequence shown in SEQ ID NO:2. By adopting the takifugu obscurus SNP molecular markers, takifugu obscurus breeding parents which grow well and resist low temperature can be effectively selected, and thus tetraodontidae variety improving and industry developing are promoted. The invention further provides a takifugu obscurus genetic quality judgement system. Therefore, workloads of breeding selection can be reduced, and the work efficiency is improved.

The invention discloses an artificial crossing cultivation method of seawater takifugu species. The method comprises screening, cultivating and artificial propagation of parent takifugu, incubating, fingerling cultivating and other processes, and is characterized in that on the basis of fully grasping the breeding ecological environment and gonad growth rules of takifugu obscurus and takifugu flavidus, the takifugu flavidus is used as a female parent and the relatively large takifugu obscurus is used as a male parent to simulate a natural ecological environment; mature gonad and ovulation of the parent takifugu are realized by hormone control; an artificial insemination method is employed to realize hybridization among the takifugu species. Synchronously, in the hybridization process, the ecology management is enhanced; and therefore, the oxytocic rate of the hybridization reaches 95 percent, the fertilization rate, the hatchability rate and the opening rate all reach above 80 percent, and the aberration rate of the fingerlings is below 2.0 thousandth.

According to the present invention, provided is a production method of a crossbreed of Takifugu obscurus and Takifugu rubripes, which comprises the following steps: extracting ova and semen from female Takifugu obscurus and male Takifugu rubripes respectively; performing artificial fertilization to obtain fertilized ova; and incubating the fertilized ova and breeding the same. According to the present invention, a new crossbreed of Takifugu obscurus and Takifugu rubripes produced by the production method has an advantage of rapid growth rate, thereby allowing the promotion of existing puffer culture industry declining due to low production rate and low economical efficiency.

Disclosed is a Takifugu obscurus fry breading method through the application of small fertilized eggs. The Takifugu obscurus fry breading method through the application of the small fertilized eggs is composed of steps of hatching of fertilized egg, fry putting, inflation, temperature control, water quality control, pond dividing and bait feeding and is characterized in that the small fertilized eggs are put in a hatching pond with a conical bottom, wherein the density of the small fertilized eggs is 80000-100000 entities per cubic meter; the hatching pond is inflated in a still water mode, the water temperature is 21-22 DEG C, the illumination is 800-1000x and water is changed one time each day; newly-hatched fry are moved to a fry breeding pond after being temporarily bred in the hatching pond for two days, the density of the put fry is 10000 entities per cubic meter, and bottom suction, pollution discharge and water changing are performed on the fourth day, pond dividing is performed one time when the newly-hatched fry are bred for 18 and 25 days respectively; initial bait and SS type freshwater rotifers are fed when the fry are bred for 2-4 days, small freshwater rotifers are fed when the fry are bred for 5-7 days, middle-sized freshwater rotifers are fed when the fry are bred for 8-11 days, and the sizes of the fed creature bait increases as the fry grow; when the total length of the fry is 0.9-1.8 cm, the fry are moved to a pond from the breeding pond.

Disclosed is a large-scale fingerling production technique for takifugu obscurus in brackish water. The technique is characterized in that newly hatched takifugu obscurus larvae are transferred to a fingerling breeding pool after being temporarily fed for 2-3 days in a hatching pool, the density of the fingerlings is 5,000-10,000 per cubic meter, and the fingerlings are bred in fresh water; natural brackish water is gradually added until the salinity reaches 5-8 parts per thousand; water is added until the water level reaches 120cm on the third to fourth day after the fingerlings are put into the pool; bottom suction is carried out every day from the fourth to fifth day after the fingerlings are put into the pool and two thirds of water is changed every other day; two thirds of water is changed every day from the eighth to tenth day after the fingerlings are put into the pool; the fingerlings are separated and placed in different pools from the thirteenth to fourteenth day after the fingerlings are put into the pool and the salinity is gradually reduced until the brackish water is changed into fresh water; the density of air dispersion stones is 1.5 per square meter and is reduced to 1 per square meter in the later stage; the water temperature for fingerling placing is 21.5 plus or minus 0.5 DEG and is increased by 0.1 DEG every other 24 hours until being increased to 24.5 plus or minus 0.5 DEG; rotifers are fed to the fingerlings from the first to eighth day after the fingerlings are put into the pool; fairy shrimp larvae are fed to the fingerlings from the fifth to thirteenth day after the fingerlings are put into the pool; and freshwater cladocerans are fed to the fingerlings from the thirteenth to fourteenth day after the fingerlings are put into the pool.

