WO2023060671A1 - 一种中药多糖饲料添加剂及其应用 - Google Patents

一种中药多糖饲料添加剂及其应用 Download PDF

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WO2023060671A1
WO2023060671A1 PCT/CN2021/128432 CN2021128432W WO2023060671A1 WO 2023060671 A1 WO2023060671 A1 WO 2023060671A1 CN 2021128432 W CN2021128432 W CN 2021128432W WO 2023060671 A1 WO2023060671 A1 WO 2023060671A1
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polysaccharide
parts
hours
chinese medicine
preparation
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French (fr)
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许启太
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海南今珠农业发展有限公司
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    • 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
    • A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • 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
    • A23K20/10—Organic substances
    • A23K20/142—Amino acids; Derivatives thereof
    • A23K20/147—Polymeric derivatives, e.g. peptides or proteins
    • 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
    • A23K20/10—Organic substances
    • A23K20/158—Fatty acids; Fats; Products containing oils or fats
    • 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
    • A23K20/10—Organic substances
    • A23K20/163—Sugars; Polysaccharides
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
    • Y02P60/87—Re-use of by-products of food processing for fodder production

Definitions

  • the invention relates to the technical field of feed additives, in particular to a traditional Chinese medicine polysaccharide feed additive and its preparation and application.
  • the invention provides a traditional Chinese medicine polysaccharide feed additive for preventing African swine fever, swine fever, and pig blue-ear disease, which comprises the following components in parts by weight: 5-10 parts of lactoferrin, 3-5 parts of medium-chain triglycerides 10-15 parts of beta glucan, 20-40 parts of betel nut polysaccharide, 10-30 parts of bovine herbaceous polysaccharide, 10-30 parts of astragalus polysaccharide, 20-50 parts of green grape algae polysaccharide, 20-50 parts of Bacillus subtilis fermentation product 40 servings.
  • lactoferrin 7 parts of lactoferrin, 5 parts of medium-chain triglycerides, 11 parts of ⁇ -glucan, 40 parts of green grape algae polysaccharide, 30 parts of betel nut polysaccharide, bovine herbaceous polysaccharide 20 parts, Radix Astragali polysaccharide 20 parts, Bacillus subtilis fermentation product 30 parts.
  • S1 Bacillus subtilis is expanded and cultivated twice to obtain a seed liquid, inoculated into a fermentation medium, and fermented and cultivated for 30-40 hours under the conditions of pH6.5-pH7.5, dissolved oxygen 60-70%, and 37 degrees, Then adjust the fermentation conditions to be pH4-pH4.5, dissolved oxygen 10-20%, and culture at 37 degrees for 6-10 hours, then adjust the pH of the fermentation broth to 7, and at the same time add 0.1mol of 2%-5% of the volume of the fermentation broth at a constant speed /L sodium chlorate, centrifuged and dried to get fermentation powder;
  • S3 is prepared by mixing lactoferrin, medium-chain triglyceride, ⁇ -glucan, areca nut polysaccharide, bovine herbaceous polysaccharide, astragalus polysaccharide, green staphylococcus polysaccharide, and Bacillus subtilis fermentation product.
  • the components of the fermentation medium are 50 g of glucose, 20 g of peptone, 10 g of yeast powder, 5 g of potassium dihydrogen phosphate trihydrate, 1 g of sodium chloride, 2 g of magnesium sulfate, and 1.5 g of ammonium sulfate per liter of water. .
  • the preparation method of Niu Dali polysaccharide in step S2 is: cut Niu Dali into slices, 8 times the weight of water, heat to 90°C, extract for 4 hours, filter, add 4 times the amount of water to the filter residue, extract for 1 hour, Filtrate, combine the secondary filtrates, concentrate under reduced pressure to 10% of the original volume; put the concentrated solution at room temperature, add ethanol to make the alcohol content 75%, stir while adding, let stand at 4°C for 12 hours, centrifuge to remove the precipitate, and the supernatant The liquid was filtered through a membrane to obtain a fine extract, which was concentrated under reduced pressure and dried.
  • the preparation method of areca nut polysaccharide in step S2 is: wash the fresh betel nut fruit and directly perform physical pressing, collect the juice through filtration, collect the residue, add 5 times the amount of water to the residue and extract at 80 degrees for 3 hours , filtered to obtain the extract; combine the extract and fruit juice, concentrate under reduced pressure to 20% of the original volume, add ethanol to make the ethanol content 80%, stir while adding, let stand at 4°C for 12 hours, centrifuge to obtain a precipitate, and dry the precipitate have to.
  • the preparation method of Radix Radix Polysaccharide in step S2 is as follows: slice Radix Radix Radix Radix Radix Radix, add 8 times the weight of water, heat to 95°C, extract for 4 hours, filter, add 4 times the amount of water to the filter residue, extract for 1 hour, Filtrate, combine the secondary filtrates, concentrate under reduced pressure to 15% of the original volume; put the concentrated solution at room temperature, add ethanol to make the alcohol content 75%, stir while adding, let stand at 4°C for 12 hours, centrifuge to remove the precipitate, supernatant The liquid was filtered through a membrane to obtain a fine extract, which was concentrated under reduced pressure and dried.
  • the preparation method of the green staphylococcus polysaccharide in step S2 is: wash the fresh green staphylococcus with fresh water, add 5 times the weight of water after crushing, heat to 40 degrees, extract for 8 hours, filter, and Concentrate the filtrate under reduced pressure to 20% of the original volume; cool to room temperature, add ethanol to make the alcohol content to 85%, stir while adding, let stand at 4°C for 12 hours, centrifuge to obtain a precipitate, and dry the precipitate to obtain the final product.
  • one expansion culture inoculate Bacillus subtilis into LB medium in LB medium at 30-37 degrees, 200-400rpm and cultivate for 24h, the initial pH of LB medium is 7, to obtain first-class seeds liquid;
  • Secondary expansion cultivation re-inoculate the primary seed liquid into LB medium for expansion cultivation, cultivate at 30-37°C and 200-400rpm for 12 hours, and inoculum amount is 5-10% of the volume of LB medium to obtain seed liquid.
