118 results about "Apostichopus" patented technology

The present invention provides a complete artificial mixed feed for stichopus japonicus. Said feed composition includes (by weight portion) 60-30 portions of sargassum thunbergii powder, 15-25 portions of sea-verdure powder, 15-45 portions of degummed sea-tangle powder, 15-5 portions of spirulina princes powder, 10-5 portions of fermentation yeast, 0.6-0.2 portion of polyvitamin complex, 0.8-0.4 portion of compound mineral salt and 10-60 portions of dried dredge.

The invention discloses a compound Chinese herbal preparation for treating bacterial ulcerate disease of apostichopus japonicus selenka; the components of the Chinese herbal preparation comprise the following weight ratios: 80-90% of dark plum and 10-20% of Chinese gall. After the decocting of the components by water, the preparation obtained is used for medicated bath of apostichopus japonicus selenka or sprinkled in a whole pool and has reasonable composition formula. The compound Chinese herbal preparation has the following advantages that the raw materials are prepared based on the formula for Chinese medicinal herbs, which strengthens the efficacy of components; the preparation has no toxicity for the apostichopus japonicus selenka and no influence on the normal life of the apostichopus japonicus selenka, has quite obvious effect on killing pathogens in apostichopus japonicusand selenka and can repair ulcerate tissues thoroughly; the compound Chinese herbal preparation is degradable under the natural conditions, which has no toxic and side effect and no residue in the apostichopus japonicus selenka; the compound Chinese herbal preparation has less recruitment, convenient use, simple composition, stable raw material source, cheap price and simple production process.

The invention discloses an excessive reproduction control technology of benthic macro-algae in an apostichopus japonicas culturing pond. The excessive reproduction of the benthic macro-algae in the apostichopus japonicas culturing pond can cause severe damage to the normal growth of the apostichopus japonicas. The excessive reproduction and massive death of the macro-algae can breed large amounts of pathogenic microorganisms such as bacteria, viruses and the like and consume much of the oxygen in the water, so as to cause the famine of the dissolved oxygen in the bottom water body, reduce the immunity of the apostichopus japonicas, and cause the disease outbreak and the like. Aiming at the problem of the traditional control technology and method of the benthic macro-algae, the invention provides three technical methods to control the damage caused by the excessive reproduction of the benthic macro-algae in the apostichopus japonicas culturing pond, wherein the first method comprises the step of compositely culturing the macro-algae of asparagus so as to control the excessive reproduction of the benthic macro-algae in a light shading manner; the second method comprises the step of splashing the mud with good suspension property so as to control the reproduction of the benthic macro-algae in a manner of reducing the transparency of the culturing water; and the third method comprises the step of by utilizing a shading net, adjusting the illumination so as to control the reproduction of the benthic harmful algae. The invention saves the resource cost, solves the key problem in the apostichopus japonicas culturing technology, and has the advantage of long-term market development and economic investment potential.

The invention relates to the technical field of screening and application of functional microorganisms in particular to screening of Bacillus amyloliquefaciens and application thereof in Apostichopus japonicas culture. The Bacillus amyloliquefaciens is collected on 24th December, 2015 in China Center for Type Culture Collection, abbreviated as CCTCC with the address: Wuchang Luojiashan Wuhan University Collection Center, Wuhan, Hubei, is claimed by Ludong University under CCTCC No. M2015774 and is named as Bacillus amyloliquefaciens ZYTC1-1. The Bacillus amyloliquefaciens is screened from healthy intestines of Apostichopus japonicas, has great ability to produce protease and amylase, can improve the activity of amylase in the testiness of Apostichopus japonicas, promote growth of Apostichopus japonicas and improve immunity and disease resistance of Apostichopus japonicas, and is widely applicable to the field of Apostichopus japonicas culture and aquatic feeds.

