CN111588801B - 一种预防畜禽疫病的中药分散颗粒剂及其制备方法 - Google Patents
一种预防畜禽疫病的中药分散颗粒剂及其制备方法 Download PDFInfo
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Abstract
本发明提供一种用于预防畜禽疫病的中药分散颗粒剂及其制备方法,属于兽药技术领域。所述中药分散颗粒剂主要由大黄115‑125份、甘草38‑42份、黄芩115‑125份、丹参192‑208份、刺五加38‑42份、天花粉115‑125份、青蒿36‑40份、荆芥38‑42份、麦冬115‑125份、板蓝根52‑58份、白术60‑66份、滑石115‑125份组成。本发明运用中医辨证原则,诸药配伍合理,充分发挥各药的作用,诸药共同协调增益药。本发明具有调节畜禽机体免疫功能,增强畜禽免疫应答能力,提高畜禽抵抗疫病的能力;且本发明为纯中药制剂,无抗生素残留问题,减少对环境污染的影响,安全无毒;同时本发明制备工艺简单,产品质量稳定性强,储存时间长,质量标准可控,能够量化生产,具有良好的市场推广和应用。
Description
发明领域
本发明属于兽药技术领域,具体涉及一种预防畜禽疫病的中药分散颗粒剂及其制备方法。
发明背景
近年来,我国畜牧业发展突飞猛进,畜牧业在农业总产值中比例逐年提高,逐渐成为保障食品安全和促进农民增收的支柱产业。畜禽养殖规模的扩大,疫病也随之增多,一些作业迅速、疗效确切的抗生素、激素等化学治疗药物的生产中得到广泛使用和迅速推广,对动物疾病的预防控制起着举足轻重的作用。但是化学药物的长期、大量、不合理的使用,带来的细菌的耐药性增强、畜产品药物残留过高等问题日益突出。于是有许多人开始意识到化学的缺陷和不足,把目光逐渐转向了中草药,将其作为畜禽保健品。中药具有低毒、低残留、绿色、安全等优点,并且大量研究报道中草药具有明显的抗菌、抗病毒、调节免疫及营养调理等作用,在很大程度上克服了化学药物的缺点如今正逐步应用于畜禽疫病防治中。
畜禽疫病中,新城疫和猪瘟是最为常见和最为常复发的。其中,新城疫是由病毒引起的禽类接触性传染,有急性高发特征。新城疫病毒由副黏病毒科腮腺炎病毒属病毒引起,可以通过呼吸道、消化道、眼结膜、创口等途径侵入禽体。猪瘟是世界动物卫生组织标明的A类16种法定传染病之一,具有极强的传染性和极高的死亡病率。如果在生猪养殖过程中爆发大规模的猪瘟,不仅会直接降低养殖户的养殖经济效益,还可能造成严重的食品安全、环境污染、动物疫病防控等问题。猪瘟属于黄病毒科瘟病毒属,传染性强,潜伏期普遍在五到七天,病毒适应能力较强。
畜禽病毒的种属及血清型复杂,且易突变,有的病毒还会导致免疫机制,这给疫苗研发和应用带来困难。抗病毒西药种类很少,且毒副作用较大,易发生耐药性,临床疗效十分有限。中药大多为天然的植物和矿物质入药,且经过人类反复试验、筛选,大多为对机体有益无害的物质,许多还是食药兼用的品种,尽管部分中草药有一定毒性,但经过适宜的炮制和配伍,能够减少其毒性,能够减少其毒性,适量使用是没有毒害的。中药的主要成分为生物碱、挥发油、苷类、有机酸、多糖、鞣质等生物大分子,这些大分子物质以复方形式结合,在机体中具有复杂的代谢反应,进入机体后从呼吸、生化代谢、能量转化、遗传物质、繁殖等多个环节干扰或阻断病原微生物的生存与繁殖继代,病原微生物很难对其产生抗药性,从而有效地抑制和杀灭病原微生物。
现有技术中,大多畜禽用中药处方来源不确切,并且其中药制剂以口服液和粉剂居多。其中,口服液容易酸败和涨瓶,粉剂容易结块,极大降低的中药的防治效果。并且中药散剂的生物利用度普遍偏低,普通中药颗粒和泡汤颗粒其溶解性对温度的依赖性比较高,对集约化冬季养殖受限。
发明内容
