53 results about "Halibut" patented technology

A method is provided for producing prey organisms such as Artermia and rotifers, for feeding aquacultural organisms in particular at the larval stage. The method comprises cultivating the prey organisms during at least part of their life cycle in an aqueous medium comprising at least one lipid component having a DHA content of at least 30 wt %. The enriched prey organisms preferably have a DHA content of at least 12 wt % of their total lipid content. The prey organisms are suitable feed for larvae of fish including halibut, turbot, bass, and flounder, and crustaceans and molluscs.

A composition is for feeding aquacultural prey organisms such as Artemia and rotifiers, comprising a lipid component comprising at least 25 wt % of phospholipids and providing a DHA content of at least 30 wt %. The lipid component is preferably derived from marine organisms such as fishineal, phytoplankton or zoo plankton biomass. The composition is used for providing prey organisms having a high content of highly-unsaturated fatty acids (HUFAs) suitable for aquaculture of fish including halibut, turbot, bass and crustaceans and molluses.

Two monoclonal antibodies B and C for the lymphocystics virus of bastard halibut are prepared from two hybrid tumor cells (CCTCC-C200419 and CCTCC-C200420) through conventional culture for 2-3 days to secrete a lot of said monoclonal antibodies B and C.

The invention discloses a method for establishing a bastard halibut brain tissue-derived cell system. The method comprises brain tissue primary culture, subculture, cryopreservation, recovery and identification, wherein cell culture fluid adopted is obtained by adding fetal calf serum and human basic fibroblast growth factor to basal cell culture fluid. The established bastard halibut brain cell system is represented as an epithelioid-like astrocytic cell, which can support continuous passage to provide a great amount of bastard halibut brain cells and can be directly used for research of bastard halibut functional gene. The construction method disclosed by the invention not only is expected to serve as a platform for theoretical research of bastard halibut molecule cellular level but also can be used as an ideal material for research on fish hemadenology, reproductive biology and environmental internal-secretion interfering-substance toxicology.

The trace element feed additive for sea water left-eyed flounder and right-eyed flounder has the effective components comprising Fe 23000-50000 mg / kg, Mn 4000-8000 mg / kg, Zn 25000-48000 mg / kg, Cu 2000-5000 mg / kg, Co 200-400 mg / kg, Se 100-200 mg / kg and I 50-130 mg / kg; and carrier of natural Chinese medicinal stone in the weight range of 60-80 %. Its compounding process includes calculating the added amount of organic chelates and inorganic compounds based on the required ratio of trace elements and their contents in organic chelate or inorganic compound; mixing through stirring in stainless steel stirrer and packing in gold foil bag. The trace element feed additive has obvious growth promoting effect on sea water left-eyed flounder and right-eyed flounder and less water environment pollution.

The invention relates to a positioning and marking method for embryonic-period primordial germ cells (PGCs), in particular to a bastard halibut embryonic-period primordial germ cell tracking and positioning method. The bastard halibut embryonic-period primordial germ cell tracking and positioning method includes the steps of fixing collected various periods of embryo samples of the bastard halibuts by a 4% PFA solution; using a PBS (phosphate buffer saline) solution with 50% of deionized formamide to preserve the embryo samples at the temperature of -20 DEG C, and subjecting the fixed and preserved embryo samples to oolemma removing, gradient methanol dewatering and rewatering; after rewatering, washing the various periods of embryo samples with PBS buffering liquid without RNA ( ribonucleic acid) enzyme, pre-hybridizing at the temperature of 62-65 DEG C for 2-4 hours; after hybridization, adding a hybridization solution with bastard halibut RNA probes into the various periods of embryo samples subjected to pre-hybridization for hybridizing overnight at the temperature of 62-65 DEG C; after hybridization, subjecting the various periods of embryo samples to washing, antibody incubation and rewashing, keeping away from light, and developing colors to achieve marking for tracking and positioning of the embryonic-period primordial germ cells of the bastard halibuts. The bastard halibut embryonic-period primordial germ cell tracking and positioning method has the advantages that the problems that yolks and oolemma of the samples hybridized in situ conventionally are difficult to strip and a background color is too deep after color developing detection are solved, and operation steps are simplified.

