92 results about "Sperm cryopreservation" patented technology

A practical method for freeze storage of fish sperm features that the diluent, antifreezing agent and container for the freeze storage of fish sperm chosen from many schemes, and a three-step freezing mode and a quick thawing method are designed.

The invention relates to a method for freezing and storing diluted semen of cow. Wherein, the invention is characterized in that: preparing said diluted semen into A, B, C, D deals; mixing them with right ratio; adding 5-20% pig semen into C and D deals, to improve the semen acrosome integral rate of cow semen control liquid, and reduce the density of semen, to save cost. The experiment has proved that the semen survival rate of stored diluted semen can reach 50.88-62.88%, while the average number is 57.41%, which is 15.13% higher than traditional method, and the semen acrosome integral rate can reach 50.88-53.20%, while the average number is 51.90%, which is 7.0% higher than traditional method.

The invention discloses a method for inducing cleavage gynogenesis of fish fry of cynoglossus semilaevis, which comprises the steps of cynoglossus semilaevis sperm freezing storage, ultraviolet radiation of sperms, and induction and identification of the cleavage gynogenesis of the fish fry. The cynoglossus semilaevis sperm freezing storage method is established, and a method for inhibiting the first cleavage to induce the multiplication of embryo chromosome of the gynogenesis of the cynoglossus semilaevis through hydrostatic pressure is adopted so as to screen effective dilution and freezing method for the cynoglossus semilaevis sperm freezing storage and screen the hydrostatic pressure shock starting time, hydrostatic pressure and processing time suitable for the cleavage gynogenesis of cynoglossus semilaevis; and an identification method for the gynogenesis of the fish fry of the cynoglossus semilaevis is established. In the method, the fish fry of the cynoglossus semilaevis through cleavage gynogenesis is obtained for the first time, and the inductivity of the gynogenesis of the fish fry reaches 0.25 percent; and the method has the characteristics of advancement, high efficiency, reliability and reliability, can be used for solving the problems that male cynoglossus semilaevis grows slowly, the ratio of female cynoglossus semilaevis is low and the culture cost is high, and has significant application value and wide promotion and application prospect in the aspects of breeding of cynoglossus semilaevis fry, sex control, culture of all female fry, pure line breeding and the like.

The invention discloses a trace sperm cryopreservation carrier and corresponding freezing and thawing methods thereof. The trace sperm cryopreservation carrier is in a rectangular, square or circular horizontal or groove shape and is made of a transparent plastic or transparent glass, and the size of the trace sperm cryopreservation carrier can bear 1-3 0.1-10.0 mu L freezing liquid micro drops. The freezing and thawing method comprises the following steps: after preparing the freezing liquid micro drops on the trace sperm cryopreservation carrier, inserting the trace sperm cryopreservation carrier into a sperm culture dish covered with oil, grabbing sperms by a microinjection needle, putting into the freezing liquid micro drops, taking out the cryopreservation carrier, soaking up mineral oil by virtue of an absorbent paper, rapidly freezing, directly inserting the cryopreservation carrier into an ICSI vessel when thawing, and thawing the sperms by utilizing the mineral oil with the temperature of 37 DEG C. According to the trace sperm cryopreservation carrier and the corresponding freezing and thawing method thereof disclosed by the invention, as the cryopreservation carrier is inserted into the sperm culture dish covered with oil, and the sperms are grabbed and frozen, a special moisture retention device is not needed, and evaporation of the freezing liquid micro drops is not needed to be worried about; the operation is very simple, the flexibility and reliability of assisted reproductive treatment for a patient with azoospermia are greatly improved, the sperm recovery rate can reach 100 percent, and the sperm viability is about 60 percent.

The invention discloses acipenser dabryanus sperm preserving fluid liquid and a preparing method and application. The acipenser dabryanus sperm preserving fluid liquid comprises saccharose, mycose, trihydroxy aminomethane, potassium chloride, reduced glutathione and carbinol. The acipenser dabryanus sperm preserving fluid liquid has simple components and is low in cryopreservation cost. The added reduced glutathione plays a good role in antioxygenation, well protects sperm plasma membranes and reduces damage to DNA molecules. The added mycose has the effects of preventing protein denaturation and effectively protecting sperms. By means of the acipenser dabryanus sperm preserving fluid liquid, motility of unfreezed sperms is higher than 70%, and the fertility rate reaches 45%.

