WO2021047210A1 - 一种免疫用助悬剂、疫苗悬液制备及喷滴免疫方法 - Google Patents

一种免疫用助悬剂、疫苗悬液制备及喷滴免疫方法 Download PDF

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WO2021047210A1
WO2021047210A1 PCT/CN2020/093093 CN2020093093W WO2021047210A1 WO 2021047210 A1 WO2021047210 A1 WO 2021047210A1 CN 2020093093 W CN2020093093 W CN 2020093093W WO 2021047210 A1 WO2021047210 A1 WO 2021047210A1
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vaccine
parts
immunization
spray
suspension
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PCT/CN2020/093093
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English (en)
French (fr)
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谭志坚
刘丽丹
曾莉
翁亚彪
梁正鹏
唐建红
位晓莉
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佛山市正典生物技术有限公司
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Priority claimed from CN201921490945.0U external-priority patent/CN210496889U/zh
Priority claimed from CN201910848311.6A external-priority patent/CN110448528B/zh
Application filed by 佛山市正典生物技术有限公司 filed Critical 佛山市正典生物技术有限公司
Publication of WO2021047210A1 publication Critical patent/WO2021047210A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/002—Protozoa antigens
    • A61K39/012—Coccidia antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P33/00—Antiparasitic agents
    • A61P33/02—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis

Definitions

  • This application relates to the field of biological immunity technology, for example, it relates to a suspension agent for immunization, preparation of a vaccine suspension, and a spray immunization method.
  • Chicken coccidiosis is a very serious global parasitic disease. It is caused by one or more Eimeria coccidia of the genus Eimeria in the genus Acrocomplexus, parasitizes in the intestinal mucosal epithelium of chickens, causing diarrhea, Bloody stools and growth retardation affect the production and development of chickens and even die, causing huge economic losses to the chicken industry every year.
  • anti-coccidiosis mainly depends on adding anti-coccidiosis drugs to feed or drinking water.
  • Chicken coccidiosis vaccine immunization methods include drinking water, mixing (spraying) materials, dripping, spraying, spraying and other methods.
  • the main popular immunization method is drinking water immunization. After the coccidia vaccine and the suspending agent are evenly mixed, they are divided manually. Installed in the drinking water bottle for the chicks to drink and get immune.
  • the spray immunization is to mix the vaccine and the suspension agent and spray it through the spray device
  • the droplets are attached to chicken feathers for the chickens to peck and be immunized, which is a practical method for chicken coccidiosis vaccine immunity.
  • Reference patent documents 201811552211.0 "A coccidia vaccine spray immunization adjuvant and its application” and 201710676054.3 "Squirt drip or drinking water immunization chicken coccidia vaccine stabilizer and its preparation method and application” both disclose the use of spray immunization
  • the suspending agent adjuvant
  • the effect of spray immunization is not only affected by the uniformity of the suspension of the oocysts in the vaccine suspension.
  • the uniformity of the chick's pecking vaccine suspension has a direct impact on the effect of spray immunity.
  • the purpose of this application is to propose a suspension for immunization, preparation of vaccine suspension and spray immunization method.
  • the suspension for immunization has a good suspension effect and can obtain a vaccine suspension with suitable viscosity.
  • a suspension agent for immunity the raw materials in parts by weight include: 15-40 parts by weight of sodium alginate, 5-20 parts of guar gum, 2-10 parts of blow-dried powder, and surfactant 1- 10 parts, 12-74 parts of anhydrous glucose.
  • the suspending agent for immunity its raw materials in parts by weight further include: 3-8 parts of food coloring.
  • the surfactant is a solid nonionic surfactant.
  • the surfactant is selected from one or more of sucrose ester, sodium dodecylbenzene sulfonate, solid Tween, polyethylene glycol 4000-6000, and Span 60.
  • the method for preparing a vaccine suspension using the above-mentioned immunization suspension agent includes the following steps:
  • the spray-drop immunization method of the vaccine suspension prepared by the above-mentioned method includes the following steps:
  • a spray nozzle is arranged above the conveyor belt. When the container passes the spray nozzle, the spray nozzle sprays the vaccine suspension, and the vaccine suspension drips on the chicks in the container. Attached to chicken feathers;
  • the spray volume of vaccine suspension per 100 chicks is 25-40ml.
  • the diameter of the glue drop ejected by the drip head is 1-3 mm.
  • a plurality of spray nozzles are arranged side by side, and the width of the spray nozzles arranged side by side matches the width of the container;
  • the spraying time of the dripping head matches the length of time that the conveyor belt carries the container through the dripping head.
  • the suspending agent compounded by the various components in this application is easily soluble in water to form a viscous, uniform and stable solution.
  • Mixing the coccidian vaccine diluent with the suspending agent can keep the coccidian oocysts evenly dispersed and suspended.
  • the glue droplets produced can retain moisture and have good adhesion, ensuring that the coccidia oocysts will not lose their vitality and will effectively adhere to the chicken down, so that the chickens can peck and build a uniform immunity.
  • the method for preparing a vaccine suspension of the present application has simple steps and is easy to operate.