The invention relates to manual compound feed for a sea cucumber when the sea cucumber or an apostichopus japonica is cultured in a net cage. The feed comprises components of raw materials in parts by mass as follows: 5-10 parts of sargassum thunbergii, 20-30 parts of sargassum, 5-10 parts of white fish meal, 10-20 parts of scallop skirt powder, 5-10 parts of shrimp shell meal, 5-10 parts of fermented bean meal, 20-25 parts of kelp meal, 1-2 parts of refined fish oil, 0.2-0.5 part of compound vitamin and 0.2-0.5 part of a compound trace element. The raw materials are weighed proportionally and mixed to be uniform, coarse powder is ground, the materials obtained by grinding the coarse powder are subjected to superfine grinding to 80 meshes and then bonded and extruded to form platy feed, and the feed is packed after dried. The feed can meet requirements of ingestion and nutrient of the sea cucumber under the culture condition of the net cage, and can increase the growing speed of the sea cucumber cultured in the net cage; compared with an ordinary feeding manner, the weight gain rate is improved by 78.8%; and the feed waste can be reduced, the utilization rate of the feed is improved, and the pollution of residual bait to cultivation sea water is reduced.

The invention discloses a south domestication farming method of apostichopus japonicus. The south domestication farming method comprises the following steps of: selecting farming fries with the specification of 33.3-62.5g; transporting the farming fries to a south cultivation place, loading the farming fries into a drain brush and placing in a running water device during the transportation, wherein the temperature is kept at 8-12 DEG C; hanging a suspension cage type apostichopus japonicus farming device, balancing the bottom of the suspension cage type apostichopus japonicus farming device in water with a heavy object and adding feed into the suspension cage type apostichopus japonicus farming device; transporting to the south cultivation place, then injecting seawater into the running water device till the temperature difference between the running water device and the seawater in a farming field is 1-2 DEG C; and selecting the farming fries and placing into the suspension cage type apostichopus japonicus farming device for farming. According to the south domestication farming method of the apostichopus japonicus, disclosed by the invention, the environmental advantages and the resource advantages of southern ocean are utilized, the water temperature in spring and winter of the southern ocean is higher and conductive to farming the apostichopus japonicus; the method disclosed by the invention is easy to operate and convenient to manage; and simultaneously, the apostichopus japonicus has the advantages of high survival rate, fast growth and high yield, the farming species of high-quality aquatic economic animals in the south can be increased and the economic benefits of fishermen can be increased.

The invention discloses a sex determination method for apostichopus japonicas. The method comprises the following steps: (1) taking apostichopus japonicas coelomic fluid, and respectively preparing coelomic fluid supernatant and coelomocyte breaking fluid supernatant; (2) taking the coelomic fluid supernatant to carry out SDS-PAGE (Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis), and taking the coelomocyte breaking fluid supernatant to carry out native-PAGE; (3) judging whether SDS-PAGE gel contains a 50kDa protein band or not to determine whether the apostichopus japonicas is male or not, and judging whether a phenoloxidase zymogram after the native-PAGE is carried out contains three bands or not to determine whether the apostichopus japonicas is female or not. The sex determination method has the following advantages: 1) the apostichopus japonicas is slightly harmed; 2) reagent, equipment and technical requirements are simple, and a result is visual; 3) determination speed is high, and handling capacity is high; 4) two technologies are used for respectively determining male and female apostichopus japonicas, and accuracy is 90% or above.

The invention relates to a foraging feed of a special compound feed for breeding stichopus japonicus and a preparation method of the foraging feed. The technical problems of poor disease control, high cost and the like in conventional breeding of the stichopus japonicus are solved. The foraging feed is prepared through the following steps of adding mixed microbe fluid to the following raw materials in parts by weight of 7-13 parts of soybean meal, 6-9 parts of sweet potato residues, 5-8 parts of corn steep liquor, 5-10 parts of sea mud, 3-5 parts of bran, 8-10 parts of soybean protein concentrate, 5-8 parts of yeast protein, 6-10 parts of wheat flour, 3-6 parts of shrimp powder, 3-5 parts of spirulina powder, 5-10 parts of sea spinach powder, 0.5-1.6 parts of sodium alginate, 0.5-1 part of bentonite, 4-8 parts of betaine, 0.5-4 parts of immune polysaccharide, 0.3-0.5 part of taurine, 0.5-2 parts of honeysuckle flowers, 1-3 parts of radix astragali, 0.5-1 part of gypsum, 0.8-2 parts of haws, 1-3 parts of licorice roots, 1-3 parts of activated carbon, 0.5-1.0 part of calcium propionate, 3-5 parts of complex vitamins, 0.5 part of complex enzymes and the like, and performing fermentation. The foraging feed is prepared through the following steps of mixing the raw materials, performing crushing, performing screening, performing fermentation, performing drying and the like. The foraging feed of the special compound feed is widely applied to breeding of the stichopus japonicus.