The invention relates to a method for breeding parent fish of paralichthys lethostigma. The method comprises desalination culture and salinization culture of standby parent fish as well as intensive culture of the parent fish. Specially made feed is fed during the intensive culture of the parent fish, wherein, the weight percentages of main materials in the feed are the following: 30-40 percent of Gobiidae or ammodytes, 15-20 percent of cuttlefish or sleeve-fish, 15-20 percent of sea cucumber and 20-30 percent of bastard halibut powder; the weight percentages of additives accounting for the main materials are the following: 1-2 percent of parent fish hardening agent, 0.4 percent of Vc and 0.4 percent of Ve; the feed particle size is 9mm-11mm. During light and temperature regulation and control, four bulbs are suspended over each parent fish culture tank with two bulbs of 60W and two bulbs of 150W suspended on the cross to obtain even light; water temperature is raised to 17 DEG C from 14.5 DEG C. Based on biological characteristics of wide optimum salinity of the paralichthys lethostigma, the invention makes use of F1 replacing fingerlings to culture standby parent fish in fresh water after desalination, and after salinization the standby parent fish is intensively cultured to obtain sexually mature egg-laying parent fish, thereby effectively solving the problem of high cost, restriction, poor parent quality and low survival ratio of introduced paralichthys lethostigma parent.

A bastard halibut sperm rapid cryopreservation method relates to a cryopreservation method for marine fish sperm and provides a bastard halibut sperm cryopreservation method with simple operation, rapid preservation and high sperm living rate. The present invention contains diluent, antifreeze agent and the confection of hormone cryoprotective liquid and is preserved in liquid nitrogen. The present invention is characterized in that the diluted natural seawater is used as the diluent for preserving sperms which can not activate sperms; glycol with 14g / 100ml density is adopted as the anti-freeze agent; the diluent and the antifreeze agent are used as the low-temperature hormone cryoprotective liquid for bastard halibut sperms and the low-temperature hormone cryoprotective liquid is precooled in a 4 DEG C thermostatic box; the sperm and the hormone cryoprotective liquid are evenly mixed in an eccentric pipe according to the proportion of 1:1 to 1:3; the eccentric pipe is placed below the port of a liquid nitrogen jar for balancing under -20 DEG C for 1 minutes and then rapidly thrown into the liquid nitrogen for preservation. The present invention is used for bastard halibut sperm rapid cryopreservation.

The invention relates to a detection method of a bastard halibut rhabdovirus in the aquatic livestock pathogen detection and diagnosis technical field. A real-time quantitative reverse transcription-polymerase chain reaction technology is adopted to detect the bastard halibut rhabdovirus of fish. A specific primer sequence and a fluorescence probe sequence are designed firstly and then nucleic acid of a sample to be detected is extracted; secondly, the real-time quantum reverse transcription-polymerase chain reaction is carried out, and ribonucleic acid containing target amplified fragments by in vitro transcription is used as a standard substance to draft a standard curve; finally, whether the sample to be detected contains the bastard halibut rhabdovirus is judged according to circulating valve value of the sample to be detected calculated by an apparatus software. The method of the invention can be widely applied to epidemic disease monitoring over aquatic plants of bastard halibut, weever, turbot, snapper sea bream, etc. and detection of the bastard halibut bullet-shaped virus in import and export trade. Compared with the prior art, the method of the invention is characterized by rapid detection, high efficiency, accurate quantum, strong specificity, wide detection scope, high sensitivity, etc.

A vitrifying freeze storage method for the embryos of fish, especially the bastard halibut, is disclosed, which features that the formula of vitrifying liquid suitable for the embryo of seawater fish is screened out, the embryo development stage suitable for vitrifying it is determined, a particular eluting solution is disclosed, and the step balance and the fast cooling and thawing techniques are used. Its advantages are high safety, easy operation and low cost.