  • the vaccine suspension in this viscosity range can effectively adhere to the chicken feathers without slipping, and will not completely wet the chicken feathers, prevent the chicken feathers from getting wet and form a mass, and effectively avoid the chick's stress response and typhoid fever.
  • the dripping immunization by spraying equipment can realize uniform spraying and automatic immunization. By controlling the amount of spraying, the amount of vaccine suspension ingested by the chicks can be controlled.
  • the immunization suspension, preparation method of vaccine suspension and spray-drop immunization method of the present application are applicable to a variety of coccidia vaccines, including Eimeria acervulina, Eimeria tenella, Eimeria poisoning, One type of Eimeria giant, Eimeria slow, Eimeria brucei, Eimeria precocious (monovalent vaccine) or several kinds of coccidia (multivalent vaccine), the above-mentioned coccidia vaccine Both the early-maturing strains and the wild strains are applicable to the technology of this application.
  • a suspension agent for immunity the raw materials in parts by weight include: 15-40 parts of sodium alginate, such as 15 parts, 18 parts, 20 parts, 22 parts, 25 parts, 28 parts, 30 parts, 32 parts, 35 parts, 38 parts, 40 parts, etc.; guar gum 5-20 parts, such as 5 parts, 10 parts, 15 parts, 20 parts, etc.; blow-dried powder 2-10 parts, such as 2 parts, 5 parts, 8 parts, 9 parts, 10 parts, etc.; 1-10 parts of surfactant, such as 1 part, 2 parts, 5 parts, 8 parts, 10 parts, etc.; and 12-74 parts of anhydrous glucose, such as 12 parts, 15 parts, 20 parts , 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 74, etc.
  • 15-40 parts of sodium alginate such as 15 parts, 18 parts, 20 parts, 22 parts, 25 parts, 28 parts, 30 parts, 32 parts, 35 parts, 38 parts, 40 parts, etc.
  • guar gum 5-20 parts such as 5 parts, 10 parts, 15 parts
  • Guar gum is a highly purified polysaccharide extracted from widely cultivated guar beans, with polymannose as the molecular backbone. Guar gum has the functions of thickening, water-retaining, stabilizing and cohesive suspension. Guar gum has good dispersibility and dissolves in water, forming a slightly yellowish translucent liquid with high viscosity. The more guar gum is used, the higher the viscosity of the solution after dissolution, and it is easy to block the needle when spraying droplets through the spraying device. And the swelling degree of a certain amount of guar gum increases as the dissolution time increases, and the viscosity becomes higher until it is stable after a certain period of time.
  • Sodium alginate also known as sodium alginate, kelp gum, alginate, and alginate, is a natural polysaccharide carbohydrate extracted from kelp.
  • Sodium alginate has strong hydrophilicity and can be dissolved in cold water. After dissolution, the solution has high viscosity and high transparency, forming a very viscous and uniform solution.
  • the formed solution has softness, uniformity and stability that are difficult to obtain by other analogs. And other excellent characteristics, it has a strong protective colloid effect.
  • the amount of sodium alginate is too large, it can form a gel, which has poor fluidity and is not suitable for spraying; if the amount is too small, it is difficult to control the uniformity and stability of the aqueous solution compounded with guar gum.
  • Blow-dried powder is a desiccant that prevents sodium alginate, guar gum, surfactants and anhydrous dextrose from absorbing moisture and deteriorating during storage.
  • anhydrous dextrose can be quickly dissolved in water.
  • carbohydrate substance which provides nutritional supplement to chicken body, and can also be used as a dispersion aid to increase the viscosity of the dispersion medium, and is adsorbed on the surface of the suspended particles to increase its hydrophilicity, which is beneficial to reduce the sedimentation speed of the particles. And coalesce to increase the stability of the suspension.
  • Guar gum and sodium alginate are used as the main thickeners and stabilizers. Within the ratio range defined in this application, the viscosity of the prepared vaccine suspension is maintained in the range of 40-100mPa.s, and within the range of 1-5 Stable within hours.
  • Surfactants are used as solubilizers and wetting agents to reduce the surface tension of the glue droplets and facilitate attachment to chicken feathers;
  • Anhydrous dextrose and blow-dried powder are used as auxiliary materials. Anhydrous dextrose can supplement nutrition and improve taste, and blow-dried powder can keep the suspension loose. Does not agglomerate, increases the fluidity of the suspending agent and maintains dry conditions to extend the storage time.
  • the suspending agent compounded by the various components in this application is easily soluble in water to form a viscous, uniform and stable solution.
  • Mixing the coccidian vaccine diluent with the suspending agent can keep the coccidian oocysts evenly dispersed and suspended.
  • the glue droplets produced can retain moisture and have good adhesion, ensuring that the coccidia oocysts will not lose their vitality and will effectively adhere to the chicken down, so that the chickens can peck and build a uniform immunity.
  • the suspending agent for immunization, and its raw materials in parts by weight further include: 3-8 parts of food coloring.
  • Food coloring acts as a coloring agent, which can attract chicks to peck.
  • the food coloring is selected from one of carmine, indigo, brilliant blue, grass green, amaranth, lemon yellow, tartrazine, sunset yellow, and white.
  • the food coloring of each color can produce an equal prompt for the chickens to eat. effect.
