CN105010304A - Prescription composition of pig antiviral semen diluent - Google Patents
Prescription composition of pig antiviral semen diluent Download PDFInfo
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- CN105010304A CN105010304A CN201410164618.1A CN201410164618A CN105010304A CN 105010304 A CN105010304 A CN 105010304A CN 201410164618 A CN201410164618 A CN 201410164618A CN 105010304 A CN105010304 A CN 105010304A
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Abstract
The invention discloses a prescription composition of a pig antiviral semen diluent. Based on every 1000ml of double distilled water, the pig antiviral semen diluent contains 15.50g of glucose, 11.65g of trisodium citrate, 2.35g of EDTA sodium salt, 1.75g of sodium bicarbonate, 1.00g of polyvinyl alcohol, 5.50g of tricarboxymethyl amino methane, 4.10g of citric acid, 0.07g of cysteine, 1.00g of vitamin C, 0.10g of enrofloxacin and 50ml of an antiviral agent. According to the embodiment of the invention, the diluent of the prescription composition of the pig antiviral semen diluent has obvious killing, inhibition and obstructing effects on pig reproduction and respiration syndrome virus (PRRSV) and is capable of obviously improving the survival rate and preservation time of semen during in vitro preservation and the litter size of a mated sow.
Description
Technical field
The invention belongs to field of livestock production.Specifically, the present invention relates to the formula composition of the antiviral semen diluent of pig in Artificial Insemination Technology of Swine.
Background technology
The artificial insemination of pig is the former seminal fluid of boar that will collect, and through quality inspection, after dilution and the sport technique segment such as constant temperature preservation, to be impregnated object propagation technique in the genital tract manually seminal fluid being input to the sow that oestruses to reach dam.Technology of artificial insemination is the effective animal nutrition improving good species boar availability and sow production performance, technology of artificial insemination apply the industrialization and intensivization development that facilitate pig-breeding.Pig industry developed country is as the U.S., Denmark, Sweden, France etc., and all large-scale application technology of artificial insemination in Swine Production, artificial insemination propagation technique popularity rate reaches 95%.
Semen diluent is artificial preparation chemicals used when being diluted by former for the boar collected seminal fluid in swine artificial insemination process, and preparation semen diluent is the most key link of artificial insemination work.The object of semen diluent preparation is energy needed for supplementary sperm in vitro activity, maintain acid-base value when seminal fluid osmotic pressure is constant, the external constant temperature of regulation and control seminal fluid is preserved, there is provided sperm in vitro constant temperature basis for the survival environmental condition, semen diluent and the former seminal fluid of boar in proportion dilution mixture form test-tube seminal fluid main body.Semen diluent is the core technology embodying swine artificial insemination quality level, domestic market does not also have commercial antiviral semen diluent, rarely has the research of pig highly efficient anti-virus semen diluent aspect to report abroad.Research and the development of antiviral semen diluent will effectively improve sperm in vitro holding time and sperm quality, suppress the expansion of pathogenic microbes in seminal fluid numerous, strengthen sow reproductive efficiency and the healthy production capacity of pig industry, effectively promote China's swine artificial insemination levels of audit quality level.
On the basis of the general actual sperm dilution formula of liquid adopted of current pig industry artificial insemination, be main Testing index with sperm in vitro constant temperature holding time and sperm motility rate, optimize and improve sperm dilution formula of liquid composition, improving Semen routine time, sperm storage quality and the antibacterial level of seminal fluid.Technology of artificial insemination is owing to decreasing the contact of boar and sow, the probability that epidemic disease is imported into can be reduced, but control and prevention of disease work is not carried out in stock boar breeding process, boar health can not ensure, then the much virus by seminal fluid propagation, bacterium and other pathogene just can infect sow, cause pregnant sow miscarriage occurs or gives birth to stillborn foetus and mummy tire etc., thus cause pig farm serious financial consequences.Research shows, stock boar, after infection pig blue-ear disease, pseudoabies, swine fever, Porcine circovirus desease or Japanese B encephalitis etc., can detect correlated virus in seminal fluid.
Therefore, the formula composition of the antiviral semen diluent of pig needs research further.
Summary of the invention
The present invention is intended to one of solve the problems of the technologies described above at least to a certain extent.For this reason, one object of the present invention be to propose a kind of have porcine reproductive and respiratory syndrome virus (PRRSV) significantly kill, suppress and the formula composition of the antiviral semen diluent of pig of blocking effect.
In one aspect of the invention, the present invention proposes the formula composition of the antiviral semen diluent of a boar.According to embodiments of the invention, based on the DDW of every 1000 milliliters, the antiviral semen diluent of described pig contains:
15.50g glucose;
11.65g trisodium citrate;
The EDETATE SODIUM salt of 2.35g;
1.75g sodium bicarbonate;
1.00g polyvinyl alcohol;
The tricarboxylic aminomethane of 5.50g;
4.10g citric acid;
The cysteine of 0.07g;
1.00g vitamin C;
The Enrofloxacin of 0.10g; And
50ml antivirotic.
The dilution formed according to the formula of the antiviral semen diluent of the pig of the embodiment of the present invention has the porcine reproductive and respiratory syndrome virus (PRRSV) in artificial insemination pig semen used significantly to be killed, suppresses and blocking effect, and can significantly improve survival rate, holding time that sperm in vitro preserves and award the nest litter size of joining sow.
In addition, the formula composition of the antiviral semen diluent of pig according to the above embodiment of the present invention can also have following additional technical characteristic:
In some embodiments of the invention, described antivirotic contains: Ribavirin, bupleurum extract, isatis root extract, glutinous rehmannia extract, Baical Skullcap root P.E, capejasmine extract, forsythia suspense extraction and Radix Scrophulariae extract.Thus, the antiviral effect of Diluent of Pig's Spermatic Fluid can be significantly improved.
In some embodiments of the invention, described bupleurum extract is prepared through the following steps: (1) utilizes 10 premium on currency to carry out immersion 12 hours to 5 kilograms of radix bupleuri, then carries out decocting and concentrating and filtration treatment successively; (2) ethanol being 70% by the filtrate obtained in step (1) and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours; And (3) are by concentrated for the filtrate heating obtained in step (2), and be that 1:2 carries out turning molten process and filtering by gained concentrate and water according to volume ratio, evaporation drying process is carried out to obtained filtrate, to obtain described bupleurum extract.Thus, effectively bupleurum extract can be isolated from radix bupleuri.