The invention discloses a semi-sliding-tongue halibut female astopic AFLP segment and PCR authenticating method of genetic sex, which comprises the following steps: recycling female astopic AFLP segment; cloning AFLP segment; analyzing sequence; authenticating semi-sliding-tongue halibut through PCR method; establishing to separate and clone semi-sliding-tongue halibut female astopic AFLP segment; obtaining female astopic DNA segment; designing astopic primer; transmitting AFLP mark into SCAR mark; creating PCR technology for genetic sex authentication of semi-sliding-tongue halibut.

A flounder and flatfish engineering pond recirculating aquaculture system belongs to the technical field of marine aquaculture. The system comprises a water storage pond, a back water treatment pond, a coarse filtration pond, a conjoined combination aquaculture pond, a water feeding and drainage system, an oxygen supply device and a water quality monitoring system. On the one hand, the system still uses a traditional earthen pond aquaculture mode for culturing flounder and flatfish to grow in an environment similar to the wild so as to improve the quality of farmed flounder and flatfish. Meanwhile, engineering and systematic arrangement and layout are performed on the outdoor culture earthen pond to achieve factory and information management of flounder and flatfish pond culture, the output of farmed flounder and flatfish such as bastard halibut can reach to 1000-2000kg / mu (one mu equals about 1.667square meters), increasing 5-10 times of the output of traditional large single pond culture, and the culture efficiency and economic and ecological benefits are greatly improved.

The invention provides a method for promoting apoptosis of testis germ cells of bastard halibuts. The method comprises the following steps: (a) primarily breeding bastard halibuts in a reproductive period at a first high temperature until the semen of the bastard halibuts is completely digested; (b) secondarily breeding the bastard halibuts bred primarily at a second high temperature, and injecting 1,4-butanediol dimethanesulfonate into the bastard halibuts at the second breeding stage. The method has the good apoptosis effect on the testis germ cells of the bastard halibuts, and meanwhile thesurvival rate of the bastard halibuts is 85% or above.

A method of rearing aquatic species is provided, comprising feeding the aquatic species during at least part of the larval and / or post-larval stage with prey organisms, such as Aremia or rotifers, having in their total lipid content a DHA content of at least 12 wt %. The method is suitable for the rearing of fish species such as halibut, turbot, bass, bream and flounder.

The invention relates to an mRNA in-situ hybridization technology, in particular to a method suitable for bastard halibut gonad frozen section mRNA in-situ hybridization. The method includes the steps that bastard halibut gonad is immobilized with an excess PFA solution with the concentration of 4% and permeated with an excess methanol solution with the concentration of 100% in sequence and then is preserved for a long term; the sample which is immobilized and then preserved is subjected to rehydration and 20-30% saccharose sedimentation, then is subjected to OCT embedding and frozen section preparing and is dried for 2-3 h at 55 DEG C, and a frozen section is obtained to serve as an in-situ hybridization sample; the sample is washed for 3-5 min with excess DEPC water to remove an OCT embedding agent in the sample frozen section; the sample frozen section is subjected to washing, proteinase K digestion and immobilization, then is subjected to pre-hybridization and then is mixed with a hybridization solution containing a bastard halibut RNA probe at 50-70 DEG C for overnight hybridization; after hybridization treatment, washing, antibody incubation, rewashing, light-proof color development, termination color development, re-immobilization and section encapsulation are carried out, and thus expression localization of bastard halibut gonad related genes at the mRNA level is achieved. By means of the method, the problem that direct gonad in-situ hybridization of bastard halibut gonadal tissue is difficult is solved, and expression localization of bastard halibut related genes in gonad at the mRNA level can be clearly described.