  • the surfactant is a solid nonionic surfactant.
  • Non-ionic surfactants have high stability in the solution and are not easily affected by strong electrolytes, inorganic salts, acids and alkalis. They can effectively reduce the surface tension of the liquid, so that the surface tension of the sprayed droplets is reduced, and it is easy to adhere to the chicken feathers and not easy to slip off. , For chickens to peck.
  • the selection of solid surfactants is easy to mix with other powdered components evenly, ensuring that the subpackaged suspending agent components are consistent and the quality is uniform.
  • the surfactant is selected from one or more of sucrose ester, sodium dodecylbenzene sulfonate, solid Tween, polyethylene glycol 4000-6000, and Span 60. These types of surfactants are highly hydrophilic, can also be used as solubilizers, wetting agents, and dispersing/suspending agents, and are easily soluble in water.
  • the method for preparing a vaccine suspension using the above-mentioned immunization suspension agent includes the following steps:
  • the viscosity of the vaccine suspension is 40-100mPa.s, such as 40mPa.s, 50mPa .s, 60mPa.s, 70mPa.s, 80mPa.s, 90mPa.s, 100mPa.s, etc.
  • the method for preparing the vaccine suspension is simple in steps and easy to operate.
  • the vaccine suspension in this viscosity range can effectively adhere to the chicken feathers without slipping, and will not completely wet the chicken feathers, prevent the chicken feathers from getting wet and form a mass, and effectively avoid the chick's stress response and typhoid fever.
  • immunizing suspending agent is added per liter of vaccine diluent, for example, 10g, 11g, 12g, 13g, 14g, or 15g of immunizing suspending agent is added per liter of vaccine diluent.
  • a vaccine suspension with a viscosity in the range of 40-100 mPa ⁇ s can be obtained.
  • the spray-drop immunization method of the vaccine suspension prepared by the above-mentioned method includes the following steps:
  • the spray nozzle is arranged above the conveyor belt.
  • the spray nozzle sprays the vaccine suspension, and the vaccine suspension drips on the chicks in the container.
  • the dripping immunization by spraying equipment can realize uniform spraying and automatic immunization.
  • the amount of vaccine suspension ingested by the chicks can be controlled.
  • the diameter of the glue droplets ejected by the spray nozzle is 1-3 mm, for example, 1 mm, 1.2 mm, 1.5 mm, 1.8 mm, 2 mm, 2.2 mm, 2.5 mm, 2.8 mm, 3 mm, and so on.
  • the glue droplet can be well attached to the chicken feathers, so that the chicks can effectively peck and prevent the feathers from being wetted in a large area.
  • a plurality of spray nozzles are arranged side by side, and the width of the spray nozzles arranged side by side matches the width of the container; the spraying time of the spray nozzles matches the length of the conveyor belt carrying the container through the spraying head .
  • the chicks are placed in a cuboid container with an opening at the top of the container.
  • the vaccine suspension can be evenly spread in the container, so that there are glue drops on each chick, which is more conducive to the uniform pecking of the vaccine by the chicks. immunity.
  • Examples 1-5 are suspensions for immunization, methods for preparing vaccine suspensions, and immunization methods.
  • the method for preparing a vaccine suspension of Examples 1-5 includes the following steps:
  • Example 1 Example 2
  • Example 3 Example 4
  • Example 5 Vaccine diluent 8L 8L 8L 8L 8L Immunization Suspending Agent 80g 90g 110g 100g 120g
  • the spray-drop immunization method of embodiment 1-5 includes the following steps:
  • a spray nozzle is arranged above the conveyor belt. When the container passes the spray nozzle, the spray nozzle sprays the vaccine suspension, and the vaccine suspension drips on the chicks in the container. Attached to chicken feathers.
  • the spray volume of vaccine suspension per 100 chicks is 25-40ml.
  • the diameter of the glue drop ejected by the drip head is 1-3mm.
  • a plurality of spray nozzles are arranged side by side, and the width of the spray nozzles arranged side by side matches the width of the container; the spraying time of the spray nozzles matches the length of the conveyor belt carrying the container through the spraying head.
  • the suspending agent of this comparative example is composed of 15 g of xanthan gum, 2 g of sodium alginate, 3 g of sodium polyacrylate, a water-stable compound, 0.2 g of lemon yellow, 0.2 g of neotame, 2 g of ginseng leaf extract, and 77.6 g of soluble starch. After mixing the ingredients of the suspension formulation uniformly in proportions, slowly add 1L of tap water and stir evenly until the liquid has a uniform color, and add the chicken coccidia vaccine to prepare a vaccine suspension.
  • the spray-drop immunization method of Comparative Example 3 is the same as that of Example 3.
  • the immunization suspension and the method for preparing the vaccine suspension of this comparative example are basically the same as those in Example 3. The difference is that in the immunization method, the vaccine suspension is sprayed with a drip-tip, and the chickens are manually pushed The container passes through the spray nozzle, and the spray drop volume of the vaccine suspension per 100 chicks is 30 ml, and the glue drop diameter sprayed by the spray nozzle is 3 mm.