In some embodiments of the invention, described isatis root extract is prepared through the following steps: (1) utilizes 10 premium on currency to carry out immersion 12 hours to 5 kilograms of Radix Isatidis, then carries out decocting and concentrating and filtration treatment successively; (2) ethanol being 70% by the filtrate obtained in step (1) and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours; And (3) are by concentrated for the filtrate heating obtained in step (2), and be that 1:2 carries out turning molten process and filtering by gained concentrate and water according to volume ratio, evaporation drying process is carried out to obtained filtrate, to obtain described isatis root extract.Thus, effectively isatis root extract can be isolated from Radix Isatidis.
In some embodiments of the invention, described glutinous rehmannia extract is prepared through the following steps: (1) utilizes 10 premium on currency to carry out immersion 12 hours to 5 kilograms of glutinous rehmannia, then carries out decocting and concentrating and filtration treatment successively; (2) ethanol being 70% by the filtrate obtained in step (1) and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours; And (3) are by concentrated for the filtrate heating obtained in step (2), and be that 1:2 carries out turning molten process and filtering by gained concentrate and water according to volume ratio, evaporation drying process is carried out to obtained filtrate, to obtain described glutinous rehmannia extract.Thus, effectively glutinous rehmannia extract can be isolated from glutinous rehmannia.
In some embodiments of the invention, described Baical Skullcap root P.E is prepared through the following steps: (1) utilizes 10 premium on currency to carry out immersion 12 hours to 5 kilograms of roots of large-flowered skullcap, then carries out decocting and concentrating and filtration treatment successively; (2) ethanol being 70% by the filtrate obtained in step (1) and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours; And (3) are by concentrated for the filtrate heating obtained in step (2), and be that 1:2 carries out turning molten process and filtering by gained concentrate and water according to volume ratio, evaporation drying process is carried out to obtained filtrate, to obtain described Baical Skullcap root P.E.Thus, effectively Baical Skullcap root P.E can be isolated from the root of large-flowered skullcap.
In some embodiments of the invention, described capejasmine extract is prepared through the following steps: (1) utilizes 10 premium on currency to carry out immersion 12 hours to 5 kilograms of cape jasmines, then carries out decocting and concentrating and filtration treatment successively; (2) ethanol being 70% by the filtrate obtained in step (1) and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours; And (3) are by concentrated for the filtrate heating obtained in step (2), and be that 1:2 carries out turning molten process and filtering by gained concentrate and water according to volume ratio, evaporation drying process is carried out to obtained filtrate, to obtain described capejasmine extract.Thus, effectively capejasmine extract can be isolated from cape jasmine.
In some embodiments of the invention, described forsythia suspense extraction is prepared through the following steps: (1) utilizes 10 premium on currency to carry out immersion 12 hours to 5 kilograms of capsules of weeping forsythia, then carries out decocting and concentrating and filtration treatment successively; (2) ethanol being 70% by the filtrate obtained in step (1) and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours; And (3) are by concentrated for the filtrate heating obtained in step (2), and be that 1:2 carries out turning molten process and filtering by gained concentrate and water according to volume ratio, evaporation drying process is carried out to obtained filtrate, to obtain described forsythia suspense extraction.Thus, effectively forsythia suspense extraction can be isolated from the capsule of weeping forsythia.
In some embodiments of the invention, described Radix Scrophulariae extract is prepared through the following steps: (1) utilizes 10 premium on currency to carry out immersion 12 hours to 5 kilograms of radix scrophulariaes, then carries out decocting and concentrating and filtration treatment successively; (2) ethanol being 70% by the filtrate obtained in step (1) and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours; And (3) are by concentrated for the filtrate heating obtained in step (2), and be that 1:2 carries out turning molten process and filtering by gained concentrate and water according to volume ratio, evaporation drying process is carried out to obtained filtrate, to obtain described Radix Scrophulariae extract.Thus, effectively Radix Scrophulariae extract can be isolated from radix scrophulariae.
In some embodiments of the invention, described antivirotic is by by Ribavirin dilution, bupleurum extract dilution, isatis root extract dilution, glutinous rehmannia extract dilution, Baical Skullcap root P.E dilution, capejasmine extract dilution, forsythia suspense extraction dilution and Radix Scrophulariae extract dilution mix and obtain, wherein, described Ribavirin dilution is by utilizing the glucose injection of 5wt% described Ribavirin dilution 10 ~ 1000 times to be obtained, described bupleurum extract dilution is by utilizing the glucose injection of 5wt% described bupleurum extract to be diluted 10 times and obtain, described isatis root extract dilution is by utilizing the glucose injection of 5wt% described isatis root extract to be diluted 10 times and obtain, described glutinous rehmannia extract dilution is by utilizing the glucose injection of 5wt% described glutinous rehmannia extract to be diluted 10 times and obtain, described Baical Skullcap root P.E dilution is by utilizing the glucose injection of 5wt% described Baical Skullcap root P.E to be diluted 10 times and obtain, described capejasmine extract dilution is by utilizing the glucose injection of 5wt% described capejasmine extract to be diluted 10 times and obtain, described forsythia suspense extraction dilution is by utilizing the glucose injection of 5wt% described forsythia suspense extraction to be diluted 10 times and obtain, described Radix Scrophulariae extract dilution is by utilizing the glucose injection of 5wt% described Radix Scrophulariae extract to be diluted 10 times and obtain.
In some embodiments of the invention, based on every 1000 milliliters of described DDWs, in described antivirotic, described Ribavirin dilution, described bupleurum extract dilution, described isatis root extract dilution, described glutinous rehmannia extract dilution, described Baical Skullcap root P.E dilution, described capejasmine extract dilution, described forsythia suspense extraction dilution and described Radix Scrophulariae extract dilution are respectively 1 milliliter.Thus, the antiviral effect of anti-virus diluent can be significantly improved.
Additional aspect of the present invention and advantage will part provide in the following description, and part will become obvious from the following description, or be recognized by practice of the present invention.
Embodiment
Be described below in detail embodiments of the invention, be intended to for explaining the present invention, and can not limitation of the present invention be interpreted as.
In one aspect of the invention, the present invention proposes the formula composition of the antiviral semen diluent of a boar.According to embodiments of the invention, based on the DDW of every 1000 milliliters, the antiviral semen diluent of pig contains: the glucose of 15.50g; 11.65g trisodium citrate; The EDETATE SODIUM salt of 2.35g; 1.75g sodium bicarbonate; 1.00g polyvinyl alcohol; The tricarboxylic aminomethane of 5.50g; 4.10g citric acid; The cysteine of 0.07g; 1.00g vitamin C; The Enrofloxacin of 0.10g; And 50ml antivirotic.Particularly, the DDW above component being fully dissolved in 1000ml is antiviral semen diluent after sterilization treatment.
