CN104161182B - 一种抗鹅病毒性疾病的饲料添加剂、制备方法及应用 - Google Patents
一种抗鹅病毒性疾病的饲料添加剂、制备方法及应用 Download PDFInfo
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Abstract
本发明公开了一种抗鹅病毒性疾病的饲料添加剂、制备方法及应用,所述饲料添加剂包括以下重量份数的组分:抗鹅病毒性疾病卵黄抗体冻干粉2?5份、黄芪多糖5?10份、当归多糖5?10份、枸杞多糖10?20份、甘草多糖10?30份。本发明的抗鹅病毒性疾病饲料添加剂为抗鹅病毒性疾病卵黄抗体冻干粉和由黄芪多糖、当归多糖、枸杞多糖、甘草多糖组成的复合多糖复配而成,添加在日常饲料中,可以提高机体免疫力和抗病能力。实验结果表明,添加本发明的饲料添加剂后,预防鹅病毒性疾病的效果显著提高,明显优于单独添加卵黄抗体冻干粉或复合多糖的效果。
Description
技术领域
本发明涉及一种抗鹅病毒性疾病的饲料添加剂,同时还涉及该饲料添加剂的制备方法及应用,属于饲料领域。
背景技术
随着社会的发展和人民生活水平的提高,越来越多人将目光投向食品安全问题。我国是鹅的生产和消费大国,养鹅产业规模日益扩大,但是由于饲养条件的限制,鹅疾病的传播较为普遍。鹅病毒性疾病是鹅疾病中较为常见的疾病,常见的鹅病毒性疾病有小鹅瘟等。
目前常见的防治鹅病毒性疾病的方法较多,主要是使用西药、接种免疫、中草药防治。其中西药使用的泛滥给鹅产品食用带来了极大的饮食隐患,药物残留已经成为影响鹅产品安全的主要因素之一。接种免疫早期效果较好,但是需要多次接种,操作麻烦,成本较高,容易造成雏鹅产生应激反应;而且疫苗对于保藏条件要求较高,需要在2-8℃条件下冷藏,室温条件下一段时间则失去效果。中草药饲料添加剂也是目前的研究方向之一,但是由于中草药的种类较多,相同的中药材对不同的禽畜疾病的作用差异很大,且中草药对疾病的免疫效果极其有限,因此其应用也具有一定的局限性。注射卵黄抗体也是较为常见的一种方法,但是单一注射卵黄抗体的免疫效果有时候不能满足实际需求。
发明内容
本发明的目的是提供一种抗鹅病毒性疾病的饲料添加剂,可以有效预防小鹅瘟等病毒性疾病。
为了实现以上目的,本发明所采用的技术方案是提供一种抗鹅病毒性疾病的饲料添加剂,所述饲料添加剂包括以下重量份数的组分:抗鹅病毒性疾病卵黄抗体冻干粉2-5份、黄芪多糖5-10份、当归多糖5-10份、枸杞多糖10-20份、甘草多糖10-30份。
所述抗鹅病毒性疾病卵黄抗体冻干粉包括以下重量份数的组分:抗鹅病毒性疾病卵黄抗体93-96份、防腐剂0.01-0.02份、灭活剂0.2-0.3份、维生素C0.5-1份、聚乙烯吡咯烷酮0.5-1份、甘氨酸0.5-1份、山梨醇1-2份、葡萄糖1-2份。
所述灭活剂为甲醛。
所述防腐剂为硫柳汞。
本发明将抗鹅病毒性疾病卵黄抗体和多种辅料进行复配,并通过调整辅料的成分、控制其用量,大大延长了卵黄抗体的保存时间,保证了卵黄抗体在较长一段时间内的活性。
本发明的目的还在于提供一种抗鹅病毒性疾病的饲料添加剂的制备方法。
本发明所采用的技术方案还在于提供一种抗鹅病毒性疾病的饲料添加剂的制备方法,包括以下步骤:
