41 results about "Fugu rubripes" patented technology

The invention provides a method for identifying fugu rubripes parentage by using a molecular marker method as well as microsatellite primers and a kit used for the molecular marker method. Selected 44 microsatellite primers are stable in pure PCR amplification reaction, clear in amplified fragment and high in polymorphism. By using the technology, different families of mixed fugu rubripes individuals can be distinguished, so that different families of the fugu rubripes individuals can be quickly and accurately identified, and the technology has great application value for breed conservation, propagation and breeding selection of fugu rubripes.

The invention relates to a method for transporting live fugu rubripes at a low temperature. The method includes the steps of conducting temporary cultivation at a low temperature, and packing each fugu rubripe, namely, loading each fugu rubripe which is in a quiet state into a single body packaging net, loading the single body packaging nets into single body packaging bags containing seawater with the water temperature ranging from 7 DEG C to 8 DEG C, the salinity ranging from 2.5% to 3.2%, the pH value ranging from 7.8 to 8.3 and the oxygen content ranging from 10mg / L to 12mg / L, and loading the plurality of sealed single body packaging bags into a thermal insulation box, wherein the mass ratio of the seawater to the fugu rubripes is 1:1.5-2. The method has the advantages that the survival rate of the transported fugu rubripes is 100%, and the longest transportation time can be 18 hours.

The invention discloses compound feed for improving muscle quality of fugu rubripes, amd belongs to the field of marine fish compound feed. The formula of the compound feed comprises the following rawmaterials by mass percent: 42 to 48% of white fish meal; 4 to 5% of yeast; 4 to 6% of krill powder; 4 to 5% vital gluten; 3 to 5% of corn gluten meal; 6 to 8% of soybean meal; 13.1 to 14.5% of wheatflour; 5.5 to 6% of fish oil; 1.5% of phospholipid; 1.5% of calcium dihydrogen phosphate; 1% of vitamin mixture; 1% of mineral mixture; 4.8 to 6.4% of functional additive. Compared with fresh trash fish, the special compound feed disclosed by the invention significantly improves the growth, muscle nutritional value and muscle quality of the fugu rubripes under the urban aquaculture conditions, reduces the dependence on the fresh trash fish in the aquaculture production, and reduces the waste of wild fishery resources and pollution of the cultured marine environment.

The invention discloses a preparation method of a recombinant fugu rubripe interferon gamma protein. The method has advantages of simple operation, low cost, high purity of the recombinant protein and high yield, and is suitable for industrial production. The preparation method comprises steps of: extracting the total RNA of fresh liver of fugu rubripes and performing inverse transcription to synthesize cDNA of a first strand; performing RT-PCR amplification by adopting the cDNA of the first strand as a template, with the upstream primer being N[co] I:5'-CATGCCATGGGCTCCTACATCCCAGTAGAAATG-3', and with the downstream primer being X[ho] I:5'-CCGCTCGAGATGACCTCGAATCGACATCT-3'; establishing a clone vector, expressing the vector, transferring into an expression bacterial, and cultivating the bacterial strain to obtain strain; subjecting the collected strain to enlarging cultivation, inducing with IPTG and collecting the bacterial strain; and smashing the bacterial strain, lysing, purifying, and collecting purified liquid.

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 temperature and light regulation and control method for maturing of fugu rubripe parent fish, and belongs to the technical field of fish fry breeding. The temperature and light regulation and control method comprises the following steps that a fugu rubripe parent fish rearing and culturing system which is free of natural light irradiation and is provided with a green spectrum irradiation lamp, a full spectrum irradiation lamp, a water temperature control system and a circulating water treatment system is constructed; sexually mature fugu rubripe parent fish is selected; in a parent fish rearing system without natural light irradiation, environmental conditions of natural reproduction of fugu rubripes is simulated, and a water temperature and an irradiation period of the irradiation lamp are arranged; mature parent fish individuals are selected according to the behavior characteristics and the morphological characteristics; and artificial spawning induction and insemination are carried out. According to the temperature and light regulation and control method for maturing of the fugu rubripe parent fish, the circulating aquaculture system is adopted for feeding the fugu rubripe parent fish, matched automatic temperature control equipment and intelligent light environment regulation and control equipment are adopted for temperature and light regulation and control, gonad development of the parent fish can be effectively promoted, the spawning time of the parent fish can be precisely regulated and controlled, and technical support is provided for stable production of fugu rubripe fries.