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 discloses a sperm cryopreservation method of Charybdisjaponica, which is characterized by comprising the steps of material drawing, sperm activity identification, cooling balance, first-time precooling, second-time precooling, freezing and preserving and the like. Aiming at the biological characteristics of Charybdisjaponica sperm, the invention screens out a cryopreservation method suitable for Charybdisjaponica sperm, which can effectively preserve Charybdisjaponica sperm for a long term. After the Charybdisjaponica sperm is preserved in liquid nitrogen for one year by utilizing the method of the invention, the livability of unfrozen sperm can reach over 70%. The method has no need of expensive apparatus, simple operation, low cost and good effect, provides source guarantee of sperm for the artificial insemination of Charybdisjaponica, and can be used for other research fields such as hybridization breeding, germplasm preference, gynogenesis, species conservation and the like of Charybdisjaponica.

The invention discloses an efficient grouper sperm cryopreservation solution and a use method thereof, and belongs to the technical field of fish germplasm cryopreservation. The efficient grouper sperm cryopreservation solution is prepared from diluent and an antifreeze agent; the diluent is an aqueous solution and is prepared from, by mass, 10-20 g / L of glucose, 7.5-8.5 g / L of sodium chloride and 0.6-0.7 g / L of potassium chloride; the antifreeze agent is 1,2-propylene glycol, and the volume concentration of the antifreeze agent in the cryopreservation solution is 10-20%. The use method of the cryopreservation solution for preserving grouper sperms comprises the seven steps of diluent preparation, antifreeze agent preparation, cryopreservation solution precooling, sperm dilution, balancing and subpackaging, cryopreservation, unfreezing and fertilization. The cryopreservation solution is simple in component, low in toxicity, easy to prepare, easy to master by breeding technicians and high in freezing activity, high universality is achieved, and the grouper sperm cryopreservation survival rate is increased.

The invention provides cryopreservation solution of horse sperms and a freezing method thereof. The freezing method comprises the steps of firstly, applying mare milk and yolk low-density lipoprotein after special treatment to the cryopreservation solution of horse sperms at the same time; adding the cryopreservation solution by sub-steps in the freezing process; and controlling cooling, lower liquid nitrogen fumigation height and shorter fumigation time. The quality of freezing the horse sperms is obviously improved by the technology; the artificial insemination efficiency of frozen semen of the horse is greatly improved.

The invention discloses an application of a polysaccharide substance in preparation of a dilution solution for freeze preservation of goat sperms and a preparation method of the dilution solution. Thepolysaccharide substance comprises one or more selected from the group consisting of tremella polysaccharide, lentinan, auricularia polysaccharide and cordyceps polysaccharide. The invention also provides the dilution solution for freeze preservation of the goat sperms. The dilution solution comprises the polysaccharide substance, a nutrient substance, a buffer substance and the like. The dilution solution for freeze preservation of the goat sperms provided by the invention has the effects of improving a sperm motility rate, an acrosome and plasma membrane integrity rate after freeze preservation, and significantly improving goat reproductive performance and economic benefits.

The invention discloses a cryogen for Pacific cod sperm preservation, which is characterized in that: the cryogen is mixed with a freeze diluent and an antifreeze in a ratio of 9:1 (volume ratio), wherein the components of the freeze diluent The contents are: NaCl: 42-48mM, KHCO3: 2.5-3.5mM, CaCl2: 2.5-3.5mM, Glucose: 35-45mM, Tris: 1.5-2.5mM, Glutathione: 1.5-2.5mM, Bovine Serum Albumin: 1.5-2.5g / L, the pH value range of the frozen diluent is: 6.7-6.9; the antifreeze agent in the frozen agent is dimethyl sulfoxide (DMSO). As well as a method for cryopreserving Pacific cod sperm by using the cryogen. The freezing agent and the preservation method can preserve Pacific cod sperm for a long time, and can ensure that the sperm viability can still reach more than 80% after thawing.