The invention relates to a compound feed for aquaculture animals, particularly relates to an epinephelus akaara compound feed, and further relates to a preparation method for the epinephelus akaara compound feed. The epinephelus akaara compound feed disclosed by the invention is composed of the following various components in parts by mass: 20-30 parts of fish meal, 10-18 parts of euphausia superba meal, 10-18 parts of clam worm meal, 6-10 parts of kelp meal, 6-10 parts of red bean meal, 4-8 parts of cottonseed meal, 4-8 parts of sweet potato meal, 4-8 parts of rapeseeds, 4-6 parts of lucerne meal, 2-4 parts of houttuynia cordata, 1-3 parts of dandelion, 1-3 parts of dry ginger powder, 1-2 parts of shell powder, 1-2 parts of compound multivitamins, 2-4 parts of fish oil and 1-3 parts of soybean oil. The epinephelus akaara compound feed disclosed by the invention has the advantages of being low in feed coefficient, high in specific growth rate of bastard halibut after feeding bastard halibut, and high in survival rate.

The immunological growth promoter for sea water left-eyed flounder and right-eyed flounder has the effective components comprising xylose oligomer 28000-46000 mg / kg, bile acid 21000-38000 mg / kg, bacillus 15000-33000 mg / kg, lactobacillus 13000-25000 mg / kg, beta-glucosan 22000-35000 mg / kg, mannan 16000-25000 mg / kg; and carrier of common starch in the weight range of 10-50 %. Its compounding process includes calculating the added amount of the ingradients based on the effective components; and mixing through stirring in stainless steel stirrer. The immunological growth promoter for sea water left-eyed flounder and right-eyed flounder has obvious growth promoting effect and can stabilize water environment pollution.

The invention provides a preparation method of a fermented traditional Chinese medicine for preventing and treating intestinal diseases of aquatic products. The fermented traditional Chinese medicineis also called a bacterial medicine, water is added into a traditional Chinese medicine composition, compound fermentation strains are inoculated, and fermentation is performed to obtain the fermentedtraditional Chinese medicine. The invention further provides the corresponding fermented traditional Chinese medicine. The fermented traditional Chinese medicine has the functions of internally strengthening digestive absorption, nourishing stomach and protecting liver and externally resisting mushroom poisons and killing pathogens, and is used for preventing and treating the intestinal diseasesin aquaculture and applicable to prevention and treatment of the intestinal diseases of special aquaculture animals such as various saltwater fishes, freshwater fishes, shrimps, crabs, sea cucumbers,turbots, bastard halibuts, large yellow croakers, snappers, sea eels, globefishes, rice field eels, soft-shelled turtles and ictalurus punctatus. The fermented traditional Chinese medicine has good effects on the intestinal diseases such as white intestine diseases, empty stomach, white discharge and hangover caused by bacteria, viruses and antibiotics, and particularly has special effects on fatal diseases of the shrimps caused by enteritis, white discharge, empty intestines and empty stomach.

The invention relates to a marine fish embryo cell culture technology, in particular to an establishing method of a myosatellite cell system of bastard halibut during an embryonic period. The establishing method comprises the following steps: carrying out in-vitro separation on an embryo during a bastard halibut kirschner capsule formation period, removing an egg membrane, dispersing cells, and carrying out primary culture and secondary culture, thus establishing the myosatellite cell system during the embryonic period. The form of the myosatellite cell system, established by the invention, ofthe bastard halibut is fusiform or spindle-shaped mononuclear cells; myofiber can be differentiated to form by culturing for 5 days or above within a single generation or carrying out induction of anexogenous factor (horse serum). The myosatellite cell system is capable of supplying a large amount of myosatellite cells, and GFD (Green Fluorescent Protein) transfection and cynoglossus semilaevisspleen and kidney necrosis virus challenge assay detection verify that the myosatellite cells can be normally transfected or infected, so that the myosatellite cell system not only can be directly used for researching functional genes related to muscle growth and development of fishes and providing research materials for exploring a molecular mechanism for induction, proliferation and differentiation of muscle cells of the fishes, but also is capable of providing a platform for improved research and application of muscle characters in bastard halibut breeding.