1)将鹅病毒性疾病病毒增殖并灭活制备成疫苗,免疫健康鸡群制得高免蛋,将高免蛋经过消毒、酸化处理、离心处理、配制、冻干得到抗鹅病毒性疾病卵黄抗体冻干粉;
2)将抗鹅病毒性疾病卵黄抗体冻干粉与其余组分混合即得。
所述抗鹅病毒性疾病的饲料添加剂的制备方法,具体包括以下步骤:
1)将鹅病毒性疾病病毒增殖并灭活制备成疫苗,每间隔7天对健康产蛋鸡群进行免疫,共免疫三次,第二次免疫后开始检测抗体效价,当抗体效价达到1:256时,收集高免蛋,将收集的高免蛋进行清理、消毒,晾干备用;
2)分离高免蛋卵黄,加入等体积的无菌水,充分搅拌,得到卵黄抗体溶液;将卵黄抗体溶液放入60-65℃水浴锅内均匀加热30min,然后加入pH=4.9-5.2、温度为4-6℃的酸化水溶液,酸化水溶液加入量为卵黄抗体溶液体积的6倍,混匀,静置沉淀,取上清液,15000rpm离心20min,得到含有卵黄抗体的上清液;
3)用0.45μm的滤膜对步骤2)所得上清液过滤,截流分子量大于200KD的大分子物质,再将滤液经0.22μm滤膜过滤,留滤液;
4)将滤液用超滤装置浓缩,使卵黄抗体溶液浓缩至2.5mL/500羽份;
5)向步骤4)所得卵黄抗体溶液中加入灭活剂、防腐剂、维生素C、聚乙烯吡咯烷酮、甘氨酸、山梨醇、葡萄糖,并在﹣80℃条件下充分冷冻;
6)将步骤5)预冷的卵黄抗体溶液转入冻干机制得抗鹅病毒性疾病卵黄抗体冻干粉;
7)将抗鹅病毒性疾病卵黄抗体冻干粉与黄芪多糖、当归多糖、枸杞多糖、甘草多糖混合即得。
本发明的目的还在于提供一种饲料添加剂在制备抗鹅病毒性疾病的饲料中的应用。
本发明所采用的技术方案还在于提供一种饲料添加剂在制备抗鹅病毒性疾病的饲料中的应用,所述饲料添加剂的加入量为饲料总重量的3-7%。
本发明所述的鹅病毒性疾病为常见的鹅病毒性疾病,如小鹅瘟、鹅的鸭瘟、鹅的副粘病毒病、禽流感、禽霍乱、鹅大肠杆菌性腹膜炎、鹅沙门氏菌病。
卵黄抗体是一种免疫球蛋白,由于机体受到外界抗原的刺激而产生的一种应激产物,可以和相应抗原结合,消除病因。
本发明免疫采用是健康的鸡群作为免疫对象,减少毒株返祖的可能性,同样可以使抗体效价快速达到1:256。本发明将抗鹅病毒性疾病卵黄抗体和多种辅料进行复配,并通过调整辅料的成分、控制其用量,大大延长了卵黄抗体的保存时间,保证了卵黄抗体在较长一段时间内的活性。
多糖具有提高机体免疫能力,增强机体的适应能力和抵抗能力、抗菌、抗病毒、调节机体代谢等作用。其中,
黄芪多糖是黄芪的主要有效成分,黄芪,甘,温。归肺、脾经。益气升阳,固表止汗,利水消肿,脱毒生肌。主治一切气虚血亏之证。主要作用于肾利尿作用,为促进细胞中DNA合成,加速肝分化增殖的作用,提高淋巴细胞的增殖和白细胞诱生干扰素能力的增强,间接提高和促进机体免疫力功能。
当归多糖是当归的主要有效成分,当归,甘、辛、苦、温。归肝、心、脾经。补血活血,调经止痛,润燥滑肠。主治血虚诸证,月经不调,经闭,痛经,崩漏,虚寒腹痛,痿痹,肌肤麻木,肠燥便难,赤痢后重,痈疽疮疡,跌扑损伤。主要作用于心肝脏器调节气血,调节机体免疫功能、具有抗癌作用。