The invention discloses a method for establishing a giant grouper sperm freezer and assisting grouper distant hybridization breeding. The sperm freezer establishing method comprises the steps of preparing sperm diluent, preparing sperm cryopreservation liquid, freezing and storing sperms and establishing the sperm freezer. The grouper distant hybridization breeding method comprises the steps of establishing the giant grouper sperm freezer, establishing saladfish breeding groups, cultivating saladfish parent fish and performing reproductive regulation, performing artificially spawning breeding, fertilizing the frozen sperms and performing incubation, and cultivating fries. According to the method, the first giant grouper sperm freezer is established in China, reproductive isolation of the groupers caused by factors, such as geographical distribution, optimum temperature range and breeding time, is overcome, and distant hybridization breeding of the grouper groups is possible, so that distant hybridization breeding varieties are cultivated.

The invention provides a human sperm cryopreservation liquid, a human sperm cryopreservation method, and a human sperm cryopreservation and recovery method. The cryopreservation liquid comprises a yolk liquid, glycerol, antibiotics, a refrigeration buffer solution, and quercetin or derivative thereof. The human sperm cryopreservation and recovery method comprises the steps of using the human sperm cryopreservation liquid, performing cold balance at a temperature of 4 DEG C, and then performing refrigeration at a temperature of minus 80 DEG C, and next, transferring to a liquid nitrogen tank for cryopreservation; and performing unfreezing and recovering in a water bath box at a temperature of 37 DEG C before use. The sperm cryopreservation liquid, added with the quercetin at a proper concentration, can effectively protect sperms and can remarkably improve survival rate and motility of revived sperms in the subsequent sperm recovery process, so that occurrence of sperm ROS and apoptosis can be reduced; and therefore, the human sperm cryopreservation liquid has application potential in the assisted reproductive technology.

The invention relates to an artificial hybridization breeding method for groupers high in growth speed, and belongs to the technical field of aquaculture. The method comprises the steps of parent selection, cage culture of parent red-spotted groupers, industrial culture of parent epinephelus tukula, sexual transformation of the epinephelus tukula, freeze preservation of seminal fluid of the epinephelus tukula, spawning induction of the parent red-spotted groupers, artificial insemination and hatching of fertilized eggs. By using the sperm freeze preservation technology, an epinephelus tukula sperm bank is built, reproductive isolation between the two kinds of groupers is broken through, and a new excellent hybrid variety which is high in growth speed is obtained. The problem is solved thatmale parent epinephelus tukula produces fewer sperms, and consequently, production is hindered, the productivity is greatly improved, and grouper breeding work is facilitated.

The present invention relates to a method for collecting high-concentration seminal fluid of scallop. Said method includes the following several procedures; incubation of high-quality parent scallop, preparation of suction head for sucking seminal fluid, collection of high-concentration seminal fluid and detecting vitality of collected spermatozoa. Said invention also provides the concrete requirements and steps of above-mentioned every procedure.

The invention relates to a pomfret sperm cryopreservation method including the steps as follows: selecting sexually matured parental pomfrets to gather fine-quality sperm; selecting suitable base liquid and anti-freezing liquid; mixing the sperm with the anti-freezing liquid by the volume ratio of 1:1 and then preserving the mixture on ice for 4-5 min; after reducing the temperature to minus 150 DEG C according to a three-step temperature reduction method, staying for 5 min; directly placing the mixture in liquid nitrogen for preservation; in defreezing, staying a freeze pipe in liquid nitrogen steam for 2 min, rapidly placing the freeze pipe in water bath of 28 DEG C to defreeze for 90 s and then placing the freeze pipe at room temperature to dissolve completely; and activating the defrozen sperm with sea water, wherein the rapid swimming time of the sperm is 57.41 s averagely, and the sperm viability (percent of the sperms moving rapidly accounting for all sperms) is 83.62% averagely. The fertilization rate of the pomfrets is greatly enhanced.