The invention belongs to the field of molecular biology DNA labeling technology and application, and in particular relates to Apostichopus japonicas express sequence tag EST micro-satellite label screening development and application. The invention provides Apostichopus japonicas AjE101 micro-satellite specific DNA primers, a polymerase chain reaction (PCR) reaction system by utilizing the primers, and a detection method for an Apostichopus japonicas AjE101 micro-satellite DNA label. The detection method comprises the following steps of: extracting genome of Litopenaeus vannamei Boone; designing the specific primers at two ends of an Apostichopus japonicas AjE101 micro-satellite DNA core sequence; and performing PCR amplification on genome DNA of different groups of the Litopenaeus vannamei Boone or individuals in the group by using the primers, analyzing products, and determining the genome of each individual so as to obtain an Apostichopus japonicas polymorphism genetic variation map. The invention is mainly applied to Apostichopus japonicas germ plasm resource and genetic diversity analysis, cular population genetics, construction of genetic maps and research of functional genes.

The invention discloses a PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) method for identifying seven sea cucumber species. The PCR-RFLP method includes the steps of firstly extracting DNA (deoxyribonucleic acid) modules of sea cucumbers; subjecting specificity fragment of a CO I (coenzyme I) gene to PCR amplification to obtain a fragment about 690bp; secondarily subjecting amplified product to restriction enzyme digestion; and finally subjecting the enzyme-digested product to electrophoresis detection. Different sea cucumbers obtain stripes in different numbers and sizes, and selected specificity incision enzymes respectively include: Californian paracanthurus cucumber: SSP I, North Atlantic Ocean cucumaria: EcoR V, apostichopus japonicas: SSP I, thelenota ananas: EcoR V, acaudina molpadioidea: Sca I, isostichopusbadionotus: Pvu II, and actinopyga mauritiana: EcoR V and SSP I. The PCR-RFLP method overcomes the defects that a conventional morphological identification method is prone to affecting in accuracy, is heavy in workload and long in consumption time and the like. The PCR-RFLP method can be completed by only one-time PCR reaction, one-time enzyme digestion reaction and one-time polypropylene gel electrophoresis, and is simple to operate, short in parting time and good in specificity.

The invention discloses a stichopus japonicus oligopeptide cross-linked microcapsule, and a preparation method and application thereof. According to the invention, sodium hyaluronate, modified starch, maltodextrin and the like serve as cross-linking agents; a cross-linking core material and a cross-linking functional wall material are prepared firstly; then, the cross-linking core material and the cross-linking functional wall material are respectively subjected to two-step cross-linking embedding; therefore, the microcapsule has a three-dimensional structure which is formed by cross-linking a core material, pre-embedding a basic wall material and then embedding the cross-linking functional wall material from inside to outside in sequence; the three-dimensional structure is more stable, so that the external environment resistance of the prepared stichopus japonicus oligopeptide cross-linked microcapsule in the invention is remarkably improved; therefore, the oral bioavailability is improved; the shelf life is prolonged; and in addition, as the stichopus japonicus oligopeptide is embedded, the microcapsule also greatly improves the sensory flavour characteristics of the stichopus japonicus oligopeptide.

