52 results about "Takifugu rubripes" patented technology

A composite feed for redfin puffer is prepared from 15 raw materials including anabarilius powder, concentrated soybean protein, soybean dregs, chicken powder, beer yeast, alpha-starch, soybean lecithin, etc. Its advantages are rich nutrients, high biological utilization rate and survival rate, and less pollution to water.

The invention discloses a primer and a method for quickly detecting gender difference of juvenile fishes of fugu rubripes. A pair of outer PCR (Polymerase Chain Reaction) primers which respectively amplify gender difference gene sequences and an inner PCR primer which contains mispaired basic groups aiming at mutation sites are designed aiming at G-C mononucleotide mutation sites existing on gender differential genes of male and female individuals of fugu rubripes. By means of the primer, different amplification results can be obtained by different gender DNA (Deoxyribonucleic Acid) samples of fugu rubripes, so that the gender difference of fugu rubripes can be judged. The primer disclosed by the invention is low in cost, simple to operate, quick and accurate and suitable for popularization and application, can quickly identify gender of juvenile fishes of fugu rubripes on a basis of extracting genome DNA of fugu rubripes one year old, and has important application value to accelerate the unisexuality breeding process of fugu rubripes and PCR detection method research in the field of molecular biology.

The invention discloses a rapid cultivation feed for takifugu rubripes. The rapid cultivation feed comprises the following components in parts by weight: 20-40 parts of domestic fish meal, 10-20 parts of shrimp powder, 10-20 parts of meat and bone meal, 1-10 parts of chicken oil, 10-20 parts of corn starch, 1-5 parts of soybean lecithin, 0.1-5 parts of an oyster shell extract, 0.1-8 parts of cylindrotheca fusiformis powder, 2-6 parts of sodium alginate, 1-15 parts of functional additive components, 0.1-3 parts of multivitamins and 0.1-5 parts of multiminerals. A preparation method of the rapid cultivation feed disclosed by the invention comprises the following steps: (1) manufacturing raw materials; (2) synthesizing a biological material; (3) converting to a cooked material; (4) puffing and granulating; (5) drying; (6) spraying and coating; and (7) performing vacuum drying. According to the feed disclosed by the invention, by scientific matching, the body color of adult takifugu rubripes can be improved, the growth can be promoted, and the enrichment of tetrodotoxin can be reduced.

The invention belongs to the field of biotechnologies, and particularly relates to a fugu rubripes nervous necrosis virus capsid protein egg yolk antibody and an application of the egg yolk antibody. The egg yolk antibody is prepared by a method comprising the following steps that: a fugu rubripes nervous necrosis virus capsid protein gene is recombined to a GST (Glutathione S-Transferase) fusion gene expression vector pGEX-5x-1; a pGEX-5x-1-CP recombinant plasmid is constructed and transformed into escherichia coli, and induces a fusion gene to express; a GST-CP recombinant protein is obtained by extraction and purification with an inclusion body purification technology; a hen is immunized by the GST-CP recombinant protein; a hen egg is collected; and the egg yolk antibody is extracted from the hen egg. The prepared egg yolk antibody can effectively inhibit a fugu rubripes nervous necrosis virus, is high in titer, has good immune protection efficacy, and can serve as a nervous necrosis virus vaccine to be added to fish feed in a juvenile fish period of fish.

The invention discloses a cage culture method of takifugu rubripes. In the cage culture process of the takifugu rubripes, fasting of one day and feeding of continuous six days is a cycle and feeding two times per day to satiation during the feeding period, so that the labor is saved, thereby reducing the investment of labor cost. The food conversion ratio of the takifugu rubripes is high with refeeding after starvation. Compared with the fish pond with regular feed, the bait investment is reduced, and the growth speed is faster than that of the single body with continuous feed per day.

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.

The invention discloses a method for preparing a tetrodotoxin positive quality control sample. The method comprises the steps of selecting muscular tissues from fresh fugu rubripes, homogenizing, and measuring the content of tetrodotoxin in the muscular tissues to verify whether the muscular tissues contain the tetrodotoxin; if the muscular tissue is a positive sample, diluting the positive sample containing toxin with a negative sample and a dispersant, mixing uniformly, continuously measuring the content of the toxin in the sample, packaging the sample after the sample meets requirements, preserving at an appropriate temperature, and performing uniformity verification and stability analysis on the sample; if the muscular tissue is a negative sample, adding a tetrodotoxin standard solution into the negative samples, adding a dispersant, mixing uniformly, measuring the content of the tetrodotoxin in the sample, packaging the sample after the sample meets requirements, preserving at an appropriate temperature, and performing uniformity verification and stability analysis on the sample. The quality control sample is simple to operate, and has enough stability and uniformity and long preservation time, so that the quality control sample can be used for laboratory quality control.