The invention discloses a Ptychidio jordani sperm cryopreservation method, which comprises four steps, namely semen collection, semen dilution, sperm cryopreservation, and sperm thawing. The semen ofPtychidio jordani is collected through an artificial abdomen squeezing manner, is then mixed with a diluent, and is mixed with an antifreeze agent to obtain a sperm preservation solution, with the diluent being one of a D-17 diluent, a Hank's solution and a Ringer's solution for fish, and the antifreeze agent being one of methanol, dimethyl sulfoxide and dimethyl sulfoxide; then the sperm preservation solution is injected into a cryopreservation tube, and the tube is immersed into liquid nitrogen for long-term storage; when to be in use, the cryopreservation tube containing the sperm preservation solution is taken out of the liquid nitrogen, and then immediately thaw in a water bath with a temperature of 37 DEG C, and the obtained Ptychidio jordani sperms are used for artificial insemination. The method can store the sperms of the Ptychidio jordani for long time and ensures the survival rates of sperms after thawing and resuscitation.

The present invention relates to method of increasing sperm and freeze preserving sperm for fish, and is especially the method of increasing sperm and freeze preserving sperm of verasper variegates as marine fish. The method includes selecting male verasper variegates capable of extruding out milk white sperm, injecting HCG in the amount of 150 IU / kg to abdominal cavity to increase sperm, adding antifreeze fluid to the obtained sperm and pre-cooling at 0-4 deg c for 9-11 min, sectionally lowering the temperature to ¿C180 deg c and maintaining in liquid nitrogen. During defrosting, the frozen sperm is first set inside warm water at 43 deg c to slight thawing and then defrosted at room temperature to obtain sperm with fertilizing capacity. The present invention is significant in breeding verasper variegates as one kind of rare marine fishes, germ plasm maintenance, genetic diversity and other aspects.

The invention belongs to the technical field of biology, and particularly relates to a sperm cryopreservation liquid and application in sperm cryopreservation of bostrichthys sinensis. Thesperm cryopreservation liquid comprises asperm diluent and an anti-freezing protective agent, wherein the sperm diluent comprises sodium chloride, sodium dihydrogen phosphate, sodium bicarbonate, potassium chloride, calcium chloride dihydrate, D-glucose and magnesium sulfate heptahydrate, and the anti-freezing protective agent comprises dimethyl sulfoxide, bovine serum albumin and trehalose, and the sperm cryopreservation liquid is used for cryopreserving bostrichthys sinensis sperms and is small in damage. The invention further provides a cryopreservation method of bostrichthys sinensis sperms, wherein the method is simple, convenient and rapid, is used for cryopreservation of the bostrichthys sinensis sperms, is high in sperm motility after recovery and high in in-vitro fertilization rate, and can be used for protection of bostrichthys sinensis germplasm resources and large-scale artificial propagation and production.

The invention discloses an acipenser schrenckii cryopreservation fluid and a long-term acipenser schrenckii sperm cryopreservation method. The pH of each liter of acipenser schrenckii cryopreservationfluid is 7.5-8.5. The acipenser schrenckii cryopreservation fluid is prepared from 20-30 mM of sucrose, 0.1-0.5 mM of potassium chloride, 20-40 mM of Tris-HCl, 5-10 g / L of bovine serum albumin, 2.5-10mM of reduced glutathione and antifreeze components and the balance ultra-pure water, wherein the antifreeze components are, by volume, one of 10% of methanol, 15% of DMSO, the mixture of 15% of DMSOand 10% of fresh egg yolk, 15% of propylene glycol and the mixture of 15% of propylene glycol and 10% fresh egg yolk mixture. The components of the acipenser schrenckii cryopreservation fluid are clear and definite, can protect sperms against damage caused by ice crystal, oxidization and the like, the fertilization rate can reach 45.87% after cryopreserved sperms are unfrozen and revive, healthyfry can be hatched, and the acipenser schrenckii cryopreservation fluid has the advantages of being simple, convenient, easy to operate and capable of completing cryopreservation without precious instruments when being applied to long-term acipenser schrenckii sperm cryopreservation.

The invention relates to a method for cryopreserving and activating lutjanus argentimaculatus sperm. The method comprises the steps of collecting, diluting, cooling and freezing the lutjanus argentimaculatus sperm and is characterized in that: the cryopreservation comprises the steps of: collecting the lutjanus argentimaculatus sperm, diluting and balancing the sperm by using a diluent and an anti-freeze agent, and cryopreserving the sperm by a fragmental and gradual cooling method; and the activation method comprises the steps of: activating the sperm in the unfrozen sperm by using seawater,and performing quality detection on the sperm and comprehensive quantitative analysis on the dynamic and static characteristics of the sperm so as to accurately evaluate the sperm quality and obtain the lutjanus argentimaculatus sperm with high survival rate and strong vigor; therefore, precious wild resources and excellent biological species can be effectively stored and can be fetched at any time as necessary.