为解决现有技术不足之处,本发明的目的在于提供一种用于预防畜禽疫病的中药分散颗粒剂及其制备方法。
为实现本发明目的,本发明的技术方案如下:
首先,本发明提供一种用于预防畜禽疫病的中药分散制剂,所述中药分散制剂包括以下重量份数的原料:大黄、甘草、黄芩、丹参、刺五加、天花粉、青蒿、荆芥、麦冬、板蓝根、白术、石膏。
进一步地,作为一种用于预防畜禽疫病的中药分散制剂的优化,所述重量份数的原料为:大黄115-125份、甘草38-42份、黄芩115-125份、丹参192-208份、刺五加38-42份、天花粉115-125份、青蒿36-40份、荆芥38-42份、麦冬115-125份、板蓝根52-58份、白术60-66份、滑石115-125份。
进一步地,作为一种用于预防畜禽疫病的中药分散制剂的优化,所述重量份数的原料为:大黄120份、甘草40份、黄芩115份、丹参204份、刺五加38份、天花粉118份、青蒿36份、荆芥42份、麦冬122份、板蓝根55份、白术65份、滑石115份。
进一步地,作为一种用于预防畜禽疫病的中药分散制剂的优化,所述中药分散制剂还包括辅料,所述辅料由以下重量份数原料组成:聚乙烯吡咯烷酮140-150份、乳糖290-300份、碳酸氢钠60-70份、柠檬酸70-80份、猪乳香0.5-1份。
另一方面,本发明提供一种预防畜禽疫病的中药分散制剂的制备方法,按照以下步骤制备:
(1)按照重量份数大黄、甘草、黄芩、丹参、刺五加、天花粉、青蒿、荆芥、麦冬、板蓝根、白术、滑石,分别准确称取原料;
(2)将步骤(1)中草药原料除滑石外全部投入中药提取机中,随后加入各原料重量份数之和份数的10-15倍量水,煎煮30-40min后加入滑石,继续煎煮30-40min,过滤收集滤液,在药渣中再次加入第一次总份数的8-10倍水,继续煎煮60-70min,过滤收集滤液,将两次收集的滤液合并,进行喷雾干燥,得到混合物A;
(3)按照重量份数聚乙烯吡咯烷酮、乳糖、碳酸氢钠、柠檬酸、猪乳香,分别准确称取,其中乳糖、碳酸氢钠和柠檬酸分别粉碎过100-200目药筛;将上述各原料混合得到混合物B;
(4)将混合物A和混合物B投入到混合机组,混合45min,得到混合物C;
(5)将混合物C投入到颗粒剂干法压片机中,经过压片、粉碎、过1号药筛、整粒,即得本发明预防畜禽疫病的中药分散颗粒剂。
本发明中所涉及中草药药理作用
大黄:味苦,性寒,归胃、肝二经;有泻热通常,凉血解毒,逐淤通经等功效;具有导泻、利胆、保肝、抗溃疡、抗菌、抗病毒等作用。
甘草:味甘,性平,归心、肺、脾、胃经;有补脾益气,清热解毒,去痰止咳,缓急止痛,调和诸药等功效;常用于脾胃虚弱,倦怠乏力,心悸气短,咳嗽痰多,痈肿疮毒等病症,能够缓解药物毒性、烈性。
黄芩:味苦,性寒,归脾、肺经;其含有黄芩苷、黄芩素、汉黄芩苷、汉黄芩素,黄芩新素Ⅰ 、Ⅱ ,去甲汉黄芩素、挥发油、苯乙醇糖苷类等,能清热燥湿、泻火解毒、止血、安胎、降血压、杀菌、杀病毒、杀真菌等。黄芩含有的黄芩苷、黄芩素有良好的抗氧化功能,人医用黄芩治疗缺血性中风(脑梗)及帕金森氏病,兽医用其治疗口蹄疫、水疱病,清肺热,消除呼吸道感染。
丹参:味苦、性微寒,归心、肝二经;对细胞因子、抗体及免疫复合物、免疫细胞有免疫调节作用,具有改善微循环,保肝抗炎,活血化瘀,安神宁心,止痛,扩张血管,降低血压等多种功效。
刺五加:味甘、微苦、性温;归脾、肺、心、肾经;具有益气健脾、补肾安神的功效,能够治疗脾肺气虚证、肾虚腰膝酸痛证、心脾不足、失眠等。
天花粉:味甘、微苦,性微寒;归肺、胃经;具有清热泻火,生津止渴,消肿排脓的功效;常用于治疗热病烦渴,肺热燥咳,内热消渴,疮疡肿毒等病症。
青蒿:味苦、辛,性寒;具有清虚热,除骨蒸,解暑热,截疟,退黄的功效;常用于温邪伤阴、阴虚发热、疟疾寒热、湿热黄疸等病症;青蒿有抗疟成分。青蒿素的发现是中国对人类的伟大贡献。兽医将青蒿用于动物弓形体病、附红细胞体病和病毒病的治疗。