The invention provides a composite microecologics capable of improving growth rate of stichopus japonicus and application of the composite microecologics. The composite microecologics comprises bacillus subtilis, saccharomyces cerevisiae, cellulase, pectinase, xylanase, neutral protease, solid fermentation lactic acid bacteria and a carrier, wherein the bacterium content of the bacillus subtilis is greater than or equal to 1* 108 per g, the bacterium content of the saccharomyces cerevisiae is greater than or equal to 1* 109 per g, the content of the cellulase is greater than or equal to 1,000 U/g, the content of the pectinase is greater than or equal to 1,200 U/g, the content of the xylanase is greater than or equal to 1,500 U/g, the content of the neutral protease is greater than or equal to 12,000 U/g, the bacterium content of the solid fermentation lactic acid bacteria is greater than or equal to 200 mg/g, and the content of the carrier is greater than or equal to 500 mg/g. The composite microecologics can effectively improve the utilization efficiency of bait by stichopus japonicus and promote the growth of stichopus japonicus through adding zymin; the added fungicide can restrain reproduction of pathogenic bacteria in a water body and reduce the occurrence probability of diseases such as skin melting, head shaking, swollen mouth and viscus discharging; the composite microecologics has the advantages of safety, efficiency, specifity, no residue, no side effect and the like.

The invention discloses a complete formula feed for young apostichopus japonicas and a preparation method of the complete formula feed. The complete formula feed is prepared from the following components in parts by weight: 15 to 25 parts of sweet potato root tuber powder, 10 to 15 parts of sargassum thunbergii powder, 15 to 25 parts of sargassum powder, 25 to 35 parts of degummed kelp powder, 2 to 5 parts of spirulina platensis powder, 8 to 12 parts of yeast powder, 1 to 2 parts of enzymed soy albumen, 1 to 3 parts of composite vitamins, 1 to 2 parts of composite mineral salt, and 0.5 to 1 part of compound enzyme. According to the complete formula feed for the young apostichopus japonicas, sweet potatoes are used as formula components of a feed of the young apostichopus japonicas, so that the application amount of macroalgae such as sargassum thunbergii is effectively reduced, the dependency on the macroalgae is reduced, and excessive development and utilization of macroalgae resources are avoided.

The invention relates to a method for identifying apostichopus japonicus by means of a specific peptide fragment group, and a group of marker peptides capable of being used for identifying apostichopus japonicus when used alone or otherwise. The marker peptides are low-homology polypeptide fragments of apostichopus japonicus protein, wherein the low-homology polypeptide fragments are exclusive to apostichopus japonicus with a length of 5-30 amino acids. Even though it is found that there are plants or bacteria having the same protein, the marker peptides are exclusive to apostichopus japonicus by comparison with thelenota ananas, acaudina molpadioidea, isostichopus badionotus and holothuria Mexicana.

The invention discloses a feed additive for stichopus japonicus, a preparation method and a compound feed containing the feed additive. The feed additive for the stichopus japonicus comprises the following raw materials: rhubarb, baical skullcap root, Chinese gall, honeysuckle flower, wild chrysanthemum, indigowoad root and a synergistic agent. The preparation method comprises the following stepsof: weighing the raw materials in a weight ratio; crushing all the raw materials; and putting all the crushed raw materials into a mixer sequentially, and mixing uniformly. The compound feed containing the additive comprises 1 to 3 weight percent of additive. The technical scheme provided by the embodiment of the invention has the advantages that: the additive is prepared from Chinese medicinal herbs, does not have medicinal tolerance and toxic or side effects, can improve the growth speed of the stichopus japonicus obviously, improve the immunological activity of the stichopus japonicus and enhance the disease-resistant capacity of the stichopus japonicus, and has extremely high resistance to vibrio splendidus serving as main pathogenic bacteria. The stichopus japonicus is difficult to sicken, the survival rate is improved obviously, and the production benefit is improved obviously.

The invention discloses a method for quickly detecting pathogen of skin ulcerative syndrome in Apostichopus on site and a kit thereof. In the method, two pairs of primers, particular operation steps and the kit aiming at the vibrio splendidus are designed by using a loop-mediated isothermal amplification (LAMP) method. The DNA sequences of the reaction primer are as follows: VS-F3 is 5'-TGGTCCCCCCATATTCAGAC; VS-B3 is 5'-AAGTACCCCGAGGAAGAGAA; VS-FIP is 5'-GCCCCGTAACCGACACATCATTTTAGGATATCACGTGTCCCGC; and VS-BIP is 5'-GGCACTTTCCAGAGCCTTCGTTTTTAGATTCCGAAAGTAGCGGC. By using the method, expensive instrument and equipment or long detecting time is not required, the pathogen can be detected even less pathogen exists. The method has the advantages of low cost, accurate detection and the like, is convenient to operate, real-time monitor can be performed, and the epidemic situation can be discovered in time, the incidence trend of the epidemic disease is reduced, the pathogen can be prevented from distributing in early stage, and more loss brought to raisers is avoided.

