CN1709403A - 以黄芪为主方的动物免疫增强剂 - Google Patents
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Abstract
本发明公开了一种以黄芪为主方的动物免疫增强剂,该增强剂的配方组分(按重量计)为:黄芪10~15份、黄连5~10份、黄芩10~15份、连翘20~25份和大青叶15份。本发明的以黄芪为主方的动物免疫增强剂,成本低,能提高动物免疫能力、改善其生长性能。
Description
技术领域
本发明涉及一种动物免疫增强剂的配方,特别是一种以黄芪为主方的动物免疫增强剂的配方。
背景技术
为了防止所饲养的动物生病,提高动物的抗病能力,饲养员一般会在饲料中添加一定比例的抗生素。但是这样不但会导致动物产生抗药性,而且还会使抗生素残留在动物体内,作为食物链最上端的人如果直接或间接食用了此种动物,会对身体健康造成极大的隐患。因此人们迫切希望有一种纯天然的健康、无毒副作用的动物免疫增强剂,以提高动物的抗病能力。
天然药物中的草药是人类最早应用的物质,也是最早被应用的饲料添加剂。我国有12807种天然物中草药资源,而其中只有10%为常用,仍需深入研究利用。黄芪是中草药中“扶正固本”类补益药物,能滋虚扶弱,调理肠道微生态平衡,提高机体非特异性免疫能力。但黄芪为主方的动物饲料免疫增强剂的却未见相关报道。
发明内容
针对现有技术中存在的不足之处,本发明提供一种成本低、能提高动物免疫能力、改善其生长性能的以黄芪为主方的动物免疫增强剂。
本发明为达到以上目的,是通过这样的技术方案来实现的:提供一种以黄芪为主方的动物免疫增强剂,该增强剂的配方组分(按重量计)为:黄芪10~15份、黄连5~10份、黄芩10~15份、连翘20~25份和大青叶15份。
本发明的以黄芪为主方的动物免疫增强剂,选用的都是常见原料,因此能最大限度控制生产成本。特别是原料之一的黄芪,具有在我国资源丰富、价格低廉、安全无毒的特点;以其为原料制成的动物免疫增强剂作为一种饲料添加剂使用,能提高动物免疫功能,在改善动物肠道微生态平衡的同时还可补充饲料蛋白作用,因此能改善动物生长性能。本发明的以黄芪为主方的动物免疫增强剂,适宜动物长期服用,不会使动物产生抗药性,可全部或大部分替代饲料中的抗生素。
本发明的以黄芪为主方的动物免疫增强剂对动物免疫能力的增强、和对动物肠道微生态平衡的改善功能可通过以下的实验方法来证明。
1、选取60只体重相近的生长肥育猪,随机分为2组(对照组和试验组),每组3个重复,每个重复10只。在试验组所食用的饲料中添加了1%重量比的本发明的以黄芪为主方的动物免疫增强剂,食用普通饲料的称为对照组。试验预饲7天,正试期30天,每日记录采食量。
2、试验结束时屠宰取样,分别称重内脏器官和免疫器官,计算脏器系数;采集血样制备血清,采用免疫浊度法试剂盒测定血清抗体IgA、IgG、IgM,补体C3、C4,ELISA方法试剂盒测定血清中细胞因子IL-1、IL-2含量,在CHEM-5半自动生化分析仪上测定;取试验猪静脉血10ml,加入事先加有1%无菌肝素的离心管中,带回实验室进行细胞培养和NBT检测。
3、采集新鲜血液和脾脏,采用MTT法进行外周血淋巴细胞培养和脾脏淋巴细胞增殖试验,测定T/B淋巴细胞增殖效果:
1)取2.0ml无菌抗凝血与等量Hanks液混合,稀释悬浮细胞。将细胞悬液小心缓慢加在与血液等量(2.0ml)的淋巴细胞分离液上,使血液重叠于分层液上,室温水平离心2000rpm 20min。除去最上面的血浆,用毛细管将血浆和分层液之间较薄、但比较清楚的乳白色淋巴细胞层吸至含5ml Hanks液的试管中,混匀后1000r/min离心10min,清洗两次,洗去残余的淋巴细胞分离液。用台盼蓝染色检测细胞活力>95%并计数。用含20%胎牛血清的细胞培养液将细胞调整为2×106个细胞/ml。
2)96孔细胞培养板每孔加入单细胞悬液100ul和100ul PHA,每组设置5个重复孔,同时设置培养液空白对照,放5%CO2、37℃细胞培养箱静置培养48h,终止培养前4小时每孔加入5mg/ml四甲基偶氮唑盐(MTT)溶液20ul继续培养4小时后每孔再加入DMSO溶液100ul,溶解蓝黑色甲臜沉淀,之后用酶联免疫检测仪(BIO-RAD)检测每孔OD570nm。试验结果用刺激指数SI来表示:
SI=试验组OD570/空白OD570
4、脾脏细胞培养试验ConA(Sigma):
1)无菌取猪脾脏,置于无血清RPMI-1640培养液(含青霉素和链霉素)平皿中,反复冲洗,用针桶栓小心将脾压碎,经400目不锈钢滤网过滤,用培养液清洗两次(1000rpm/min,5分钟),用含20%胎牛血清的RPMI-1640培养液制成细胞悬液。用含20%胎牛血清的RPMI-1640培养液调整脾淋巴细胞数为6×106个细胞/ml。
2)细胞接种:用96孔平板培养,每孔加上述细胞悬液100ul、另加ConA或PHA,每组设置5个重复孔,同时设置培养液空白对照,放5%CO2、37℃细胞培养箱静置培养48h。
3)MTT法测细胞增殖:于终止培养前4小时每孔加5mg/mlMTT溶液20ul,继续放培养箱培养4小时后每孔再加入DMSO溶液100ul,在微型震荡器上震荡10分钟,之后用酶联免疫检测仪(BIO-RAD)570nm波长下检测每孔OD570值。试验结果用细胞增殖率百分率来表示:
增殖率(%)=(试验组OD570-空白OD570)/空白OD570×100%
5、选择新鲜血液采用NBT还原法测定嗜中性白细胞还原能力:取上述抗凝血0.1ml,加入等量的NBT应用液,放入12×75mm塑料管混匀,将管口盖上,试管可放37度温箱,其间振摇一次。将上述液体取出摇匀,用毛细滴管吸取1滴于玻片的一端做推片,要求推出尾部,推片要厚薄适宜,立即用吹风机吹干,甲醇固定1-2min,吹干,用1%沙黄水溶液染色5min,自来水冲洗,油镜观察,计数。