The invention discloses a method for culturing primary gill cells of takifugu obscurus. The method comprises the following steps: (1) taking a gill tissue of the takifugu obscurus, soaking with a soaking solution containing double-resistant penicillin and streptomycin, cleaning with a cleaning solution, and cutting gill filaments; (2) digesting the gill filaments by using trypsin, filtering, centrifuging and taking precipitates; and then removing red blood cells by using a red blood cell lysate, and centrifuging to obtain cell precipitates; and (3) adding a complete culture solution into the cell precipitates, blowing the cells, supplementing the complete culture solution, and culturing. Compared with the prior art, the method provided by the invention is applied to culture of the primarygill cells of the takifugu obscurus, tissues can be effectively separated, the cell yield is high after gill digestion treatment, the cultured cells grow well, the physiological state is stable, the experimental research standard is achieved, the research approach of various disease problems possibly occurring in takifugu obscurus culture production can be fundamentally provided, and the bottleneck of research of the takifugu obscurus is broken through.

The invention discloses a puffing feed using marine microalgae as main raw materials, for takifugu obscurus, and a making method of the puffing feed, and belongs to the field of feeds. According to the puffing feed and the making method thereof disclosed by the invention, the nutritional requirements of the takifugu obscurus are analyzed in advance, nannochloropsis oculata powder, chlorella powder, and spirulina powder in the feed are rich in EPA and DHA, which are favorable for the health and the rapid growth of the takifugu obscurus, the compounding ratio is scientific, the nutrition is balanced, the nutritional requirements of the takifugu obscurus can be met, and the feed can promote the food intake and the growth of the takifugu obscurus; the feed is rich in fragrance, and good in a food-calling effect, the sizes of particles of the feed conform to the foraging requirement of the takifugu obscurus, the quality guarantee period is long, and the immunity of fish can also be improved; the feed is high in ageing degree, and good in water resistance; few feeds are wasted, the feed coefficient is low, pollution to water is small, the use is convenient, and the feed is convenient for fish breeding in a scaled, industrialized and mechanized manner.

The invention relates to a staged population selective breeding method of a Takifugu obscurus non-toxic pedigree, which is used for selecting unrelated artificial freshwater aquaculture parent fishes according to the population selecting standard and the individual selecting standard. The selective breading stage comprises a preoviposition period, an artificial reproduction period, a larva period, a postlarva period, a fishseed period, a commercial fish period and a prematuration period. The method comprises the following steps of: selecting in the continuous culturing period of three to four years according to the selecting standard of each stage to selectively breed individuals not containing tetrodotoxin; stabilizing the non-toxicity character generation by generation through at least two continuous selective breeding periods to eliminate the possibility of hereditarily carrying the tetrodotoxin, and then selectively breeding non-toxic populations with rapid growth and strong stress resistance; and judging whether the whole fishes and highly toxic organs comprising livers and ovaries of commercial fishes in various specifications are toxic or not, and selecting and preserving the non-toxic populations. The method has the advantages of simple and easy execution and rapid effect taking and is suitable for selectively breeding the populations in any scales of fish fry fields.

The invention relates to an artificial propagation method for takifugu obscurus in winter. The method comprises the following steps of parent fish greenhouse cultivation pool building, parent fish cultivation, parent fish spawning induction, artificial fertilization, fertilized egg hatching and alevin cultivation. According to the artificial propagation method for the takifugu obscurus in winter, the cultivation time of fry from propagation in the current year to overwintering is prolonged, the overwintering cultivation link of the takifugu obscurus is avoided, and the artificial propagation method has the advantages of meeting the market requirement, increasing the cultivation benefit and reducing the cultivation cost.

