43 results about "Acrosome formation" patented technology

The invention provides a frozen semen diluent formula for flocks and herds, and the formula of each 100mL of diluent comprises the following components: 1.35-1.55g of citric acid, 2.4-2.6g of trihydroxymethyl aminomethane (tris), 1.0-1.2g of glucose, 4.5-6.8ml of glycerol, 18.0-21ml of yolk, 4.5-7.5ml of bovine serum albumin (BSA), 0.45-0.75ml of vitamin B12, 50000-100000IU of benzylpenicillin potassium, 50000-100000IU of streptomycin sulfate and the balance of distilled water. The formula provided by the invention is low in cost, simple and convenient in preparation and using and operation steps, capable of effectively improving the sperm motility, the acrosome integrity rate and the plasmalemma integrity rate, and capable of reducing the sperm aberration rate.

The invention discloses an antifreeze used for freezing and preserving boar semens as well as preparation and application thereof. The antifreeze used for freezing and preserving the boar semens, which is provided by the invention, contains pigeon egg yolk and concretely comprises the following components in parts by volume: 82 to 90 of freezing base liquid and 10 to 18 of pigeon egg yolk. The antifreeze used for freezing and preserving the boar semens, which is provided by the invention, has simple preparation method, easy material acquirement, reliable effect and easy popularization, is suitable for insemination after long-time preservation and long-distance transportation after the artificial insemination of the frozen semens of the tubules of boars and the ultra-low temperature freezing of the semens of the boars, can obviously enhance the survival rate of the semens, the integrity of the acrosomes and the integrity of the plasmalemma after the boar semens are frozen and unfrozen, and keeps the insemination activity of the boar semens.

The invention relates to preserving liquid for sperm, in particular to the preserving liquid for swine sperm, and belongs to the technical field of animal reproduction. The preserving liquid for the swine sperm comprises the following components: 25.5 to 35.0g / l of anhydrous dextrose, 5.1 to 7.0 g / l of trisodium citrate, 1 to 10 g / l of sodium bicarbonate, 2 to 4g / l of EDTA, 1 to 3g / l of potassium chloride, 2.4 to 3.4g / l of citric acid, 8.5 to 9.5g / l of trihydroxymethylaminomethane, 0.26 to 1g / l of amikacin and 0.2 to 1g / l of vitamin C. Through detection, in the swine sperm preserved by the preserving liquid, the sperm viability reaches 0.60 to 0.85 and the integrity rate of sperm acrosome reaches 90 percent, which excess the requirement of the current artificial fertilization.

The invention discloses a novel frozen semen diluent used for improving the freeze-thaw quality and fertilization rate of frozen bovine semen and a preparation method thereof. Every 100ml of the prepared frozen semen diluent contains 0.9-1.2g of sodium citrate, 3-5g of sucrose, 18-22ml of yelk, 4-7ml of glycerin, 6-9mg of vitamin E, 500-700mg of vitamin C, 35mg of spectinomycin and 80 thousand units of gentamicin. The invention adds VE and VC into the semen diluent for the first time, so as to increase the motility rate and acrosome integrity of the frozen sperm as well as the activity of antioxidant enzyme in seminal plasma, thus improving the quality and fertilization rate of the freeze-thaw sperm.

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 relates to a freezing diluent for a seminal fluid of livestock. Every 100 ml of the freezing diluent comprises components as follows: 2.71 g of trihydroxymethyl aminomethane, 1.4 g of citric acid, 1.0 g of monosaccharide, 5-20 ml of fresh yolk, 0-10 ml of a permeable protecting agent, 0.1 million IU of penicillin, 0.1 million IU of streptomycin, 0.05-5 g of polyvinyl alcohol and the balance of ultrapure water. The common high-molecular polymer polyvinyl alcohol serving as an ice crystal inhibitor replaces natural antifreeze protein and is applied to the cryopreservation research of the seminal fluid of livestock for the first time. The survival rate of unfrozen sperms is about 75%, the activity is 50% or higher, the acrosome completeness is 65% or higher, the plasma membrane completeness is about 50%, and the non-return rate after artificial insemination is 70% or higher.

The present invention provides compositions and methods useful for regulating fertilization and for use as a contraceptive based on the discovery herein of an oocyte specific protein, SAS1R (Sperm Acrosomal SLLP1 Receptor), which is a sperm protein receptor. Six SAS1R variants, including the full length SAS1R, were identified. mSLLP1 and SAS1R co-localized to oocytes and to acrosomes of acrosome-reacted sperm. Interactions between mSLLP1 and SAS1R were demonstrated by far-western analysis, in a yeast two-hybrid system under stringent selection conditions, and by immunoprecipitation of SAS1R by anti-mSLLP1 as well as the converse. Purified recombinant SAS1R was found to have protease activity, to inhibit fertilization in-vitro, and to induce an immune response in females. Together, the results suggest SAS1R is a proteolytically active, oocyte and early embryo specific oolemmal metalloprotease receptor for the sperm intra-acrosomal ligand SLLP1 and is a target for regulating fertilization and as a contraceptive.