  • the beneficial effect of the present invention is: a kind of traditional Chinese medicine polysaccharide feed additive provided by the present invention, main component contains lactoferrin, medium-chain triglyceride, beta glucan, traditional Chinese medicine polysaccharide additive and Bacillus subtilis fermentation product, synergistic effect, Can significantly improve the immunity of pigs, mixed with pig feed for pig breeding, internal adjustment and external sparseness, can improve the non-specific immunity and specific immunity of pigs, thereby reducing swine fever, African swine fever, and highly pathogenic swine fever The harm of virus infection such as ear disease to the body, increase the body's disease resistance, and achieve the preventive effect.
  • the invention provides a traditional Chinese medicine polysaccharide additive for preventing African swine fever, which comprises the following components in parts by weight: 5-10 parts of lactoferrin, 3-5 parts of medium-chain triglycerides, 10-5 parts of beta glucan 15 parts, 20-40 parts of betel nut polysaccharide, 10-30 parts of bovine polysaccharide, 10-30 parts of red qi polysaccharide, 20-50 parts of green staphylococcus polysaccharide, 20-40 parts of Bacillus subtilis fermentation product.
  • the preparation method of the above-mentioned traditional Chinese medicine polysaccharide additive comprises the following steps:
  • Bacillus subtilis is expanded and cultivated twice to obtain the seed solution, specifically:
  • Primary expansion culture Inoculate Bacillus subtilis into LB medium at 30-37°C, 200-400rpm and cultivate for 24h, the initial pH of LB medium is 7, to obtain the first-grade seed liquid;
  • Secondary expansion cultivation re-inoculate the primary seed liquid into LB medium for expansion cultivation, cultivate at 30-37°C and 200-400rpm for 12 hours, and inoculum amount is 5-10% of the volume of LB medium to obtain seed liquid.
  • the components of the fermentation medium are 50 g of glucose, 20 g of peptone, 10 g of yeast powder, 5 g of potassium dihydrogen phosphate trihydrate, 1 g of sodium chloride, 2 g of magnesium sulfate, and 1.5 g of ammonium sulfate per liter of water.
  • the preparation method of the betel nut polysaccharide is as follows: wash the fresh betel nut fruit and directly perform physical pressing, collect the juice through filtration, collect the residue, add 5 times the amount of water to the residue and extract at 80 degrees for 3 hours, and filter to obtain the extract; combine the extract and juice, Concentrate under reduced pressure to 20% of the original volume, add ethanol to make the ethanol content 80%, stir while adding, let stand at 4°C for 12 hours, centrifuge to obtain a precipitate, and dry the precipitate to obtain the final product.
  • Niu Dali polysaccharide The preparation method of Niu Dali polysaccharide is as follows: cut Niu Dali into slices, add 8 times the weight of water, heat to 90°C, extract for 4 hours, filter, add 4 times the amount of water to the filter residue, extract for 1 hour, filter, combine the secondary filtrates, and concentrate under reduced pressure to the original Volume 10%; put the concentrated solution at room temperature, add ethanol to make the alcohol content 75%, stir while adding, stand at 4°C for 12 hours, centrifuge to remove the precipitate, filter the supernatant through a membrane to obtain a fine extract, concentrate under reduced pressure After that, dry it.
  • the preparation method of Radix Radix Polysaccharides is as follows: slice Radix Radix Radix Radix Radix Radix Radix, add 8 times the weight of water, heat to 95°C, extract for 4 hours, filter, add 4 times the amount of water to the filter residue, extract for 1 hour, filter, combine the secondary filtrates, and concentrate under reduced pressure to the original Volume 15%; put the concentrated solution at room temperature, add ethanol to make the alcohol content 75%, stir while adding, let stand at 4°C for 12 hours, centrifuge to remove the precipitate, filter the supernatant through a membrane to obtain a fine extract, concentrate under reduced pressure After that, dry it.
  • the preparation method of grape algae polysaccharide is as follows: wash fresh green staphylococcus with fresh water, add 5 times the weight of water after crushing, heat to 40 degrees, extract for 8 hours, filter, and concentrate the filtrate under reduced pressure to 20% of the original volume; cool to room temperature, Add ethanol to make the alcohol content to 85%, stir while adding, let stand at 4°C for 12 hours, centrifuge to obtain a precipitate, and dry the precipitate.
  • lactoferrin medium chain triglyceride, ⁇ -glucan, betel nut polysaccharide, bovine polysaccharide, astragalus polysaccharide, green staphylococcus polysaccharide, and Bacillus subtilis fermentation product to obtain the product.
  • the traditional Chinese medicine polysaccharide additive in this embodiment includes the following components by weight: 7 parts by weight of lactoferrin, 5 parts by medium chain triglycerides, 11 parts by beta glucan, 30 parts by betel nut polysaccharide 20 parts of bovine herbaceous polysaccharide, 20 parts of red qi polysaccharide, 40 parts of green staphylococcus polysaccharide, 30 parts of Bacillus subtilis fermentation product.
  • the preparation method of the above-mentioned traditional Chinese medicine polysaccharide additive comprises the following steps:
  • Bacillus subtilis is expanded and cultivated twice to obtain the seed solution, specifically:
  • Primary expansion culture inoculate Bacillus subtilis into LB medium and culture in LB medium at 35°C and 300rpm for 24 hours, the initial pH of LB medium is 7, and obtain the primary seed liquid;
  • Secondary expansion cultivation re-inoculate the primary seed liquid into LB medium for expansion cultivation, cultivate at 35 degrees and 300 rpm for 12 hours, and the inoculum amount is 10% of the volume of LB medium to obtain a seed liquid.
  • the components of the fermentation medium are 50 g of glucose, 20 g of peptone, 10 g of yeast powder, 5 g of potassium dihydrogen phosphate trihydrate, 1 g of sodium chloride, 2 g of magnesium sulfate, and 1.5 g of ammonium sulfate per liter of water.
  • Wash the fresh betel nut fruit and directly perform physical pressing collect the juice through filtration, collect the residue, add 5 times the amount of water to the residue and extract at 80 degrees for 3 hours, filter to obtain the extract; combine the extract and juice, and concentrate under reduced pressure to the original volume of 20 %, add ethanol to make the ethanol content to 80%, stir while adding, let stand at 4°C for 12 hours, centrifuge to obtain a precipitate, and dry the precipitate.