The invention discloses a nutriology method for regulating fat deposition in muscle of Takifugurubripes, and belongs to the field of nutrition feed of aquatic animals. According to the method, a certain amount of astaxanthin is added to the feed of Takifugurubripes, and the content of methionine is controlled, so that the fat content in muscle of bred Takifugurubripes is raised or reduced. Throughthe method, directional regulation of the fat content in the muscle of Takifugurubripes is made possible, and the method is environmentally friendly, does not produce toxic by-products at all, does not affect food safety, and has cost within a controllable range.

The invention provides a method for quickly cultivating Takifugu bimaculatus, and relates to a method for cultivating fishes. The method comprises the following steps: S1. selecting female Takifugu bimaculatus and adult male Takifugu rubripes whose weights are larger than 500 g as parent fishes every February, cultivating the parent fishes in a cement pond full of sea water, and feeding fresh oyster or shelled shrimp to the parent fishes to promote fully development of sexual glands; S2. injecting oxytocic to the parent fishes when the temperature of water in the indoor cement pond naturally rises up to more than 20 DEG C in the spring, putting injected parent fishes in a spawning pond, keeping the environment quiet so that the two kinds of parent fish can develop to adult fish at the same time; S3. manually collecting spawns, carrying out dry artificial insemination and hatching the oosperm to prelarva; S4. cultivating the prelarva in a cultivating pond until the average length of fry is larger than 2.0 cm; and S5. cultivating the fry in an earth pond until to the next winter, to achieve commercial fishes whose average weight is 400g. The period of cultivation can be shortened by one year.

The invention relates to a takifugu rubripes biological preservative as well as a preparation method and application thereof, and the method comprises the following steps: step 1), unfreezing frozen fish skin, cleaning, pulping, then adding water, and boiling at 100-121 DEG C; and 2) cooling to 65-75 DEG C, carrying out ultrasonic treatment, and adding compound protease to carry out enzymolysis, thereby obtaining the product. The fish skin enzymatic hydrolysate biological preservative provided by the invention is applied to ice-coated takifugu rubripes; after takifugu rubripes meat is frozen for 12 months, the product quality is kept at a relatively good level, and sensory evaluation keeps 94% or above of fresh takifugu rubripes meat and is remarkably superior to pure water ice-coated takifugu rubripes meat.

The invention discloses a gene for preparing a recombinant fugu rubripes interleukin 2 protein by eukaryotic expression and a preparation method, wherein the protein can be efficiently expressed and an expression product has obvious biological activity. The preparation method comprises the following steps of: cloning a gene as shown in SEQ ID NO: 1 into a prokaryotic expression vector pPICZalphaA to construct an eukaryotic expression plasmid pPICZalphaA-IL-2; electrically converting the eukaryotic expression plasmid into a pichia pastoris GS115 competent cell and screening a positive clone; inoculating the screened positive clone to a BMGY liquid culture medium to be cultured in an amplified manner till OD600nm is equal to 2-6, and centrifugally collecting thalli; and extracting the protein after methanol inducible expression to obtain the recombinant fugu rubripes interleukin 2 protein. Verified by MTT, the recombinant protein has activity of promoting CTLL-2 cell proliferation.

The invention discloses a SNP molecule mark for identifying individual gender of Takifugu rubripes. The SNP molecule mark for identifying individual gender of Takifugu rubripes is characterized in that a DNA sequence is shown in SEQ ID NO: 1, and a base N of the 121st position and the 262th position in the sequence is C or G. The identification method perform according to the following steps: a Takifugu rubripes genome DNA is used as a template, PCR amplification is performed by upstream primer and downstream primer, a PCR product is obtained; the DNA sequences of the upstream primer and the downstream primer is respectively shown in SEQ ID NO: 2 and SEQ ID NO: 3; sequencing is performed on the obtained PCR product, the individual that the 121st position and the 262th position of the measured DNA sequence are simultaneously double peaks is a Takifugu rubripes male fish, and the individual that the 121 position and the 262th position of the measured DNA sequence are simultaneously a single peak is a Takifugu rubripes female fish.

