33 results about "Pelodiscus" patented technology

The invention relates to a male trait induction method for pelodiscus sinensis in embryonic periods. The male trait inducer is characterized in that through dissolving an aromatase inhibitor and an adjuvant into alcohol with the purity of 92-98%, the inducer with an aromatase inhibitor concentration of 50-150 mu g / mu l and an adjuvant concentration of 50-150 mu g / mu l is obtained; and the adjuvant is selected from testosterone, methyltestosterone or testosterone propionate. According to the induction method provided by the invention, the male induction rate can be more than 95% just through spraying the inducer two times at the sex determination key time point in embryonic development, and meanwhile, the normal development and low drug residues of pelodiscus sinensis gonads are ensured; and the induction method provided by the invention is easy to operate, low in cost, and remarkable in economic benefit, thereby being a sex control technology which is worthy of popularization in the existing pelodiscus sinensis breeding.

The invention discloses pelodiscus sinensis sex identification specific primers and a method. The two pairs of primers in a sex identification primer system are a pair of female specific primers and apair of positive reference primers which can be amplified by using male and female genomic DNA. Among the primers, the female specific primers can only stably amplify a 196 bp band in the female genomic DNA, and the band cannot be amplified in the male genomic DNA; the positive reference primers can amplify a 482 bp band in the male and female genomic DNA. Thus, in a PCR reaction system, the female genomic DNA can amplify two bands of 196 bp and 482 bp, while the male can only amplify a band of 482 bp. The use of the primers facilitates the correct identification of the sex of pelodiscus sinensis, especially the sex identification of the pelodiscus sinensis in the embryonic stage which is difficult to distinguish through naked eyes. The primers lay the foundation for the study of the pelodiscus sinensis, and also provide an important tool for the breeding and artificial breeding of the pelodiscus sinensis.

The invention relates to a manufacture method of continuous paraffin sections for gonad of pelodiscus sinensis in embryonic period and application of the manufacture method in sex determination. The manufacture method comprises the following steps: (1) fixing a sample, (2) dying the gonad, (3) carrying out dehydration and transparentizing procedure, (4) absorbing paraffin and embedding, (5) cutting into sections and flatting sections, and (6) dying and mounting. The manufacture method is characterized in that the gonad is first dyed in the step (2); sections are subjected twice flatting by adopting water bath and high-temperature paraffin in the step (5). Compared with the prior art, the manufacture method has the advantages that the gonad is dyed first before being embedded, the gonads of a plurality of individuals are embedded in one paraffin block, determination is carried out through a manner of continuous tissue sections, and then the result accuracy is ensured. The manufacture method is simple to operate and convenient to popularize and apply. By utilizing the manufacture method, sex ratio of the pelodiscus sinensis in the embryonic period can be rapidly and accurately subjected to statistics.

The invention discloses a pelodiscus sinensis breeding and egg-spawning system under simulative natural ecological environment, and the system comprises a culture pond, wherein the edge of the culture pond is provided with an egg-spawning room in which a sand pit is arranged, a heating panel is tiled on the sand pit bottom, an insulation board is covered with a ceramic title layer on which sands from Han River are totally paved, an atomization nozzle is arranged above the sand pit and the edge of the sand pit is provided with a baffle, a hole is arranged on one side wall of the egg-spawning room facing towards the culture pond and connected with a wood ladder extended under the water in the culture pond. The system has the characteristics of high survival rate, quick growth rate and fine meat quality, the simulative natural ecological environment of sands from Han River is configured in the egg-spawning room so as to improve the egg-spawning and hatching rate, the cultured pelodiscus sinensis has the high quality and nutritive value, the edible taste and color is the same as that of the uncultivated testudinate, according to market conditions, the pelodiscus sinensis is widely accepted by consumers. The system has the advantages of great market capacity, wide prospect, and greater market value and social value.