  • Slice Radix Radix Radix Radix Radix Radix Radix add 8 times the weight of water, heat to 95°C, extract for 4 hours, filter, add 4 times the amount of water to the filter residue, extract for 1 hour, filter, combine the secondary filtrates, concentrate under reduced pressure to 15% of the original volume; put the concentrated solution in At room temperature, add ethanol to make the alcohol content to 75%, stir while adding, let stand at 4°C for 12 hours, centrifuge to remove the precipitate, filter the supernatant through a membrane to obtain a fine extract, concentrate under reduced pressure, and dry.
  • the traditional Chinese medicine polysaccharide additive of this embodiment includes the following components by weight: 5 parts of lactoferrin, 5 parts of medium-chain triglycerides, 10 parts of ⁇ -glucan, 40 parts of betel nut polysaccharide, 30 parts of bovine herbaceous polysaccharide, 30 parts of polysaccharide, 50 parts of green grape algae polysaccharide, 20 parts of Bacillus subtilis fermentation product.
  • step S2 is as follows:
  • the traditional Chinese medicine polysaccharide additive of this embodiment includes the following components by weight: 10 parts of lactoferrin, 3 parts of medium-chain triglycerides, 15 parts of ⁇ -glucan, 20 parts of betel nut polysaccharide, 10 parts of bovine herbaceous polysaccharide, and 10 parts of polysaccharide, 20 parts of green staphylococcus polysaccharide, 40 parts of Bacillus subtilis fermentation product.
  • step S2 is as follows:
  • the traditional Chinese medicine polysaccharide additive in this comparative example includes the following components by weight: 30 parts of betel nut polysaccharide, 20 parts of bovine polysaccharide, 20 parts of red qi polysaccharide, 40 parts of green grape algae polysaccharide, and 30 parts of Bacillus subtilis fermentation product.
  • each component is the same as in Example 1, except that lactoferrin, medium-chain triglyceride and ⁇ -glucan are not added.
  • Example 1 The difference between this comparative example and Example 1 is that the fermentation conditions of Bacillus subtilis are different, which are as follows: the seed solution is inoculated into the fermentation medium and fermented under the conditions of pH7, dissolved oxygen 70%, 37 degrees, and stirring speed of 300rpm for 43 Hours, and in the last 7 hours of the fermentation process, 0.1mol/L sodium chlorate of 4% of the fermentation broth volume was added at a constant speed; during the period, dissolved oxygen and pH value were not changed.
  • the fermentation conditions of Bacillus subtilis are different, which are as follows: the seed solution is inoculated into the fermentation medium and fermented under the conditions of pH7, dissolved oxygen 70%, 37 degrees, and stirring speed of 300rpm for 43 Hours, and in the last 7 hours of the fermentation process, 0.1mol/L sodium chlorate of 4% of the fermentation broth volume was added at a constant speed; during the period, dissolved oxygen and pH value were not changed.
  • Example 1 The difference between this comparative example and Example 1 is that the fermentation conditions of Bacillus subtilis are different, and the specific difference is that calcium chlorate is not added in the latter part of the fermentation.
  • polysaccharides in this comparative example are as follows: 20 parts of bovine polysaccharides, 20 parts of red root polysaccharides, and 40 parts of green grape algae polysaccharides.
  • polysaccharides in this comparative example are as follows: 30 parts of betel nut polysaccharide, 20 parts of red qi polysaccharide, and 40 parts of green grape algae polysaccharide.
  • polysaccharides in this comparative example are as follows: 30 parts of betel nut polysaccharide, 20 parts of bovine polysaccharide, and 40 parts of green grape algae polysaccharide.
  • polysaccharides in this comparative example are as follows: 30 parts of betel nut polysaccharide, 20 parts of bovine polysaccharide, and 20 parts of red qi polysaccharide.
  • the polysaccharide traditional Chinese medicines prepared in Examples 1 to 3 and Comparative Examples 1 to 7 were prepared using DMEM with 30% ultrapure water added, and the preparation concentration was 2g/100ml to obtain test solutions 1-6.
  • Vero cells grown in the 96-well plate were treated with the above test solution at a concentration of 80 ⁇ M, and each test solution was repeated in 4 wells.
  • the adjuvant (lactoferrin, medium-chain triglyceride, ⁇ -glucan) in the application of the present invention itself has no effect on the virus, but it has no effect on the virus in live pigs.
  • the body can play the role of transporting effective substances, so that the pig serum can have an inhibitory effect on the virus.
  • the swine fever virus detection was carried out on this batch of pigs, and the kit used the swine fever virus/foot-and-mouth disease virus universal (CSFV/FMDV-U) nucleic acid kit of the test organism to ensure that the batch of pigs were all negative before the test.
  • CSFV/FMDV-U swine fever virus/foot-and-mouth disease virus universal
  • Select 120 heads as the experimental group administer food mixed with food for two weeks, use the traditional Chinese medicine polysaccharide additive prepared in Example 1-3 at 2% of the daily weight of the ration, and switch to the basal ration after two weeks.
  • Select 40 as the control group feed the same amount of basal food normally.
  • the traditional Chinese medicine polysaccharide additive provided by the present invention also has a good preventive effect on African swine fever (ASFV).
  • ASFV African swine fever
  • CSFV section on swine fever
  • the inventor has also experimentally carried out a re-raising test for pig blue-ear disease:
  • the batch of pigs was tested for porcine PRRS virus, and the kit used the classic type (PRRSV-C) nucleic acid kit of Shanghai Bike Biotechnology Co., Ltd. to ensure that the batch of pigs were all negative before the test.
  • PRRSV-C classic type nucleic acid kit of Shanghai Bike Biotechnology Co., Ltd.
  • Select 10 as the control group feed the same amount of basal food normally.
  • the Chinese medicine polysaccharide additive provided by the present invention also has a certain effect for preventing pig blue-ear disease, and 10 live pigs of the control group are all infected with this virus, while the additives prepared in Examples 1 to 3 have an infection rate that is significantly lower than that of the contrast Group.