The dilution formed according to the formula of the antiviral semen diluent of the pig of the embodiment of the present invention has porcine reproductive and respiratory syndrome virus (PRRSV) significantly to be killed, suppresses and blocking effect, and can significantly improve living spermatozoa percentage and fertility of sow.
Specifically describe, antiviral semen diluent has following effect in swine artificial insemination: one is the volume that antiviral semen diluent effectively can expand the former seminal fluid of pig, rationally effectively utilize outstanding stock boar seminal fluid, improve the seminal fluid availability of boar, accelerate and expand the genetic progress of merit; Two is that antiviral semen diluent can be when sperm in vitro constant temperature is preserved and supplies metabolic nutrient component, improves sperm in vitro and preserves quality and holding time, so that seminal fluid can carry out long distance transportation to enterprise of raising pigs, expand high-quality seminal fluid range of application; Three is that antiviral semen diluent can ensure sperm quality, removes seminal fluid off quality by seminal fluid quality check process, eliminates the stock boar of semen quality difference, to improve sow conception rate; Four is by adding antimicrobial medicine in seminal fluid, suppresses and stops germ numerous with the expansion of virus in seminal fluid, blocking the path that pathogenic microbes is propagated by seminal fluid, effectively improve the swinery general level of the health.
According to embodiments of the invention, the composition of antivirotic is also not particularly limited, according to a particular embodiment of the invention, antivirotic can contain: Ribavirin, bupleurum extract, isatis root extract, glutinous rehmannia extract, Baical Skullcap root P.E, capejasmine extract, forsythia suspense extraction and Radix Scrophulariae extract.Thus, the antiviral effect of Diluent of Pig's Spermatic Fluid can be significantly improved.
According to embodiments of the invention, the separate mode of bupleurum extract is also not particularly limited, according to a particular embodiment of the invention, bupleurum extract can be prepared through the following steps: utilize 10 premium on currency to carry out immersion 12 hours to 5kg radix bupleuri, then extract under 95 degrees Celsius after 4 hours and filter, the ethanol being 70% by obtained filtrate and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours, and obtained filtrate is carried out at 80 degrees celsius heating and be concentrated into 2 liters of liquid, be that 1:2 carries out turning molten process and filtering by obtained concentrate and water according to volume ratio, rotary evaporation drying process is carried out to obtained filtrate, thus dry dry matter can be obtained, be bupleurum extract (containing radix bupleuri active component).
According to embodiments of the invention, the separate mode of isatis root extract is also not particularly limited, according to a particular embodiment of the invention, isatis root extract can be prepared through the following steps: utilize 10 premium on currency to carry out immersion 12 hours to 5kg Radix Isatidis, then extract under 95 degrees Celsius after 4 hours and filter, the ethanol being 70% by obtained filtrate and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours, and obtained filtrate is carried out at 80 degrees celsius heating and be concentrated into 2 liters of liquid, be that 1:2 carries out turning molten process and filtering by obtained concentrate and water according to volume ratio, rotary evaporation drying process is carried out to obtained filtrate, thus dry dry matter can be obtained, be isatis root extract (containing Radix Isatidis active component).
According to embodiments of the invention, the separate mode of glutinous rehmannia extract is also not particularly limited, according to a particular embodiment of the invention, glutinous rehmannia extract can be prepared through the following steps: utilize 10 premium on currency to carry out immersion 12 hours to 5kg glutinous rehmannia, then extract under 95 degrees Celsius after 4 hours and filter, the ethanol being 70% by obtained filtrate and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours, and obtained filtrate is carried out at 80 degrees celsius heating and be concentrated into 2 liters of liquid, be that 1:2 carries out turning molten process and filtering by obtained concentrate and water according to volume ratio, rotary evaporation drying process is carried out to obtained filtrate, thus dry dry matter can be obtained, be glutinous rehmannia extract (containing active component of Radix Rehmanniae).
According to embodiments of the invention, the separate mode of Baical Skullcap root P.E is also not particularly limited, according to a particular embodiment of the invention, Baical Skullcap root P.E can be prepared through the following steps: utilize 10 premium on currency to carry out immersion 12 hours to the 5kg root of large-flowered skullcap, then extract under 95 degrees Celsius after 4 hours and filter, the ethanol being 70% by obtained filtrate and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours, and obtained filtrate is carried out at 80 degrees celsius heating and be concentrated into 2 liters of liquid, be that 1:2 carries out turning molten process and filtering by obtained concentrate and water according to volume ratio, rotary evaporation drying process is carried out to obtained filtrate, thus dry dry matter can be obtained, be Baical Skullcap root P.E (containing root of large-flowered skullcap active component).
According to embodiments of the invention, the separate mode of capejasmine extract is also not particularly limited, according to a particular embodiment of the invention, capejasmine extract can be prepared through the following steps: utilize 10 premium on currency to carry out immersion 12 hours to 5kg cape jasmine, then extract under 95 degrees Celsius after 4 hours and filter, the ethanol being 70% by obtained filtrate and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours, and obtained filtrate is carried out at 80 degrees celsius heating and be concentrated into 2 liters of liquid, be that 1:2 carries out turning molten process and filtering by obtained concentrate and water according to volume ratio, rotary evaporation drying process is carried out to obtained filtrate, thus dry dry matter can be obtained, be capejasmine extract (containing cape jasmine active component).
According to embodiments of the invention, the separate mode of forsythia suspense extraction is also not particularly limited, according to a particular embodiment of the invention, forsythia suspense extraction can be prepared through the following steps: utilize 10 premium on currency to carry out immersion 12 hours to the 5kg capsule of weeping forsythia, then extract under 95 degrees Celsius after 4 hours and filter, the ethanol being 70% by obtained filtrate and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours, and obtained filtrate is carried out at 80 degrees celsius heating and be concentrated into 2 liters of liquid, be that 1:2 carries out turning molten process and filtering by obtained concentrate and water according to volume ratio, rotary evaporation drying process is carried out to obtained filtrate, thus dry dry matter can be obtained, be forsythia suspense extraction (containing capsule of weeping forsythia active component).
According to embodiments of the invention, the separate mode of Radix Scrophulariae extract is also not particularly limited, according to a particular embodiment of the invention, Radix Scrophulariae extract can be prepared through the following steps: utilize 10 premium on currency to carry out immersion 12 hours to 5kg radix scrophulariae, then extract under 95 degrees Celsius after 4 hours and filter, the ethanol being 70% by obtained filtrate and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours, and obtained filtrate is carried out at 80 degrees celsius heating and be concentrated into 2 liters of liquid, be that 1:2 carries out turning molten process and filtering by obtained concentrate and water according to volume ratio, rotary evaporation drying process is carried out to obtained filtrate, thus dry dry matter can be obtained, be Radix Scrophulariae extract (containing radix scrophulariae active component).