The invention discloses a pelodiscus sinensis fodder containing low-protein fish meal added with methionine hydroxy analog-Ca. The raw material of the pelodiscus sinensis fodder comprises white fish meal, soy protein concentrate, fermented soybean meal, extruded soybean, chicken powder, squid powder, beer yeast, alpha-starch, gluten flour, zeolite powder, multivitamin, a compound mineral substance, choline chloride, glycine betaine, sodium dihydrogen phosphate and methionine hydroxy analog-Ca. According to the pelodiscus sinensis fodder disclosed by the invention, the methionine hydroxy analog-Ca is taken as the additive and is added into the pelodiscus sinensis fodder containing low-protein fish meal, so that the amino acid balance in the fodder is effectively kept, the growth promotion effect is excellent and the nutritional requirement for the growth of the pelodiscus sinensis is met. Compared with the traditional pelodiscus sinensis fodder taking fish meal as the main protein source, the pelodiscus sinensis fodder containing low-protein fish meal added with methionine hydroxy analog-Ca has the advantages that the protein content of the fodder is less than or equal to 44% and the additive amount of the fish meal is less than or equal to 26%. The fodder provided by the invention is capable of saving the dosage of the fish meal in the pelodiscus sinensis fodder and lowering the cost of the fodder, is ecological and environmentally friendly, and is capable of increasing the comprehensive breeding benefit.

The invention discloses a PCR detection method for identifying germplasms of four populations of Pelodiscus sinensis, a primer group and a kit. The primer group comprises primers of SEQ No.1 / SEQ No.4, SEQ No.2 / SEQ No.5 and SEQ No.3 / SEQ No.6. The kit comprises a 2*TaqPCR premixed liquid, a 5U / mul enzyme BglI and 10*digestion buffer liquid A, a 5U / mul HpaII and 10*digestion buffer liquid B, a 5U / mul ClaI and 10*digestion buffer liquid C, 10mg / ml BSA, a contrast DNA template and a 10pM amplimer. The method comprises the following steps: carrying out PCR amplification of extracted DNA through using the primer group, digesting to obtain a digestion product, carrying out agarose gel electrophoresis, and comparing the obtained electrophoresis result with a standard electrophoretogram to obtain a result. The detection method has the advantages of simplicity, good reaction stability and repeatability, and cost saving.

The invention relates to an integrated incubation system for pelodiscus sinensis eggs. The integrated incubation system comprises an insulated house, and is characterized in that a plurality of heaters are arranged on the ground of the insulated house; an atomization spray head is arranged on a suspension girder of the insulated house; convection windows are arranged at the upper parts of the vertical walls of the insulated house; shutter-type fans arranged inside the convection windows are connected with a frequency converter; a water pool is arranged in the center of the insulated house; incubation racks are arranged on the two sides of the water pool; a humidity sensor is arranged on the top of each incubation rack; a plurality of layers of incubation boxes are arranged on each incubation rack; a plurality of rows of pelodiscus sinensis egg brackets are arranged in each incubation box; clean sand is arranged among the pelodiscus sinensis egg brackets; a temperature sensor is buried in the clean sand; a plurality of layers of launching crawling ladders corresponding to the incubation boxes are arranged on each incubation rack; the launching crawling ladders are connected with the edge of the water pool. The integrated incubation system has the characteristics of high incubation rate, short incubation time and the like. The system is low in investment, easy to mount, large in market capacity and wide in prospect, and has huge market and social values.

The invention relates to a young pelodiscus sinensis constant-temperature culture system, which comprises a water pond and a greenhouse, wherein the water pond is covered by the greenhouse, and the bottom part of the water pond is provided with clean sand. The pelodiscus sinensis constant-temperature culture system is characterized in that the greenhouse consists of a north air blocking wall, a support skeleton and a light transmitting plastic film, wherein the support skeleton comprises inclined surface skeletons and vertical surface skeletons, the light transmitting plastic film is covered on the inclined surface skeletons and the vertical surface skeletons, a plurality of rows of ultraviolet lamps are arranged under the support skeleton, the air blocking wall is provided with a ventilation window, the ventilation window is provided with a louver type fan, a plurality of bait tables are arranged in the water pond, the top part of each bait table is in a step shape, the bottom part of the water pond is provided with an aeration spray head, the upper part of the water pond is provided with a water inlet hole, the water inlet hole is connected with the water feeding end of a natural gas boiler through a water inlet electromagnetic valve, the lower part of the side wall of the water pond is provided with a water drainage hole, the water drainage hole is connected with the water inlet end of the natural gas boiler through a water drainage electromagnetic valve and a purifier, and the purifier is arranged at the exterior of the pool. The young pelodiscus sinensis constant-temperature culturing system has the characteristics that the survival rate is improved, the occurrence risk of diseases is reduced, the use amount of medicines is reduced, and the like.