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Abstract

一种中药多糖饲料添加剂及制备方法和应用，该添加剂由下述重量份的成份组成：乳铁蛋白5-10份、中链甘油三酯3-5份，β-葡聚糖10-15份、槟榔多糖20-40份、牛大力多糖10-30份、红芪多糖10-30份、绿葡萄藻多糖20-50份和枯草芽孢杆菌发酵产物20-40份。该添加剂用于生猪养殖，可预防猪瘟、非洲猪瘟、高致病性猪蓝耳病等烈性病毒性传染病。

Description

一种中药多糖饲料添加剂及其应用 技术领域
本发明涉及饲料添加剂技术领域，具体涉及一种中药多糖饲料添加剂及其制备和应用。
背景技术
中国是世界公认的生猪养殖大国，如何有效预防疫病，是我国养猪生产面临的一个主要问题，其中猪瘟、非洲猪瘟、高致病性猪蓝耳病等烈性病毒性传染病是危害养殖业的大敌。我国自2018年8月爆发非洲猪瘟以来，迅速蔓延到全国，造成极其惨重的损失，一场始料不及的非瘟造成各种肉类蛋白大幅涨价，直接导致2019年中国的CPI比同期上升0.5％，极大影响了十几亿人口的生活水平。后非洲猪瘟时代，防控非洲猪瘟与产业稳定发展需同时进行。虽然国家出台了支持生猪发展的一系列优惠政策，但由于没有疫苗及药物的防控保障，仍然存在不敢养、养不活的问题，难以消除非洲猪瘟给养猪业带来的威胁和恐惧。在病毒肆虐而缺乏有效防控疫苗和药物的情况下，急需研究以中药及天然成分为主的猪瘟防控饲料产品。
发明内容
本发明提供一种用于预防非洲猪瘟、猪瘟、猪蓝耳病的中药多糖饲料添加剂，包括如下重量份的各组分：乳铁蛋白5-10份、中链甘油三酯3-5份，β葡聚糖10-15份、槟榔多糖20-40份、牛大力多糖10-30份、红芪多糖10-30份、绿葡萄藻多糖20-50份、枯草芽孢杆菌发酵产物20-40份。
其中优选的是，包括如下重量份的各组分：乳铁蛋白7份、中链甘油三酯5份，β葡聚糖11份、绿葡萄藻多糖40份、槟榔多糖30份、牛大力多糖20份、红芪多糖20份、枯草芽孢杆菌发酵产物30份。
并提供上述中药多糖饲料添加剂的制备方法，包括如下步骤：
S1将枯草芽孢杆菌经两次扩大培养得种液，将种液接种至发酵培养基中于pH6.5-pH7.5、溶解氧60-70％、37度条件下发酵培养30-40小时，再调整发酵条件为pH4-pH4.5、溶解氧10-20％、37度培养6-10小时后将发酵液回调pH至7，并同时匀速流加发酵液体积2％-5％的0.1mol/L氯酸钠，离心、烘干得发酵粉；
S2制备槟榔多糖、牛大力多糖、红芪多糖、绿葡萄藻多糖；
S3将配方量的乳铁蛋白、中链甘油三酯、β葡聚糖、槟榔多糖、牛大力多糖、红芪多糖、绿葡萄藻多糖、枯草芽孢杆菌发酵产物进行混合即得。
上述任一方案中优选的是，发酵培养基组份为每升水中含葡萄糖50g、蛋白胨20g、酵母粉10g、三水合磷酸二氢钾5g、氯化钠1g、硫酸镁2g、硫酸铵1.5g。
上述任一方案中优选的是，步骤S2中牛大力多糖的制备方法为：将牛大力切片，8倍重量水量，加热至90℃，提取4h，过滤，滤渣加4倍量水，提取1h，过滤，合并二次滤液，减压浓缩至原体积10％；浓缩液放至室温，加入乙醇使含醇量为75％，边加边搅拌，4℃静置12小时，离心除去沉淀，上清液经膜过滤得精提液，减压浓缩后，干燥即得。
上述任一方案中优选的是，步骤S2中槟榔多糖的制备方法为：将新鲜槟榔果洗净直接进行物理压榨，经过过滤收集果汁，收集残渣，向残渣加5倍水量80度下提取3小时，过滤得提取液；合并提取液和果汁，减压浓缩至原体积20％，加入乙醇使乙醇含量为80％，边加边搅拌，4℃静置12小时，离心得沉淀，沉淀烘干即得。
上述任一方案中优选的是，步骤S2中红芪多糖的制备方法为：将红芪切片，8倍重量水量，加热至95℃，提取4h，过滤，滤渣加4倍量水，提取1h，过滤，合并二次滤液，减压浓缩至原体积15％；浓缩液放至室温，加入乙醇使含醇量为75％，边加边搅拌，4℃静置12小时，离心除去沉淀，上清液经膜过滤得精提液，减压浓缩后，干燥即得。
上述任一方案中优选的是，步骤S2中绿葡萄藻多糖的制备方法为：将新鲜的绿葡萄藻淡水洗净，破碎后加入5倍重量水量，加热至40度，提取8h,过滤，将滤液减压浓缩至原体积20％；冷却至室温，加入乙醇使含醇量为85％，边加边搅拌，4℃静置12小时，离心得沉淀，沉淀烘干即得。
上述任一方案中优选的是，一次扩大培养：将枯草芽孢杆菌接种至LB培养基中LB培养基中30-37度、200-400rpm条件下培养24h，LB培养基初始pH7，得一级种子液；
二次扩大培养：将一级种子液再次接种至LB培养基中扩大培养，30-37度、200-400rpm条件下培养12h，接种量为LB培养基体积5-10％,得种液。
本发明的有益效果为：本发明提供的一种中药多糖饲料添加剂，主要成分含有乳铁蛋白、中链甘油三酯、β葡聚糖、中药多糖添加剂和枯草芽孢杆菌发酵产物，协同增效，能显著提高猪的机体免疫力，拌食猪饲料用于生猪养殖，内调外疏，能提高生猪的非特异免疫和特异性免疫，从而降低猪瘟、非洲猪瘟、高致病性猪蓝耳病等病毒感染对机体的危害，增加机体抗病能力，达到预防效果。
具体实施方式
下面对本发明实施例中的技术方案进行清楚、完整地描述。在下面的描 述中阐述了很多具体细节以便于充分理解本发明，但是本发明还可以采用其他不同于在此描述的其它方式来实施，本领域技术人员可以在不违背本发明内涵的情况下做类似推广，因此本发明不受下面公开的具体实施例的限制。
本发明提供一种用于预防非洲猪瘟的中药多糖添加剂，其包括如下重量份的各组分：乳铁蛋白5-10份、中链甘油三酯3-5份，β葡聚糖10-15份、槟榔多糖20-40份、牛大力多糖10-30份、红芪多糖10-30份、绿葡萄藻多糖20-50份、枯草芽孢杆菌发酵产物20-40份。
上述中药多糖添加剂的制备方法包括如下步骤：
1.将枯草芽孢杆菌经两次扩大培养得种液，具体为：
一次扩大培养：将枯草芽孢杆菌接种至LB培养基中30-37度、200-400rpm条件下培养24h，LB培养基初始pH7，得一级种子液；
二次扩大培养：将一级种子液再次接种至LB培养基中扩大培养，30-37度、200-400rpm条件下培养12h，接种量为LB培养基体积5-10％,得种液。
2.将种液接种至发酵培养基中于pH6.5-7.5、溶解氧60-70％、37度条件下发酵培养30-40小时，搅拌转速300rpm，再加入1mol/L盐酸调整发酵条件为pH4、溶解氧10-20％、37度培养6-10小时后加入1mol/氢氧化钠将发酵液回调pH至7,搅拌转速500rpm，并同时匀速流加发酵液体积2％-5％的0.1mol/L氯酸钠；