Parasites on the body surface of fishes are removed without causing side effects by orally administering bithionol to fishes in an amount of 50 to 250 mg/kg of body weight per day. Bithionol is orally administered for 1 to 3 days every day or every other day. This method is suitable for fishes of the order Perciformes, Pleuronectiformes and Tetraodontiformes such as amberjack, yellow tail, Seriola lalandi, Seriola rivoliana Valenciennes, cobia, common horse mackerel, Japanese horse mackerel, Japanese seaperch, Pagrus major, Zebrasoma flavescens, Epinephelus akaara, Epinephelus bruneus, Japanese flounder, Verasper moseri, Takifugu rubripes,Oplegnathus fasciatus, Oplegnathus punctatus, Scomber japonicus, threadsail filefish, Scophthalmus aquosus and tilapia, which are parasitized with Neobenedenia girellae. When removing parasites on the body surface of fishes, feed for fish culture supplemented with a necessary amount of bithionol can be used.

The invention discloses a gene capable of being efficiently expressed for preparing recombination fugu rubripe interleukin-2 and a preparation method. The expression product of the gene has the obvious biological activity. The method comprises the following steps that the gene shown in SEQ ID NO:1 is cloned to a prokaryotic expression vector pET-32a, and prokaryotic expression plasmid is constructed; the prokaryotic expression plasmid is transformed into escherichia coli BL21(DE3), positive clones are screened out, and recombinant escherichia coli BL21(DE3) is constructed; LB is used as a basic culture medium, and amplification culture is performed on the recombinant escherichia coli BL21(DE3); an IPTG inductive agent is added into an amplification culture for induction culture, and thalli is collected through centrifuging; ultrasonication is performed on the collected thalli, a supernate is taken after centrifuging is performed, and a crude recombination fugu rubripe interleukin-2 protein product is obtained.

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 discloses a method for obtaining takifugu rubripes androgenesis haploids. The steps of the method are as follows: (a) the spawning of male takifugu rubripes and female takifugu rubripes with well-developed gonads is induced, the sperms of the male takifugu rubripes and the ova of the female takifugu rubripes are respectively taken, and the sperms and the ova are mixed to form fertilized ova; (b) the androgenesis of the takifugu rubripes is induced by cold shock. The method adopts the sperms of the male takifugu rubripes and the ova of the female takifugu rubripes to form the fertilized ova; cold shock is used for processing (30 to 60 minutes of immersion in ice water of 2 DEG C to 4 DEG C 5 to 8 minutes after fertilization), the egg nuclei and second polar bodies of the takifugu rubripes are induced to release together, and thereby fertilized ova which only have sperm nuclei are formed. Because the genetic inactivation of ova is not required, a complex operation procedure is avoided, the method is cheap and easy to operate, the problem that conventional physiochemical induction can injure fertilized ova is solved, the androgenesis rate is increased, and the method is suitable for use in mass production.

The invention relates to a method for promoting gonad development of male takifugu rubripes, and belongs to the technical field of takifugu rubripes breeding. From September or October, an LED light source is arranged above the culture water surface of 2.5-age and above takifugu rubripes, the LED light source is full-spectrum white light, the light period is 12-hour illumination and 12-hour darkness, and the culture period is 90-150 days. By creating a specific light environment, the gonad development of the male takifugu rubripes is promoted, the gonad development time is shortened, and the economic benefits of breeding enterprises are improved.

The invention discloses SNP primers for screening takifugu rubripes seedlings and a screening method, wherein the human and material resources can be saved, the seed selection cost is reduced, and good characters of filial generations are guaranteed. The sequences of the SNP primers are shown as SEQ ID No.1 and SEQ ID No.2; and the seedling screening method comprises the following steps: extracting the genome DNA of the to-be-detected takifugu rubripes; with the obtained genome DNA as a template, performing PCR amplification on the forward primer and the reverse primer to obtain a PCR product; and performing sequencing and genotyping of the obtained PCR product and screening out the takifugu rubripes of which the DNA sequence is shown as SEQ ID No.3 and the genotype of the 145th site from the 5' end is homozygous TT.

The invention provides an SNP site related to rapid growth of takifugu rubripes and a fry screening application capable of reducing fry selection cost and guranteeing good filial generation characters. The SNP site is located at the 162nd site of a nucleotide sequence as shown in SEQ ID NO: 1, wherein the base of the SNP site is C or G. The SNP site provided by the invention can be used for breeding takifugu rubripes individuals with rapid growth potential. According to the invention, through analysis of the correlation between the allelic SNP sit genotype and the takifugu rubripes growth characters, the SNP site related to the growth characters is found to exists at the 162 base of the nucleotide sequence, as shown in the SEQ ID NO: 1, of the takifugu rubripes, wherein the values of the weight and the body length of individuals with a CC homozygous genotype are significantly higher than the phenotype values (p (0.05)) of the growth characters of the individuals with a GG genotype. Meanwhile, through screening of an SSCP technology, excellent individuals with the CC genotype can be screened out through different electrophoretic banding patterns. Thus, the method provided by the invention can be used in production to screen the individuals with the site genotype of CC type as parents for breeding and large-scale breeding of large-sized fries.