枸杞多糖是枸杞的主要有效成分,枸杞,甘,平。归肝、肾、肺经。养肝,滋润,润肺。主治肝肾亏虚,头晕目眩,目视不清,腰膝酸软,阳痿遗精,虚劳咳嗽,消渴引饮。主要作用于肝肾等脏器,可调节机体免疫功能、能有效抑制肿瘤生长和细胞突变、具有延缓衰老、抗脂肪肝、调节血脂和血糖、促进造血功能等方面的作用。
甘草多糖是甘草的主要有效成分,甘草,甘,平。归脾、胃、心、肺经,主要作用于脾脏,补益气,清热解毒,祛痰止咳,缓急止痛,调和诸药。用于脾胃虚弱,倦怠乏力,心悸气短,咳嗽痰多,缓解药物毒性。
以上几种中草药多糖覆盖了机体的心、肝、脾、肺、肾、脾等脏器,保证了机体脏器之间的平衡运转,加强机体的免疫能力调节机能,是机体健康的保证,使用本发明的复合多糖效果明显优于单一使用。
本发明又将抗鹅病毒性疾病卵黄抗体冻干粉和由黄芪多糖、当归多糖、枸杞多糖、甘草多糖组成的复合多糖复配,制备得到抗鹅病毒性疾病饲料添加剂,具有针对性强,抗体效价高,生物活性良好的优点,能够起到有效预防鹅病毒性疾病性疾病的作用,且在常温下保存很长时间仍能保持活性。本发明的饲料添加剂无毒副作用,无残留危害,安全有效,针对鹅病毒性疾病,尤其是对小鹅瘟的预防效果显著。
本发明将抗鹅病毒性疾病卵黄抗体冻干粉和复合多糖合理搭配组合,添加在日常饲料中,可以提高机体免疫力和抗病能力。实验结果表明,饲喂添加本发明的饲料添加剂后,雏鹅的抗病毒性疾病能力明显提高,其抗鹅病毒性疾病的效果明显优于单独添加卵黄抗体冻干粉或复合多糖的效果。
本发明制备方法简单,易于操作,便于工业化推广。
具体实施方式
实施例1
本实施例的抗小鹅瘟卵黄抗体冻干粉,包括以下重量份数的组分:抗小鹅瘟卵黄抗体93份、硫柳汞0.01份、甲醛0.3份、维生素C 0.5份、聚乙烯吡咯烷酮1份、甘氨酸1份、山梨醇1份、葡萄糖2份。
本实施例的抗小鹅瘟的饲料添加剂,包括以下重量份数的组分:抗小鹅瘟卵黄抗体冻干粉4份、黄芪多糖5份、当归多糖10份、枸杞多糖20份、甘草多糖30份。
本实施例的抗小鹅瘟的饲料添加剂的制备方法,包括以下步骤:
1)分离毒株制备疫苗:将小鹅瘟病毒增殖并灭活制备成疫苗,备用;
2)免疫健康鸡群获得蛋:每隔7天对健康鸡免疫一次,共免疫三次,第一次免疫健康鸡群接种灭活疫苗1.5mL,第二次、第三次免疫接种加量1倍,第二次免疫后开始检测抗体效价,当抗体效价达到1:256时,收集高免蛋,将收集的高免蛋进行清理、消毒,晾干备用;
3)酸化处理:分离高免蛋卵黄,加入等体积的无菌水,充分搅拌,得到卵黄抗体溶液;将卵黄抗体溶液放入63℃水浴锅内均匀加热30min,然后加入pH=5、温度为5℃的酸化水溶液,酸化水溶液加入量为卵黄抗体溶液体积的6倍,混匀,静置沉淀,取上清液,15000rpm,离心20min,得到含有卵黄抗体的上清液;
4)用0.45μm的滤膜对步骤3)所得上清液过滤,截流分子量大于200KD的大分子物质,再将滤液经0.22μm滤膜过滤,留滤液;
5)将滤液用超滤装置浓缩,使卵黄抗体溶液浓缩至2.5mL/500羽份;
6)向步骤5)所得卵黄抗体浓缩溶液中加入甲醛、硫柳汞、维生素C、聚乙烯吡咯烷酮、甘氨酸、山梨醇、葡萄糖;
7)预冻:将浓缩后的卵黄溶液放置到-80℃条件下,充分冷冻;
8)将步骤7)所得预冷卵黄抗体溶液转入冻干机制得抗小鹅瘟卵黄抗体冻干粉;
9)将抗小鹅瘟卵黄抗体冻干粉与黄芪多糖、当归多糖、枸杞多糖、甘草多糖混合即得。