其中，发酵培养基组份为每升水中含葡萄糖50g、蛋白胨20g、酵母粉10g、三水合磷酸二氢钾5g、氯化钠1g、硫酸镁2g、硫酸铵1.5g。
3.将发酵液离心、50度烘干得发酵粉；
4.制备槟榔多糖、牛大力多糖、红芪多糖、绿葡萄藻多糖；
槟榔多糖的制备方法为：将新鲜槟榔果洗净直接进行物理压榨，经过过滤收集果汁，收集残渣，向残渣加5倍水量80度下提取3小时，过滤得提取液；合并提取液和果汁，减压浓缩至原体积20％，加入乙醇使乙醇含量为80％，边加边搅拌，4℃静置12小时，离心得沉淀，沉淀烘干即得。
牛大力多糖的制备方法为：将牛大力切片，8倍重量水量，加热至90℃，提取4h，过滤，滤渣加4倍量水，提取1h，过滤，合并二次滤液，减压浓缩至原体积10％；浓缩液放至室温，加入乙醇使含醇量为75％，边加边搅拌，4℃静置12小时，离心除去沉淀，上清液经膜过滤得精提液，减压浓缩后，干燥即得。
红芪多糖的制备方法为：将红芪切片，8倍重量水量，加热至95℃，提取4h，过滤，滤渣加4倍量水，提取1h，过滤，合并二次滤液，减压浓缩至原体积15％；浓缩液放至室温，加入乙醇使含醇量为75％，边加边搅拌，4℃静置12小时，离心除去沉淀，上清液经膜过滤得精提液，减压浓缩后，干燥即得。
葡萄藻多糖制备方法为：将新鲜的绿葡萄藻淡水洗净，破碎后加入5倍重量水量，加热至40度，提取8h,过滤，将滤液减压浓缩至原体积20％；冷却至室温，加入乙醇使含醇量为85％，边加边搅拌，4℃静置12小时，离心得沉淀，沉淀烘干即得。
5.将配方量的乳铁蛋白、中链甘油三酯、β葡聚糖、槟榔多糖、牛大力多糖、红芪多糖、绿葡萄藻多糖、枯草芽孢杆菌发酵产物进行混合即得。
材料来源说明，枯草芽孢杆菌，北京中科质检生物技术有限公司ID183710，产品编号ZKCCT-020；其余材料均为市售，不再进一步说明。
实施例1
本实施例中的中药多糖添加剂包括如下重量份的各组分：包括如下重量份的各组分：乳铁蛋白7份、中链甘油三酯5份，β葡聚糖11份、槟榔多糖30份、牛大力多糖20份、红芪多糖20份、绿葡萄藻多糖40份、枯草芽孢杆菌发酵产物30份。
上述中药多糖添加剂的制备方法包括如下步骤：
1.将枯草芽孢杆菌经两次扩大培养得种液，具体为：
一次扩大培养：将枯草芽孢杆菌接种至LB培养基中LB培养基中35度、300rpm条件下培养24h，LB培养基初始pH7，得一级种子液；
二次扩大培养：将一级种子液再次接种至LB培养基中扩大培养，35度、300rpm条件下培养12h，接种量为LB培养基体积10％,得种液。
2.将种液接种至发酵培养基中于pH7、溶解氧70％、37度、搅拌转速300rpm条件下发酵培养35小时，再加入1mol/L盐酸调整发酵条件为pH4、溶解氧10％、37度、搅拌转速500rpm培养8小时后加入1mol/氢氧化钠将发酵液回调pH至7，并同时匀速流加发酵液体积4％的0.1mol/L氯酸钠；
其中，发酵培养基组份为每升水中含葡萄糖50g、蛋白胨20g、酵母粉10g、三水合磷酸二氢钾5g、氯化钠1g、硫酸镁2g、硫酸铵1.5g。
3.将发酵液离心、50度烘干得发酵粉；
4.制备槟榔多糖、牛大力多糖、红芪多糖、绿葡萄藻多糖；
将新鲜槟榔果洗净直接进行物理压榨，经过过滤收集果汁，收集残渣，向残渣加5倍水量80度下提取3小时，过滤得提取液；合并提取液和果汁，减压浓缩至原体积20％，加入乙醇使乙醇含量为80％，边加边搅拌，4℃静置 12小时，离心得沉淀，沉淀烘干即得。
将牛大力切片，8倍重量水量，加热至90℃，提取4h，过滤，滤渣加4倍量水，提取1h，过滤，合并二次滤液，减压浓缩至原体积10％；浓缩液放至室温，加入乙醇使含醇量为75％，边加边搅拌，4℃静置12小时，离心除去沉淀，上清液经膜过滤得精提液，减压浓缩后，干燥即得。
将红芪切片，8倍重量水量，加热至95℃，提取4h，过滤，滤渣加4倍量水，提取1h，过滤，合并二次滤液，减压浓缩至原体积15％；浓缩液放至室温，加入乙醇使含醇量为75％，边加边搅拌，4℃静置12小时，离心除去沉淀，上清液经膜过滤得精提液，减压浓缩后，干燥即得。
将新鲜的绿葡萄藻淡水洗净，破碎后加入5倍重量水量，加热至40度，提取8h,过滤，将滤液减压浓缩至原体积20％；冷却至室温，加入乙醇使含醇量为85％，边加边搅拌，4℃静置12小时，离心得沉淀，沉淀烘干即得。S5将配方量的乳铁蛋白、中链甘油三酯、β葡聚糖、绿葡萄藻多糖、枸杞多糖、甘草多糖、枯草芽孢杆菌发酵产物进行混合即得。
实施例2
本实施例的中药多糖添加剂包括如下重量份的各个组份：乳铁蛋白5份、中链甘油三酯5份，β葡聚糖10份、槟榔多糖40份、牛大力多糖30份、红芪多糖30份、绿葡萄藻多糖50份、枯草芽孢杆菌发酵产物20份。
制备方法与实施例一的不同之处在于步骤S2如下：
将种液接种至发酵培养基中于pH6.5、溶解氧60％、37度条件下发酵培养30小时，再调整发酵条件为pH4、溶解氧15％、37度培养10小时后将发酵液回调pH至7，并同时匀速流加发酵液体积2％的0.1mol/L氯酸钠。
实施例3
本实施例的中药多糖添加剂包括如下重量份的各个组份：乳铁蛋白10份、中链甘油三酯3份，β葡聚糖15份、槟榔多糖20份、牛大力多糖10份、红芪多糖10份、绿葡萄藻多糖20份、枯草芽孢杆菌发酵产物40份。
制备方法与实施例一的不同之处在于步骤S2如下：
将种液接种至发酵培养基中于pH7.5、溶解氧70％、37度条件下发酵培40小时，再调整发酵条件为pH4.5、溶解氧20％、37度培养7小时后将发酵液回调pH至7，并同时匀速流加发酵液体积4％的0.1mol/L氯酸钠。
对比例1
本对比例中的中药多糖添加剂按重量份包括如下组份：槟榔多糖30份、牛大力多糖20份、红芪多糖20份、绿葡萄藻多糖40份、枯草芽孢杆菌发酵产物30份。
各个组份的制备方法同实施例一，与实施例一不同之处在于未加入乳铁蛋白、中链甘油三酯和β葡聚糖。
对比例2
本对比例中的中药多糖添加剂各个组份与实施例一相同。
本对比例与实施例一的区别在于枯草芽孢杆菌的发酵条件不同，其具体如下：将种液接种至发酵培养基中于pH7、溶解氧70％、37度、搅拌转速300rpm条件下发酵培养43小时，并于发酵过程最后7小时匀速流加发酵液体积4％的0.1mol/L氯酸钠；期间并未改变溶解氧以及pH值。
对比例3
本对比例的中药多糖添加剂各个组份与实施例一相同。