The invention belongs to the field of molecular biology and the technical field of genetic engineering, and in particular discloses apostichopus japonicus peptidoglycan recognition protein with sterilization activity as well as a preparation method and application of the apostichopus japonicus peptidoglycan recognition protein. The apostichopus japonicus peptidoglycan recognition protein AJ-PGRP is obtained for the first time by utilizing an in-vitro recombination expression technology; the apostichopus japonicus peptidoglycan recognition protein (AJ-PGRP) is shown as an amino acid sequence in a sequence table of SEQ ID NO. 1. The recombinant protein has agglutination effect on staphylococcus aureus and escherichia coli, and has inhibition effect on the staphylococcus aureus, the escherichia coli, vibro splendidus and bacillus subtilis, and has potential application value in the aspects of developing novel broad-spectrum antibacterial drugs, feed additives and the like.

The invention provides a feeding system and method for apostichopus japonicas industrial aquaculture. The feeding system comprises a culture pond, a feed trough, a feed guiding pipe and a sprayer. The feeding system is characterized in that the feed trough is arranged on one side of the culture pond, the bottom of the feed trough is higher than the culture pond, the bottom of the feed trough is connected with the feed guiding pipe, the tail end of the feed guiding pipe is connected with the sprayer, an electromagnetic valve and a timer are installed at the joint of the feed trough and the feed guiding pipe, and the electromagnetic valve and the timer are connected with a control system. A spraying pipe is held manually or driven by a movable frame to move at constant speed form one end to the other end along the position above the culture pond, and apostichopus japonicas feed in the feed trough is drained into the culture pond below the spraying pipe through the feed guiding pipe based on the siphon principle. The mechanical degree is high, the culture efficiency can be improved, the labor intensity can be reduced, diseases can be reduced, the product quality can be improved, and comprehensive culture benefits can be increased.

The invention discloses an apostichopus japanicus ITGB gene, coding protein, a cloning method of the ITGB gene and a construction method of recombinant apostichopus japanicus ITGB genetically engineered bacteria. The apostichopus japanicus ITGB gene is characterized in that the apostichopus japanicus ITGB gene sequence is shown as SEQIDNO.1; the cloning method of the ITGB gene comprises the steps that nested primers of RACE are designed according to the sequence of an expression sequence tag (EST) which is homologous with the ITGB gene, and the full length of the gene is amplified through an RACE technology; the apostichopus japanicus ITGB protein sequence is shown as SEQIDNO.2, and the apostichopus japanicus ITGB protein is amplified through primers containing a BamH I locus and a Not I locus respectively; a target gene obtained through cloning is inserted into a carrier to obtain recombinant plasmid, induction expression is conducted on the recombinant plasmid, purification renaturation is conducted, and then the genetically engineered bacteria are obtained. The construction method of the recombinant apostichopus japanicus ITGB genetically engineered bacteria has the advantages that the obtained genetically engineered bacteria have high combining capacity with endotoxin lipopolysaccharide, and the combining effect with peptidoglycan and mannan is weak.