According to embodiments of the invention, the preparation method of antivirotic is also not particularly limited, according to a particular embodiment of the invention, antivirotic can obtain by Ribavirin dilution, bupleurum extract dilution, isatis root extract dilution, glutinous rehmannia extract dilution, Baical Skullcap root P.E dilution, capejasmine extract dilution, forsythia suspense extraction dilution and Radix Scrophulariae extract dilution being mixed.According to embodiments of the invention, the preparation method of Ribavirin dilution is also not particularly limited, and according to a particular embodiment of the invention, Ribavirin dilution 10 ~ 1000 times can obtain by utilizing the glucose injection of 5wt% by Ribavirin dilution.According to embodiments of the invention bupleurum extract dilution, isatis root extract dilution, glutinous rehmannia extract dilution, Baical Skullcap root P.E dilution, capejasmine extract dilution, the dilution mode of forsythia suspense extraction dilution and Radix Scrophulariae extract dilution is also not particularly limited, according to a particular embodiment of the invention, bupleurum extract can be diluted 10 times by utilizing the glucose injection of 5wt% and obtain by bupleurum extract dilution, isatis root extract can be diluted 10 times by utilizing the glucose injection of 5wt% and obtain by isatis root extract dilution, glutinous rehmannia extract can be diluted 10 times by utilizing the glucose injection of 5wt% and obtain by glutinous rehmannia extract dilution, Baical Skullcap root P.E can be diluted 10 times by utilizing the glucose injection of 5wt% and obtain by Baical Skullcap root P.E dilution, capejasmine extract can be diluted 10 times by utilizing the glucose injection of 5wt% and obtain by capejasmine extract dilution, forsythia suspense extraction can be diluted 10 times by utilizing the glucose injection of 5wt% and obtain by forsythia suspense extraction dilution, Radix Scrophulariae extract can be diluted 10 times by utilizing the glucose injection of 5wt% and obtain by Radix Scrophulariae extract dilution.According to embodiments of the invention, above-mentioned adopted dilution mode is also not particularly limited, according to a particular embodiment of the invention, gradient dilution mode can be adopted Ribavirin, bupleurum extract, isatis root extract, glutinous rehmannia extract, Baical Skullcap root P.E, capejasmine extract, forsythia suspense extraction and Radix Scrophulariae extract carry out dilution process, thus Ribavirin dilution can be obtained, bupleurum extract dilution, isatis root extract dilution, glutinous rehmannia extract dilution, Baical Skullcap root P.E dilution, capejasmine extract dilution, forsythia suspense extraction dilution and Radix Scrophulariae extract dilution.It should be noted that, above-mentioned dilution all aseptically operates.
According to embodiments of the invention, Ribavirin dilution, bupleurum extract dilution, isatis root extract dilution, glutinous rehmannia extract dilution, Baical Skullcap root P.E dilution, capejasmine extract dilution, the consumption of forsythia suspense extraction dilution and Radix Scrophulariae extract dilution is also not particularly limited, according to a particular embodiment of the invention, based on the DDW of 1000 milliliters, in antivirotic, Ribavirin dilution, bupleurum extract dilution, isatis root extract dilution, glutinous rehmannia extract dilution, Baical Skullcap root P.E dilution, capejasmine extract dilution, forsythia suspense extraction dilution and Radix Scrophulariae extract dilution can be independently 1 milliliter.Inventor finds, drug concentration is too high, and sperm tolerance is poor, thus affects living spermatozoa percentage and time-to-live.It should be noted that, above-mentioned Ribavirin dilution, bupleurum extract dilution, isatis root extract dilution, glutinous rehmannia extract dilution, Baical Skullcap root P.E dilution, capejasmine extract dilution, forsythia suspense extraction dilution and Radix Scrophulariae extract dilution pipette in the centrifuge tube of 10 milliliters with sterile working method pipette respectively, and shake 10 minutes in vortex oscillator, then seal up for safekeeping with ampoule bottle is aseptic subpackaged, room temperature preservation.
It should be noted that, other composition in the antiviral semen diluent of pig except antivirotic, mixes in proportion with sterile working method within 12 hours all before use.
Below with reference to specific embodiment, present invention is described, it should be noted that, these embodiments are only descriptive, and do not limit the present invention in any way.
Embodiment
Composition of raw materials:
The citric acid of the polyvinyl alcohol of the EDETATE SODIUM salt of the glucose of 15.50g, the trisodium citrate of 11.65g, 2.35g, the sodium bicarbonate of 1.75g, 1.00g, the tricarboxylic aminomethane of 5.50g, 4.10g, the cysteine of 0.07g, the vitamin C of 1.00g, the Enrofloxacin of 0.10g, 50ml antivirotic.
Preparation method:
Each composition above is fully mixed in the DDW of 1000ml according to above composition.
Evaluate:
1, respectively on anti-porcine reproductive and respiratory syndrome virus (PRRSV) effect of the antiviral semen diluent of embodiment gained pig, detect on sperm quality impact with on fertility of sow impact.
2, evaluation index and method of testing:
The detection of A, anti-porcine reproductive and respiratory syndrome virus (PRRSV) effect:
1) test material
Marc145 cell, PK15 cell, calf serum (positive pole bio tech ltd, Shanghai), DMEM culture fluid (Bo Gu bio tech ltd, Shanghai), 0.25% trypsase (GIBCOGO company of the U.S.), Chinese medical extract concentrate, MTT (tetrazolium bromide), dimethyl sulfoxide (DMSO), mycillin liquid, PBS (phosphate buffer).
2) test apparatus
Superclean bench SCW-HS (the grand auspicious purification Science and Technology Ltd. in Suzhou), constant water bath box, inverted microscope (LWD300-38LT, Shanghai Ce Wei photoelectricity technology corporation, Ltd.), CO2gas incubator (Beijing Hong Datianju testing equipment Co., Ltd), centrifuge (Hangzhou Zang Han Science and Technology Ltd.), blake bottle, 96 porocyte culture plates, microplate reader (Thermo company).
3) virus titer measures
PRRSV titer determination: cultivate Marc145 cell according to a conventional method, become unicellular through 0.25% Trypsin Induced, cell number is adjusted to 2 × 10
5individual/ml, adds Marc145 cell suspension 100 μ L in every hole, and 5% CO2gas incubator is cultivated.PRRSV DMEM culture fluid does continuous 10 times of dilutions in 1.5mL Ep pipe.By 10
-1-10
-10virus dilution liquid inoculation monolayer cell culture plate, every dilution factor 8 hole, 100 μ L are inoculated in every hole.If 2 rows compare (100 μ l growth-promoting media+100 μ l cell suspension), put 37 DEG C, in 5% CO2gas incubator, cultivate 10 days.