The invention relates to an antifreeze agent for diluting pig sperm, pertaining to the technical field of animal breeding. The antifreeze agent is prepared by N-acetyl-D-dextrosamine, hyaluronic acid, amido-sodium-lauryl sulfuric ester, glucose, trihydroxymethyl aminomethane, sodium citrate, sodium bicarbonate, potassium chloride, bactericide, distilled water and the like, wherein, the N-acetyl-D-dextrosamine can stimulate the emulsification function of the amido-sodium-lauryl sulfuric ester, enhances the protection function of the amido-sodium-lauryl sulfuric ester and yolk, the hyaluronic acid and the amido-sodium-lauryl sulfuric ester can effectively inhibit the damage rate of freezing and thawing sperm plasma membrane and DNA and improve the quality of freezing and thawing sperm, the glucose is the energy source for metabolism in vitro of the sperm, and the sodium citrate can maintain the osmotic pressure balance between diluted antifreeze agent environment and the internal environment of the sperm. The antifreeze agent for diluting pig sperm can effectively prolong the survival time of pig sperms for ultra-low temperature cryopreservation, and improve the liveliness of the freezing and thawing pig sperms, the integrity of plasma membrane and the integrity of acrosome.

A natural vegetable feed additive for stud ram relates to a feed additive for stud ram, which solves the problems of body situation reducing, libido and semen quality reducing of the ram during the service period, not high conception rate of ewe and service life reducing of the ram during the service period. The invention employs the prescription of the following components, which namely consists of Epimedium, dodder, actinolite, campanumaea pilosula, Rehmannia Glutinosa and Charred Triplet. The product of the invention enhances the sexual behavior of the ram, and remarkably improves the indexes like the sperm viability, sperm density, and ejaculatory amount, does not have side effects to the cloud characteristic of the sperm, the pH, the sperm abnormal rate, and the abnormal rate of acrosome, and can simultaneously prolong the service life of the stud ram by 1 to 1.5 years.

The invention discloses a culture-medium of pig semen. Per 100ml of the culture-medium contains the following components: 5-20 g of lactose, 0.005-0.05 g of N-acetyl-D-glucosamine, 0.05-0.5 g of DHA, 1.0-5.0 ml of glycerine, 0.2-1.5 ml of OEP, 10-40 ml of yolk and 50,000-200,000 IU of penicillin and / or streptomycin. The invention also discloses a method for processing the pig semen. Proved by detection, the activity of the pig semen processed by the method reaches 0.3-0.6, and the acrosome integrity exceeds 50 percent, thereby completely conforming to the requirement of artificial fertilization at present.

This invention discloses one sperm acrosin activity quantifying testing agent, which comprises depressor, buffer liquid, terminal liquid and bottom liquid, wherein the said bottom liquid comprises N-ª

The invention provides a fluorescent staining method of evaluating sperm motility of a tree shrew. The fluorescent staining method comprises the following steps: (1) preparing a TALP-HEPES sperm rinse; (2) collecting sperms; (3) washing the sperms; (4) staining; and (5) detecting by a microscope. According to the fluorescent staining method provided by the invention, by utilization of the characteristics that Hoeches 333422 and PI are different in reaction to dead and alive sperms and can be excited by a laser with a same wavelength, fluorescent staining is carried out on sperms of the tree shrew, and the sperm motility can be accurately detected without needing to switch a light source; in addition, as the two dyes respectively emit red and blue fluorescence after being excited by the laser, a dye which emits green fluorescence can also be introduced to detect the integrity of sperm acrosomes while the motility is detected, and the sperm motility and the integrity of the sperm acrosomes of the tree shrew are accurately detected by virtue of a fluorescence microscope or a flow cytometer.

The present invention belongs to the technical field of immunoassay, and more particularly relates to a method for analyzing and identifying the mammal Y sperm typing by using an identification protein. According to the present invention, by using immunofluorescence, immunoblotting and flow cytometry analysis, the gene protein TSPY expressed and positioned at the Y sperm acrosome part in the Y sperm is determined, and can be used as the Y sperm identification antigen protein in the separation of the X sperm and the Y sperm of mammal; the Y sperm identification protein TSPY can be used as the Y sperm identification antigen protein in the separation of the X sperm and the Y sperm in the mammal semen, wherein the mammal comprises cow, sheep, pigs, rabbits, horses and the like, and does not comprises humans; and the Y sperm identification protein has advantages of accuracy, rapidness, simpleness, low cost, simple equipment, low harm on sperm and the like in the separation of the X sperm and the Y sperm, and is suitable for animal production.