  • the immunization suspension, the method for preparing vaccine suspension, and the spray-drop immunization method of this comparative example are basically the same as those in Example 3. The difference is that, in the spray-drop immunization method, the droplet ejected by the spray-drop head has a larger diameter. , Showing a connection trend.
  • Example 1-5 and Comparative Examples 1-5 were applied.
  • the suspending agent was sprayed on 1-day-old chicks, 100 chicks in each group. Thirty minutes after spraying, randomly select 20 chickens from each group for necropsy to observe whether the mouth, esophagus, crop, and glandular stomach of each chicken have the color of food coloring. If there is, it proves that the chickens have eaten the coccidia vaccine droplets. .
  • Table 1 The test results of each group of chicks are shown in Table 1.
  • the vaccine suspensions of Examples 1-5 can be sprayed steadily during the spraying process, and the sprayed droplets are of uniform size, and are more effectively attached to chicken feathers and are pecked off by chickens and are not easy to wet the hairs.
  • the results of the necropsy in Table 1 show that 20 chickens randomly selected from the groups of Examples 1 to 5 have the color of food coloring observed in the mouth and esophagus, indicating that each chicken has pecked vaccine droplets. Tests have proved that the vaccine droplets sprayed with the suspending agent of the application can effectively attach to chicken feathers and have a suitable size, which is more conducive to the feeding of the chicks, thereby improving the immune effect.
  • Comparative Example 1 had fewer droplets attached to the feathers during the spraying process, and some of them slipped from the chicks, and the feathers that fell on the chicks were more wet. In the later observation, 3 chickens suffered from breathing difficulties. , Diarrhea and other symptoms. The effective attachment amount of the vaccine suspensions in Comparative Example 2 and Comparative Example 3 on chicken feathers is worse than that of the present application. Comparative Example 4 was immunized by spraying manually. The vaccine suspension was not evenly sprayed and it was difficult to immunize uniformly. In Comparative Example 5, the gel droplets for spraying immunization were too large, and it was difficult to spray evenly on each chick, and it could not be uniformly immunized. And some chicks attach too much glue to cause stress response.