The invention relates to a pelodiscus sinensis heart cell continuous cell line which is preserved in the common microorganism center of China Committee for Culture Collection of Microorganisms and has a preservation number of CGMCC No.10597, and further relates to an establishing method and ultra-low-temperature cryopreservation method of the pelodiscus sinensis heart cell continuous cell line. The establishing method includes the following steps of firstly, conducting primary culture, wherein a pelodiscus sinensis heart is cut into tissue small pieces after being preprocessed, trypsin is inoculated into a culture bottle after being digested at the room temperature, and subculture is not started until cells are fully laid on 90% or more of the bottle bottom; secondly, conducting subculture, wherein air is blown on the bottle bottom through a trypsin digesting method to make adherent cells fall down so that subculture can be conducted. By means of the established pelodiscus sinensis heart cell continuous cell line, the research on viral diseases of pelodiscus sinensis and other turtle animals and the research on related therapy drugs for instance the analysis of chromosome and the research of iridescent virus can be conducted on the level of molecular cells, and the requirements for pelodiscus sinensis germplasm resource conservation and theoretical research and application can be met.

The invention relates to a method for purifying collagen derived from pelodiscus sinensis. The method is characterized by comprising the steps of soaking, fat removal and enzymolysis; in an enzymolysis process, carinas of the pelodiscus sinensis, of which impurities are removed, are put into acetic acid with the concentration of 0.5mol / L, wherein the ratio of the carinas to the acetic acid is 1:25, pepsase enzymatic hydrolysate is added for enzymolysis treatment, and extraction is carried out at 4-10 DEG C for 24-48 h, so that a pelodiscus sinensis carina collagen coarse extracting solution is obtained; in a salting-out purification process, the coarse extracting solution is dissolved into a Tris-HCL buffer solution containing NaCl, the mixed solution is centrifuged to obtain supernatant, NaCl is added into the supernatant, salting-out is carried out, centrifugation is carried out again, and precipitates and the supernatant are respectively collected; and precipitates are dissolved into acetic acid with the concentration of 0.5 mol / L, and freeze-drying is carried out after dialysis, so that the pelodiscus sinensis collagen freeze-dried product is obtained. Compared with the prior art, the method provided by the invention has the advantages that collagen extracted through enzymolysis is purified through a salting-out purification method, so that no environment pollution is realized, the extraction efficiency is high and the obtained product has good biological characteristics.

The invention relates to a Pelodiscus sinensis DNA fingerprint identification method adopting the technical scheme that high-purity genome DNA is quickly extracted from Pelodiscus sinensis muscle, and then a random amplified primer and an amplified reaction procedure are screened and built. The method comprises the following specific steps: (1) muscle pretreatment comprising drying temperature, moisture content and the like; (2) genome DNA extraction condition optimization, such as water bath time and temperature, centrifugal speed, time and the like; (3) a PCR amplified reaction system and a program establishment comprising a template, a primer and enzyme addition and the like, annealing temperature, time and cycle time design and the like; (4) under the optimal amplified condition, screening different random primers and building the molecular markers of Pelodiscus sinensis in different areas. Compared with the common shape identification technology, the technology is quick and accurate, is not affected by external feeding environmental condition, can quickly identify the Pelodiscus sinensis parent with different regional characteristics and lays the foundation for successively carrying out hybridization breeding.

The invention relates to a separated nucleic acid molecule having a nucleotide sequence shown as SEQ ID NO:1 or an antisense sequence of the SEQ ID NO:1. The nucleic acid molecule is derived from pelodiscus sinensis picornavirus strain CNSR2011. The nucleic acid molecule can be used for preparing primers, probes or kits, wherein the primers, the probes and the kits are used for detecting the pelodiscus sinensis picornavirus. The nucleic acid molecule can also be used for preparing a vaccine of the pelodiscus sinensis picornavirus, and an anti-pelodiscus sinensis picornavirus antibody.