Novel parathyroid hormone polypeptides and biologically active fragments thereof are disclosed along with nucleic acid molecules encoding same. In particular, parathyroid hormone polypeptides and biologically active fragments (and encoding nucleic acid molecules) derived from fish species (eg Japanese pufferfish (Fugu rubripes)) are disclosed. Such polypeptides and fragments may be used for treatment of diseases associated with abnormal calcium homeostasis (eg osteoporosis, osteopenia, Paget's disease, bone cancer, hyperparathyroidism, hypoparathyroidism, hypercalcemia, psoriasis and other skin-related conditions).

The invention relates to a method for promoting survival and growth of fugu rubripes from fertilized eggs to larvae and juveniles by using an LED light source, belonging to the technical field of industrial aquaculture. With the method provided by the invention, the LED light source is arranged on the water surface; in the cultivation process of fertilized eggs of the fugu rubripes, the LED lightsource is in a full spectrum; in the cultivation process of larvae and juveniles of the fugu rubripes, the LED light source is green light with a wavelength of 525-530 nm or yellow light with a wavelength of 590-595 nm. According to the invention, through regulation and controlling of illumination parameters in a fry cultivation period, the survival rate of fries is increased, and growth of the fries is promoted, so production efficiency and economic benefit of industrial fry cultivation of the fugu rubripes can be improved.

The invention discloses a SNP (Single Nucleotide Polymorphism) primer for screening fugu rubripes seedlings and a screening method, which are capable of saving manpower and material resources, lowering seed screening cost and guaranteeing excellent characters of the filial generation. The sequence of the SNP primer is as shown in SEQ ID NO.1 and SEQ ID NO.2. The seedling screening method comprises the following steps: extracting the genomic DNA of to-be-detected fugu rubripes; taking the acquired genomic DNA as a template; performing PCR amplification with upstream primer and downstream primer, thereby obtaining a PCR product; sequencing and genetically typing the obtained PCR product; screening the fugu rubripes with DNA sequence as shown in SEQ ID NO.3 and the No.591 locus gene started from 5' end being homozygosis TT.

The invention discloses a lighting adjustment and control method capable of promoting surviving, growth and development of young fish of fugu rubripes, and belongs to the technical field of aquaculture. According to requirements of the fugu rubripes at the stage of young fish for light spectrum and light intensity, light environment can be adjusted and controlled for the stage of young fish, and in an early development stage of the fugu rubripes, the light spectrum and light intensity can be adjusted and controlled, so that the light environment of the fugu rubripes can be improved, the growthand development situation of the young fish of the fugu rubripes can be promoted, and further the productivity level and the economic benefits of breeding enterprises can be increased. The lighting system used by the lighting adjustment and control method comprises a single light spectrum LED light source and a controller for adjusting light intensity.

The invention discloses a functional feed for relieving winter syndromes of fugu rubripes and belongs to the field of aquatic feed. The functional feed is based on existing fugu rubripes compound feed, and a part of wheat gluten, corn gluten meal and soy protein concentrate powder are replaced with fish meal and hydrolyzed fish protein, so that the content of protein in the fugu rubripes feed is reduced by two percentage points and is 46%; at the same time, the use amount of vegetable oil in a formula is reduced, so that the content of fat in the fugu rubripes feed is reduced by two percentagepoints and is 9%. Compared with minced trash fishes, the functional feed has the advantages that the liver antioxidant capacity is improved under the condition that the fugu rubripes have no significant differences during growth, the incidence rate of the winter syndromes is significantly reduced by 19%, and the survival rate is increased. At the same time, the feed formula also saves feed protein and fat resources, reduces the breeding cost and ensures the yield and benefit of breeding of fugu rubripes over-wintering fry.

The invention relates to a fugu rubripe triploid induction method based on hydrostatic pressure, and belongs to the field of genetic breeding of fugu rubripes. The fugu rubripe triploid induction method based on hydrostatic pressure includes the following steps that sperms of mature male fugu rubripes and mature ova of female fugu rubripes are taken respectively; the sperms are added into the mature ova, and sea water is added to obtain fertilized ova; the fertilized ova are transferred to a pressurization chamber of a hydrostatic press, and the pressure of 30-80 MPa is applied for 2-10 min; the fertilized ova are transferred to an incubator for incubation after hydrostatic pressure treatment. The method has high success rate of triploid induction, high incubation rate and simple operation, time is saved, the practicability and the operability are good, and the method can be used as an industrial application measure.

The invention relates to a microsatellite marker related to low temperature resistance of fugu rubripes and application thereof, and belongs to the field of fish DNA marking technologies and application, particularly screening of low temperature resistance molecular markers of fugu rubripes and molecular marker-assisted breeding by utilizing the markers. A microsatellite marker (fms108) related tolow temperature resistance of the fugu rubripes is screened by using a BSA-SSR technology, and the primer sequences of the microsatellite marker (fms108) are shown by SEQUENCE LISTING 1 and SEQUENCELISTING 2. Breeding parents can be selected according to the marker, so that the low-temperature resistance of offspring is improved.