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Abstract

提供了一种免疫用助悬剂，其以重量份计的原料包括：海藻酸钠15-40份、瓜尔胶5-20份、吹干粉2-10份、表面活性剂1-10份、无水葡萄糖12-74份。相应的，还提供了采用上述免疫用助悬剂制备疫苗悬液的方法，以及喷滴免疫方法。所述各组分复配而成的助悬剂易溶于水形成粘稠、均一、稳定的溶液，将球虫疫苗稀释液与助悬剂混合能保持球虫卵囊均匀分散悬浮，喷出的胶滴能保持水分且附着性良好，保证球虫卵囊活力不丧失且有效粘附在鸡绒毛上，供鸡啄食而建立均匀免疫力。制备疫苗悬液的方法步骤简单、易操作。通过喷滴设备进行喷滴免疫，能实现均匀喷滴和自动化免疫，通过控制喷滴量控制雏鸡摄入疫苗悬液的量。

Description

一种免疫用助悬剂、疫苗悬液制备及喷滴免疫方法 技术领域
本申请涉及生物免疫技术领域，例如涉及一种免疫用助悬剂、疫苗悬液制备及喷滴免疫方法。
背景技术
鸡球虫病是一种危害极为严重的全球性寄生虫病，由顶复门孢子纲艾美耳属的一种或多种艾美耳球虫寄生于鸡肠道黏膜上皮内，引起腹泻、血便、生长迟缓等影响鸡只生产发育甚至死亡，每年都给养鸡业造成巨大经济损失。目前，抗球虫病主要依靠在饲料或饮水中添加抗球虫药物。但由于球虫耐药性问题愈来愈严重，且上市的球虫疫苗安全性和有效性已逐步获得市场认可，使用球虫疫苗预防鸡球虫病成为新趋势。鸡球虫疫苗免疫方法包括饮水、拌(喷)料、滴口、喷雾、喷滴等方法，目前主要推广使用的免疫方法为饮水免疫，将球虫疫苗与助悬剂混合均匀后，人工分装在饮水壶内供雏鸡饮用而得以免疫。饮水免疫虽然较拌料、滴口免疫等方法操作简便，但是其应用范围存在一定的局限性，特别是大规模集约化养鸡场由于饲养员数量配比少，不适宜用工作量较大的饮水免疫。孵化场和大规模集约化养鸡场适宜使用劳动强度小的喷雾、喷滴等免疫方法，传统喷雾免疫喷出的雾滴过小，即便附着在羽毛上也容易蒸发且部分雾滴悬浮在空气中造成浪费，影响雏鸡啄食摄入充足的球虫疫苗，因此传统的喷雾免疫方法不适于鸡球虫病疫苗的免疫，喷滴免疫是将疫苗与助悬剂混合后经喷滴设备喷出液滴附着在鸡毛上供鸡啄食而得以免疫，是一种切实可行的鸡球虫疫苗免疫方法。参考专利文献201811552211.0《一种球虫疫苗喷滴免疫用佐剂及其应用》和201710676054.3《喷滴或饮水免疫用鸡球虫疫苗稳定剂 及其制备方法和应用》均公开了用于喷滴免疫的助悬剂(佐剂)，该助悬剂(佐剂)可使得鸡球虫疫苗卵囊均匀悬浮，然而喷滴免疫的效果不仅受疫苗悬液中卵囊悬浮均匀程度的影响，每只雏鸡啄食疫苗悬液量的均匀问题更直接影响喷滴免疫效果。若喷出的疫苗悬液胶滴无法有效附着在鸡毛上大量滑落则部分雏鸡可能无法啄食摄入充足的疫苗卵囊，或者大量的胶滴附着在鸡毛上即使被鸡啄食后鸡毛易湿成团，容易引起雏鸡寒冷、应激等副反应，这些问题均影响鸡球虫疫苗免疫效果，有待进一步解决。
发明内容
本申请的目的在于提出一种免疫用助悬剂、疫苗悬液制备及喷滴免疫方法，该免疫用助悬剂具有较好的悬浮效果，能获得粘度适宜的疫苗悬液。
为达此目的，本申请采用以下技术方案：
第一方面，一种免疫用助悬剂，其以重量份计的原料包括：海藻酸钠15-40份、瓜尔胶5-20份、吹干粉2-10份、表面活性剂1-10份、无水葡萄糖12-74份。
可选地，免疫用助悬剂，其以重量份计的原料还包括：食用色素3-8份。
可选地，表面活性剂为固体非离子表面活性剂。
可选地，表面活性剂选自蔗糖酯、十二烷基苯磺酸钠、固体吐温、聚乙二醇4000-6000、司盘60中一种或多种。
第二方面，采用上述的免疫用助悬剂制备疫苗悬液的方法，包括以下步骤：
(1)、将用于喷滴免疫的疫苗加水稀释，制成疫苗稀释液；
(2)、将免疫用助悬剂加入不断搅拌的疫苗稀释液中，搅拌至完全溶解，混合均匀，获得疫苗悬液；疫苗悬液的粘度为40-100mPa.s。
可选地，每升疫苗稀释液加入10-15g免疫用助悬剂。
第三方面，上述方法制成的疫苗悬液的喷滴免疫方法，包括以下步骤:
将盛有雏鸡的容器放置在输送带上，输送带上方设置有喷滴头，当容器经过喷滴头时，喷滴头喷出疫苗悬液，疫苗悬液滴落在容器中的雏鸡上并附着在鸡毛上；
每100只雏鸡的疫苗悬液喷滴量为25-40ml。
可选地，喷滴头喷出的胶滴直径为1-3mm。
可选地，喷滴头并排设置有多个，并排设置的喷滴头宽度与容器的宽度相匹配；
喷滴头的喷滴时长与输送带携带容器经过喷滴头的时长相匹配。
本申请的有益效果包括：