The invention discloses a feed for improving calipash yield of pelodiscus sinensis. The feed for improving the calipash yield of the pelodiscus sinensis is prepared from the following materials: bean meal, corn flour, silkworms, bamboo worms, tenebrio molitor, wine lees, ootheca mantidis, herbs of shorthorned epimedium, hawthorn fruits, scutellaria baicalensis, radix isatidis, catechu, vetiver herbs, pelargonium geraniums, black algae, hydrocotyle vulgaris, fern root powder, banana peels, konjac gum, muskmelons, peanut shells, wheat bran, attractant and water. The ingredients of the feed disclosed by the invention are rationally matched, so that nutrition requirements and eating habits of the pelodiscus sinensis are satisfied; thus, the feed not only promotes digestions and absorptions of the pelodiscus sinensis, but also reduces food coefficients, decreases water pollution as well as improves disease resistances and growth rates of the pelodiscus sinensis. The calipash yields of the pelodiscus sinensis fed on the feed are averagely improved by 30-40%, so that the fed has obvious economic benefits and environmental benefits.

The invention discloses an MSTN (Myostatin) gene molecular marker selection method for growth characteristics of pelodiscus sinensis. The MSTN gene molecular marker selection method comprises the following steps: taking an initiation codon of one SNP (Single Nucleotide Polymorphism) site on a first intron sequence in a pelodiscus sinensis MSTN gene as a starting point T2195C; screening a breedingmarker of an MSTN gene which is tightly correlated with the growth characteristics from a pelodiscus sinensis colony and screening and constructing a pelodiscus sinensis breeding colony with the breeding marker from a breeding colony of the pelodiscus sinensis. By adopting the method disclosed by the invention, the accuracy of selecting the growth characteristics of the pelodiscus sinensis can beimproved, a foundation is laid for developing molecular marker auxiliary breeding of the pelodiscus sinensis and the reference is provided.

The invention discloses a seedling feeding method for artificial breeding pelodiscus sinensis. The method includes the following steps of: (1) selecting seedlings; (2) disinfecting; (3) releasing seedlings reasonably; and (4) feeding. The seedling feeding method for artificial breeding pelodiscus sinensis enables pelodiscus sinensis seedlings to adapt to the environment of an artificial breeding pool as soon as possible, and the growth is faster.

The invention discloses a LAMP detection kit and detection primers for pathogenic bacteria of bacterial mumps of pelodiscus sinensis; the kit includes specific primers, a reaction buffer liquid, Bst DNA polymerase and a fluorescent dye. The specific primers are composed of external primers, internal primers and ring primers. The kit has the advantages of strong specificity, high sensitivity and convenient operation, can make up for the current situation that mumps occurs frequently in the culture process of the pelodiscus sinensis but the pathogenic bacteria cannot be confirmed, and is suitable for screening and prevention of bacterial mumps of the pelodiscus sinensis.

The invention relates to an all-male pelodiscus sinensis fry breeding technology, in particular to a short-period efficient all-male pelodiscus sinensis fry breeding method. The method comprises the steps of pelodiscus sinensis egg temperature control hatching, pelodiscus sinensis egg estrogen treatment, young pelodiscus sinensis greenhouse breeding, pseudo-female pelodiscus sinensis screening andparent pelodiscus sinensis pond breeding, wherein the pelodiscus sinensis egg hatching temperature is controlled to be 28+ / -1 DEG C in the pelodiscus sinensis egg temperature control hatching; the pelodiscus sinensis egg estrogen treatment is that an estradiol solution is dropwise added to the eggshell surface closest to the embryo position in each of 15-18 time periods of pelodiscus sinensis embryo hatching; and a fluorescent quantitative PCR technology is adopted in the pseudo-female pelodiscus sinensis screening. According to the method, a sex hormone dropwise adding and pseudo-female pelodiscus sinensis induction technology is adopted, the operation is easy, the conversion rate is high, the sexual maturity period is greatly shortened through the young pelodiscus sinensis greenhouse breeding, and pseudo-female pelodiscus sinensis screened out through the fluorescent quantitative PCR technology are normally mated with original male pelodiscus sinensis to obtain all-male pelodiscus sinensis fries. The all-male pelodiscus sinensis fry breeding method provided by the invention is short in period and high in commercialization degree, and economic benefits can be remarkably improved.