试验结果如下表所示:
表1为本发明对杜长大断奶仔猪日增重、料重比和腹泻率的影响;
表2为本发明对杜长大断奶仔猪外周血淋巴细胞转化率的影响;
表3为本发明对杜长大断奶仔猪脾脏淋巴细胞增殖的影响;
表4为本发明对杜长大断奶仔猪血清IgG、IgA和IgM及C3、C4水平的的影响;
表5为本发明对杜长大断奶仔猪血清中IL-1和IL-2水平的影响;
表1
对照组 | 试验组 | |
始重(kg)终重(kg)日增重(g)料重比腹泻率(%) | 21.94±1.9745.13±0.38773.1±57.02.03±0.296.37±0.31 | 21.82±0.5147.93±1.15870.3±28.61.90±0.062.73±3.23 |
与对照组相比,试验组日增重12.5%(P<0.05)、料重比6.4%(p>0.05)、腹泻率降低133%(P<0.05)
表2
对照组 | 试验组 | |
Con诱导(SI)LPS诱导(SI) | 1.06±0.081.11±0.17 | 1.27±0.081.26±0.06 |
与对照组相比,试验组在Con诱导下外周血T-淋巴细胞转化率分别提高了19.8%(P<0.05)。在Lps诱导B-淋巴细胞转化率提高了13.5%但差异不显著。
表3
对照组 | 试验组 | |
Con诱导(%)LPS诱导(%) | 84.37±28.2849.82±13.62 | 153.8±40.02101.9±22.09 |
与对照组相比,试验组在Con诱导下脾脏细胞转化率分别提高了82.3%(P<0.05)。在LPS诱导下脾脏细胞转化率提高了105%(P<0.05)。
表4
对照组 | 试验组 | |
IgG(g/l)IgA(g/l)IgM(g/l)C3(g/l)C4(g/l) | 4.41±0.1680.529±0.0070.63±0.090.173±0.0180.021±0.006 | 4.81±0.3100.658±0.0170.68±0.060.201±0.0170.024±0.004 |
与对照组相比,试验组仔猪血清中IgG、C3和C4水平分别提高了9.1%、16.2%和14.2%但差异均不显著,使血清中IgA水平提高了24.4%(P<0.05),使血清中IgM水平降低了8.6%但差异不显著。
表5
对照组 | 试验组 | |
IL-1(pg/ml)IL-2(pg/ml) | 8.48±1.3913.95±1.18 | 19.33±1.8927.28±1.18 |
与对照组相比,试验组血清中IL-1α水平提高了128%(P<0.05),使血清中IL-2水平提高了95.6%(P<0.05)。
具体实施方式
实施例1、一种以黄芪为主方的动物免疫增强剂,按下述配比称取原料(kg):
黄芪12、黄连8、黄芩13、连翘22和大青叶15。
将上述原料简单加以混合。
实施例2、一种以黄芪为主方的动物免疫增强剂,按下述配比称取原料(kg):
黄芪10、黄连10、黄芩10、连翘25和大青叶15。
将上述原料简单加以混合。
实施例3、一种以黄芪为主方的动物免疫增强剂,按下述配比称取原料(kg):
黄芪15、黄连5、黄芩15、连翘20和大青叶15。
将上述原料简单加以混合。
最后,还需要注意的是,以上列举的仅是本发明的若干个具体实施例。显然,本发明不限于以上实施例,还可以有许多变形。本领域的普通技术人员能从本发明公开的内容直接导出或联想到的所有变形,均应认为是本发明的保护范围。
Claims (1)
1、以黄芪为主方的动物免疫增强剂,其特征是该增强剂的配方组分(按重量计)为:黄芪10~15份、黄连5~10份、黄芩10~15份、连翘20~25份和大青叶15份。
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Cited By (2)
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CN102940717A (zh) * | 2012-12-05 | 2013-02-27 | 邓凯伟 | 一种治疗牛口蹄疫的中药 |
CN108272910A (zh) * | 2018-03-20 | 2018-07-13 | 长沙小新新能源科技有限公司 | 一种对抗病毒的兽药及其制备方法 |
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CN1216541C (zh) * | 2003-07-03 | 2005-08-31 | 中国农业科学院饲料研究所 | 一种禽类饲料添加剂 |
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Publication number | Priority date | Publication date | Assignee | Title |
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CN102940717A (zh) * | 2012-12-05 | 2013-02-27 | 邓凯伟 | 一种治疗牛口蹄疫的中药 |
CN108272910A (zh) * | 2018-03-20 | 2018-07-13 | 长沙小新新能源科技有限公司 | 一种对抗病毒的兽药及其制备方法 |
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