4) antiviral agent matter sample is to Marc145 cytotoxic assay
Had digestive transfer culture Marc145 cell according to a conventional method, regulate cell density with cell growth medium, on 96 porocyte culture plates, every hole adds 100 μ L Marc145 cell suspensions, culture plate is placed in 5% CO2gas incubator to cultivate, treats that Marc145 cell grows up to monolayer adherence cell.After 8 groups of Chinese medical extract cell maintenance mediums do serial dilution, by 10
-1-10
-7dilution adds to have been cultivated in the culture plate of cell monolayer, and every hole adds 100ul drug dilution liquid, and each dilution gradient repeats 5 holes, if cell control well (not drug containing), put 5% CO2gas incubator, observe CPE situation every day, to determine that medicine is to cell maximal non-toxic concentration.After cultivating 96h, suck supernatant, after rushing 2-3 time with PBS, add 20 μ LMTT solution (5mg/ml, i.e. 0.5%MTT), cultivate 4 hours, culture fluid in hole, place to go, avoid during absorption encountering cell as far as possible.Every hole adds dimethyl sulfoxide (DMSO) (DMSO) 150ul, puts low-speed oscillation 15min on shaking table, makes DMSO dissolve bluish violet crystallization first a ceremonial jade-ladle, used in libation.If control wells (cell maintenance medium, cell, culture fluid, MTT, dimethyl sulfoxide (DMSO)), zeroing hole (medium, MTT, dimethyl sulfoxide (DMSO)), surveys light absorption value with enzyme-linked immunosorbent assay instrument at OD570nm place.
5) antiviral agent matter sample is tested the killing action of PRRSV
By 100TCID
50virus (PRRSV, the maintenance medium of 2% NBCS) dilution drug sample different from equal-volume mixes, 37 DEG C of effect 2h.Had digestive transfer culture Marc145 cell according to a conventional method, adjustment cell concentration, every hole inoculation 100ul cell suspension, be cultured to after cell covers with individual layer and discard medium, three times are washed, the mixed liquor of virus and Chinese medical extract after access effect 2h, 100ul/ hole with PBS, repeat 4 holes, separately establish cell controls and virus control.37 DEG C, 5% carbonic acid gas is cultured to virus control and occurs CPE.Suck supernatant, after rushing 2-3 time with PBS, add 20 μ LMTT solution (5mg/ml, i.e. 0.5%MTT), cultivate after 4 hours, remove culture fluid, every hole adds dimethyl sulfoxide (DMSO) 150ul, puts shaking table low-speed oscillation 15min, and first a ceremonial jade-ladle, used in libation crystal is dissolved.Zeroing hole (medium, MTT, dimethyl sulfoxide (DMSO)) is set simultaneously.The light absorption value in each hole is measured at enzyme-linked immunosorbent assay instrument OD570nm place.
6) antiviral agent matter sample is tested the inhibitory action of PRRSV
Had digestive transfer culture Marc145 cell according to a conventional method, adjustment cell concentration, every hole inoculation 100ul cell suspension, is cultured to after cell covers with individual layer and adds different dilution drug sample, 100ul/ hole, repeat 4 holes.Cultivate 2h, discard liquid and add 100TCID for 37 DEG C
50virus liquid, 100ul/ hole, separately establishes cell controls and virus control.37 DEG C, 5% carbonic acid gas is cultured to virus control and occurs CPE.Suck supernatant, after rushing 2-3 time with PBS, add 20 μ LMTT solution (5mg/ml, i.e. 0.5%MTT), cultivate after 4 hours, remove culture fluid, every hole adds dimethyl sulfoxide (DMSO) 150ul, puts shaking table low-speed oscillation 15min, and first a ceremonial jade-ladle, used in libation crystal is dissolved.Zeroing hole (medium, MTT, dimethyl sulfoxide (DMSO)) is set simultaneously.The light absorption value in each hole is measured at enzyme-linked immunosorbent assay instrument OD570nm place.
7) antiviral agent matter sample is tested the blocking effect of PRRSV
Had digestive transfer culture Marc145 cell according to a conventional method, adjustment cell concentration, every hole inoculation 100ul cell suspension, is cultured to after cell covers with individual layer, adds 100TCID
50virus liquid (PRRSV, the maintenance medium of 2% NBCS), 100ul/ hole, repeats 4 holes.After putting 37 DEG C of cultivation 2h, discard virus liquid and add different dilution drug sample, 100ul/ hole, separately establish cell controls and virus control.37 DEG C, 5% carbonic acid gas is cultured to virus control and occurs CPE.Suck supernatant, after rushing 2-3 time with PBS, add 20 μ LMTT solution (5mg/ml, i.e. 0.5%MTT), cultivate after 4 hours, remove culture fluid, every hole adds dimethyl sulfoxide (DMSO) 150ul, puts shaking table low-speed oscillation 15min, and first a ceremonial jade-ladle, used in libation crystal is dissolved.Zeroing hole (medium, MTT, dimethyl sulfoxide (DMSO)) is set simultaneously.The light absorption value in each hole is measured at enzyme-linked immunosorbent assay instrument OD570nm place.
B, the detection that sperm quality is affected:
Take former for the boar collected seminal fluid to semen diluent workshop fast, after weighing former ejaculate volume quality, determine dilution ratio.First slowly inject isometric former seminal fluid with the antiviral semen diluent of isothermal, leave standstill balance 15 minutes, by binocular phase-contrast microscopy sperm motility rate, according to motility rate and former seminal fluid volume determination thinner ratio.According to thinner ratio, former seminal fluid is diluted to ratio volume further, leaves standstill towel parcel after carrying out equilibrium at room temperature and put into sperm Special constant temperature Refrigerator store.
The sperm that can carry out straight ahead motion at microscope 37 DEG C of temperature is called effective sperm, and the vigor of sperm refers to that effective sperm accounts for the percentage of total number of sperm.The ability of sperm straight ahead is the main manifestations of the normal and membrane integrity of sperm metabolism, only has effective sperm just to have to be combined with sow ovum and the ability of being fertilized.Sperm motility rate is that semen quality evaluates most important index.Generally. normally carrying out test-tube seminal fluid sperm viability should lower than 0.7, and namely microexamination has the sperm being not less than 70% to do rectilinear motion.
C, antiviral semen diluent detect the impact of pig fertility:
The antiviral sperm dilution formula of liquid that application the present invention obtains, respectively in the piggery of Hainan sieve Ox Mountain herding parent company and Hainan Farm herding Group Co., Ltd, to Duroc, long white, great Bai tri-kinds carried out the evaluation test of efficient semen diluent artificial insemination practical application effect, while and male and female pig natural breeding mode compare test, observe its impact on the reproductive performance such as conception rate, litter size, the difference of more antiviral semen diluent reproductive performance such as conception rate, litter size between 3 kinds.