A method of rearing aquatic species is provided, comprising feeding the aquatic species during at least part of the larval and / or post-larval state with prey organisms, such as Artemia or rotifers, having in their total lipid content a DHA content of at least 12 wt %. The method is suitable for the rearing of fish species such as halibut, turbot, bass, bream and flounder. The total lipid content of the prey organisms further including highly unsaturated fatty acids comprising 18:3, 18:4, 20:4, 20:5 (EPA) and 22:5 fatty acids. Also the prey organisms include 5 to 35 wt % phospholipids of the total lipid content, wherein the total lipid content is at least 20 wt % on a dry weight basis.

The invention relates to a bastard halibut dead end sequence, in particular to a bastard halibut primordial germ cell dead end gene and application thereof. The bastard halibut Dead end gene is shown as SEQ ID No.1. siRNA of the bastard halibut dead end gene is siRNA of 20-50 bp. A sequence of Dead end (Dnd) is acquired in bastard halibut, and the siRNA synthesized by using the sequence as a target gene has growth promoting effect, so that the sequence and related siRNA have extremely high value in application of promoting growth of fishes including seawater fishes, and a basic technical guarantee is provided for study related to growth promoting of the fishes including the seawater fishes. In addition, the gene sequence is a sequence unique to bastard halibut, so that growth of the seawater fishes like bastard halibut can be promoted.

The invention belongs to the technical field of marine organism cell culture and specifically relates to a construction method for a normal and whitening skin cell line of bastard halibut. The method comprises the following steps: firstly preparing a cell culture fluid, taking normal and whitening skin tissues of the bastard halibut as materials, rinsing and cutting, then directly putting into a 25cm<2> culture flask filled with a defined amount of cell culture fluid, and culturing at 24 DEG C; exchanging the cell culture fluid every 5 days, and adopting 0.25% trypsin for digesting cells for sub-culturing when the original generation cell grows into a single layer; and sub-culturing for one time per 8-10 days, and successfully constructing a cell line when the tenth generation is sub-cultured and the serum content in the cell culture fluid is reduced from 20% to 10%. The cell line of bastard halibut, acquired according to the method, is in a fiber form, more than 40 generations can be continuously sub-cultured, and the cell line can be directly applied to the etiologic characteristic research, vaccine research and functional gene research. The construction method is suitable for constructing the skin cell lines of the other fishes.

Two monoclonal antibodies B and C for the lymphocystics virus of bastard halibut are prepared from two hybrid tumor cells (CCTCC-C200419 and CCTCC-C200420) through conventional culture for 2-3 days to secrete a lot of said monoclonal antibodies B and C.

The invention provides an inducing method of bastard halibut allogynogenetic tetraploid, and belongs to the technical field of a marine organism breeding technique. The inducing method includes stepsof performing ultraviolet irradiation on fresh red sea bream sperm to inactivate; mixing the inactivated red sea bream sperm with fresh bastard halibut ovum, mixing the obtained mixed fluid of sperm and ovum with seawater at 15-19 DEG C for 3 minutes; then transferring the mixed fluid of sperm and ovum to the seawater at 0-3 DEG C to perform cold shock treatment for 40-50 minutes; cultivating theprocessed mixed fluid of sperm and ovum for 45-71 minutes; selecting floating spawns from the mixed fluid of sperm and ovum and placing the spawns in an environment with pressure of 550-750 kg / cm2 toprocess for 5-7 minutes; performing the flowing hatching on the floating spawns after the static pressure treatment. The method applies the cold shock and static water pressure method to perform chromosome multiplying treatment on the ovum activated by inactivated heterologous sperm, and realizes the objective that the induced tetraploid only contains maternal inheritance information; meanwhile, the method has high inductivity.

A method is provided for producing prey organisms such as Artermia and rotifers, for feeding aquacultural organisms in particular at the larval stage. The method comprises cultivating the prey organisms during at least part of their life cycle in an aqueous medium comprising at least one lipid component having a DHA content of at least 30 wt %. The enriched prey organisms preferably have a DHA content of at least 12 wt % of their total lipid content. The prey organisms are suitable feed for larvae of fish including halibut, turbot, bass, and flounder, and crustaceans and molluscs.