The invention discloses a method for acquiring ecological early breeding seeds of acanthogobius ommaturus Richardson, and relates to technologies for acquiring early seeds for early breeding for acanthogobius ommaturus Richardson. The method includes selecting parent fish; controlling the breeding quantities; selecting heat-insulation sheds of ponds; cultivating the parent fish through the winters; collecting fish fries. The method is characterized in that the parent fish with the weight ranging from 100 grams to 250 grams is selected in periods from November to December, and is cultivated in the ponds for takifugu rubripes through the winters, nylon films are constructed to form heat-insulation greenhouses on the upper sides of the ponds, the areas of the ponds preferably range from 3mu to 8mu, the ratio of female parent fish to male parent fish is 1:1, 10-12 pieces of parent fish is bred in each mu of the ponds under the control, 3-5 kilograms of live exopalaemon carinicauda are placed in each mu in winter cultivation periods and are used as fodders for the parent fish, water is changed regularly, an excellent water quality condition needs to be kept, 1.2-meter water levels of the ponds are kept, and the salinity of the water in the ponds keeps ranging from 6%o to 15%o; the fish fries are sampled after 20th of March in the next year, growth conditions of the fish fries are inspected, and the fish fries are timely collected and distributed when the sizes of the growing fish fries range from 4 centimeters to 6 centimeters.

The invention discloses a synchronous gonad maturing method of female fugu obscurus and male fugu obscurus. According to the method, female fugu obscurus and male fugu obscurus which are large in size, strong in physique and great in gonad development condition are adopted as parents, composite vitamins are added into feed for nutrient reinforcement, and protein feed control is carried according to the sexual maturity; illumination different in intensity is adopted in a breeding process of the parent fishes, gonad development of the two parent fishes is facilitated through water temperature control, water flow stimulation and other approaches, one-time physiologic maturation pre-acceleration control is combined, and the problem that female fugu obscurus and male fugu obscurus are not synchronous in gonad development is solved. The quality of eggs and sperms during crossbreeding of female fugu obscurus and male fugu obscurus is improved, and therefore the spawning induction rate and fertility rate of the parent fishes are increased.

The invention relates to a method for accelerating feeding habit conversion of early fries of takifugu rubripes, and belongs to the technical field of marine fish culture. The method comprises the steps that from the fifteenth day or the sixteenth day after incubation, compound feed and biological bait are fed in turn, the feeding frequency of the compound feed is gradually increased, the feedingfrequency of the biological bait is gradually decreased, and only the compound feed is fed from the thirty-second day or the thirty-third day or the thirty-fourth day after incubation. According to the feeding rule of the early fries of the takifugu rubripes, a combined feeding mode is adopted, compound feed with different particle sizes is gradually used for replacing traditional biological baits, feeding habit conversion of the early fries of the takifugu rubripes is completed, the expenditure cost of the baits is reduced, and breeding operation is simplified, so that the production efficiency and economic benefits of industrial fry cultivation of takifugu rubripes are improved, meanwhile, emission of cultivation pollutants is reduced, and environmental protection is facilitated.

The invention discloses a crossbreeding method for female fugu rubripes and male takifugu obscures. The method comprises the following steps: by taking female fugu rubripes and male takifugu obscureswhich are large in size and health and strong in physique as parents, putting composite vitamins for enhancing nutrition into feeds; in a breeding season, implementing an artificial spawning and fishfertilization method according to a certain ratio, and incubating in a special incubation facility; after fish fries are incubated, feeding with animal biological baits such as rotifera, artemia, cladocerans and copepods, vegetal baits such as chlorella and microorganism baits such as ocean rhodotorula to start feeding, and gradually feeding with artificial composite feeds; in a growth period of hybridized pufferfish, and breeding by using a gradual thinning method, thereby finally obtaining the hybridized pufferfish which grows at a speed which is 30% faster than that of the takifugu obscuresand is delivered to the market 3-6 months ahead. The hybridized pufferfish has meat more delicious than that of the fugu rubripes, and the taste of white juice hybridized pufferfish is approximate tothat of the takifugu obscures.