荆芥:味辛、苦,性微温,归肺、肝经;具有解表祛风、透疹止血等功效;荆芥中主要有效成分为挥发油及黄酮类化合物,具有明显的发汗、解热、抗炎、镇痛作用;荆芥中的有效成分通过抑制相关致炎因子含量,缩短其细胞生长周期,进而发挥抗菌、抗病毒、抗肿瘤作用。
麦冬:味甘、微苦,性微寒,归心、肺、胃经;具有养阴生津,润肺止咳等功效;麦冬的化学成分主要包括甾体皂苷类、高异黄酮类、多糖类等,具有降血糖、保护心血管系统,增强免疫力、抗皮肤衰老、抗炎、抗肿瘤等药理作用。
板蓝根:性寒,味苦,归心、胃经;有清热解毒、凉血、利咽的功效,对瘟、毒、痈、斑、丹毒及风热症效果好。其含靛兰、靛玉红、β-谷甾醇、黑芥子甙、靛甙、色胺酮、棕榈酸、蔗糖、1-硫氰酸-2-羟基丁-3-烯及多种氨基酸,靛甙是抗病毒的主要成分。板蓝根具有较强的抗病毒、抗菌、增强免疫力、抗内毒素、抗癌和解热等作用。
白术:性苦、甘,性温,归脾、胃经;具有健脾益气,燥湿利水,止汗,安胎等功效;常用于脾虚食少、泄泻便溏、胎动不安等症;其主要化学成分 有挥发油、多糖、内脂类等,具有抗肿瘤、抗炎调节消化系统等作用。
滑石:味甘,性寒,归膀胱、肺、胃经;具有利尿通淋、清热解暑的功效;常用于热淋、石淋、尿热涩痛、湿热水泻等病症。
有益效果
本发明提供了一种用于预防畜禽的中药分散颗粒剂及其制备方法。本发明为纯中药制剂,主要由大黄、甘草、黄芩、丹参、刺五加、天花粉等十一味中草药材组成,本发明运用中医辨证原则,选用药材,诸药配伍合理,充分发挥各药的作用,诸药共同协调增益药。本发明具有调节畜禽机体免疫功能,增强畜禽免疫应答能力,提高畜禽抵抗疫病的能力;且本发明为纯中药制剂,无抗生素残留问题,减少对环境污染的影响,安全无毒;同时本发明制备工艺简单,产品质量稳定性强,储存时间长,质量标准可控,能够量化生产,具有良好的市场推广和应用价值。
具体实施方式
实施例1
一种用于预防畜禽疫病的中药分散颗粒剂,按照以下步骤制备而成:
(1)按照重量份数大黄120份、甘草40份、黄芩115份、丹参204份、刺五加38份、天花粉118份、青蒿36份、荆芥42份、麦冬122份、板蓝根55份、白术65份、滑石115份,分别准确称取原料;
(2)将步骤(1)中草药原料除滑石外全部投入中药提取机中,随后加入各原料重量份数之和份数的15倍量水,煎煮40min后加入滑石,继续煎煮40min,过滤收集滤液,在药渣中再次加入第一次总份数的10倍水,继续煎煮70min,过滤收集滤液,将两次收集的滤液合并,进行喷雾干燥,得到混合物A;
(3)按照重量份数聚乙烯吡咯烷酮140份、乳糖290份、碳酸氢钠60份、柠檬酸70份、猪乳香0.5份,分别准确称取,其中乳糖、碳酸氢钠和柠檬酸分别粉碎过100目药筛;将上述各原料混合得到混合物B;
(4)将混合物A和混合物B投入到混合机组,混合45min,得到混合物C;
(5)将混合物C投入到颗粒剂干法压片机中,经过压片、粉碎、过1号药筛、整粒,即得本发明预防畜禽疫病的中药分散颗粒剂。
本发明诸药配伍合理,充分发挥各药的作用,诸药共同协调增益药效,本发明以中药理论,选用药材,对药材进行组方,本发明中由于实验数据较多,对方剂进行多次调整并进行实验,在此仅列出多组疗效较高的组方并与本发明配方进行对比。
配方1:大黄120份、黄芩115份、丹参204份、刺五加38份、天花粉118份、青蒿36份、荆芥42份、板蓝根55份、白术65份、滑石115份
配方2:大黄120份、甘草40份、黄芩115份、丹参204份、刺五加38份、天花粉118份、青蒿36份、荆芥42份、麦冬122份、滑石115份
配方3:大黄120份、甘草40份、丹参204份、刺五加38份、天花粉118份、荆芥42份、麦冬122份、板蓝根55份、白术65份、滑石115份