实施例2
本实施例的抗小鹅瘟卵黄抗体冻干粉,包括以下重量份数的组分:抗小鹅瘟卵黄抗体95份、硫柳汞0.02份、甲醛0.2份、维生素C 1份、聚乙烯吡咯烷酮0.5份、甘氨酸0.5份、山梨醇1.5份、葡萄糖1.5份。
本实施例的抗小鹅瘟的饲料添加剂,包括以下重量份数的组分:抗小鹅瘟卵黄抗体冻干粉5份、黄芪多糖8份、当归多糖7份、枸杞多糖15份、甘草多糖10份。
本实施例的抗小鹅瘟的饲料添加剂的制备方法同实施例1,不同之处在于各组分的用量不同。
实施例3
本实施例的抗小鹅瘟卵黄抗体冻干粉,包括以下重量份数的组分:抗小鹅瘟卵黄抗体96份、硫柳汞0.015份、甲醛0.25份、维生素C 0.8份、聚乙烯吡咯烷酮0.8份、甘氨酸0.8份、山梨醇2份、葡萄糖1份。
本实施例的抗小鹅瘟的饲料添加剂,包括以下重量份数的组分:抗小鹅瘟卵黄抗体冻干粉2份、黄芪多糖10份、当归多糖5份、枸杞多糖10份、甘草多糖20份。
本实施例的抗小鹅瘟的饲料添加剂的制备方法同实施例1,不同之处在于各组分的用量不同。
对比例1
本对比例的抗小鹅瘟的饲料添加剂,包括以下重量份数的组分:抗小鹅瘟卵黄抗体96份、硫柳汞0.015份、甲醛0.25份、维生素C 0.8份、聚乙烯吡咯烷酮0.8份、甘氨酸0.8份、山梨醇2份、葡萄糖1份。
对比例2
本对比例的抗小鹅瘟的饲料添加剂,包括以下重量份数的组分:黄芪多糖10份、当归多糖5份、枸杞多糖10份、甘草多糖20份。
实验例1
本发明饲料添加剂的质量检测
实验方法:将本发明的饲料添加剂保存在37℃培养箱中,保存24个月,分别1周、2周、1月、2月、4月、8月、12月、16月、20月、24月后,用琼脂扩散(AGP)法,测定每个时间点取样品的卵黄抗体效价变化,结果见表1。
表1 常温放置冻干粉不同时间段效价的变化(项目:DHV)
组别 | 1周 | 2周 | 1月 | 2月 | 4月 | 8月 | 12月 | 16月 | 20月 | 24月 |
实施例1 | 1:32 | 1:32 | 1:32 | 1:32 | 1:32 | 1:16 | 1:16 | 1:16 | 1:16 | 1:16 |
实施例2 | 1:32 | 1:32 | 1:32 | 1:32 | 1:32 | 1:32 | 1:16 | 1:16 | 1:16 | 1:16 |
实施例3 | 1:32 | 1:32 | 1:32 | 1:32 | 1:32 | 1:32 | 1:16 | 1:16 | 1:16 | 1:16 |
卵黄抗体质量检测:
[支原体检验]按《中国兽药典》进行,无支原体生长。
[外源病毒检验]按《中国兽药典》进行,符合规定。
[安全检验]用7日龄雏鹅5只,肌肉注射5羽份,观察2周,应全部健活,注射部位没有病变。
[保质期]根据常温效价检测实验结果,可以存放2年。
实验例2
本发明的饲料添加剂抗小鹅瘟病的效果分析
本实验例所用普通饲料为市售常规饲料。
实验方法:选取7日龄雏鹅700只,其中100只作为攻毒组隔离饲养,100只作为空白组隔离饲养,200只作为对照组隔离饲养,300只作为试验组隔离饲养。攻毒组和空白组均饲喂普通饲料,对照组分为2组,每组100只,分别饲喂添加量7%对比例1-2饲料添加剂+普通饲料,试验组分为3组,每组100只,隔离饲养,分别饲喂添加量7%实施例1-3饲料添加剂+普通饲料。饲养方式采用自由采食,自由饮水,24h后,取试验组、对照组和攻毒组的雏鹅肌注接种稀释后GPV病毒液,记录120h患病情况,结果如表2所示。