3, testing result:
The testing result of A, the antiviral semen diluent of pig anti-porcine reproductive and respiratory syndrome virus (PRRSV) effect:
(1) the antiviral semen diluent sample of pig is that 125 μ g/ml show suppression PRRSV virus function in concentration;
(2) the antiviral semen diluent sample of pig is that 125 μ g/ml have significantly directly kill activity to PRRSV virus in concentration;
(3) pig antiviral semen diluent sample is that the blocking effect of 500 μ g/ml to PRRSV virus is obvious in concentration.
B, the antiviral semen diluent of pig affect testing result to sperm quality:
(1) pig semen sample is added in the antiviral semen diluent of pig with 500 μ g/ml amounts, and within 24 hours, living spermatozoa percentage is 0.80;
(2) pig semen sample is added in the antiviral semen diluent of pig with 500 μ g/ml amounts, and within 48 hours, living spermatozoa percentage is 0.75;
(3) pig semen sample is added in the antiviral semen diluent of pig with 500 μ g/ml amounts, and within 72 hours, living spermatozoa percentage is 0.70;
(4) pig semen sample is added in the antiviral semen diluent of pig with 500 μ g/ml amounts, and within 96 hours, living spermatozoa percentage is 0.60.
C, the antiviral semen diluent of pig affect testing result to pig fertility:
1) antiviral semen diluent affects Duluke's sow fertility
Table 1 artificial insemination is on duroc conception rate, litter size impact
Note: same row same letter mark difference is not significantly (P>0.05)
As known from Table 1, natural mating with contrast the Insemination effect that dilution carries out and compare, the conception rate of result display Duluke's sow group, number born per litter affect difference all significantly (P>0.05), but compare with natural mating reproductive effect, antiviral semen diluent is used for manually awarding the conception rate of rear Duluke's sow and litter size and productivity and improves Be very effective (P < 0.05), antiviral semen diluent compares with normal control semen diluent reproductive performance reproductive performances such as the conception rate of Duluke's sow and litter sizes, the conception rate of Duluke's sow, litter size significant difference (P < 0.05), show that antiviral semen diluent effect is significantly higher than the reproductive performance of common semen diluent and the natural mating of male and female pig.Antiviral semen diluent produces direct impact to the artificial insemination effect of Duluke's sow, nest average litter size is than another more than 2 groups 1, reach 9.86, average many 1.03 of the healthy litter size that weans, shows that antiviral semen diluent artificial insemination Be very effective is better than common semen diluent.
The healthy litter size that weans is the important indicator of sow Population Health.The healthy litter size that weans is high, and illustrate that the sodium selenite that every sow provides is many, piglet postnatal mortality is low, and Population Health level is high, and the breeding disorder disease risks of pig is low, and whole swinery is in production status in good health.As can be seen from Table 1, apply the nest litter size average out to 9.86 of antiviral semen diluent Duluke's sow, average healthy nest number pigs weaned is 9.1, and difference is not remarkable, illustrate that antiviral semen diluent is applied to swine artificial insemination and effectively can improves wean sodium selenite number, piglet lethality significantly reduces by 11.3% than common semen diluent, the corresponding raising of swinery health degree.
Find in process of the test that the common semen diluent of application carries out in swine artificial insemination process, control group has that 7% part sow is out of heat, not become pregnant phenomenon after oestrus delay or rutting.Sow 3d after weaned piglet just starts rutting under normal circumstances.Reason is as follows:
(1) seminal fluid is subject to courses of infection.Seminal fluid is in gatherer process and sperm dilution link, and a large amount of microbes invades seminal fluid and utilizes various nutrient propagation in seminal fluid further, and the seminal fluid carrying pathogenic microorganism enters gential tract of dam through artificial insemination and then causes brood sow to return feelings or miscarriage;
(2) Reproduction Disorder.Award and join the virus that sow itself carries Reproduction Disorder, the numerous propagation of expansion of virus causes sow genital system diseases, causes sow to return feelings and miscarriage;
(3) sow suffers from reproduction organic disease.The reproductive diseases that various inflammation and infected by microbes cause can make brood sow occur habitual abortion.
The experimental group sow of antiviral semen diluent returns feelings rate and is reduced to 1%, illustrate that antiviral semen diluent effectively can improve sow and return the reproductive diseases such as feelings and miscarriage symptom, reduce pathogenic microorganism seminal fluid in artificial insemination procedures to pollute, apply antiviral semen diluent to raising sow conception rate, keep brood sow Population Health level to have important function.
2) antiviral semen diluent affects Landrace fertility
Table 2 artificial insemination affects Landrace conception rate and litter size
Note: same row same letter mark person difference is not significantly (P>0.05)
As can be seen from Table 2, Landrace uses the conception rate of natural mating and common dilution control group, number born per litter difference all remarkable (P>0.05), but antiviral semen diluent Insemination effect is than control group and the numerous rate of natural breeding high (P < 0.05), antiviral efficient semen diluent wean sodium selenite number every nest pig many 1 average than control group, illustrates that antiviral semen diluent effectively can improve sow reproductive performance.
The natural breeding of male and female pig is compared with control group result of the test, the conception rate of female Landrace group, number born per litter affect difference all significantly (P>0.05), illustrate that swine artificial insemination can replace the modes of reproduction of male and female pig natural breeding completely.Antiviral semen diluent compares with normal control semen diluent reproductive performance reproductive performance impacts such as the conception rate of female Landrace and litter sizes, the conception rate of female Landrace, litter size significant difference (P < 0.05), show that antiviral semen diluent effect is significantly higher than the reproductive performance of common semen diluent and the natural mating of male and female pig.Antiviral semen diluent produces direct impact to the artificial insemination effect of female Landrace, nest average litter size is than another more than 2 groups 0.9, reach 11.1, average many 2 of the healthy litter size that weans, shows that antiviral semen diluent artificial insemination Be very effective is better than common semen diluent.
As can be seen from table 2 simultaneously, apply the nest litter size average out to 11.1 of antiviral semen diluent female Landrace, average healthy nest number pigs weaned is 11.1, between the two and difference is not remarkable.The average litter size of normal control semen diluent is 10.8, and the sodium selenite number that weans is 9.1, illustrate that antiviral semen diluent is applied to swine artificial insemination and effectively can improves wean sodium selenite number, piglet lethality significantly reduces by 21.8% than common semen diluent, and antiviral semen diluent can improve swinery health degree effectively.