本对比例与实施例一的区别在于枯草芽孢杆菌的发酵条件不同，具体区别在于发酵后段并未流加氯酸钙。
对比例4
本对比例中多糖种类以及用量如下：牛大力多糖20份、红芪多糖20份、绿葡萄藻多糖40份。
其余组份与实施例一相同。
对比例5
本对比例中多糖种类以及用量如下：槟榔多糖30份、红芪多糖20份、绿葡萄藻多糖40份。
其余组份与实施例一相同。
对比例6
本对比例中多糖种类以及用量如下：槟榔多糖30份、牛大力多糖20份、绿葡萄藻多糖40份。
其余组份与实施例一相同。
对比例7
本对比例中多糖种类以及用量如下：槟榔多糖30份、牛大力多糖20份、红芪多糖20份。
其余组份与实施例一相同。
取上述实施例以及对比例制得的中药多糖添加剂验证对猪瘟(CSFV)的预防效果，包括如下四个验证试验：
验证试验一
试验步骤：
1.将悬浮细胞以1000细胞(100μL)每孔的密度加入96孔板，培养20h。
2.取实施例1至3和对比例1至7制得的多糖中药使用添加30％超纯水的DMEM进行配置，配置浓度2g/100ml，得测试液1-6。
3.将上述测试液分别对96孔板中生长的Vero细胞进行处理，处理浓度80μM，每个测试液4孔重复。
4.用新鲜培养基将病毒稀释到200TCID50/100μL。
5.将50μL猪瘟病毒稀释液加入96孔板中，培养72小时后观察记录每孔CPE程度(％)试验结果如下表所示：将悬浮细胞以1000细胞(100μL)每孔的密度加入96孔板，培养20h，加入80μM、30％超纯水的DMEM，后接种50μL病毒稀释液培养72小时，6孔重复作为对照。
6.记录统计，结果见表1。
验证试验二
1.选取220头阴性成年壮猪，其中200头每20头一组，分别喂食实施例1至3以及对比例1-7制得的中药多糖添加剂，给药组按基础粮的2％重量配给实施例一制得中药多糖添加剂，喂食两周，20头作为对照喂食基础粮。
2.取试验生猪血液样本离心获血清5ml。
3.将含有经过20h培养悬浮细胞(1000细胞/100μL每孔)的96孔板中加入上述上清80μM。
4.用新鲜培养基将猪瘟病毒稀释到200TCID50/100μL。
5.将50μL病毒稀释液加入96孔板中，培养72小时后观察记录每孔CPE程度(％)。
6.试验结果见表1。
表1猪瘟验证试验一、验证试验二
试验一 CPE 试验二 CPE
实施例1 3％ 实施例1 2％
实施例2 6％ 实施例2 4％
实施例3 4％ 实施例3 5％
对比例1 5％ 对比例1 95％
对比例2 95％ 对比例2 91％
对比例3 92％ 对比例3 95％
对比例4 4％ 对比例4 78％
对比例5 7％ 对比例5 85％
对比例6 3％ 对比例6 83％
对比例7 4％ 对比例7 74％
对照 96％ 对照 97％
结合表1能够得出：
1.实施例1-3制得的中药多糖添加剂在验证试验一和二中均具有良好的病毒抑制效果。
2.由对比例1在两个验证试验的结果可以看出，本发明申请中的辅料(乳铁蛋白、中链甘油三酯，β葡聚糖)本身对病毒并无效果，但其在生猪体内能够起到转运有效物质的作用，使得生猪血清能够对病毒具有抑制效果。
3.对比例2-3在验证试验中能够看出，当改变发酵条件以及流程后，本实施例提供的中药多糖添加剂并不具备抑制病毒的效果。
4.由对比例4-7在验证试验一中的测试结果能够看出多糖添加物对病毒抑制效果并无影响，但在验证试验二中能够明显看出，这四种多糖无论缺少哪一种，其效果均明显不如实施例1至3制备的多糖，其可能在于多种多糖的配合作用能内调外疏，提高生猪的非特异免疫和特异性免疫，使生猪代谢循环加快。
验证试验三
将验证试验二中喂食实施例1至3制得的中药多糖添加剂生猪改喂基础 粮，14d、21d、28d后取血清再次重复验证试验二中的测试，结果见表2.
表2添加剂预防时效试验
试验三 14d(CPE) 21d(CPE) 28d(CPE)
实施例1 7％ 6％ 61％
实施例2 3％ 6％ 58％
实施例3 6％ 10％ 72％
由表2数据可知，在停喂后其药效依然能够维持至少21天，因此在日常预防过程中，可以施行每间隔21天进行一次持续14天的混药喂食。
验证试验四
1.选择160头壮猪进行复养试验，地点海南某猪场，此前曾发生猪瘟。
2.对该批生猪进行猪瘟病毒检测，其中试剂盒使用莼试生物的猪瘟病毒/口蹄疫病毒通用型(CSFV/FMDV-U)核酸试剂盒，确保试验前该批生猪均为阴性。
3.选120头作为实验组：给药拌食两周，使用实施例1-3制得中药多糖添加剂按日喂粮重量的2％使用，两周后改喂基础粮。
选40头作为对照组：正常喂食同量基础粮。
4.于试验开始后7d、14d、28d统计生猪状态，并于36d进行病毒检测统计存活率，结果见表3。
表3猪瘟复养试验
Figure PCTCN2021128432-appb-000001
由验证试验4可知，在使用了实施例1至3制得的多糖中药后，相对于对照组，试验组明显获得了对猪瘟病毒的抵抗力。
取上述实施例以及对比例制得的中药多糖添加剂验证对非洲猪瘟(ASFV)的预防效果，进行复养试验，试验步骤与方法猪瘟验证试验四相同，试剂盒使用上海研尊非洲猪瘟病毒核测试剂盒(荧光PCR法)-YZP963652，结果见表4.
表4非洲猪瘟复养试验
Figure PCTCN2021128432-appb-000002
从表4可知，本发明提供的中药多糖添加剂对非洲猪瘟(ASFV)也具有良好的预防效果，对于数据结果分析参照猪瘟(CSFV)部分。
本发明人还实验性的进行了一次对于猪蓝耳病的复养试验：
1.选择70头壮猪进行复养试验，地点海南某猪场，此前曾发生猪蓝耳病。
2.对该批生猪进行猪蓝耳病病毒检测，试剂盒使用上海帛科生物技术有限公司猪蓝耳病病毒经典型(PRRSV-C)核酸试剂盒，确保试验前该批生猪均为阴性。
3.选60头作为实验组：给药拌食两周，使用实施例1-3制得中药多糖添加剂按日喂粮重量的2％使用，两周后改喂基础粮。
选10头作为对照组：正常喂食同量基础粮。
4.于试验开始后5d、10d、15d统计生猪状态，并于15d进行病毒检，结果见表7。
表7
组 症状(5d) 症状(10d) 症状(15d) 病毒检测阳性(15d) 阳性率％
实施例1 1 3 6 6 30％
实施例2 1 3 5 6 30％
实施例3 0 5 8 9 45％
对照 4 9 10 10 100％
由表7可以发现，本发明提供的中药多糖添加剂对于预防猪蓝耳病也具有一定效果，对照组的10头生猪均感染该病毒，而实施例一直三制备的添加剂则感染率明显低于对照组。