配方4:大黄120份、甘草40份、黄芩115份、天花粉118份、青蒿36份、荆芥42份、麦冬122份、板蓝根55份、白术65份、滑石115份
配方5:甘草40份、黄芩115份、丹参204份、刺五加38份、天花粉118份、青蒿36份、麦冬122份、板蓝根55份、白术65份、滑石115份
配方6:大黄120份、甘草40份、黄芩115份、丹参204份、刺五加38份、天花粉118份、青蒿36份、荆芥42份、麦冬122份、板蓝根55份、白术65份
本发明配方:大黄120份、甘草40份、黄芩115份、丹参204份、刺五加38份、天花粉118份、青蒿36份、荆芥42份、麦冬122份、板蓝根55份、白术65份、滑石115份
试验方法:将270只1日龄AA肉鸡随机分为9组,分别为A—I组,每组3个重复,每个重复10只鸡。其中A组为空白组,饲喂基础日粮,正常饮用水;B组为模型组,饲喂基础日粮,正常饮用水;C—I组为试验组,分别饲喂基础日粮和质量分数为20%的按照实施例1的制备方法得到的配方1-配方6及本发明配方的中药分散颗粒剂溶液,中药分散颗粒剂溶液在10-30日龄饲喂,其余时间饲喂正常饮用水。试验期为35天,饲养条件和环境保持一样。对B-I组的7日龄进行新城疫首免,新城疫Ⅳ弱毒苗2倍量点眼滴鼻;对B-I组的21日龄进行新城疫再免,新城疫Ⅳ弱毒苗4倍量饮水。饲养至35日龄,停料和饮水12小时后,计算成活率。结果如表1所示。
表1不同配方中药分散颗粒对感染新城疫肉鸡成活率的影响(只、%)
类别 | A组 | B组 | C组 | D组 | E组 | F组 | G组 | H组 | I组 |
动物数 | 30 | 30 | 30 | 30 | 30 | 30 | 30 | 30 | 30 |
存活数 | 30 | 23 | 27 | 25 | 24 | 26 | 24 | 28 | 29 |
死亡数 | 0 | 7 | 3 | 5 | 6 | 4 | 6 | 2 | 1 |
成活率 | 0 | 76.7<sup>a</sup> | 90.0<sup>b</sup> | 83.3<sup>b</sup> | 80.0 | 86.7<sup>b</sup> | 80.0 | 93.3<sup>b</sup> | 96.7<sup>b</sup> |
注: 与A组比较a P<0. 01; 与B组比较b P<0. 05。
从表1结果可知,与B组相比较,C—I组对感染新城疫肉鸡的生活率有积极的影响作用,说明配方1-6和本发明中药颗粒对感染新城疫肉鸡有一定的治疗作用。但是,配方1-6其中都缺少本发明中药颗粒中的个别中药原料,其疗效并没有本发明中药颗粒效果显著。与B组相比,I组即本发明中药颗粒组成活率明显升高(P<0.05),且也远高于其他配方制剂组的成活率。以上结果可以说明,本发明中药颗粒可以明显降低感染新城疫肉鸡的死亡率,提高成活率,具有生命保护作用,发明的配方中药颗粒选用大黄、甘草、黄芩等十一味中草药,应用中医辨证施治原则合理配伍而得,充分发挥各药的作用,诸药共同协调增益药效,配方中各成分缺一不可。
为进一步说明本发明中药分散颗粒的稳定性、毒性及对畜禽特异性免疫功能的影响,以下提供动物试验作为说明。以下所涉及本发明中药分散颗粒剂皆按照实施例1制备得到。
1、稳定性研究
试验方法:按照实施例1制备得到本发明中药分散颗粒剂,按市售包装,于温度25±2℃,相对湿度为60±10%条件下,放置24个月,分别于0、3、6、9、12、18、24个月末取样检测,结果如表2所示。
表2 本发明中药分散颗粒剂长期试验测定结果
时间(月) | 形状 | 溶解性 | 水分(%) | 含量(%) |
0 | 棕黄色颗粒 | 符合规定 | 1.8 | 99.5 |