The invention discloses a separation and in-vitro culture method for pelodiscus sinensis ovary cells. The separation and in-vitro culture method comprises the following steps: (1) selecting sexually immature female pelodiscus sinensis; sterilizing and taking out in-vitro ovary tissues; washing the in-vitro ovary tissues with a phosphate buffering solution and washing; putting the in-vitro ovary tissues into a container; (2) carrying out digestion treatment on the washed in-vitro ovary tissues through adopting collagenase and pancreatin, and centrifuging and collecting the cells; (3) putting the collected cells into a treated cell culture container and adding a complete culture medium for culturing; (4) after the cells are proliferated and fully spread on the bottom of the cell culture container, digesting by utilizing the pancreatin and carrying out subculture or freezing preservation to obtain the pelodiscus sinensis ovary cells. The method disclosed by the invention has novel steps, is simple and convenient and has high repeatability; a pelodiscus sinensis ovary cell line with a high survival rate can be obtained.

The invention discloses a method for fast identifying a Qingxi black strain and a Japanese strain in pelodiscus sinensis. The method for fast identifying the Qingxi black strain and the Japanese strain in pelodiscus sinensis is characterized in that skin tissue RNA of to-be-identified pelodiscus sinensis is extracted, reverse transcription is conducted to obtain cDNA, with the cDNA as a template,PCR amplification is conducted, an amplification forward primer sequence is shown in SEQ ID No.1 in the description, a reverse primer sequence is shown in SEQ ID No.2 in the description, if a 374-bp amplification product can be obtained from amplification, the to-be-identified pelodiscus sinensis is the Japanese strain, and if no 374-bp amplification products can be obtained, the to-be-identifiedpelodiscus sinensis is the Qingxi black strain. The method for fast identifying the Qingxi black strain and the Japanese strain in pelodiscus sinensis can be used as a method for identifying the Qingxi black strain and the Japanese strain in pelodiscus sinensis for genetic breeding, especially breeding of a black character, of pelodiscus sinensis.

The invention discloses an energy-saving water supply system for a pelodiscus sinensis aquaculture greenhouse. The system comprises the aquaculture greenhouse, a heating water storage system, a waterstorage temperature adjusting pool and a plurality of greenhouse water supply systems. The heating water storage system is arranged in the aquaculture greenhouse, the water storage temperature adjusting pool is arranged in the aquaculture greenhouse, and the greenhouse water supply systems are fittingly communicated with the water storage temperature adjusting pool. The heating water storage system comprises a water feeding pipe, a water feeding control three-way valve and a water delivery pipe, the water feeding control three-way valve is communicated with the water feeding pipe arranged in the aquaculture greenhouse, and the water delivery pipe matches with the water feeding control three-way valve. The water delivery pipe is laid on a black antioxidant plastic film layer and penetratesa roof of the aquaculture greenhouse to be laid above the roof of the aquaculture greenhouse in a continuously bent form, and the tail end of the water delivery pipe penetrates the roof of the aquaculture greenhouse to be positioned above the interior of the aquaculture greenhouse. The water storage temperature adjusting pool comprises a ground source heat pump air conditioner and a temperature display element. The ground source heat pump air conditioner is fittingly connected with the water feeding control three-way valve, and the temperature display element displays the temperature of the heating water storage system.