The invention relates to a preparation method of a soya lecithin frozen diluent. Firstly, TCG liquid is prepared, soya lecithin is added to the preheated prepared TCG liquid in a weight percentage of1-10%, and the soya lecithin and the TCG liquid are uniformly stirred and then subjected to ultrasonic treatment to obtain a mixed solution; and finally, the mixed solution is subjected to high-pressure homogenization treatment to obtain the clear and transparent soya lecithin frozen diluent with stable properties and no precipitation after being placed for a long time. The soya lecithin frozen diluent prepared by the preparation method of the invention is used for cryopreservation of sheep semen after adding glycerol, which can improve the solubility of the soya lecithin and increase the stability of the solution by greatly reducing the particle size of the soya lecithin in the frozen diluent and has good effect on sperm cryopreservation.

The invention belongs to the technical field of sperm freezing, and particularly relates to a Hexagrammos otakii sperm productive cryopreservation method. The method comprises the following steps: after collecting sperms, uniformly mixing seminal fluid with a cryoprotectant, and pre-cooling in a refrigerator at 4 DEG C for 5 minutes; freezing a straw at -20 DEG C for 10 minutes, putting the straw into a straw sleeve, and putting the straw sleeve into an aluminum cryopreservation strip; putting the cryopreservation strip into a pail of a liquid nitrogen tank, putting the pail on the liquid nitrogen surface of the liquid nitrogen tank, cooling for 10 minutes under the condition of -160 DEG C to -180 DEG C, and then putting the cryopreservation strip linto liquid nitrogen for preservation; wherein the cryopreservation liquid is prepared by taking boiled and cooled sterilized natural seawater as a diluent and ethylene glycol with the final concentration of 15% (v / v) as a protective agent. Based on production requirements, the invention provides a simple and batch productive sperm cryopreservation method, and provides technical support for selective breeding of improved varieties of hexagrammos otakii and fishery production; according to the method, the sperm anti-freezing liquid is simple in proportioning, the freezing procedure is relatively convenient and fast, and the requirement for cryopreservation of productive sperms can be met.

A dairy cow sperm cryopreservation method relates to sperm preservation, in particular to a milk cow sperm cryopreservation method which has a protective effect on dairy cow sperm freezing and can improve the freezing effect. The cow sperm cryopreservation method is characterized in that the preservation method is to put the cow sperm into a low-temperature cryopreservation solution added with dove egg yolk for low-temperature preservation, and utilize the low-density lipoprotein contained in the dove egg yolk to inhibit the cow sperm Protective effect, reducing mortality in dairy cows after semen recovery. The dairy cow sperm cryopreservation method of the present invention adopts pigeon egg yolk to prepare, and pigeon egg yolk has a better protective effect on dairy cow sperm. Under this cryopreservation solution, dairy cow frozen sperm recovery motility, motility recovery rate and sperm quality The membrane integrity rate is higher than other cryopreservation solutions that have been reported to add different species of yolk; the use of this preservation solution can improve the freezing effect of dairy cow semen, and it is easy to popularize in practical production.

The invention relates to urechis unicinctus sperm cryopreservation liquid which comprises urechis unicinctus sperms, preservatives and antifreeze agents. A preparation method of the cryopreservation liquid includes the steps: collecting sperms; identifying sperm activity; diluting the sperms; reducing temperature, and balancing; firstly pre-cooling; secondly pre-cooling; preserving; unfreezing; eluting and the like. According to the cryopreservation liquid, the urechis unicinctus sperms can be effectively preserved for a long time and are preserved in liquid nitrogen, and activation rate of unfrozen sperms can reach 70% or more. According to the preparation method, expensive instruments are omitted, and the preparation method is simple to operate, low in cost and good in effect, provides sperm source guarantee for artificial insemination operations of urechis unicinctus and can be used for other research fields such as crossbreeding, germplasm selection, gynogenesis and species protection of the urechis unicinctus.