3 | 棕黄色颗粒 | 符合规定 | 1.8 | 99.5 |
6 | 棕黄色颗粒 | 符合规定 | 1.9 | 99.5 |
9 | 棕黄色颗粒 | 符合规定 | 1.9 | 99.4 |
12 | 棕黄色颗粒 | 符合规定 | 1.9 | 99.4 |
18 | 棕黄色颗粒 | 符合规定 | 2.1 | 99.3 |
24 | 棕黄色颗粒 | 符合规定 | 2.1 | 99.3 |
以上结果可以说明,本发明中药分散颗粒剂于温度25±2℃,相对湿度为60±10%条件下,放置24个月,分别于0、3、6、9、12、18、24个月末取样检测,所有检测指标没有明显变化,说明本发明中药分散颗粒剂稳定性好,可长期保存。
2、急性毒性试验
2.1 半数致死量(LD50)测定
取健康昆明小鼠12只,体重(16±1)g,随机分成高、中、低3个剂量组,每组4只,雌雄各半,分笼饲养。试验前禁食12 h,不限制饮水。经口灌服本发明中药分散颗粒剂,高、中、低剂量组灌服剂量分别为5000 mg/kg BW、1000 mg/kg BW、200 mg/kg BW,灌胃后正常饲喂,观察7天,认真记录各组小鼠临床症状及死亡情况。若高剂量组出现死亡,则利用简化寇式法测定半数致死量(median lethal dose,LD50);若高剂量组小鼠未出现死亡,说明本品毒性较低,则改用最大耐受量(Maximal tolerable dose,MTD)分析其急性毒性。
2.2 最大耐受量(MTD)试验
取健康昆明小鼠40只,体重(16±1)g,随机分成高、中、低3个试验组和生理盐水对照组,每组10只,雌雄各半,分笼饲养。试验前禁食12 h,不限制饮水。由于本发明中药分散颗粒剂水溶解液最大浓度和小鼠最大灌胃体积的限制,最大给药量为60000 mg/kg BW,设置高、中、低试验组灌胃剂量分别为60000 mg/kg BW、30000 mg/kg BW、15000 mg/kg BW,生理盐水对照组灌服相同体积的生理盐水,灌胃后正常饲喂,观察7天。观察项目指标有:小鼠行为动作变化,呼吸变化,皮肤和被毛变化,进食饮水量变化,精神状态等。记录各组小鼠死亡情况,以未出现死亡的最高剂量为最大耐受量(MTD),并计算最大耐受量倍数。
计算公式:
最大耐受量倍数=(小鼠最大耐受量/小鼠平均体重)×(小鼠平均体重/畜禽每次用量)
2.3 脏器指数的测定
将MTD试验小鼠在试验结束后将所有小鼠处死,剖检,分离心脏、肾脏、肝脏和脾脏,称重各脏器重量,记录并计算各脏器指数。
计算公式:脏器指数=脏器重量/小鼠体重
2.4 数据处理
用SPSS 17.0对数据进行处理,数据用平均数±标准差表示。
2.5 结果与分析
在LD50测定试验中,各组小鼠在灌胃7天后,均未出现死亡,故进行MTD试验。MTD试验中,各组小鼠在灌胃7天后,小鼠死亡情况见表3,小鼠各脏器指数见表4。
表3各组小鼠在MTD试验中死亡情况
组别 | 动物数量(只) | 给药剂量(mg/kg BW) | 死亡数(只) |
高剂量组 | 10 | 60000 | 0 |
中剂量组 | 10 | 30000 | 0 |
低剂量组 | 10 | 15000 | 0 |
生理盐水对照组 | 10 | — | 0 |
表4 MTD试验小鼠主要脏器指数
组别 | 心脏指数 | 肝脏指数 | 肾脏指数 | 脾脏指数 |
高剂量组 | 6.57±1.31 | 43.6±5.23 | 14.5±2.14 | 2.48±0.12 |
中剂量组 | 7.12±2.06 | 42.8±4.62 | 14.7±3.08 | 3.14±0.67 |
低剂量组 | 6.75±1.49 | 43.1±5.20 | 15.6±2.27 | 2.71±0.82 |