Claims (10)

  1. 一种中药多糖饲料添加剂，其特征在于，包括如下重量份的各组分：乳铁蛋白5-10份、中链甘油三酯3-5份，β葡聚糖10-15份、槟榔多糖20-40份、牛大力多糖10-30份、红芪多糖10-30份、绿葡萄藻多糖20-50份、枯草芽孢杆菌发酵产物20-40份。
  2. 根据权利要求1所述的一种中药多糖饲料添加剂，其特征在于，包括如下重量份的各组分：乳铁蛋白7份、中链甘油三酯5份，β葡聚糖11份、槟榔多糖30份、牛大力多糖20份、红芪多糖20份、绿葡萄藻多糖40份、枯草芽孢杆菌发酵产物30份。
  3. 根据权利要求1或2所述中药多糖饲料添加剂的制备方法，其特征在于，包括如下步骤：
    S1将枯草芽孢杆菌经两次扩大培养得种液，将种液接种至发酵培养基中于pH6.5-pH7.5、溶解氧60-70％、37度条件下发酵培养30-40小时，再调整发酵条件为pH4-pH4.5、溶解氧10-20％、37度培养6-10小时后将发酵液回调pH至7，并同时匀速流加发酵液体积2％-5％的0.1mol/L氯酸钠，离心、烘干得发酵粉；
    S2制备槟榔多糖、牛大力多糖、红芪多糖、绿葡萄藻多糖；
    S3将配方量的乳铁蛋白、中链甘油三酯、β葡聚糖、槟榔多糖、牛大力多糖、红芪多糖、绿葡萄藻多糖、枯草芽孢杆菌发酵产物进行混合即得。
  4. 根据权利要求3所述中药多糖饲料添加剂的制备方法，其特征在于，发酵培养基组份为每升水中含葡萄糖50g、蛋白胨20g、酵母粉10g、三水合磷酸二氢钾5g、氯化钠1g、硫酸镁2g、硫酸铵1.5g。
  5. 根据权利要求3所述中药多糖饲料添加剂的制备方法，其特征在于， 步骤S2中槟榔多糖的制备方法为：将新鲜槟榔果洗净直接进行物理压榨，经过过滤收集果汁，收集残渣，向残渣加5倍水量80度下提取3小时，过滤得提取液；合并提取液和果汁，减压浓缩至原体积20％，加入乙醇使乙醇含量为80％，边加边搅拌，4℃静置12小时，离心得沉淀，沉淀烘干即得。
  6. 根据权利要求3所述中药多糖饲料添加剂的制备方法，其特征在于，步骤S2中牛大力多糖的制备方法为：将牛大力切片，8倍重量水量，加热至90℃，提取4h，过滤，滤渣加4倍量水，提取1h，过滤，合并二次滤液，减压浓缩至原体积10％；浓缩液放至室温，加入乙醇使含醇量为75％，边加边搅拌，4℃静置12小时，离心除去沉淀，上清液经膜过滤得精提液，减压浓缩后，干燥即得。
  7. 根据权利要求3所述中药多糖饲料添加剂的制备方法，其特征在于，步骤S2中红芪多糖的制备方法为：将红芪切片，8倍重量水量，加热至95℃，提取4h，过滤，滤渣加4倍量水，提取1h，过滤，合并二次滤液，减压浓缩至原体积15％；浓缩液放至室温，加入乙醇使含醇量为75％，边加边搅拌，4℃静置12小时，离心除去沉淀，上清液经膜过滤得精提液，减压浓缩后，干燥即得。
  8. 根据权利要求3所述中药多糖饲料添加剂的制备方法，其特征在于，步骤S2中绿葡萄藻多糖的制备方法为：将新鲜的绿葡萄藻淡水洗净，破碎后加入5倍重量水量，加热至40度，提取8h,过滤，将滤液减压浓缩至原体积20％；冷却至室温，加入乙醇使含醇量为85％，边加边搅拌，4℃静置12小时，离心得沉淀，沉淀烘干即得。
  9. 根据权利要求3所述中药多糖饲料添加剂的制备方法，其特征在于，
    一次扩大培养：将枯草芽孢杆菌接种至LB培养基中LB培养基中30-37度、200-400rpm条件下培养24h，LB培养基初始pH7，得一级种子液；
    二次扩大培养：将一级种子液再次接种至LB培养基中扩大培养，30-37度、200-400rpm条件下培养12h，接种量为LB培养基体积5-10％,得种液。
  10. 一种权利要求1或2所述的中药多糖饲料添加剂用于预防猪瘟、非洲猪瘟、猪蓝耳病。
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113907208A (zh) * 2021-11-04 2022-01-11 淮阴工学院 一种预防仔猪腹泻的饲料添加剂及其制备方法和应用