The invention discloses a temperature and light regulation and control method for out-of-season breeding of fugu rubripes parent fishes and application, and belongs to the technical field of factory-like circulating water cultivation of the fugu rubripes parent fishes. The breeding period of the parent fish of fugu rubripes is concentrated from March to May, and the production of fries is limited by seasons. By simulating the illumination period and the annual change period of the water temperature, the change rate of the annual period is accelerated from one time of 12 months to one time of 6 months, gonad development of the parent fish is promoted to be advanced, and breeding and egg laying in winter are achieved. The fry obtained by out-of-season breeding is easy to be subjected to low-temperature masculinization induction.

The invention discloses a preparation method for a fugu rubripes uronema marinum disease vaccine. The preparation method comprises the following steps that: picking up a single uronema marinum to be inoculated to an L-15 complete medium to be subjected to pure culture; then, carrying out enlargement cultivation; adding 100 mu l / L of formalin to kill larvae; carrying out centrifugation for 10 min at 4000rpm by a low-speed centrifugal machine to collect sediments; adding into the L-15 complete medium to prepare inactivation larva liquid of which the density is 2 million / ml; collecting in a centrifuge tube of 1.5ml, and preserving at the temperature of -80 DEG C. Before immunization is carried out, dead larvae are unfrozen on ice, are broken by electromagnetic wave and are mixed with equivalent Freund's adjuvant or Freund's incomplete adjuvant, and the mixed liquid is oscillated to emulsion to prepare the fugu rubripes uronema marinum disease vaccine. Since an intraperitoneal injection way is adopted, the immunity of the fugu rubripes for the uronema marinum disease can be improved.

The present invention provides a method for guiding genetic breading of red-fin blowfish by utilizing microsatellite pleiomorphy analysis result. Said method includes the following steps: (1), according to phenotypic character selecting parental red-fin blowfish for propagation; (2), utilizing microsatellite marker to detect parental DNA, statistically analyzing and calculating individual similarity and genetic distance; (3), according to the individual genetic distance guiding parental one-to-one grouping propagation; and (4), making progeny breeding proparation and cultivation.

The invention discloses a fugu-rubripe-oil oral solution for improving sexual hypofunction. According to the raw material formula, the 1,000 parts of fugu-rubripe-oil oral solution comprise 4-6 parts of fugu rubripe oil, 10-15 parts of herba epimedii, 8-12 parts of angelica sinensis, 10-15 parts of the roots of kudzu vine, 15-20 parts of acanthopanax roots, 10-13 parts of prepared rehmannia roots, 8-12 parts of the bark of eucommia, 10-15 parts of fiveleaf gynostemma herbs, 10-16 parts of fructus lycii, 10-18 parts of cynomorium songaricum, 5-10 parts of flatstem milkvetch seeds, 5-10 parts of the roots of bidentate achyranthes, 10-15 parts of liquorice and 10-15 parts of common macrocarpium fruits. The ultrasonic-wave assisted extraction process, the volatile oil distilling process, the water decocting process and the ethanol extraction process are adopted for preparing. The fugu-rubripe-oil oral solution has the advantages of being remarkable in sexual-hypofunction improvement effect, safe, free of side effects, convenient to drink and the like.

The invention discloses a method for freeze-preservation and recovery of uronema marinum isolated from fugu rubripes. The method comprises the following steps of using an L-15 complete culture mediumto culture the uronema marinum to the exponential growth phase, exponential phase and adult worm phase, and using the L-15 complete culture medium to resuspend until the density is 200000 uronema marinum per 1ml; sucking an adult worm solution of uronema marinum, adding into a freeze-preservation tube, adding 40vol% sterilizing glycerin and 10vol% dimethyl sulfoxide, uniformly mixing, quickly putting the freeze-preservation tube into a programmable gradient cooling cell freeze-preservation box, staying overnight at the temperature of -80 DEG C, and putting into liquid nitrogen, so as to preserve for a long time; fetching the freeze-preservation tube out of the liquid nitrogen, quickly putting into a water bath pot, treating for 2min in a water bath with the temperature of 37 DEG C, centrifuging for 5min at the rotation speed of 4000rpm, discarding a supernatant, adding the L-15 complete culture medium to resuspend, transferring into a porous plate, and culturing for three days in a constant-temperature culture box with temperature of 20 DEG C, so as to complete recovery.