The invention relates to a feed additive for preventing and treating streptococcicosis of bastard halibuts and also relates to a preparation method of the feed additive. The feed additive is composed of, by mass, 27-33 parts of honey suckles, 20-27 parts of onions, 20-27 parts of earth from the east wall, 19-25 parts of mentha haplocalyx, 19-25 parts of roots of rosa roxbunghii, 19-25 parts of gleditsia sinensis, 16-24 parts of cirsium setosum, 16-23 parts of lycoris radiata, 16-23 parts of caesalpinia minax, 15-20 parts of caesalpinia sappan, 14-21 parts of cortex cercis chinensis, 14-21 parts of oxalis corymbosa, 12-19 parts of hydnocarpus anthelmintica, 11-18 parts of flos albizzia julibrissin, 10-18 parts of rhizoma chuanxiong, 10-17 parts of cactus, 9-17 parts of flower of kudzu vines, 9-16 parts of fructus perillae, 7-12 parts of rhizoma bolbostemmae and the like. The feed additive can effectively prevent and treat the streptococcicosis of bastard halibuts.

The invention relates to the field of molecular biology, specifically to preparation and application of expression plasmids of calmodulin and calreticulin of bastard halibut, specifically to application of the plasmids pCNCam and pCNCrt expressing calmodulin and calreticulin to antibacterial infection. When fish is injected with the expression plasmids of calmodulin and calreticulin, the fish has improved capability of resisting bacterial infection.

The invention discloses an artificial culture method for bastard halibuts. The artificial culture method comprises the following specific steps: (1) selecting an inner bay with smaller stormy waves and abundant fish baits; (2) selecting uniform specifications of offspring seeds with the overall length of more than 8 cm and the color and luster of approximately natural fish body color, and rejecting black and white individuals; (3) putting 800 bastard halibuts in every square meter at the beginning, wherein 15 bastard halibuts are put in every square meter when the weight of each bastard halibut is 800g, and the coverage rate of a stocking area is 80%-90% in a common water pool; (4) feeding by using trash fish, feeding four to six times every day in a young fish period, feeding three to four times every day when the weight of each bastard halibut is about 100g, and feeding twice every day when the weight of each bastard halibut is 400 g; (5) feeding after the separation of the bastard halibuts two to three times generally in the period that the length of the bastard halibuts is 8-10 cm, and feeding after the separation according to growth situation when the length of the bastard halibuts is 10-20 cm. Through the manner, according to the artificial culture method for the bastard halibuts disclosed by the invention, the manner of seaside captivity is adopted, and a good growing environment complemented with a scientific cultivation technique is provided for the bastard halibuts, so that the stress resistance is increased, and the survival rate is greatly improved.

The invention relates to a bastard halibut ecological marking method for release counting. The bastard halibut ecological marking method for release counting comprises the steps that firstly, bastard halibut spawns are distributed; secondly, when nutrient-enriched rotifer starts to be put for feeding when fry are 3 days old, wherein a nutrient enriching agent in the rotifer is added at the following quantity that 15 g of the nutrient enriching agent is used for every 100 millions pieces of rotifer when the fry are 3-11 days old, 25 g of the nutrient enriching agent is used for every 100 millions pieces of rotifer when the fry are 12-20 days old, and 15 g of the nutrient enriching agent is used for every 100 millions pieces of rotifer when the fry are older than 20 days; thirdly, when the fry are 12 days old, nutrient-enriched artemia starts to be put for feeding; fourthly, when the fry are 15-19 days old, compound feed starts to be put for feeding; the ecological marked-type bastard halibut is obtained and used for release bastard halibut counting study, wherein the nutrient enriching agent in the second step and the third step is prepared from DHA, EPA, vitamins and amino acid. By means of the method, the death rate is not increased, growth is not influenced, the maintaining time is long, markers are not likely to drop, the technological cost is low, quantization operation is easy, re-caught bastard halibut is easy to recognize, and wide application and popularization prospects are achieved.