In addition, Landrace reproductive performance is significantly higher than Duroc, this is mainly between kind caused by hereditary difference, Duroc is mainly used for the hybridization of paternal terminal, and long white main and Large White to hybridize production binary terminal maternal, but Landrace antiviral semen diluent effect is identical with duroc, namely antiviral semen diluent is than control group and male and female pig natural breeding reproductive performance Be very effective.
3) antiviral semen diluent affects Large White fertility
Table 3 artificial insemination is on the impact of Large White conception rate and litter size
Note: same row same letter mark person difference is not significantly (P>0.05)
As can be seen from Table 3, the reproductive performance difference such as conception rate, number born per litter of the natural breeding of Large White sow and common dilution control group is not significantly (P>0.05), antiviral semen diluent insemination reproductive performance is than control group and natural breeding reproductive performance significant difference (P < 0.05), the wean nest sodium selenite number every nest pig many 2 average than control group of antiviral semen diluent large white sow, illustrates that antiviral semen diluent effectively can improve sow reproductive performance.
The natural breeding of male and female pig is compared with control group result of the test, the reproductive performance difference such as conception rate, number born per litter of large white sow group all not significantly (P>0.05), further illustrates the breeding breeding wony that swine artificial insemination can replace the natural breeding of male and female pig completely.Antiviral semen diluent compares with normal control group reproductive performance reproductive performance impacts such as the conception rate of large white sow and litter sizes, the conception rate of large white sow, litter size significant difference (P < 0.05), show that antiviral semen diluent effect is significantly higher than the reproductive performance of normal control semen diluent and the natural mating of male and female pig.Antiviral semen diluent produces directly effectively impact to the artificial insemination effect of large white sow, nest average litter size is than another more than 2 groups 1, reach 11.2, average many 2 of the healthy litter size that weans, shows that antiviral semen diluent artificial insemination Be very effective is better than common semen diluent.
As can be seen from table 3 simultaneously, apply the nest litter size average out to 11.2 of antiviral semen diluent large white sow, average healthy nest number pigs weaned is 11.5, between the two and difference is not remarkable, illustrate that antiviral semen diluent awards that to join sow piglet lethality low, the healthy rate of piglet is high.The average litter size of normal control semen diluent is 10.6, and the sodium selenite number that weans is 9.1, and between piglet birth to weaning period, lethality is up to 14%, illustrates that the piglet general level of the health is low.Antiviral semen diluent is applied to swine artificial insemination can improve wean sodium selenite number effectively, and piglet lethality significantly reduces by 23.2% than common semen diluent, and result shows that antiviral semen diluent can improve swinery health degree effectively.
Apply antiviral semen diluent, normal control semen diluent and the natural breeding litter size analysis to 3 product boars, 6-15 interval is nearly all dropped in the interval of individual litter size.This shows the brood sow for single body, control group artificial insemination and the Reproduction of natural breeding to sow not obvious, the nest litter size that the natural breeding of male and female pig can reach, use artificial insemination can reach too, but artificial insemination is because of the raising of boar utilization rate, thus the stock boar raised can significantly reduce, so artificial insemination efficiency exceeds more than 10 times than the natural breeding of male and female pig, can be the improvement of producing and bringing higher economic benefit and swinery hereditary quality.Antiviral semen diluent is applied to swine artificial insemination, the nest litter size of brood sow and number born alive ideal in 3 pig variety result of the tests, common semen diluent artificial insemination effect is relatively poor.Very large economic benefit difference is also there is in 3 pig varieties in use artificial insemination and natural mating 2 kinds of breeding wonies.
In process of the test, result display removes 3 pig variety pigs and uses efficient semen diluent to carry out artificial insemination, the index such as conception rate, average litter size of its brood sow is significantly higher than outside the natural breeding of male and female pig, also find that 3 its degrees of variation of pig variety male and female pig natural breeding reproductive performance are large simultaneously, nest litter size is unstable, what have is joined female Litter Size in Pigs up to 17, and what have is joined female Litter Size in Pigs only 4, illustrates that natural mating boar semen difference in quality is larger.May mainly the natural breeding stock boar number of animals raised be large, feeding environment condition and dietary nutritional standard can not be unified, semen quality can not be unified to check, cause individual difference between breeding boar larger, the further reproductive efficiency affecting sow, quality of boar semen is also by the impact of hereditary difference between individuality in addition.
Result of the test shows simultaneously, Duroc, Landrace, Large White use between the reproductive performance such as conception rate, average litter size of artificial insemination breeding variant, Duroc average litter size is significantly lower than reproductive performance that is in vain long and large white sow, and between Landrace reproductive performance and Large White, difference is not remarkable.
In the description of this specification, specific features, structure, material or feature that the description of reference term " embodiment ", " some embodiments ", " example ", " concrete example " or " some examples " etc. means to describe in conjunction with this embodiment or example are contained at least one embodiment of the present invention or example.In this manual, identical embodiment or example are not necessarily referred to the schematic representation of above-mentioned term.And the specific features of description, structure, material or feature can combine in an appropriate manner in any one or more embodiment or example.
Although illustrate and describe embodiments of the invention above, be understandable that, above-described embodiment is exemplary, can not be interpreted as limitation of the present invention, those of ordinary skill in the art can change above-described embodiment within the scope of the invention when not departing from principle of the present invention and aim, revising, replacing and modification.
Claims (11)
1. the formula composition of the antiviral semen diluent of a boar, it is characterized in that, based on every 1000 milliliters of DDWs, the antiviral semen diluent of described pig contains:
15.50g glucose;
11.65g trisodium citrate;
The EDETATE SODIUM salt of 2.35g;
1.75g sodium bicarbonate;
1.00g polyvinyl alcohol;
The tricarboxylic aminomethane of 5.50g;
4.10g citric acid;
The cysteine of 0.07g;
1.00g vitamin C;
The Enrofloxacin of 0.10g; And
50ml antivirotic.
2. the formula composition of the antiviral semen diluent of pig according to claim 1, it is characterized in that, described antivirotic contains: Ribavirin, bupleurum extract, isatis root extract, glutinous rehmannia extract, Baical Skullcap root P.E, capejasmine extract, forsythia suspense extraction and Radix Scrophulariae extract.
3. the formula composition of the antiviral semen diluent of pig according to claim 2, it is characterized in that, described bupleurum extract is prepared through the following steps:
(1) utilize 10 premium on currency to carry out immersion 12 hours to 5 kilograms of radix bupleuri, then carry out decocting and concentrating and filtration treatment successively;
(2) ethanol being 70% by the filtrate obtained in step (1) and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours; And
(3) by concentrated for the filtrate heating obtained in step (2), and be that 1:2 carry out turn molten process and filter with water according to volume ratio by gained concentrate, evaporation drying process carried out to obtained filtrate, to obtain described bupleurum extract.
