CN110800875A - Preparation method of additive for improving nonspecific immunity of laying hens - Google Patents
Preparation method of additive for improving nonspecific immunity of laying hens Download PDFInfo
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- 230000036039 immunity Effects 0.000 title claims abstract description 29
- 239000000654 additive Substances 0.000 title claims abstract description 24
- 230000000996 additive effect Effects 0.000 title claims abstract description 24
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 241001075517 Abelmoschus Species 0.000 claims abstract description 38
- 208000010359 Newcastle Disease Diseases 0.000 claims abstract description 37
- 210000003837 chick embryo Anatomy 0.000 claims abstract description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 24
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 21
- 239000012530 fluid Substances 0.000 claims abstract description 21
- 150000004676 glycans Chemical class 0.000 claims abstract description 21
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 21
- 239000005017 polysaccharide Substances 0.000 claims abstract description 21
- 229960005486 vaccine Drugs 0.000 claims abstract description 18
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims abstract description 17
- 108010073771 Soybean Proteins Proteins 0.000 claims abstract description 10
- 238000004108 freeze drying Methods 0.000 claims abstract description 9
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims abstract description 7
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims abstract description 7
- 235000010413 sodium alginate Nutrition 0.000 claims abstract description 7
- 239000000661 sodium alginate Substances 0.000 claims abstract description 7
- 229940005550 sodium alginate Drugs 0.000 claims abstract description 7
- 239000008223 sterile water Substances 0.000 claims abstract description 7
- 238000007865 diluting Methods 0.000 claims abstract description 5
- 239000008098 formaldehyde solution Substances 0.000 claims abstract description 5
- 238000007710 freezing Methods 0.000 claims abstract description 5
- 230000008014 freezing Effects 0.000 claims abstract description 5
- 230000002779 inactivation Effects 0.000 claims abstract description 5
- 238000005406 washing Methods 0.000 claims abstract description 5
- 239000000706 filtrate Substances 0.000 claims description 28
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 16
- 238000003756 stirring Methods 0.000 claims description 14
- 238000002386 leaching Methods 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 12
- 238000002156 mixing Methods 0.000 claims description 12
- 239000011259 mixed solution Substances 0.000 claims description 11
- 239000000243 solution Substances 0.000 claims description 9
- 239000004744 fabric Substances 0.000 claims description 8
- 238000001914 filtration Methods 0.000 claims description 8
- 238000010438 heat treatment Methods 0.000 claims description 8
- 238000011081 inoculation Methods 0.000 claims description 8
- 238000001816 cooling Methods 0.000 claims description 4
- 230000000415 inactivating effect Effects 0.000 claims description 4
- 210000001161 mammalian embryo Anatomy 0.000 claims description 4
- 239000002244 precipitate Substances 0.000 claims description 4
- 238000007789 sealing Methods 0.000 claims description 4
- 229940001941 soy protein Drugs 0.000 claims description 4
- 239000002504 physiological saline solution Substances 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims 1
- 229940071440 soy protein isolate Drugs 0.000 claims 1
- 235000019710 soybean protein Nutrition 0.000 abstract description 6
- 206010033799 Paralysis Diseases 0.000 abstract description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 abstract description 3
- 102000039446 nucleic acids Human genes 0.000 abstract description 3
- 108020004707 nucleic acids Proteins 0.000 abstract description 3
- 150000007523 nucleic acids Chemical class 0.000 abstract description 3
- 210000002966 serum Anatomy 0.000 abstract description 3
- 210000002784 stomach Anatomy 0.000 abstract description 2
- 239000011248 coating agent Substances 0.000 abstract 1
- 238000000576 coating method Methods 0.000 abstract 1
- 230000009466 transformation Effects 0.000 abstract 1
- 102000016943 Muramidase Human genes 0.000 description 6
- 108010014251 Muramidase Proteins 0.000 description 6
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 6
- 239000004325 lysozyme Substances 0.000 description 6
- 229960000274 lysozyme Drugs 0.000 description 6
- 235000010335 lysozyme Nutrition 0.000 description 6
- 108010074328 Interferon-gamma Proteins 0.000 description 5
- 102000008070 Interferon-gamma Human genes 0.000 description 5
- 102000004889 Interleukin-6 Human genes 0.000 description 5
- 108090001005 Interleukin-6 Proteins 0.000 description 5
- 229960003130 interferon gamma Drugs 0.000 description 5
- 229940100601 interleukin-6 Drugs 0.000 description 5
- 230000008901 benefit Effects 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 210000001035 gastrointestinal tract Anatomy 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- 206010020601 Hyperchlorhydria Diseases 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 230000001804 emulsifying effect Effects 0.000 description 2