The invention discloses an acrosome reaction kit, which comprises tissue slide preserving fluid, a fluorescent conjugate and a fluorescent sealing solid liquid, wherein each 100 ml of the tissue slide preserving fluid contains 0.5ml of glutaraldehyde, 5ml of methanol, 2ml of ethanol, 0.2g of polyvinylpyrrolidone (PVP), 1ml of formalin and the balance of purified water. The kit can carry out a sperm-induced acrosome reaction and detection; and if calcium ion induction is not carried out, the kit can carry out the acrosome reaction condition evaluation and the acrosome integrity determination. By the tissue slide preserving fluid, the test process for the sperm-induced acrosome reaction and detection is carried out by two parts, samples and test results which are obtained in the first part can be preserved and the final test result cannot be influenced if the operations of the second part are carried out in the subsequent 72 hours, so that the work intensity can be relieved. The fluorescence of a fluorescent staining system is strong and cannot be attenuated after being stabilized for 15 days, so that the detection result is more stable and is easy to interprete and the repeatability of the result can be ensured. The kit and a determination method can be clinically applied and popularized.

The invention discloses a preparation method of a porcine frozen semen diluent. The diluents comprises the following components by weight, and the preparation steps of the diluent includes: S1: material taking, S2: pH value adjustment, S3: fresh egg yolk adding, S4: centrifugation, S5: standing, and S6: filtration. The inositol compound employed in the invention can effectively reduce the formation of ice crystals formed in a semen freezing process, reduce the mechanical injury of ice crystals on sperms, and finally improve the motility rate, vitality, acrosome integrity, plasmolemma integrityand other technical indexes of porcine frozen semen, wherein penicillin and streptomycin mainly play the role of sterilization and microbial growth; monosaccharide is used as the main energy substance; citric acid and trihytdroxymethyl aminomethane interact to mainly serve as a buffer solution for decreasing sperm motility, thereby prolonging sperm lifetime and maintaining the fertilizing capability.

The invention relates to an ultralow temperature cryopreservation method of macaca primate semen, belonging to an animal breeding method and comprising the following steps: dissolving Tris, TES, lactose, glucose, raffinose, sodiumpenicillinG, streptomycin sulphate and the like in water, centrifuging, taking supernate, preparing the supernate into semen diluent, adding glycerol to prepare deicing fluid, diluting the semen by using diluent, cooling the temperature to 4 DEG C, mixing with the dicing fluid precooled to 4 DEG C with the same volume, subpackaging into frozen straws, cooling by utilizing liquid nitrogen steam, and placing in liquid nitrogen at the temperature of minus 196 DEG C for preservation. When in use, the semen is defrosted and revived. The method in the invention can improve the revived motion range, the survival time, the plasma membrane and acrosome completeness and the fertility rate of the frozen sperm of the macaca primate.

The invention provides a cryoprotectant suitable for cryopreservation of human spermatozoa. The cryoprotectant mainly comprises the following components: sodium citrate, trihydroxymethyl aminomethane,N-[tris(hydroxymethyl)metyl]-2-aminopropanesulfonic acid, glycerinum and mycose. The provided cryoprotectant does not have ingredients of animal origin such as yolk, and thus, potential biological safety hazards are avoided. Compared with the prior art, the cryoprotectant has the advantages that if the human spermatozoa is subjected to cryopreservation by the cryoprotectant, the recovery can be better after the human spermatozoa is unfrozen, the number of recovered forward moving spermatozoa is large, the forward moving ability is good, meanwhile, the spontaneous acrosome reaction rate and DNA fragmentation rate of the recovered spermatozoa are low, namely, the number of spermatozoa with complete acrosome is large, nuclear injury of the spermatozoa is small, and protecting ability to functions of the spermatozoa is better.