4. the formula composition of the antiviral semen diluent of pig according to claim 2, it is characterized in that, described isatis root extract is prepared through the following steps:
(1) utilize 10 premium on currency to carry out immersion 12 hours to 5 kilograms of Radix Isatidis, then carry out decocting and concentrating and filtration treatment successively;
(2) ethanol being 70% by the filtrate obtained in step (1) and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours; And
(3) by concentrated for the filtrate heating obtained in step (2), and be that 1:2 carries out turning molten process and filtering by gained concentrate and water according to volume ratio, evaporation drying process is carried out to obtained filtrate, to obtain described isatis root extract.
5. the formula composition of the antiviral semen diluent of pig according to claim 2, it is characterized in that, described glutinous rehmannia extract is prepared through the following steps:
(1) utilize 10 premium on currency to carry out immersion 12 hours to 5 kilograms of glutinous rehmannia, then carry out decocting and concentrating and filtration treatment successively;
(2) ethanol being 70% by the filtrate obtained in step (1) and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours; And
(3) by concentrated for the filtrate heating obtained in step (2), and be that 1:2 carry out turn molten process and filter with water according to volume ratio by gained concentrate, evaporation drying process carried out to obtained filtrate, to obtain described glutinous rehmannia extract.
6. the formula composition of the antiviral semen diluent of pig according to claim 2, it is characterized in that, described Baical Skullcap root P.E is prepared through the following steps:
(1) utilize 10 premium on currency to carry out immersion 12 hours to 5 kilograms of roots of large-flowered skullcap, then carry out decocting and concentrating and filtration treatment successively;
(2) ethanol being 70% by the filtrate obtained in step (1) and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours; And
(3) by concentrated for the filtrate heating obtained in step (2), and be that 1:2 carry out turn molten process and filter with water according to volume ratio by gained concentrate, evaporation drying process carried out to obtained filtrate, to obtain described Baical Skullcap root P.E.
7. the formula composition of the antiviral semen diluent of pig according to claim 2, it is characterized in that, described capejasmine extract is prepared through the following steps:
(1) utilize 10 premium on currency to carry out immersion 12 hours to 5 kilograms of cape jasmines, then carry out decocting and concentrating and filtration treatment successively;
(2) ethanol being 70% by the filtrate obtained in step (1) and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours; And
(3) by concentrated for the filtrate heating obtained in step (2), and be that 1:2 carry out turn molten process and filter with water according to volume ratio by gained concentrate, evaporation drying process carried out to obtained filtrate, to obtain described capejasmine extract.
8. the formula composition of the antiviral semen diluent of pig according to claim 2, it is characterized in that, described forsythia suspense extraction is prepared through the following steps:
(1) utilize 10 premium on currency to carry out immersion 12 hours to 5 kilograms of capsules of weeping forsythia, then carry out decocting and concentrating and filtration treatment successively;
(2) ethanol being 70% by the filtrate obtained in step (1) and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours; And
(3) by concentrated for the filtrate heating obtained in step (2), and be that 1:2 carry out turn molten process and filter with water according to volume ratio by gained concentrate, evaporation drying process carried out to obtained filtrate, to obtain described forsythia suspense extraction.
9. the formula composition of the antiviral semen diluent of pig according to claim 2, it is characterized in that, described Radix Scrophulariae extract is prepared through the following steps:
(1) utilize 10 premium on currency to carry out immersion 12 hours to 5 kilograms of radix scrophulariaes, then carry out decocting and concentrating and filtration treatment successively;
(2) ethanol being 70% by the filtrate obtained in step (1) and volumetric concentration carries out equal-volume and mixes and carry out alcohol precipitation process and filter after 24 hours; And
(3) by concentrated for the filtrate heating obtained in step (2), and be that 1:2 carry out turn molten process and filter with water according to volume ratio by gained concentrate, evaporation drying process carried out to obtained filtrate, to obtain described Radix Scrophulariae extract.
10. the formula composition of the antiviral semen diluent of pig according to claim 2, it is characterized in that, described antivirotic is by obtaining the mixing of Ribavirin dilution, bupleurum extract dilution, isatis root extract dilution, glutinous rehmannia extract dilution, Baical Skullcap root P.E dilution, capejasmine extract dilution, forsythia suspense extraction dilution and Radix Scrophulariae extract dilution
Wherein,
Described Ribavirin dilution is by utilizing the glucose injection of 5wt% described Ribavirin dilution 10 ~ 1000 times to be obtained,
Described bupleurum extract dilution is by utilizing the glucose injection of 5wt% described bupleurum extract to be diluted 10 times and obtain,
Described isatis root extract dilution is by utilizing the glucose injection of 5wt% described isatis root extract to be diluted 10 times and obtain,
Described glutinous rehmannia extract dilution is by utilizing the glucose injection of 5wt% described glutinous rehmannia extract to be diluted 10 times and obtain,
Described Baical Skullcap root P.E dilution is by utilizing the glucose injection of 5wt% described Baical Skullcap root P.E to be diluted 10 times and obtain,
Described capejasmine extract dilution is by utilizing the glucose injection of 5wt% described capejasmine extract to be diluted 10 times and obtain,
Described forsythia suspense extraction dilution is by utilizing the glucose injection of 5wt% described forsythia suspense extraction to be diluted 10 times and obtain,
Described Radix Scrophulariae extract dilution is by utilizing the glucose injection of 5wt% described Radix Scrophulariae extract to be diluted 10 times and obtain.
The formula composition of the antiviral semen diluent of 11. pig according to claim 10, it is characterized in that, based on every 1000 milliliters of described DDWs, in described antivirotic, described Ribavirin dilution, described bupleurum extract dilution, described isatis root extract dilution, described glutinous rehmannia extract dilution, described Baical Skullcap root P.E dilution, described capejasmine extract dilution, described forsythia suspense extraction dilution and described Radix Scrophulariae extract dilution are respectively 1 milliliter.
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| CN105325401A (en) * | 2015-11-10 | 2016-02-17 | 天津市畜牧兽医研究所 | Fresh-pig-semen preserving long-acting dilution powder, preparing method thereof and application thereof |
| CN105918311A (en) * | 2016-07-01 | 2016-09-07 | 江苏省家禽科学研究所 | Livestock seminal fluid disinfectant, preparation method thereof and application |
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| CN114081891A (en) * | 2020-08-24 | 2022-02-25 | 山西好医生药业有限公司 | Preparation process and application of five-vitamin glucose |
| CN113545336A (en) * | 2021-07-19 | 2021-10-26 | 杨凌职业技术学院 | Pig semen purifying agent |
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Application publication date: 20151104 |