- 238000005187 foaming Methods 0.000 description 2
- 230000002496 gastric effect Effects 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- 210000004400 mucous membrane Anatomy 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 230000001018 virulence Effects 0.000 description 2
- 241000287828 Gallus gallus Species 0.000 description 1
- 102000015696 Interleukins Human genes 0.000 description 1
- 108010063738 Interleukins Proteins 0.000 description 1
- 239000003674 animal food additive Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000010429 evolutionary process Effects 0.000 description 1
- BCQZXOMGPXTTIC-UHFFFAOYSA-N halothane Chemical compound FC(F)(F)C(Cl)Br BCQZXOMGPXTTIC-UHFFFAOYSA-N 0.000 description 1
- 108010046721 halothane dehalogenase Proteins 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
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- 238000012986 modification Methods 0.000 description 1
- 230000037125 natural defense Effects 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 239000000273 veterinary drug Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Birds (AREA)
- Animal Husbandry (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention provides a preparation method of an additive for improving the nonspecific immunity of laying hens, which comprises the following steps: diluting Newcastle disease LaSota strain, inoculating the diluted strain in chick embryo, incubating, collecting to obtain chick embryo allantoic fluid of Newcastle disease vaccine, adding formaldehyde solution for inactivation to obtain inactivated chick embryo allantoic fluid of Newcastle disease vaccine, dissolving soybean protein isolate in sterile water, adding sodium alginate, glycerol, okra polysaccharide and the inactivated chick embryo allantoic fluid of Newcastle disease vaccine, then dropwise adding calcium chloride solution, washing with sterile normal saline, pre-freezing, and freeze-drying to obtain the additive for improving the nonspecific immunity of laying hens. The isolated soybean protein mainly has the function of protecting the nucleic acid of the Newcastle disease LaSota strain, the inactivated Newcastle disease LaSota strain can avoid the risks of the virulent return of a live vaccine, the immune paralysis and the strong transformation of a strain, and the non-specific immunity in the serum of the laying hen is obviously improved by effectively coating the live vaccine with the okra polysaccharide and effectively passing through the stomach.
Description
Technical Field
The invention belongs to the technical field of feed additives, and particularly relates to a preparation method of an additive for improving nonspecific immunity of laying hens.
Background
The non-specific immunity of the laying hens is a natural defense function gradually established in the long-term phylogenetic and evolutionary processes of organisms, and has the characteristics of rapid identification (within 96 h), broad spectrum and hereditary interspecific difference.
According to the traditional method, the okra leaf powder is added into daily ration of the laying hens, so that the production performance and the egg quality of the laying hens can be improved, and meanwhile, the cecal intestinal microbial flora and the intestinal tissue structure of the laying hens can be improved to a certain extent. The okra middle leaf powder contains a plurality of components, different components contain different solvents, for example, the polysaccharide is dissolved in water, a special process is needed, after the polysaccharide is extracted, the dry powder leaves are difficult to take advantage of the polysaccharide in the digestive tract of the laying hens.
Disclosure of Invention
The invention aims to solve the technical problem that the invention aims at overcoming the defects of the prior art and provides a preparation method of an additive for improving the nonspecific immunity of laying hens, wherein soybean protein isolate in the method has emulsifying property, foaming property, gelling property and thermal stability, and mainly plays a role in protecting the nucleic acid of a Newcastle disease LaSota strain from gastric hyperacidity, the inactivated Newcastle disease LaSota strain can avoid the risks of strong virulence and immune paralysis of a live vaccine caused by using the Newcastle disease LaSota strain for a long time in a large quantity, and the risk of strong virus strain transfer, the inactivated Newcastle disease LaSota strain is effectively coated by okra polysaccharide to effectively transgastric, so that the contents of nonspecific immune interferon-gamma, interleukin-6 and lysozyme in the serum of the laying hens can be obviously improved, the using effect of the laying hens in the actual breeding process is obvious, in addition, the selection of the okra polysaccharide can avoid the influence of the basic nutrition of the feed by adding excessive okra dry leaves, meanwhile, the okra polysaccharide also has the advantages of repairing digestive tract mucous membranes, resisting gram-negative bacteria and the like, keeps higher resistance of the laying hens and achieves the effect of preventing epidemic diseases.