生理盐水对照组 | 6.44±1.17 | 43.5±4.51 | 15.2±2.51 | 2.83±0.54 |
从表3结果可知,试验组、对照组小鼠均未出现死亡,确定发明分散颗粒剂对小鼠最大耐受量即为最大给药量60000mg/kg BW,按照最大耐受量倍数公式计算最大耐受量倍数为150倍,表明本发明分散颗粒剂在正常用量范围内,对畜禽安全性较高,实际无毒级药物。
观察小鼠结果显示,灌胃操作0.5h后,试验组小鼠呼吸频率加快,被毛竖立,身体微颤;生理盐水对照组小鼠呼吸频率加快,呆滞。1.5h后,试验组小鼠蜷缩成堆,精神萎靡,身体震颤消失;生理盐水对照组小鼠开始进食,动作迟缓。1d后,高剂量组小鼠开始进食饮水,精神状态好转,粪便不成形;中、低剂量组和生理盐水对照组小鼠各指标恢复正常。2d后,高剂量组小鼠粪便及其他指标均恢复正常。观察7d各组小鼠均未出现死亡,处死剖检各主要脏器,未见脏器异常。小鼠主要脏器指数结果显示,本发明分散颗粒剂高、中、低剂量组小鼠主要脏器指数与生理盐水对照组相比差异不显著,表明分散颗粒未引起小鼠脏器改变。以上结果说明,本发明中药分散颗粒制剂安全无毒,对机体没有损害,可以放心在养殖中使用。
3 本发明中药分散颗粒剂对畜禽特异性免疫功能的影响
3.1 材料
3.1.1 试验动物:180只1日龄AA肉鸡;30头30日龄三元断奶猪仔。
3.1.2 试验药品:质量分数为20%的本发明中药分散颗粒剂溶液;盐酸左旋咪唑片。
3.2试验方法
3.2.1肉鸡的分组和免疫器官指数、新城疫抗体效价的测定
1日龄AA肉鸡180只,随机分为3组,既Ⅰ组为空白对照组(生理盐水),Ⅱ组为盐酸左旋咪唑组(10mg/kg),Ⅲ组为本发明中药分散颗粒剂组(质量分数为20%的水溶液),每组3个重复,每个重复10只。各组在7日龄进行新城疫首免,新城疫Ⅳ弱毒苗2倍量点眼滴鼻;21日龄进行新城疫再免,新城疫Ⅳ弱毒苗4倍量饮水。各组在5-30日龄按剂量饮水给药,其余时间饲喂正常饮用水,试验期为42天,饲养条件和环境保持一样。在饲养至32日龄和42日龄时,每组随机选10只鸡,每只翅静脉采血1mL,静置,收集血清,用于检测新城疫的抗体效价。检测方法参照国标鸡新城疫抗体检测技术GBT 16550-2008)进行操作。在饲养至42日龄时,放血致死全部鸡只,取法氏囊、胸腺、脾脏,剔除脂肪,称重,记录。计算法氏囊、胸腺、脾脏指数。
免疫器官指数(g/kg)=免疫器官重量(mg)/动物体重(g)
3.2.2三元断奶猪仔的分组及猪瘟疫苗免疫效果
30日龄三元断奶猪仔30只,随机分为三组,既Ⅰ组为空白对照组(生理盐水),Ⅱ组为盐酸左旋咪唑组(10mg/kg),Ⅲ组为本发明中药分散颗粒剂组(质量分数为20%的水溶液),每组10只。32日龄开始各组按剂量饮水给药,连续喂养15天,各组分别在35日龄时注射猪瘟活疫苗正常剂量免疫接种。仔猪于疫苗接种后的第36、40、45、50、55日龄分别对各组仔猪耳静脉采血5mL,静置,收集血清,采用酶标分析仪检测猪瘟血清抗体水平。
3.3结果与讨论
3.3.1本发明中药分散颗粒剂对肉鸡免疫器官的影响
各组肉鸡免疫器官的检测结果见表5。Ⅱ组和Ⅲ组的法氏囊指数、脾脏指数和胸腺指数均显著高于Ⅰ组(P<0.05),并且Ⅲ组的各个免疫器官指数相较于Ⅱ组也有些许提高,差异显著(P<0.05)。
表5本发明中药分散颗粒剂对肉鸡免疫器官的影响
组别 | 法氏囊指数 | 脾脏指数 | 胸腺指数 |
Ⅰ组 | 1.62±0.01<sup>a</sup> | 1.16±0.03<sup>a</sup> | 1.80±0.10<sup>a</sup> |
Ⅱ组 | 1.74±0.02<sup>b</sup> | 1.24±0.02<sup>b</sup> | 2.26±0.04<sup>b</sup> |