The invention discloses a method for carrying out anesthesia and blood drawing on Takifugu obscurus, which comprises the following operating steps of: firstly, adding pood water into a container; regulating the water temperature into 15 to 20 DEG C; adding clove oil into the container to prepare anesthetic solution with the weight ratio of the water to the clove oil of 99.98:0.02; placing the Takifugu obscurus into the anesthetic solution, wherein the anesthesia time is 2 to 5 minutes; drawing blood by a syringe at the caudal vein, wherein the blood drawing time is 1 to 1.2 minutes; and after drawing blood, transferring the Takifugu obscurus into a temporary culturing pood for revival. According to the invention, aiming at the physiological property of the Takifugu obscurus, the method is successfully used for carrying out anesthesia and blood drawing on the Takifugu obscurus, has low cost, is simple and convenient to operate, has small damage to the fish body and has a large blood drawing quantity.

The invention discloses an SNP molecular marker combination for identifying species of puffers. The SNP molecular marker combination comprises an SNP molecular marker combination for identifying takifugu bimaculatus, an SNP molecular marker combination for identifying takifugu flavidus, an SNP molecular marker combination for identifying takifugu niphobles, an SNP molecular marker combination for identifying takifugu rubripes and an SNP molecular marker combination for identifying takifugu obscurus. The invention further discloses a method for identifying the species of the puffers, the species of puffer fries and the species of parents of hybridized puffers. The species of the puffers are identified, so that the situation that detected puffers belong to the species of legitimated edible puffers can be guaranteed, and the benefits of breeding households and food safety are guaranteed; and the species of the puffer hybrids are identified, so that the information of parent species of the bred puffer groups can be clearly traced. The SNP molecular markers are large in number, the genome coverage rate is high, the subtyping operation is simple and quick, and the subtyping accuracy rate is high.

The invention discloses an SNP molecular marker for distinguishing Takifugu fasciatus Zhongyang No. 1 from common Takifugu fasciatus and application of the SNP molecular marker. The sequence of the SNP molecular marker is shown as SEQ ID NO: 1 (derived from a CIRP gene sequence of Takifugu fasciatus); and the SNP molecular marker is located at the 66th site, namely an SNP site, of the sequence shown as SEQ ID NO: 1, and the basic group of the SNP site is A or T. The SNP site disclosed by the invention is remarkably related to a low temperature resistance character, and can be used for distinguishing a new Takifugu fasciatus variety Zhongyang No. 1 group from a common Takifugu fasciatus group. The SNP site can also be effectively applied to early identification fry of a new Takifugu fasciatus variety Zhongyang No. 1 and common Takifugu fasciatus. A method provided by the invention is accurate in operation, and is simple and practical.

The invention discloses a specific primer and method for identifying takifugu obscurus, takifugu rubripes and hybrid takifugu fluviatilis. The specific primer comprises a forward primer and a reverseprimer, and nucleotide sequences of the forward primer and the reverse primer are respectively shown as SEQ ID NO.1-2. The identification method comprises the following steps of firstly, collecting apuffer fish sample to be identified, and extracting genome DNA; then taking the genomic DNAs of the three puffer fishes as templates, and performing PCR amplification on the templates by using the primer; and finally, performing electrophoresis on amplification products and performing identifying according to strip difference. The method for identifying takifugu obscurus, takifugu rubripes and hybrid takifugu fluviatilis by using the specific primer does not need complex steps such as microsatellite computer sequencing, can quickly and accurately distinguish the three kinds of fishes only according to the number and positions of strips, and has the advantages of good stability, strong practicability, simplicity in operation, high accuracy and the like.

The invention provides a method for manufacturing a feed for control of Takifugu obscurus fatty liver, and in particular relates to the fish farming field, and steps are as follows: first, raw materials are prepared, wherein the raw materials comprise 40% by weight of baked pearl mussel meat, 23% by weight of wheat bran, 8% by weight of soybean meal, 20% by weight of an eel feed, 6% by weight of green bran, 0.75% by weight of aquatic vitamins, 0.75% by weight of trace element minerals and 1.5% by weight of an aquatic feed binder; the baked pearl mussel meat, the wheat bran, the soybean meal and the green bran are mixed by a mixer in ratio and crushed by a crusher to obtain a mixture; the mixture, the eel feed, the aquatic vitamins, the trace element minerals and the aquatic feed binder are evenly mixed by a mixer according to the ratio of the raw materials and bagged and sealed to obtain an initial material; and the initial material and water are mixed and stirred evenly to obtain the feed. The feed can effectively control the Takifugu obscurus fatty liver, and Takifugu obscures with small liver and tasty meat can be fed.