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117322536A (zh) * 2023-10-31 2024-01-02 宁夏大洋饲料科技有限公司 一种提高犊牛机体免疫力的预混合饲料及其制备方法

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104911120A (zh) * 2015-03-11 2015-09-16 浙江理工大学 一种枯草芽孢杆菌的发酵生产工艺
CN105168807A (zh) * 2015-10-16 2015-12-23 大连民族大学 治疗猪蓝耳病的中药组合物及其制备方法
CN111150787A (zh) * 2020-01-21 2020-05-15 许启太 一种中兽药预混剂及其制备方法
CN111544447A (zh) * 2020-05-27 2020-08-18 海南今珠农业发展有限公司 一种中药多糖、制备方法及其应用
CN112546166A (zh) * 2020-12-07 2021-03-26 湖南优久农牧有限公司 一种有效防治猪瘟的中药饲料添加剂及其生产方法
CN112691172A (zh) * 2019-10-23 2021-04-23 李大伟 预防非洲猪瘟的一种新型添加剂发酵中药

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103315191B (zh) * 2013-07-18 2014-09-03 广南(湛江)家丰饲料有限公司 一种猪用免疫增强剂及其制备方法
CN103355499B (zh) * 2013-07-29 2014-07-30 广南(湛江)家丰饲料有限公司 改善猪肉品质和提高饲料利用率的饲料添加剂及其制备
CN105985916A (zh) * 2015-01-28 2016-10-05 河南惠通天下动物药业有限公司 一种复合微生物制剂及其制备方法

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104911120A (zh) * 2015-03-11 2015-09-16 浙江理工大学 一种枯草芽孢杆菌的发酵生产工艺
CN105168807A (zh) * 2015-10-16 2015-12-23 大连民族大学 治疗猪蓝耳病的中药组合物及其制备方法
CN112691172A (zh) * 2019-10-23 2021-04-23 李大伟 预防非洲猪瘟的一种新型添加剂发酵中药
CN111150787A (zh) * 2020-01-21 2020-05-15 许启太 一种中兽药预混剂及其制备方法
CN111544447A (zh) * 2020-05-27 2020-08-18 海南今珠农业发展有限公司 一种中药多糖、制备方法及其应用
CN112546166A (zh) * 2020-12-07 2021-03-26 湖南优久农牧有限公司 一种有效防治猪瘟的中药饲料添加剂及其生产方法

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113907208A (zh) * 2021-11-04 2022-01-11 淮阴工学院 一种预防仔猪腹泻的饲料添加剂及其制备方法和应用
CN113907208B (zh) * 2021-11-04 2024-02-23 淮阴工学院 一种预防仔猪腹泻的饲料添加剂及其制备方法和应用

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