本申请中各组分复配而成的助悬剂易溶于水形成粘稠、均一、稳定的溶液，将球虫疫苗稀释液与助悬剂混合能保持球虫卵囊均匀分散悬浮，喷出的胶滴能保持水分且附着性良好，保证球虫卵囊活力不丧失且有效粘附在鸡绒毛上，供鸡啄食而建立均匀免疫力。
本申请的制备疫苗悬液的方法步骤简单、易操作。该粘度范围的疫苗悬液能有效附着在鸡羽毛上不滑落，也不会全部打湿鸡羽毛，防止鸡毛湿成团，有效避免雏鸡应激反应和伤寒。通过喷滴设备进行喷滴免疫，能实现均匀喷滴和自动化免疫，通过控制喷滴量控制雏鸡摄入疫苗悬液的量。
本申请的免疫用助悬剂、制备疫苗悬液方法和喷滴免疫方法适用于多种球虫疫苗，包括堆型艾美耳球虫、柔嫩艾美耳球虫、毒害艾美耳球虫、巨型艾美耳球虫、和缓艾美耳球虫、布氏艾美耳球虫、早熟艾美耳球虫的一种(单价苗) 或几种球虫(多价苗)，上述球虫疫苗的早熟株和野生株均适用本申请的技术。
具体实施方式
下面结合附具体实施方式进一步说明本申请的技术方案。
为达此目的，本申请采用以下技术方案：
一种免疫用助悬剂，其以重量份计的原料包括：海藻酸钠15-40份，例如15份、18份、20份、22份、25份、28份、30份、32份、35份、38份、40份等；瓜尔胶5-20份，例如5份、10份、15份、20份等；吹干粉2-10份，例如2份、5份、8份、9份、10份等；表面活性剂1-10份，例如1份、2份、5份、8份、10份等；和无水葡萄糖12-74份，例如12份、15份、20份、25份、30份、35份、40份、45份、50份、55份、60份、65份、70份、74份等。
瓜尔胶为从广泛种植的瓜尔豆中提取的一种高纯化多糖，以聚甘露糖为分子主链。瓜尔胶具有增稠、保水、稳定及粘结悬浮的作用。瓜尔胶分散性好，遇水即溶，形成略发黄的半透明高粘稠度液体。瓜尔胶使用量越多，溶解后溶液粘稠度越高，经过喷滴设备喷出液滴时易堵塞针头。且一定量的瓜尔胶随着溶解时间延长溶胀程度越高，黏度随之变高直至一定时间后才稳定。
海藻酸钠又名褐藻酸钠、海带胶、褐藻胶、藻酸盐，是由海带中提取的天然多糖碳水化合物。海藻酸钠亲水性强，在冷水中能溶解，溶解后溶液粘度大，透明度高，形成非常粘稠的均匀的溶液，形成的溶液具有其他类似物难于获得的柔软性、均一性、稳定性及其他优良特性，具有很强的保护胶体的作用。海藻酸钠用量过大可形成凝胶体，流动性差，不适合喷滴；用量过小，与瓜尔胶复配而成的水溶液均一性和稳定性较难以控制。
吹干粉是一种干燥剂，防止海藻酸钠、瓜尔胶、表面活性剂和无水葡萄糖 在保存期间吸潮变质。无水葡萄糖作为一种甜味剂，可快速溶于水中。它也是一种糖类物质，给予鸡体营养补充，也可作为分散助剂，增加分散媒介的粘度，并被吸附于混悬微粒的表面增加其亲水性，有利于减小微粒的沉降速度和聚结，增加混悬剂的稳定性。
瓜尔胶和海藻酸钠作为主要的增稠剂、稳定剂，在本申请限定的配比范围内，使得制成的疫苗悬液黏度维持在40-100mPa.s范围内，且在1-5小时内即趋于稳定。表面活性剂作为增溶剂、湿润剂，使得胶滴表面张力下降利于附着在鸡毛上；无水葡萄糖和吹干粉作为辅料，无水葡萄糖能补充营养改善口味，吹干粉能保持助悬剂松散不结块，增加助悬剂的流动性以及维持干燥条件延长保存时间。
本申请中各组分复配而成的助悬剂易溶于水形成粘稠、均一、稳定的溶液，将球虫疫苗稀释液与助悬剂混合能保持球虫卵囊均匀分散悬浮，喷出的胶滴能保持水分且附着性良好，保证球虫卵囊活力不丧失且有效粘附在鸡绒毛上，供鸡啄食而建立均匀免疫力。
作为可选的实施方案，免疫用助悬剂，其以重量份计的原料还包括：食用色素3-8份。食用色素作为一种显色剂，能吸引雏鸡啄食。食用色素选取自胭脂红、靛蓝、亮蓝、青草绿、苋菜红、柠檬黄、酒石黄、日落黄、白色中的一种，各颜色的食用色素均能对雏鸡采食产生均等的提示效果。
作为可选的实施方案，表面活性剂为固体非离子表面活性剂。非离子表面活性剂在溶液中稳定性高，不易受强电解质无机盐和酸、碱的影响，可有效降低液体表面张力，使得喷滴的液滴表面张力下降易于附着在鸡绒毛上，不易滑落，供鸡啄食。选用固体类的表面活性剂易于与其他粉末状组分混合均匀，保 证分装的助悬剂组分一致，质量均匀。
作为可选的实施方案，表面活性剂选自蔗糖酯、十二烷基苯磺酸钠、固体吐温、聚乙二醇4000-6000、司盘60中一种或多种。这些种类的表面活性剂亲水性强，也可作为增溶剂、湿润剂、分散/助悬剂，易溶于水。
采用上述的免疫用助悬剂制备疫苗悬液的方法，包括以下步骤：
(1)、将用于喷滴免疫的疫苗加水稀释，制成疫苗稀释液；
(2)、将免疫用助悬剂加入不断搅拌的疫苗稀释液中，搅拌至完全溶解，混合均匀，获得疫苗悬液；疫苗悬液的粘度为40-100mPa.s，例如40mPa.s、50mPa.s、60mPa.s、70mPa.s、80mPa.s、90mPa.s、100mPa.s等。
上述制备疫苗悬液的方法步骤简单、易操作。该粘度范围的疫苗悬液能有效附着在鸡羽毛上不滑落，也不会全部打湿鸡羽毛，防止鸡毛湿成团，有效避免雏鸡应激反应和伤寒。
作为可选的实施方案，每升疫苗稀释液加入10-15g免疫用助悬剂，例如每升疫苗稀释液加入10g、11g、12g、13g、14g或15g等的免疫用助悬剂。通过限定助悬剂的加入量，能获得粘度范围在40-100mPa.s的疫苗悬液。