The invention relates to a special compound nutrient for fishes and in particular relates to a special compound micro-element nutrient for fishes for facility aquaculture of fugu rubripes and a preparation method thereof. The special compound micro-element nutrient is composed of 10-25% of a monomeric compound micro-element nutrient source material and 75-90% of a carrier in percentage by weight.The monomeric compound micro-element nutrient source material is iron, zinc, manganese, copper, cobalt, selenium and iodine; and the carrier is natural medical stones. The technical product provided by the invention has a promoting effect on growth performance of high density animals by facility aquaculture, has an obvious effect in reducing water pollution and keeping ecological balance of waterand healthy and environment-friendly breeding, and has a wide application prospect in facility aquaculture with indoor flowing water and plant circulating water of fugu rubripes in China and production of ecological clean feeds.

The invention relates to an in-vitro construction method and application of a fugu rubripes ovarian cell line. The construction method comprises the steps of (1) preparing a primary cell culture solution; (2) performing primary culture; (3) performing subculture; and (4) separating cell lines with different forms. The application comprises application of the fugu rubripes ovarian cell line to multiplication and expanding propagation of two typical fish viruses and application of the fugu rubripes ovarian cell line to transgenic research. According to the in-vitro construction method, an ovary is used as a culture and line construction object to build a biological platform for research on extraction and utilization of tetrodotoxin and safe detoxification treatment of cultured globefish, so that the conventional aquaculture industry and the emerging bio-pharmaceutical industry can be well served.

The invention discloses a SNP (Single Nucleotide Polymorphism) primer for screening fugu rubripes seedlings and a screening method, which are capable of saving manpower and material resources, lowering seed screening cost and guaranteeing excellent characters of the filial generation. The sequence of the SNP primer is as shown in SEQ ID NO.1 and SEQ ID NO.2. The seedling screening method comprises the following steps: extracting the genomic DNA of to-be-detected fugu rubripes; taking the acquired genomic DNA as a template; performing PCR amplification with upstream primer and downstream primer, thereby obtaining a PCR product; sequencing and genetically typing the obtained PCR product; screening the fugu rubripes with DNA sequence as shown in SEQ ID NO.3 and the No.591 locus gene started from 5' end being homozygosis TT.

The invention discloses a method for evaluating gastrointestinal emptying parameters of fugu rubripes, and relates to the technical field of aquaculture, and the method comprises the following steps: a1, selecting experimental fishes: randomly selecting 3000 healthy and injury-free large-specification fugu rubripes fries, and carrying out detailed evaluation on the gastrointestinal emptying mode of the fugu rubripes fries by setting a comprehensive evaluation method; gastrointestinal emptying mode research of different feeding frequencies under the condition of a small circulating water system is carried out, and an optimal mathematical model conforming to the gastrointestinal emptying characteristics of the takifugu rubripes large-scale offspring seeds is established, so that the problem that research data about the gastrointestinal emptying mode of the takifugu rubripes, especially in the large-scale offspring seed stage, is not reported yet nowadays is solved. The problems that effective parameters cannot be provided for fugu rubripes culturists, certain troubles are brought to the corresponding fugu rubripes culturists to formulate feeding strategies, and great difficulty is brought to early-stage breeding of large-size fries of fugu rubripes are solved.

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 novel method for breeding fugu rubripes fries, which relates to the technical field of aquaculture and comprises the following steps of a1, preparing a breeding environment, namely four sets of circulating water systems. The factory-like closed circulating water aquaculture system is realized by arranging a corresponding totally-closed circulating water system and comprehensively integrating advanced technologies in various modern fields, so that water resources can be greatly saved, and an aquaculture water body continuously keeps a high dissolved oxygen state and a stable water quality environment; compared with a traditional breeding mode, water resources and land resources are fully utilized, power consumption is reduced, and therefore the problems that at present, the fugu rubripes fries are mainly bred in an indoor cement pool in a flowing water mode; and the problems of extensive technology, high water resource consumption, high difficulty in controlling water quality conditions, low degree of facility and mechanization and the like are increasingly prominent, so that the survival rate of the fries is reduced, diseases are caused, and great troubles are brought to mass culture of the fugu rubripes.

The present invention relates to a production processing method of an artificial bait for fish farming, particularly to a fugu rubripes artificial bait production processing method, wherein the operation steps comprise: adding vitamin C, folic acid, vitamin B1, vitamin B2, a zinc salt, an iron salt and raw egg yolk to per 1 kg of corn starch, adding water to prepare an aqueous solution with a corn starch weight concentration of 18-25%, pasting, carrying out slurry coating on ammodytes personatus, rapidly freezing, carrying out cold preservation, thawing at a normal temperature in 20-40 min before use, and cutting into sections with a length of 1-3 cm so as to be used. With the method of the present invention, the nutrients can be completely retained on the ammodytes personatus surface, the phenomenon that the nutrients of the traditional bait are dispersed in the fishpond while only a small amount of the nutrients are absorbed by the fugu rubripes is overcome, the bait utilization rate is improved, and the cost is reduced.