The invention belongs to the technical field of fish sperm cryopreservation, and particularly discloses deep-freezing preserving fluid of loach sperms and a freezing recovery method and application thereof. The method is utilized for carrying out deep-freezing (-196 DEG C) cryopreservation on sperms of loaches, and the motility rate of the resurgent sperms reaches 72.3%. Loach eggs are fertilizedwith the resurgent sperms, the fertility rate reaches 81.5%, the hatchability reaches 84.9%, and the spersms have no remarkable differences (P>0.05) with fresh sperms. The method is large in cryopreservation amount, good in effect and easy and convenient to implement and needs no expensive instrument and equipment, the problem that during actual production, the number of male loaches is small, andthe eggs cannot be fertilized in good time can be solved, and convenience is provided for large-scale production and genetic breeding studies of loach fry.

The invention discloses a human testicular tissue suspension cryoprotectant and a preparation method thereof. The human testicular tissue suspension cryoprotectant comprises the following raw materials: sodium chloride, potassium chloride, calcium chloride, magnesium chloride, monopotassium phosphate, sodium bicarbonate, HEPES, caffeine, sodium pyruvate, pentoxifylline, glycine, glucose and sucrose. The human testicular tissue suspension cryoprotectant also comprises sodium lactate and glycerol. The invention further discloses the preparation method of the human testicular tissue suspension cryoprotectant and a freezing method adopting the human testicular tissue suspension cryoprotectant. The cryoprotectant is specially designed for a testicular tissue suspension, the testicular sperm cryopreservation and resuscitation effect can be improved, the problems that potential safety hazards exist, the cryopreservation and resuscitation effect is poor, and movable testicular sperms are not easy to find are effectively solved, and the human testicular tissue suspension cryoprotectant has quite important clinical application value.

The invention discloses a rat sperm cryopreservation agent (RS-CPA) for a SD rat and a gene engineering rat under a SD rat genetic background. The RS-CPA comprises 5% of beam milk extract as a RS-CPA main component and also comprises raffinose. Through use of the beam milk extract, the RS-CPA greatly improves resurgent sperm vitality and a fertilization rate and realizes a refrigeration recovery rate of 10% or more.

The invention belongs to the technical field of marine organisms, particularly a method for carrying out interspecies cross artificial insemination on an egg of paralichthys olivaceus and a semen of paralichthys dentatus, which is frozen for preservation. The semen of paralichthys dentatus, which is frozen for preservation at an ultralow temperature for a long time, is dissolved in water by two steps; after the semen of paralichthys dentatus is dissolved, diluent of which the volume is three times of that of the frozen semen of paralichthys dentatus is added into the frozen semen of paralichthys dentatus; after the frozen semen of paralichthys dentatus and the diluent are mixed, centrifugation is carried out for 5min; then supernate is removed to obtain frozen semen suspension with the same volume with the frozen semen of paralichthys dentatus before the frozen semen of paralichthys dentatus is diluted; the frozen semen and natural seawater are uniformly mixed according to the proportion of a volume ratio of 1:40 and the mixture is activated; after the mixture is activated, the egg of paralichthys olivaceus is injected and the mixture and the egg of paralichthys olivaceus are slightly and uniformly mixed to enable the egg of paralichthys olivaceus and the frozen semen of paralichthys dentatus to be sufficiently in contact; standing is carried out for 5 to 10min and egg washing is carried out to obtain a fertilized egg to carry out incubation. The invention establishes a cryopreservation technology for semen of paralichthys dentatus and a method for interspecies cross artificial insemination on the egg of paralichthys olivaceus, overcomes the difficulty of asynchrony of development of parental fishes of paralichthys dentatus and paralichthys olivaceus and provides a novel method and a novel idea for seawater fish cross breeding.

The invention provides cryopreservation solution of horse sperms and a freezing method thereof. The freezing method comprises the steps of firstly, applying mare milk and yolk low-density lipoprotein after special treatment to the cryopreservation solution of horse sperms at the same time; adding the cryopreservation solution by sub-steps in the freezing process; and controlling cooling, lower liquid nitrogen fumigation height and shorter fumigation time. The quality of freezing the horse sperms is obviously improved by the technology; the artificial insemination efficiency of frozen semen of the horse is greatly improved.