In order to solve the technical problems, the invention adopts the technical scheme that: a preparation method of an additive for improving the nonspecific immunity of laying hens comprises the following steps:
s1, adding water a into okra, heating to 70 ℃, preserving heat for 3 hours, carrying out primary leaching, filtering with filter cloth after leaching to obtain filtrate a, adding water b into the filtered okra residue, heating to 70 ℃ again, preserving heat for 3 hours, carrying out secondary leaching, filtering with filter cloth to obtain filtrate b, mixing the filtrate a and the filtrate b to obtain total filtrate, adding ethanol solution with the mass fraction of 95% into the total filtrate, standing for 12 hours at the temperature of 4 ℃, centrifuging for 10-20 minutes at the temperature of 4 ℃ and the rotating speed of 8000rpm, and carrying out freeze drying on the precipitate for 2 hours at the temperature of-20 ℃ to obtain okra polysaccharide;
s2, adjusting the content of the strain to be more than or equal to 4 multiplied by 108.0EID50Diluting 0.1mL of Newcastle disease La Sota strain by 10 times, inoculating the diluted strain into 9-10 days old chick embryos, wherein the inoculation amount of each embryo is 0.1mL, sealing needle holes by using candles after inoculation, incubating for 48 hours at the temperature of 37 ℃, and collecting allantoic fluid of the chick embryos of the Newcastle disease;
s3, adding a formaldehyde solution with the mass fraction of 38% -40% into the allantoic fluid of the chick embryo of the Newcastle disease vaccine obtained in S1 until the mass fraction of the formaldehyde is 0.1%, inactivating for 72 hours at the temperature of 4 ℃, and shaking for 3-4 times during the inactivation period to obtain the inactivated chick embryo allantoic fluid of the Newcastle disease vaccine;
s4, adding isolated soy protein into sterile water, stirring and dissolving, modifying at 60 ℃ for 30min, naturally cooling to room temperature, adding sodium alginate and glycerol, stirring and mixing for 10min, adding okra polysaccharide obtained in S1 and inactivated Newcastle disease vaccine chick embryo allantoic fluid obtained in S3, stirring and mixing for 5min to obtain a mixed solution a, dropwise adding a calcium chloride solution with the mass fraction of 1% into the mixed solution a for immobilization for 30min, washing with sterile physiological saline for 3 times, pre-freezing at-20 ℃ for 8-12 h, and freeze-drying at-50 ℃ for 10h to obtain the additive for improving the nonspecific immunity of the laying hens.
Preferably, the mass ratio of the okra to the water a and the mass ratio of the filtered okra residue to the water b in S1 are both 1: 10.
preferably, the volume ratio of the total filtrate to the 95% ethanol solution in the S1 is 1: 3.
preferably, the dosage ratio of the soybean protein isolate, the sterile water, the sodium alginate, the glycerol, the okra polysaccharide and the inactivated chick embryo allantoic fluid in the mixed solution a in S4 is 4 g: 100mL of: 1 g: 6 g: 10mL of: 10 mg.
Preferably, the volume ratio of the mixed solution a to the calcium chloride solution with the mass fraction of 1% in S4 is 1: 1.
preferably, the stirring rate in S4 is 150 r/min.
Preferably, the dropping rate in S4 is 5 drops/min to 8 drops/min.