上述方法制成的疫苗悬液的喷滴免疫方法，包括以下步骤:
将盛有雏鸡的容器放置在输送带上，输送带上方设置有喷滴头，当容器经过喷滴头时，喷滴头喷出疫苗悬液，疫苗悬液滴落在容器中的雏鸡上并附着在鸡毛上；每100只雏鸡的疫苗悬液喷滴量为25-40ml，例如每100只雏鸡的疫苗悬液喷滴量为25ml、28ml、30ml、32ml、35ml、38ml、40ml等。
通过喷滴设备进行喷滴免疫，能实现均匀喷滴和自动化免疫，通过控制喷滴量控制雏鸡摄入疫苗悬液的量。
作为可选的实施方案，喷滴头喷出的胶滴直径为1-3mm，例如1mm、1.2mm、1.5mm、1.8mm、2mm、2.2mm、2.5mm、2.8mm、3mm等。通过限定胶滴的直径，使胶滴能很好的附着在鸡羽毛上，使得雏鸡能有效啄食，并避免大面积打湿羽毛。
作为可选的实施方案，喷滴头并排设置有多个，并排设置的喷滴头宽度与容器的宽度相匹配；喷滴头的喷滴时长与输送带携带容器经过喷滴头的时长相匹配。一般情况下，雏鸡盛放在长方体的容器内，容器上方开口。通过限定喷滴头的宽度和针对每个容器的喷滴时间，使得疫苗悬液能均匀铺洒在容器中，使每个雏鸡身上都有胶滴，更有利于雏鸡均匀啄食疫苗，实现均匀免疫。
以下通过实施例和对比例进一步阐述本申请。
实施例1-5为免疫用助悬剂、制备疫苗悬液的方法及免疫方法。
实施例1-5的免疫用助悬剂的原料如下表所示。
Figure PCTCN2020093093-appb-000001
实施例1-5的制备疫苗悬液的方法包括以下步骤：
(1)、将用于喷滴免疫的疫苗加水稀释，制成疫苗稀释液；
(2)、将免疫用助悬剂加入不断搅拌的疫苗稀释液中，搅拌至完全溶解， 混合均匀，获得疫苗悬液；疫苗悬液的粘度为40-100mPa.s，每升疫苗稀释液加入10-15g免疫用助悬剂。
实施例1-5中免疫用助悬剂的加入量如下表所示。
配料 实施例1 实施例2 实施例3 实施例4 实施例5
疫苗稀释液 8L 8L 8L 8L 8L
免疫用助悬剂 80g 90g 110g 100g 120g
实施例1-5的喷滴免疫方法，包括以下步骤:
将盛有雏鸡的容器放置在输送带上，输送带上方设置有喷滴头，当容器经过喷滴头时，喷滴头喷出疫苗悬液，疫苗悬液滴落在容器中的雏鸡上并附着在鸡毛上。每100只雏鸡的疫苗悬液喷滴量为25-40ml。喷滴头喷出的胶滴直径为1-3mm。
喷滴头并排设置有多个，并排设置的喷滴头宽度与容器的宽度相匹配；喷滴头的喷滴时长与输送带携带容器经过喷滴头的时长相匹配。
实施例1-5的喷滴免疫方法中疫苗悬液喷滴量和胶滴直径如下表所示。
Figure PCTCN2020093093-appb-000002
对比例1
称取3g可食用色素溶解于已加入鸡球虫病活疫苗的8L水中，色素仅用作显色，制成疫苗悬液。对比例1的喷滴免疫方法与实施例3相同。
对比例2
取瓜尔胶2.5g，壳寡糖0.5g，吹干粉0.375g，食用色素0.375g，无水葡萄糖21.25g，混合均匀后投入已装有500mL球虫疫苗稀释液的烧杯中，搅拌均 匀，得具有黏性的溶液为疫苗悬液。对比例2的喷滴免疫方法与实施例3相同。
对比例3
本对比例的助悬剂由黄原胶15g、海藻酸钠2g、水稳定化合物聚丙烯酸钠3g、柠檬黄0.2g、纽甜0.2g，人参叶提取物2g，可溶性淀粉77.6g组成。将所助悬剂配方成分按比例混合均匀后，缓慢加入1L自来水中，搅拌均匀，至液体呈均一颜色，加入鸡球虫疫苗，制成疫苗悬液。对比例3的喷滴免疫方法与实施例3相同。
对比例4
本对比例的免疫用助悬剂和制备疫苗悬液的方法与实施例3基本相同，不同之处在于，免疫方法中，将疫苗悬液采用喷滴头进行喷滴，人工推动盛有雏鸡的容器经过喷滴头，每100只雏鸡的疫苗悬液喷滴量为30ml，喷滴头喷出的胶滴直径为3mm。
对比例5
本对比例的免疫用助悬剂、制备疫苗悬液的方法和喷滴免疫方法与实施例3基本相同，不同之处在于，喷滴免疫方法中，喷滴头喷出的胶滴直径较大，呈连线趋势。
将实施例1-5和对比例1-5的方法进行应用。将助悬剂分别喷于1日龄雏鸡，每组100只雏鸡。喷滴后30分钟每组随机挑选20羽鸡剖检，观察每只鸡的口腔、食道、嗉囊、腺胃是否有食用色素的颜色，若有，则证明雏鸡有食用到球虫疫苗液滴。每组雏鸡的检测结果如表1所示。
表1
Figure PCTCN2020093093-appb-000003
Figure PCTCN2020093093-appb-000004
实施例1-5的疫苗悬液在喷滴过程中均可稳定喷滴，喷出的液滴大小均匀，较有效地附着在鸡毛上被鸡啄食掉而不易湿毛。表1中的剖检结果表明，实施例1～5的组别随机挑选的20羽鸡在口腔、食道均有观察到食用色素的颜色，说明每羽鸡均有啄食到疫苗液滴。试验证明，使用本申请的助悬剂进行喷滴后的疫苗液滴能有效附着在鸡毛上，大小适宜，更利于雏鸡的采食，进而提高免疫效果。
对比例1的疫苗悬液在喷滴过程鸡毛上附着的液滴较少，部分从雏鸡身上滑落，落于雏鸡身上的则羽毛湿得较多，在后期观察中，3羽鸡出现呼吸不畅、拉稀等症状。对比例2和对比例3中的疫苗悬液在鸡毛的有效附着量差于本申请。对比例4以人工进行喷滴免疫，疫苗悬液喷滴不均匀，难以均匀免疫，对比例5中，喷滴免疫的胶滴过大，难以均匀喷滴在每个雏鸡上，不能均匀免疫，且有些雏鸡附着的胶滴过多造成应激反应。