The invention relates to a method for improving the anti-stress ability of fugu rubripes under different feed fat levels, and belongs to the field of aquatic nutrition. The method aims at the problem that stress is prone to occurring in the fugu rubripes breeding process, a series of anti-stress substances including lysolecithin, taurine and L-carnitine are comprehensively applied, the content of substances such as arachidonic acid and phosphorus elements closely related to anti-stress metabolic signal transduction in feed is controlled, and therefore the resistance of bred fugu rubripes to a series of stress conditions such as high density and high temperature is improved. According to the method, the anti-stress ability of the bred fugu rubripes can be significantly improved, and accurate regulation and control under different feed fat conditions are achieved. Under the general background that high-density intensive breeding is increasingly prevalent, the breeding success rate can be increased, the production income can be increased, and development of intensive aquaculture can be supported from the perspective of nutriology.

The invention discloses a method for preparing a marine filariasis vaccine of redfin puffer. A single marine filariasis is picked and inoculated in L-15 complete medium for purification and cultivation, and then enlarged and cultivated, and 100 μl / L of Foreria is added. Marin killed the larvae, centrifuged at 4000rpm in a low-speed centrifuge for 10 minutes to collect the precipitate, added L‑15 medium to prepare an inactivated larvae liquid with a density of 2 million / ml, collected in a 1.5ml centrifuge tube, and stored at ‑80°C. Before immunization, thaw the dead larvae on ice, crush them with electromagnetic waves, mix them with the same amount of Freund's adjuvant or Freund's incomplete adjuvant, and shake the mixture until it is in the form of emulsion, which is to prepare the marine tail silk of puffer puffer Insect vaccines. Intraperitoneal injection can improve the immunity of pufferfish against marine filariasis.

The invention discloses a gene and method for preparing recombinant fugu rubripes IFN-gamma protein. The gene has a DNA sequence as shown in SEQ ID NO:1. The method comprises the following steps: cloning the gene having a DNA sequence as shown in the SEQ ID NO:1 onto eukaryotic expression plasmids pPICZ alpha A to obtain eukaryotic expression recombinant plasmids pPICZ alpha A-IFN-gamma; electronically transforming the eukaryotic expression recombinant plasmids pPICZ alpha A-IFN-gamma into a pichia pastoris GS115 competent cell, and screening positive clone; inoculating the screened positive clone into a BMGY liquid culture medium, enlarging and culturing until OD600nm is 2-6, and centrifuging to collect strains; and extracting protein after methanol induced expression to obtain recombinant fugu rubripes IFN-gamma protein.

The invention provides a feeding method for fugu rubripes at different growth stages. The feeding method comprises the steps that in the fry stage, rotifers are splashed and fed, the feeding density of the rotifers is 10-15 / ml, and the rotifers are fed once a day; in the juvenile fish stage, artemia is splashed and fed, the feeding density of the artemia is 5-10 pieces / ml, and the artemia is fed twice a day; after the artemia is fed for one week, pellet feed is added for domestication, and small-particle floating feed is mainly used as the feed; in the juvenile fish stage, pellet feed is fed, the total feeding amount is 1%-3% of the total mass of juvenile fish, and feeding is carried out four times a day; in the adult fish stage, pellet feed is used for feeding, and the total feeding amount is 2%-3% of the total mass of adult fish; and in the parent fish stage, pellet feed is used as main food, jade squid, anchovy, squid, nereis and the like are used for auxiliary feeding, and the total feeding amount is 2%-4% of the total mass of parent fish. The bait is reasonably utilized, the breeding cost is reduced, and the bait conversion rate is high.

The invention discloses a method for inducing masculinization of fugu rubripes at a low temperature. The method comprises the following steps of: performing artificial induced spawning and insemination on healthy male and female fishes which are high in fatness and undergo good gonad development; hatching fertilized eggs under the water temperature condition of 20 DEG C for 45-75 days; and putting into water of 15-17 DEG C, and culturing for 35-65 days. Due to the adoption of the method, male fugu rubripes fries can be produced rapidly, the male rate in the fries is 65-80 percent, the average male rate is 75 percent, ultraviolet ray irradiation is not required, sex hormone is not fed, and complex operation processes are avoided; and the method is low in cost, is easy to operate, and has contribution to mass production.