Compared with the prior art, the invention has the following advantages:
the soybean protein isolate and the soybean protein isolate have emulsifying property, foaming property, gelling property and thermal stability, and mainly play a role in protecting the nucleic acid of the Newcastle disease LaSota strain from gastric hyperacidity, the inactivated Newcastle disease LaSota strain can avoid the risks of strong virulence of live vaccine, immune paralysis and virus strain transfer caused by using the Newcastle disease LaSota strain for a long time in a large quantity, the inactivated Newcastle disease LaSota strain is effectively coated by okra polysaccharide and effectively passes through the stomach, so that the contents of nonspecific immune interferon-gamma, interleukin-6 and lysozyme in the serum of laying hens can be obviously improved, the using effect of the laying hens in the actual breeding process is obvious, in addition, the okra polysaccharide is selected to avoid that the excessive okra dry leaves added in the feed influence the basic nutrition of the feed, and simultaneously, the okra polysaccharide also has the advantages of repairing digestive tract mucous membranes, resisting gram negative bacteria and the like, and keeps higher resistance of the laying hens, the effect of preventing epidemic diseases is achieved.
The present invention will be described in further detail with reference to examples.
Detailed Description
Example 1
The preparation method of the additive for improving the nonspecific immunity of the laying hens comprises the following steps:
s1, adding water a into okra, heating to 70 ℃, preserving heat for 3 hours, performing primary leaching, filtering with filter cloth after leaching to obtain filtrate a, adding water b into the filtered okra residue, heating to 70 ℃ again, preserving heat for 3 hours, performing secondary leaching, filtering with filter cloth to obtain filtrate b, mixing the filtrate a and the filtrate b to obtain total filtrate, adding ethanol solution with the mass fraction of 95% into the total filtrate, standing for 12 hours at the temperature of 4 ℃, centrifuging for 10 minutes at the temperature of 4 ℃ and the rotating speed of 8000rpm, and freeze-drying the precipitate for 2 hours at the temperature of-20 ℃ to obtain okra polysaccharide; the mass ratio of the okra to the water a and the mass ratio of the filtered okra residues to the water b are both 1: 10; the volume ratio of the total filtrate to 95% ethanol solution is 1: 3;
s2, adjusting the content of the strain to be more than or equal to 4 multiplied by 108.0EID50Diluting 0.1mL of Newcastle disease La Sota strain by 10 times, inoculating the diluted strain into 9-10 days old chick embryos, wherein the inoculation amount of each embryo is 0.1mL, sealing needle holes by using candles after inoculation, incubating for 48 hours at the temperature of 37 ℃, and collecting allantoic fluid of the chick embryos of the Newcastle disease; the New castle disease La Sota strain is purchased from Qilu animal health products Co., Ltd, approval articleThe number is veterinary medicine raw character 150253007;
s3, adding a formaldehyde solution with the mass fraction of 38% into the allantoic fluid of the chick embryo of the Newcastle disease vaccine obtained in S1 until the mass fraction of the formaldehyde is 0.1%, inactivating for 72 hours at the temperature of 4 ℃, and shaking for 3 times during the inactivation period to obtain the inactivated chick embryo allantoic fluid of the Newcastle disease vaccine;
s4, adding 4g of isolated soy protein into 100mL of sterile water, stirring and dissolving, modifying at 60 deg.C for 30min, naturally cooling to room temperature, adding 1g sodium alginate and 6g glycerol, stirring and mixing at the speed of 150r/min for 10min, adding 10mL of okra polysaccharide obtained from S1 and 10mg of inactivated chick Newcastle disease chick embryo allantoic fluid obtained from S3, stirring and mixing for 5min at the speed of 150r/min to obtain a mixed solution a, dripping 1 percent by mass of calcium chloride solution into the mixed solution a at the dripping speed of 5 drops/min for immobilization for 30min, washing for 3 times by using sterile normal saline, pre-freezing at-20 deg.C for 8h, and freeze-drying at-50 deg.C for 10h to obtain additive for improving nonspecific immunity of laying hen.