Ⅲ组 | 1.86±0.01<sup>c</sup> | 1.36±0.02<sup>c</sup> | 2.34±0.07<sup>c</sup> |
注:同一列标有不同字母表示差异显著(P<0.05)。
家禽的中枢免疫器官有骨髓、胸腺和法氏囊,该类器官诱导免疫活性细胞的产生、增殖、分化和成熟,调节外周免疫器官发育和动物整体的免疫反应。而外周免疫器官包括脾脏、骨髓和粘膜相关淋巴组织等,是淋巴细胞聚集和发生免疫应答的场所,胸腺是不成熟T细胞发育为成熟T细胞的场所,法氏囊是B细胞分化成熟的场所,脾脏含有大量的T淋巴细胞和B淋巴细胞。免疫器官是动物实现免疫防御、免疫调节的中药场所,所以免疫器官指数的大小与肉鸡的健康密切相关。从表5结果可知,本发明中药分散颗粒剂对肉鸡的免疫器官指数有积极影响,可以较为明显的提升被感染肉鸡的免疫器官指数,说明本发明中药分散颗粒剂能够很好的促进机体免疫器官的发育,增强免疫器官的免疫能力,使其具备良好的抵抗病院微生物感染的物质基础。
3.3.2本发明中药分散颗粒剂对肉鸡新城疫抗体效价的影响
32日龄肉鸡新城疫抗体效价测定结果比较,Ⅱ组和Ⅲ组的抗体效价明显高于Ⅰ组,差异及其显著(P<0.05);42日龄肉鸡新城疫抗体效价,Ⅱ组和Ⅲ组的抗体效价明显高于Ⅰ组,差异极其显著(P<0.05),且Ⅲ组的抗体效价明显高于Ⅱ组,差异显著(P<0.05)。结果详见表6
表6本发明中药分散颗粒剂对肉鸡新城疫抗体效价的影响
类别 | Ⅰ组 | Ⅱ组 | Ⅲ组 |
32日龄鸡 | 4.91±0.61<sup>a</sup> | 5.53±0.72<sup> b</sup> | 5.79±0.32<sup> b</sup> |
42日龄鸡 | 5.20±0.41<sup> a</sup> | 6.52±0.51<sup>b</sup> | 7.13±0.26<sup>b</sup> |
注:同一行标有不同字母表示差异显著(P<0.05)。
由表6可以看出,32日龄Ⅱ组和Ⅲ组中新城疫的抗体水平与Ⅰ组对比,随着疫苗的接种明显增加,其中Ⅲ组中血清中新城疫抗体水平为最高。42日龄Ⅱ组和Ⅲ组中新城疫的抗体水平仍在持续上升,且Ⅲ组中血清中新城疫抗体水平仍为最高,说明本发明中药分散颗粒剂能够对提高鸡体抗NDV血凝抑制抗体效价,而且本发明中药分散颗粒剂在维持鸡体新城疫抗体价高水平持续时间较长,优于左旋咪唑组。
3.3.3本发明中药分散颗粒剂对仔猪猪瘟疫苗免疫效果的影响
从表7可知,在试验期间,Ⅱ组和Ⅲ组的抗体水平均高于Ⅰ组,差异显著(P<0.05)。45日龄时,各组中抗体水平达到最高,其中Ⅲ组的抗体水平最高;50日龄时,各组抗体水平有不同程度的下降,但是Ⅲ组的抗体水平仍然维持在较高的水平,与Ⅰ组和Ⅱ组相比差异显著(P<0.05)。以上结果可以说明,本发明中药分散颗粒剂能够有效提高猪瘟抗体水平,并且可以保持较长的抗体高峰期维持时间。
表7本发明中药分散颗粒剂对仔猪猪瘟疫苗免疫效果的影响
日龄 | Ⅰ组 | Ⅱ组 | Ⅲ组 |
32日 | 0.35± 0.05<sup> a</sup> | 0.46±0.02<sup> b</sup> | 0.46±0.03<sup> b</sup> |
40日 | 0.42± 0.04<sup> a</sup> | 0.52±0.05<sup>b</sup> | 0.59±0.06<sup>b</sup> |
45日 | 0.49±0.07<sup> a</sup> | 0.65±0.03<sup> b</sup> | 0.72±0.04<sup> c</sup> |
50日 | 0.38±0.01<sup> a</sup> | 0.58±0.02<sup> b</sup> | 0.64±0.01<sup> c</sup> |