The invention relates to a diluent for preserving rabbit sperms at normal temperature and a preparation method thereof and belongs to the technical field of animal reproduction. The diluent is prepared by adding water into 30-40g of tris(hydroxymethyl)aminomethane, 15-20g of citric acid, 6-15g of dextrose, 0.5-1g of potassium chloride, 0.2-0.5g of glutathione and 0.05-0.1g of kanamycin until the total volume reaches 1L; and the pH value of the diluent is 6.8-7.0. By using the diluent, the rabbit sperms can be preserved for a short time, and the sperm viability can be 0.7 and the acrosome integrity rate can be 87% after the rabbit sperms are preserved by using the diluent for 48 hours at normal temperature, thus the sperm viability and the acrosome integrity rate exceed the requirements of artificial insemination of rabbits, the conception rate is up to 89.4%, and the conception rate reaches or is superior to those obtained by using the similar import diluents. Meanwhile, the diluent provided by the invention is used and stored more conveniently, and the diluent can be instant availably at the time of artificial insemination, thus being convenient and quick. The diluent is prepared simply and can be used for lowering the cost of the artificial insemination of the rabbits.

The invention discloses a nutritional antifreeze diluting agent for improving sperm in-vitro storage and a preparation method of the nutritional antifreeze diluting agent. The nutritional antifreeze diluting agent is prepared from carbohydrates, a cytomembrane stabilizing agent, amino acids, Tris, glycerin, citric acid, penicillin, disodium edetate and double distilled water, wherein the pH is 6.9-7.2, and the osmotic pressure is 300mOsM. Sperm motility, acrosome integrity and plasmolemma permeability are all improved to different extents while the sperm malformation rate is decreased, and remarkable effects are achieved. By application of the nutritional antifreeze diluting agent to boar semen, farrowing rate and live farrowing quantity of sows can be increased, and reproductive performances of the sows are improved.

The invention discloses a single ultraviolet light excited fluorescent staining method used for evaluating sperm activity of Kunming wolf dogs. The method is realized by virtue of four steps of preparing a TCG sperm washing solution, washing sperms, staining and detecting with a microscope. According to the method disclosed by the invention, two types of fluorescent dyes which can be excited by ultraviolet light of a same wavelength are adopted to perform fluorescent staining on sperms of Kunming wolf dogs, and the sperm activity can be accurately identified without switching a light source; and moreover, one of the two types of fluorescent dyes emits red fluorescent light and the other one emits blue fluorescent light after laser excitation, so that the activity is detected, a dye emitting green fluorescent light can be introduced to detect acrosome integrality. The fluorescent staining method disclosed by the invention is simple to operate, accurate in result and high in reliability, and the sperm activity of the Kunming wolf dogs and the acrosome integrality can be accurately detected by using a fluorescence microscope or a flow cytometry.

A composition of matter for increasing the motility and / or percentage of intact acrosomes (PIA) in sperm is described. The composition includes, in combination, an amount of FAA and an amount of TIMP-2, wherein the amounts are effective to increase the motility, the PIA, or the motility and PIA of sperm contacted with the composition. The composition can be used as a cryopreservation medium for sperm.

An antibody biding specifically to rat's acrosome reacted sperm is produced and hybridomas (FARS-91 and FARS-92 strains) capable of stably proliferating are obtained by fusing mouse spleen cells having a high antibody titer against rat's acrosome reacted sperm with mouse-origin myeloma cells and screening fused cells reacting strongly with rat's acrosome reacted sperm. From these hybridomas, monoclonal antibodies selectively binding to rat's acrosome reacted sperm can be obtained. Thus, a diagnostic method for evaluating fertility of rat's spermatozoa is presented.

The invention discloses a balanus seat, an acrosome, and a nail cabin linkage transmission mechanism. The balanus seat, the acrosome, and the nail cabin linkage transmission mechanism comprise the balanus seat, the acrosome, a drive mechanism, a pulling rod and a lock catch mechanism, the front end of the acrosome is sleeved with the nail cabin, and the balanus seat is arranged at the front end ofthe nail cabin; a circumcising cutter is arranged in the acrosome, the acrosome is used for driving the front end of the circumcising cutter and the front end of the nail cabin to be in abutting connection with the balanus seat under the drive of the drive mechanism, an abutting portion in abutting connection with the acrosome is arranged on the pulling rod, and the lock catch mechanism is suitable for locking the pulling rod before the acrosome does not abut on the abutting portion; the balanus seat, the acrosome, and the nail cabin linkage transmission mechanism can solve the problems in the prior art that during the circumcision process, the linking time among various steps is long, the damage to tissues is large, the circumcision efficiency is low, and resetting of the circumcising cutter and unlocking of the lock catch mechanism cannot be carried out synchronously.