The 25-week-old adult laying hens, Hailan laying hens, were 12, divided into 4 groups (groups A-D), 1 each, and 3 replicates each. The group A-group C were fed with the additive for improving the nonspecific immunity of the laying hens prepared in this example at 4 mg, 2 mg, 1 mg, respectively, and the group D was a blank control. Respectively measuring the contents of non-specific immune interferon-gamma, interleukin-6 and lysozyme in the blood of laying hens fed for 0h, 24h, 48h, 72h and 96h, wherein the measurement results are shown in table 1:
TABLE 1 content of non-specific IFN-. gamma.interleukin-6 and lysozyme in the blood of layer chicken
As can be seen from Table 1, the non-specific immunity of the laying hens can be improved by feeding the additive for improving the non-specific immunity of the laying hens, the contents of interferon-gamma, interleukin and lysozyme of groups A-C are improved compared with those of a group D of a control group, wherein the interferon-gamma of the group A is improved by 2.7 times, the interleukin-6 is improved by 2.8 times and the lysozyme is improved by 1.6 times after the group A is fed with the additive for improving the non-specific immunity of the laying hens, so that the additive for improving the non-specific immunity of the laying hens, which is prepared by the embodiment, can improve the non-specific immunity of the laying hens.
Example 2
The preparation method of the additive for improving the nonspecific immunity of the laying hens comprises the following steps:
s1, adding water a into okra, heating to 70 ℃, preserving heat for 3 hours, performing primary leaching, filtering with filter cloth after leaching to obtain filtrate a, adding water b into the filtered okra residue, heating to 70 ℃ again, preserving heat for 3 hours, performing secondary leaching, filtering with filter cloth to obtain filtrate b, mixing the filtrate a and the filtrate b to obtain total filtrate, adding ethanol solution with the mass fraction of 95% into the total filtrate, standing for 12 hours at the temperature of 4 ℃, centrifuging for 20 minutes at the temperature of 4 ℃ and the rotating speed of 8000rpm, and freeze-drying the precipitate for 2 hours at the temperature of-20 ℃ to obtain okra polysaccharide; the mass ratio of the okra to the water a and the mass ratio of the filtered okra residues to the water b are both 1: 10; the volume ratio of the total filtrate to 95% ethanol solution is 1: 3;
s2, adjusting the content of the strain to be more than or equal to 4 multiplied by 108.0EID50Diluting 0.1mL of Newcastle disease La Sota strain by 10 times, inoculating the diluted strain into 9-10 days old chick embryos, wherein the inoculation amount of each embryo is 0.1mL, sealing needle holes by using candles after inoculation, incubating for 48 hours at the temperature of 37 ℃, and collecting allantoic fluid of the chick embryos of the Newcastle disease; the new castle disease La Sota strain is purchased from Qilu animal health products Co., Ltd, and the approved literature is veterinary drug Sheng Zi 150253007;
s3, adding a formaldehyde solution with the mass fraction of 40% into the allantoic fluid of the chick embryo of the Newcastle disease vaccine obtained in S1 until the mass fraction of the formaldehyde is 0.1%, inactivating for 72 hours at the temperature of 4 ℃, and shaking for 4 times during the inactivation period to obtain the inactivated chick embryo allantoic fluid of the Newcastle disease vaccine;
s4, adding 4g of isolated soy protein into 100mL of sterile water, stirring and dissolving, modifying at 60 deg.C for 30min, naturally cooling to room temperature, adding 1g sodium alginate and 6g glycerol, stirring and mixing at the speed of 150r/min for 10min, adding 10mL of okra polysaccharide obtained from S1 and 10mg of inactivated chick Newcastle disease chick embryo allantoic fluid obtained from S3, stirring and mixing for 5min at the speed of 150r/min to obtain a mixed solution a, dripping 1 percent calcium chloride solution at the dripping speed of 8 drops/min into the mixed solution a for immobilization for 30min, then washing for 3 times by using sterile normal saline, pre-freezing at-20 deg.C for 12h, and freeze-drying at-50 deg.C for 10h to obtain additive for improving nonspecific immunity of laying hen.
The additive for improving the nonspecific immunity of the laying hens, which is prepared by the embodiment, is added into basic feed of 25-week-old halan laying hens to feed 50 laying hens, and through detection, the nonspecific immunity of each laying hen is improved, and the disease rate of the laying hens is obviously reduced in the feeding process.
The above description is only for the preferred embodiment of the present invention, and is not intended to limit the present invention in any way. Any simple modification, change and equivalent changes of the above embodiments according to the technical essence of the invention are still within the protection scope of the technical solution of the invention.