The invention provides a biological regulation technology for an apostichopus japonicus aquaculture pond by macroalgae control with siganus guttatus. According to the technical scheme, the technologyis divided into two steps: Step 1: large-sized fry culture of siganus guttatus: by overwintering culture of large-sized siganus guttatus fry in a nursery workshop, the problem that the size of siganusguttatus fry in the biological regulation of a pond is small and has weak algae density control ability and a narrow proper temperature range is solved, and survival rate and size of the siganus guttatus fry are increased; and Step 2: putting of the siganus guttatus: a proper size of siganus guttatus is selected and put into the pond according to the types and density of macroalgae in the apostichopus japonicas aquaculture pond. The invention has the following beneficial effects: by putting siganus guttatus into the Apostichopus japonicas aquaculture pond to establish a biologic chain of theapostichopus japonicas aquaculture pond, an ecological balance system is perfectly established, the number of macroalgae in the apostichopus japonicas aquaculture pond is rapidly and effectively controlled, and the effect of regulating water quality and building a proper growth environment for apostichopus japonicas is achieved; and water layers of the apostichopus japonicas aquaculture pond is effectively utilized for polyculture of apostichopus japonicas pond culture, and a new economic growth point is brought.

The invention discloses a fast quantitative detection method of apostichopus japonicas coelomic fluid virus. The method comprises the following steps: 1) extracting apostichopus japonicas coelomic fluid; 2) conducting ultrasonic disruption on coelomic fluid cells, freezing and thawing, and releasing virus particles; 3) filtering the coelomic fluid to prepare a alumina filter membrane containing virus; and 4) staining by avoiding light, flaking, observing and counting. The method provided by the invention can directly and fast detect virus in the apostichopus japonicus without killing the apostichopus japonicas; and the detection process can be completed in 5 hours, has the characteristics of simple operation, fastness and accuracy, and can be used for rapid screening of breeding apostichopus japonicus in apostichopus japonicas larval breeding.

The invention discloses bacillus amyloliquefaciens and an application thereof, and particularly relates to bacillus CQN-2 which is preserved in China General Microbiological Culture Collection Center (CGMCC) with the preservation number of CGMCC No.12420. The bacillus amyloliquefaciens disclosed by the invention is separated from healthy apostichopus japonicas intestinal tract fed in the south, strongly antagonizes bacillus of vibrio vulnificus, has an obvious antagonistic effect on pathogenic bacteria-vibrio vulnificus, and can replace an antibiotic drug for treating vibrio vulnificus.

The invention discloses a PCR detection primer and method of Vibro Splendidus and application of the PCR detection primer. The PCR detection primer is characterized by being designed according to the DNA sequences of a fur gene of Vibro Splendidus, the sequences of the designed PCR detection primer are VSFur RTF3:5'-ACAACAACCAGATTGCCAACA-3' and VSFurRTR3:5'-GATAACTTCACCGCAGTCTAAACAT-3'. The PCR detection method includes the following steps of firstly, preparing a matrix of seawater, apostichopus japonicus epidermis, apostichopus japonicus muscle or apostichopus japonicus coelomic fluid; secondly, establishing a 25-microlitre PCR reaction system; thirdly, conducting a PCR reaction, and finally conducting agarose gel electrophoresis to conduct result analysis. The PCR detection primer can be used for detecting Vibro Splendidus in seawater, apostichopus japonicus epidermis, apostichopus japonicus muscle or apostichopus japonicus coelomic fluid and has the advantages of being high in detection speed and sensitivity.

The invention provides a breeding method for pure white apostichopus japonicas.The method is characterized by comprising the steps that pure wild apostichopus japonicas bodies with the white backs and abdomens are collected and transferred into an aquaculture pond for independent isolation temporary rearing and storage; breeding base populations are screened out starting from the last ten days of April, artificial spawning, incubation and breeding are performed; when 30% of plankton larvae are developed to doliolaria, a white corrugated plate frame is put in to be used as an adhering substrate; regular water changing and feed putting are performed, the apostichopus japonicas larva color and growth development situations are recorded, poor larvae are screened out and eliminated, and the overall breeding process is performed in the white culture pond on the basis of the apostichopus japonicas color changing apatetic principle; the bred generation F1 white apostichopus japonicas is used as parents to be subjected to selfing and hybridization with stored basic populations to form a filial generation F2, and the filial generation F2 and the generation F1 are the same in breeding technological process and breeding conditions.High-strength standardized breeding is performed three times, the pure white apostichopus japonicas novel line of the generation F3 is obtained, the selecting and remaining rate is 90% or above, and the growth speed and adverse resistance characters are not significantly different from those of protospecies apostichopus japonicas.