以上结合具体实施例描述了本申请的技术原理。这些描述只是为了解释本申请的原理，而不能以任何方式解释为对本申请保护范围的限制。基于此处的解释，本领域的技术人员不需要付出创造性的劳动即可联想到本申请的其它具 体实施方式，这些方式都将落入本申请的保护范围之内。

Claims (7)

  1. 一种免疫用助悬剂，以重量份计的原料包括：海藻酸钠15-40份、瓜尔胶5-20份、吹干粉2-10份、表面活性剂1-10份、无水葡萄糖12-74份。
  2. 根据权利要求1所述的免疫用助悬剂，其中，其以重量份计的原料还包括：食用色素3-8份。
  3. 根据权利要求1所述的免疫用助悬剂，其中，所述表面活性剂为固体非离子表面活性剂。
  4. 根据权利要求1所述的免疫用助悬剂，其中，所述表面活性剂选自蔗糖酯、十二烷基苯磺酸钠、固体吐温、聚乙二醇4000-6000、司盘60中一种或多种。
  5. 一种采用权利要求1-4任一项所述的免疫用助悬剂制备疫苗悬液的方法，包括以下步骤：
    (1)、将用于喷滴免疫的疫苗加水稀释，制成疫苗稀释液；
    (2)、将所述免疫用助悬剂加入不断搅拌的所述疫苗稀释液中，其中每升所述疫苗稀释液加入10-15g所述免疫用助悬剂，搅拌至完全溶解，混合均匀，获得疫苗悬液；所述疫苗悬液的粘度为40-100mPa.s。
  6. 一种由权利要求5所述方法制成的疫苗悬液的喷滴免疫方法，包括以下步骤:
    将盛有雏鸡的容器放置在输送带上，输送带上方设置有喷滴头，当所述容器经过喷滴头时，喷滴头喷出疫苗悬液，疫苗悬液滴落在容器中的雏鸡上并附着在鸡毛上；
    其中，每100只雏鸡的疫苗悬液喷滴量为25-40ml，且所述喷滴头喷出的胶滴直径为1-3mm。
  7. 权利要求6所述的方法，其中，所述喷滴头并排设置有多个，并排设置的喷滴头宽度与容器的宽度相匹配；
    且所述喷滴头的喷滴时长与所述输送带携带容器经过所述喷滴头的时长相匹配。
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Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0682874A2 (en) * 1994-05-19 1995-11-22 Ewos Aktiebolag Bioactive feed
US20080260837A1 (en) * 2007-04-20 2008-10-23 Qpharma, L.L.C. Physically stable aqueous suspensions of active pharmaceuticals
CN102512417A (zh) * 2011-12-16 2012-06-27 南京农业大学 一种防治家畜子宫内膜炎用复方利福昔明干混悬剂及其制备方法
CN107469074A (zh) * 2017-08-18 2017-12-15 中农威特生物科技股份有限公司 一种鸡球虫病活卵囊疫苗助悬剂
CN107625959A (zh) * 2017-08-09 2018-01-26 程金水 喷滴或饮水免疫用鸡球虫疫苗稳定剂及其制备方法和应用
CN109432006A (zh) * 2018-12-18 2019-03-08 佛山市正典生物技术有限公司 一种球虫疫苗喷滴免疫用佐剂及其应用
CN110448528A (zh) * 2019-09-09 2019-11-15 佛山市正典生物技术有限公司 一种免疫用助悬剂、疫苗悬液制备及喷滴免疫方法

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0682874A2 (en) * 1994-05-19 1995-11-22 Ewos Aktiebolag Bioactive feed
US20080260837A1 (en) * 2007-04-20 2008-10-23 Qpharma, L.L.C. Physically stable aqueous suspensions of active pharmaceuticals
CN102512417A (zh) * 2011-12-16 2012-06-27 南京农业大学 一种防治家畜子宫内膜炎用复方利福昔明干混悬剂及其制备方法
CN107625959A (zh) * 2017-08-09 2018-01-26 程金水 喷滴或饮水免疫用鸡球虫疫苗稳定剂及其制备方法和应用
CN107469074A (zh) * 2017-08-18 2017-12-15 中农威特生物科技股份有限公司 一种鸡球虫病活卵囊疫苗助悬剂
CN109432006A (zh) * 2018-12-18 2019-03-08 佛山市正典生物技术有限公司 一种球虫疫苗喷滴免疫用佐剂及其应用
CN110448528A (zh) * 2019-09-09 2019-11-15 佛山市正典生物技术有限公司 一种免疫用助悬剂、疫苗悬液制备及喷滴免疫方法

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