Claims (7)
1. A preparation method of an additive for improving the nonspecific immunity of laying hens is characterized by comprising the following steps:
s1, adding water a into okra, heating to 70 ℃, preserving heat for 3 hours, carrying out primary leaching, filtering with filter cloth after leaching to obtain filtrate a, adding water b into the filtered okra residue, heating to 70 ℃ again, preserving heat for 3 hours, carrying out secondary leaching, filtering with filter cloth to obtain filtrate b, mixing the filtrate a and the filtrate b to obtain total filtrate, adding ethanol solution with the mass fraction of 95% into the total filtrate, standing for 12 hours at the temperature of 4 ℃, centrifuging for 10-20 minutes at the temperature of 4 ℃ and the rotating speed of 8000rpm, and carrying out freeze drying on the precipitate for 2 hours at the temperature of-20 ℃ to obtain okra polysaccharide;
s2, adjusting the content of the strain to be more than or equal to 4 multiplied by 108.0EID50/0.Diluting 1mL of Newcastle disease La Sota strain by 10 times, inoculating into 9-10 days old chick embryos, wherein the inoculation amount of each embryo is 0.1mL, sealing a pinhole by using a candle after inoculation, incubating for 48 hours at the temperature of 37 ℃, and collecting allantoic fluid of the chick embryos of the Newcastle disease vaccine;
s3, adding a formaldehyde solution with the mass fraction of 38% -40% into the allantoic fluid of the chick embryo of the Newcastle disease vaccine obtained in S1 until the mass fraction of the formaldehyde is 0.1%, inactivating for 72 hours at the temperature of 4 ℃, and shaking for 3-4 times during the inactivation period to obtain the inactivated chick embryo allantoic fluid of the Newcastle disease vaccine;
s4, adding isolated soy protein into sterile water, stirring and dissolving, modifying at 60 ℃ for 30min, naturally cooling to room temperature, adding sodium alginate and glycerol, stirring and mixing for 10min, adding okra polysaccharide obtained in S1 and inactivated Newcastle disease vaccine chick embryo allantoic fluid obtained in S3, stirring and mixing for 5min to obtain a mixed solution a, dropwise adding a calcium chloride solution with the mass fraction of 1% into the mixed solution a for immobilization for 30min, washing with sterile physiological saline for 3 times, pre-freezing at-20 ℃ for 8-12 h, and freeze-drying at-50 ℃ for 10h to obtain the additive for improving the nonspecific immunity of the laying hens.
2. The method for preparing the additive for improving the nonspecific immunity of laying hens as claimed in claim 1, wherein the mass ratio of okra to water a and the mass ratio of filtered okra residue to water b in S1 are both 1: 10.
3. the method for preparing the additive for improving the nonspecific immunity of laying hens as claimed in claim 1, wherein the volume ratio of the total filtrate to the 95% ethanol solution in S1 is 1: 3.
4. The method of claim 1, wherein the mixture a of S4 contains soy protein isolate, sterile water, sodium alginate, glycerol, okra polysaccharide and inactivated Newcastle disease chick embryo allantoic fluid in a ratio of 4 g: 100mL of: 1 g: 6 g: 10mL of: 10 mg.
5. The method for preparing the additive for improving the nonspecific immunity of laying hens as claimed in claim 1, wherein the volume ratio of the mixed solution a to the 1% calcium chloride solution in S4 is 1: 1.
6. the method for preparing the additive for improving the nonspecific immunity of laying hens as claimed in claim 1, wherein the stirring rate in S4 is 150 r/min.
7. The method for preparing the additive for improving the nonspecific immunity of laying hens as claimed in claim 1, wherein the dropping rate in S4 is 5 drops/min to 8 drops/min.