The invention discloses a biogas residue pelodiscus sinensis feed with an effect of promoting growth and a preparation method thereof. The feed is prepared from the following raw materials in parts by weight: 70-80 parts of biogas residues, 400-500 parts of pork liver powder, 200-300 parts of pork blood powder, 80-90 parts of corn meal, 30-40 parts of wheat middling, 60-70 parts of pine needle meal, 20-30 parts of cactus powder, 20-30 parts of hawthorn leaf powder, 10-15 parts of lard, 3-5 parts of edible salt, 2-3 parts of soybean sauce, 70-80 parts of garlic peel, 2-3 parts of cortex phellodendri, 1-2 parts of coptis chinensis, 2-3 parts of purslane, 1-2 parts of wild chrysanthemum flowers, 2-3 parts of radix sophorae flavescentis, 20-30 parts of apple juice, 20-30 parts of black fungus powder, 15-20 parts of sesame powder, 5-8 parts of rape flower powder, 0.5-1 part of lemon oil, 1-2 parts of honey, 2-3 parts of crab shell powder and 2-3 parts of chicken bone powder. The feed is comprehensive in nutrition, delicious in taste and convenient to use and has remarkable benefits; and pelodiscus sinensis grows fast, develops robustly, and has an elegant figure, a healthy body, a high meat production rate, delicious meat and rich nutrition.

The invention discloses a low temperature diluent of pig sperm based on Pelodiscus sinensis hibernating serum, which includes hibernating serum of Pelodiscus sinensis, yolk liquid of Pelodiscus sinensis, buffering salts, nutritional components and antibiotics. The low temperature diluent of pig sperm based on Pelodiscus sinensis hibernating serum of the invention is added by the hibernating serumof Pelodiscus sinensis so as to reduce the in vitro storage temperature of the pig sperm to 4 degrees Celsius, with the most suitable temperature being 6 to 8 degrees Celsius. Histological analysis ofcryopreserved sperm shows that the integrity of sperm acrosome is obviously better than that of normal temperature preservation.

The invention discloses a method for constructing an in-vitro culture and damage model of pelodiscus sinensis embryonic period skirt fibroblasts. The method is characterized in that: the cells are derived from a key period of formation of the skirt character of a special nutritional tissue part of pelodiscus sinensis; and according to the characteristics of the hierarchical structure of the skirt organization, the cells have functional specificity of generation, secretion and deposition of different types of collagen. The method mainly comprises the following steps: primary culture, subculture, cell cryopreservation and skirt fibroblast damage model construction. The skirt fibroblasts are treated with lipopolysaccharide (LPS) with the concentration of 160 [mu]g / mL for 4 hours to obtain the skirt fibroblast damage model. The method has the advantages of being high in survival rate, good in operability, strong in characteristic feature, simple and practical. The pelodiscus sinensis embryonic period skirt cell system constructed in the invention can be used for researching the generation and physiological action of the pelodiscus sinensis skirt specialized character on the molecular cell level.

The invention discloses pelodiscus sinensis bone strengthening feed. The pelodiscus sinensis bone strengthening feed is prepared from the following raw materials in parts by mass: 20 to 30 parts of fish meal, 10 to 15 parts of bean flour, 4 to 6 parts of menhaden oil, 2 to 3 parts of linseed oil, 10 to 15 parts of peanut meal, 8 to 15 parts of mung bean flour, 10 to 15 parts of chinese chive juice, 10 to 15 parts of radix astragali seu hedysari, 8 to 10 parts of moss, 5 to 10 parts of radix codonopsis, 10 to 15 parts of dry black fungus, 10 to 15 parts of rhizoma dioscoreae, 8 to 15 parts of porcine blood powder, 10 to 15 parts of colla corii asini, 8 to 10 parts of fructus jujubae, 1 to 1.5 parts of shelling hormone, 5 to 7 parts of sodium alginate, 1 to 3 parts of calcium dihydrogen phosphate, 1 to 3 parts of saccharomyces cerevisiae, 1 to 3 parts of vital gluten and 1 to 3 parts of xanthophylls. The pelodiscus sinensis bone strengthening feed disclosed by the invention contains richtraditional Chinese medicine ingredients and has a more delicious taste; fining breeding is carried out on pelodiscus sinensis through the pelodiscus sinensis bone strengthening feed, nutritive ingredients and the medicinal value of bones of the pelodiscus sinensis are increased, the bones of the pelodiscus sinensis grow more quickly, and the absorption ability of the bones of the pelodiscus sinensis is stronger, so that the self value of the pelodiscus sinensis is increased, and income of pelodiscus sinensis farmers is increased.