表2 实施例1-3的饲料添加剂抗小鹅瘟病的效果分析
从表2中可以看出,添加了本发明的饲料添加剂后,鹅抗病毒性疾病能力明显提高,抗鹅病毒性疾病的效果也明显优于单独添加卵黄抗体冻干粉或复合多糖的效果。
Claims (7)
1.一种抗鹅病毒性疾病的饲料添加剂,其特征在于:所述饲料添加剂包括以下重量份数的组分:抗鹅病毒性疾病卵黄抗体冻干粉2-5份、黄芪多糖5-10份、当归多糖5-10份、枸杞多糖10-20份、甘草多糖10-30份,所述抗鹅病毒性疾病卵黄抗体冻干粉包括以下重量份数的组分:抗鹅病毒性疾病卵黄抗体93-96份、防腐剂0.01-0.02份、灭活剂0.2-0.3份、维生素C 0.5-1份、聚乙烯吡咯烷酮0.5-1份、甘氨酸0.5-1份、山梨醇1-2份、葡萄糖1-2份。
2.根据权利要求1所述的抗鹅病毒性疾病的饲料添加剂,其特征在于:所述灭活剂为甲醛。
3.根据权利要求1所述的抗鹅病毒性疾病的饲料添加剂,其特征在于:所述防腐剂为硫柳汞。
4.一种如权利要求1所述的抗鹅病毒性疾病的饲料添加剂的制备方法,其特征在于:包括以下步骤:
1)将鹅病毒性疾病病毒增殖并灭活制备成疫苗,免疫健康鸡群制得高免蛋,将高免蛋经过消毒、酸化处理、离心处理、配制、冻干得到抗鹅病毒性疾病卵黄抗体冻干粉;
2)将抗鹅病毒性疾病卵黄抗体冻干粉与其余组分混合即得。
5.根据权利要求4所述的抗鹅病毒性疾病的饲料添加剂的制备方法,其特征在于:具体步骤以下:
1)将鹅病毒性疾病病毒增殖并灭活制备成疫苗,每间隔7天对健康产蛋鸡群进行免疫,共免疫三次,第二次免疫后开始检测抗体效价,当抗体效价达到1:256时,收集高免蛋,将收集的高免蛋进行清理、消毒,晾干备用;
2)分离高免蛋卵黄,加入等体积的无菌水,充分搅拌,得到卵黄抗体溶液;将卵黄抗体溶液放入60-65℃水浴锅内均匀加热30min,然后加入pH=4.9-5.2、温度为4-6℃的酸化水溶液,酸化水溶液加入量为卵黄抗体溶液体积的6倍,混匀,静置沉淀,取上清液,15000rpm离心20min,得到含有卵黄抗体的上清液;
3)用0.45μm的滤膜对步骤2)所得上清液过滤,截流分子量大于200KD的大分子物质,再将滤液经0.22μm滤膜过滤,留滤液;
4)将滤液用超滤装置浓缩,使卵黄抗体溶液浓缩至2.5mL/500羽份;
5)向步骤4)所得卵黄抗体溶液中加入灭活剂、防腐剂、维生素C、聚乙烯吡咯烷酮、甘氨酸、山梨醇、葡萄糖,并在﹣80℃条件下充分冷冻;
6)将步骤5)预冷的卵黄抗体溶液转入冻干机制得抗鹅病毒性疾病卵黄抗体冻干粉;
7)将抗鹅病毒性疾病卵黄抗体冻干粉与黄芪多糖、当归多糖、枸杞多糖、甘草多糖混合即得。
6.一种如权利要求1所述的饲料添加剂在制备抗鹅病毒性疾病的饲料中的应用。
7.根据权利要求6所述的应用,其特征在于:所述饲料添加剂的加入量为饲料总重量的3-7%。
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