The invention relates to a swine semen freezing protecting agent and a cryopreserving method for swine semen by using the same. The swine semen freezing protecting agent is prepared from the followingcomponents in parts by weight: 2.5 parts of glucose, 2 parts of citric acid, 3 parts of coconut oil monoethanolamide, 2.5 parts of hyaluronic acid, 1.5 parts of skimmed milk, 1.2 parts of trehalose,0.07 part of penicillin sodium, 0.2 part of streptomycin sulfate, 95 parts of double distilled water, 23 parts of fresh yolk, 3.5 parts of glycerin, 1.5 parts of astaxanthin and 5 parts of extract ofradix rehmanniae praeparata. The cryopreserving method comprises the following steps of firstly, collecting the swine semen, centrifuging, and filtering, so as to obtain the swine semen A; pouring onepart of swine semen A into an experiment bottle, and adding two parts of freezing protecting agent to thin; finally, adding into liquid nitrogen, and cryopreserving. The swine semen freezing protecting agent has the advantages that the freezing protecting agent is reasonably compounded, so that the cryopreserving effect of the swine semen is improved under the function of the components; the survival rate, acrosome integrity and plasmalemma integrity of the swine semen are improved; the insemination activity and use effect of the swine semen are guaranteed.

The invention discloses a sheep seminal fluid cryopreservation diluent and method, and relates to the technical field of livestock breeding. The diluent comprises the following components:12-29 g of glucose, 10-50 g of sucrose, 8-13 g of sodium citrate, 600 thousand to 1.20 million IU of penicillin, 600 thousand to 1.20 million IU of streptomycin and 1000 mL of distilled water. The seminal fluid and the diluent are mixed at the ratio of 1 to 4 and stored in a small battle, and the small battle is wrapped with 4 to 8 layers of gauze and then placed in a refrigerator at 0-5 DEG C. The effectivesurvival time of the diluent is not less than 100 hours; the total survival time is not less than 140 hours; the survival index is not less than 69; the acrosome integrity rate after 48 hours of preservation is not less than 72%; the deformity rate after 48 hours of preservation is not more than 18%; substances used in the preparation of the diluent are all chemical reagents, animal derived substances such as yolk or milk are not contained, the safety and reliability are achieved, the quality is excellent, the cost is low, and the preparation method of the diluent is easy.

The invention discloses a safety catch applied to a circumcising stitching instrument. The safety catch comprises a circular arc plate, assembling and disassembling plates and buckles, and is characterized in that the assembling and disassembling plates are arranged outside the two ends of the circular arc plate; the buckles are arranged inside the two ends of the circular arc plate. When the circumcising stitching instrument is not used, acrosome, a handle and other components do not perform relative movement, so that a blade and a stitching nail are prevented from being damaged, and the service life of the circumcising stitching instrument is prolonged.

The invention belongs to the technical field of sperm acrosome detection, and discloses an improved crystal violet staining method for detecting sperm acrosome. The method comprises the following steps: preparing an improved gelatin film; liquefying collected fresh seminal fluid, subjecting seminal fluid to centrifugation, discarding seminal plasma, suspending precipitates in a phosphate buffer solution, placing the buffer solution in a thermotank, evenly paving sperm suspension on the gelatin film, incubating the sperms in a thermotank with a temperature of 37 DEG C, after 30 minutes, takingout sperm suspension for microscopic examination; dropwise adding a crystal violet dyeing solution on a fixed smear, carrying out dyeing for 1 to 3 minutes, washing the smear by water to remove excessdyes; dropwise adding a Lugol iodine solution, after one minute, washing the smear by water, then dropwise adding alcohol (95%) to perform color separation, shaking the glass slide until the purple color is not faded, washing the smear by flowing water, and carrying out microscopic examination after drying. A simple and rapid dyeing medium adopted by the crystal violet staining method can enhancethe cell dyeing affinity, through a discoloring treatment, the binding strength between a fuel and a polluted object can be measured, the structural difference of cells can be discriminated, and thesituation of sperm acrosome is well displayed.

The invention discloses a method for detecting sperm acrosin activity based on electrochemistry. The method comprises the steps that electrodes modified with a sperm acrosin substrate are fully soaked with sperm acrosin standard samples with different concentrations respectively, impedances of the electrodes are measured respectively, and then a standard curve of the concentration of active sperm acrosin and the electrode impedances is established; the electrodes are fully soaked with a to-be-measured sample containing sperm acrosin with an unknown concentration, impedances of the electrodes are measured, and the concentration of the sperm acrosin in the to-be-measured sample is obtained according to the standard curve, wherein the standard samples and the to-be-measured sample all adopt a buffer solution which is suitable for enabling the sperm acrosin to achieve specific binding and hydrolyzing the sperm acrosin substrate as a solvent. The invention further discloses a kit. By means of the kit and the detection method, the activity of the sperm acrosin can be detected rapidly, reliably and sensitively at a low cost, wherein detection can be completed within about 3 min, and the limit of detection can reach 0.001 micromole.