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Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103110942A (en) * | 2012-12-18 | 2013-05-22 | 山东滨州沃华生物工程有限公司 | Preparation method of inactivated newcastle disease vaccine |
| CN103251938A (en) * | 2013-04-15 | 2013-08-21 | 内蒙古神元生物工程股份有限公司 | Industrial production process of eimeria coccidiosis attenuated vaccine |
| CN104397348A (en) * | 2014-12-26 | 2015-03-11 | 郭庆 | Preparation method of new castle disease traditional Chinese medicine feed additive |
| CN104548088A (en) * | 2014-12-22 | 2015-04-29 | 天津瑞普生物技术股份有限公司 | Method for preparing bivalent vaccine of newcastle disease virus La Sota strain and infectious bronchitis virus N-S multi-epitope protein |
| CN105214082A (en) * | 2014-07-04 | 2016-01-06 | 乾元浩生物股份有限公司 | Newcastle disease, bird flu bivalent inactivated vaccine and preparation method thereof |
| CN106074868A (en) * | 2016-07-15 | 2016-11-09 | 浙江省海洋开发研究院 | A kind of selenizing polysaccharide immunostimulant |
| CN107823641A (en) * | 2017-12-15 | 2018-03-23 | 山东农业大学 | A kind of injection-type inactivated vaccine adjuvant for animals and preparation method thereof |
| CN108114277A (en) * | 2018-02-07 | 2018-06-05 | 姜新鹏 | The microcapsule preparation method of oral killed Porcine epidemic diarrhea virus |
| CN108210919A (en) * | 2018-04-12 | 2018-06-29 | 天津瑞普生物技术股份有限公司 | A kind of preparation method of duck infectious serositis microencapsulation oral vaccine |
| CN109674748A (en) * | 2019-02-15 | 2019-04-26 | 大连三仪动物药品有限公司 | A kind of traditional Chinese medicine oral liquid and its application enhancing newcastle disease vaccine immune effect |
| CN109820834A (en) * | 2018-12-24 | 2019-05-31 | 内蒙古必威安泰生物科技有限公司 | A kind of protective agent of aftosa inactivation of viruses and the preparation method of microcapsule vaccine |
-
2019
- 2019-12-04 CN CN201911226486.XA patent/CN110800875A/en active Pending
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103110942A (en) * | 2012-12-18 | 2013-05-22 | 山东滨州沃华生物工程有限公司 | Preparation method of inactivated newcastle disease vaccine |
| CN103251938A (en) * | 2013-04-15 | 2013-08-21 | 内蒙古神元生物工程股份有限公司 | Industrial production process of eimeria coccidiosis attenuated vaccine |
| CN105214082A (en) * | 2014-07-04 | 2016-01-06 | 乾元浩生物股份有限公司 | Newcastle disease, bird flu bivalent inactivated vaccine and preparation method thereof |
| CN104548088A (en) * | 2014-12-22 | 2015-04-29 | 天津瑞普生物技术股份有限公司 | Method for preparing bivalent vaccine of newcastle disease virus La Sota strain and infectious bronchitis virus N-S multi-epitope protein |
| CN104397348A (en) * | 2014-12-26 | 2015-03-11 | 郭庆 | Preparation method of new castle disease traditional Chinese medicine feed additive |
| CN106074868A (en) * | 2016-07-15 | 2016-11-09 | 浙江省海洋开发研究院 | A kind of selenizing polysaccharide immunostimulant |
| CN107823641A (en) * | 2017-12-15 | 2018-03-23 | 山东农业大学 | A kind of injection-type inactivated vaccine adjuvant for animals and preparation method thereof |
| CN108114277A (en) * | 2018-02-07 | 2018-06-05 | 姜新鹏 | The microcapsule preparation method of oral killed Porcine epidemic diarrhea virus |
| CN108210919A (en) * | 2018-04-12 | 2018-06-29 | 天津瑞普生物技术股份有限公司 | A kind of preparation method of duck infectious serositis microencapsulation oral vaccine |
| CN109820834A (en) * | 2018-12-24 | 2019-05-31 | 内蒙古必威安泰生物科技有限公司 | A kind of protective agent of aftosa inactivation of viruses and the preparation method of microcapsule vaccine |
| CN109674748A (en) * | 2019-02-15 | 2019-04-26 | 大连三仪动物药品有限公司 | A kind of traditional Chinese medicine oral liquid and its application enhancing newcastle disease vaccine immune effect |
Non-Patent Citations (1)
| Title |
|---|
| 许明录等: "黄秋葵果皮与根的抗氧化活性比较", 《河南科技学院学报(自然科学版)》 * |
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