The invention discloses a SNP locus, a primer and a method for quickly identifying the individual gender of takifugu obscurus, wherein a base sequence of the SNP locus is shown as SEQ ID NO. 1. According to the invention, a male and female-specific SNP locus is obtained by carrying out genome analysis on male and female individuals of three independent genetic populations of the takifugu obscurus,and two pairs of male-specific PCR primers are designed for the locus for identifying the male/female gender of the takifugu obscurus individual; a fin-ray DNA extraction can be completed within 10 minutes by improving a DNA extraction method, with simple and quick operations, and through PCR amplification and agarose gel electrophoresis, the individual with DNA bands is identified as the male, otherwise, the individual is identified as the female, so that distinguishing the male from the female is easy. The method has the characteristics of high speed, high accuracy, simple and convenience in operation, low cost, etc., and the genetic gender identification of the takifugu obscurus can be finished within 2 hours by using the method.

The invention relates to the field of aquaculture, in particular to a takifugu obscures, coilia nasus schlegel and trionyx sinensis three-dimensional ecological breeding method, which comprises sevensteps of pond preparation, stocking of coilia nasus schlegel fingerlings and takifugu obscures, stocking of pseudorasbora parva and silver carps, bait feeding, daily management, and net pulling for catching. The invention fully utilizes the water space of a takifugu obscurus parent breeding pond, increases yield of the coilia nasus schlegel and trionyx sinensis under the conditions of not increasing the input, not reducing the takifugu obscures parent breeding density, and not affecting the takifugu obscurus parent gonad development, improves the water utilization rate of the pond, reduces theproduction cost, improves the economic benefit, reduces the feed cost, improves the quality of the trionyx sinensis, and makes the water quality of the pond more stable; the method of the invention is easy to operate and can be easily accepted and promoted.

The invention provides a simple fertilized egg hatching device for takifugu obscurus, and particularly relates to the technical field of aquaculture. The device comprises a hatching barrel in a pond, a ventilation pipeline, an air pump and a siphon device, wherein the hatching barrel comprises a cylindrical upper part and a conical bottom part, the upper part and the bottom part are integrally formed, an air stone is arranged in the hatching barrel, and the air stone is connected with the air pump through the ventilation pipeline; the hatching barrel is movably connected with the siphon device. According to the device, the hatching barrel is placed in the pond, the temperature in the hatching barrel is convenient to control, and egg escape and fry escape are prevented; by means of the air stone, the fertilized eggs can be overturned to prevent the fertilized eggs from being attached to the wall for a long time and prevent mold growth; meanwhile, the unfertilized dead eggs are sucked out in time, so that the situation that water mold generated and affects the hatching rate is prevented.

The invention relates to a preparation method of white sauce globefish. The preparation method comprises the following steps of: cutting open fish skin from the head of artificially-bred takifugu obscurus globefish, tearing down the fish skin, removing gut, fish membranes and gills, completely removing blood and cutting off fish liver; cutting the fish liver into thick slices, rinsing by using running water, putting the fish skin in hot water for scalding, washing away mucus, and rinsing a fish body by using the running water; adding soybean salad oil into a pot, heating, putting the fish liver, the fish skin, an ingredient bag and supplementary materials into the pot at different times, and elaborately cooking; and finally canning the cooked globefish and cooling. The preparation method can be used for relieving the fleshy mouthfeel of the globefish by using sour and hot type seasoning, removing the greasy feeling, stimulating the taste buds of people, enhancing the appetite of people, and improving the phenomenon of inappetence of a part of people. The white sauce globefish is convenient to eat and can retain nutritional components of the globefish as much as possible.