55日 | 0.34±0.02<sup> a</sup> | 0.55±0.06<sup> b</sup> | 0.59±0.06<sup> b</sup> |
注:同一列标有不同字母表示差异显著(P<0.05)。
实施例2
本实施例与实施例1的区别在于
原料(按重量份数计):大黄115份、甘草38份、黄芩115份、丹参192份、刺五加38份、天花粉115份、青蒿36份、荆芥38份、麦冬115份、板蓝根52份、白术60份、滑石115份、聚乙烯吡咯烷酮140份、乳糖290份、碳酸氢钠60份、柠檬酸70份、猪乳香0.5份。
实施例3
本实施例与实施例1的区别在于
原料(按重量份数计):大黄125份、甘草42份、黄芩125份、丹参208份、刺五加42份、天花粉125份、青蒿40份、荆芥42份、麦冬125份、板蓝根58份、白术66份、滑石125份、聚乙烯吡咯烷酮150份、乳糖300份、碳酸氢钠70份、柠檬酸80份、猪乳香1份。
实施例4
本实施例与实施例1的区别在于
原料(按重量份数计):大黄120份、甘草40份、黄芩120份、丹参200份、刺五加40份、天花粉120份、青蒿38份、荆芥40份、麦冬120份、板蓝根56份、白术64份、滑石120份、聚乙烯吡咯烷酮145份、乳糖295份、碳酸氢钠65份、柠檬酸75份、猪乳香0.8份。
实施例5
本实施例与实施例1的区别在于
原料(按重量份数计):大黄118份、甘草42份、黄芩122份、丹参196份、刺五加38份、天花粉115份、青蒿40份、荆芥42份、麦冬124份、板蓝根55份、白术62份、滑石116份、聚乙烯吡咯烷酮142份、乳糖296份、碳酸氢钠68份、柠檬酸72份、猪乳香0.6份。
实施例6
本实施例与实施例1的区别在于
原料(按重量份数计):大黄122份、甘草40份、黄芩116份、丹参202份、刺五加38份、天花粉116份、青蒿40份、荆芥39份、麦冬118份、板蓝根52份、白术66份、滑石115份、聚乙烯吡咯烷酮148份、乳糖298份、碳酸氢钠66份、柠檬酸76份、猪乳香1份。
实施例7
本实施例与实施例1的区别在于
原料(按重量份数计):大黄116份、甘草40份、黄芩124份、丹参206份、刺五加42份、天花粉122份、青蒿38份、荆芥42份、麦冬124份、板蓝根54份、白术64份、滑石116份、聚乙烯吡咯烷酮148份、乳糖292份、碳酸氢钠62份、柠檬酸78份、猪乳香0.9份。
以上各实施例仅用以说明本发明的技术方案,而非对其限制;尽管参照前述各实施例对本发明进行了详细的说明,本领域的普通技术人员应当理解:其依然可以对前述各实施例所记载的技术方案进行修改,或者对其中部分或者全部技术特征进行等同替换;而这些修改或者替换,并不使相应技术方案的本质脱离本发明各实施例技术方案的范围。
Claims (3)
1.一种用于预防畜禽疫病的中药分散制剂,其特征在于,所述中药分散制剂由以下重量份数的原料制成:大黄115-125份、甘草38-42份、黄芩115-125份、丹参192-208份、刺五加38-42份、天花粉115-125份、青蒿36-40份、荆芥38-42份、麦冬115-125份、板蓝根52-58份、白术60-66份、滑石115-125份;所述中药分散制剂还包括以下重量份数的辅料:聚乙烯吡咯烷酮140-150份、乳糖290-300份、碳酸氢钠60-70份、柠檬酸70-80份、猪乳香0.5-1份。
2.根据权利要求1所述一种用于预防畜禽疫病的中药分散制剂,其特征在于,所述中药分散制剂由以下重量份数的原料制成:大黄120份、甘草40份、黄芩115份、丹参204份、刺五加38份、天花粉118份、青蒿36份、荆芥42份、麦冬122份、板蓝根55份、白术65份、滑石115份。
3.如权利要求1至2任一项所述的一种预防畜禽疫病的中药分散制剂的制备方法,其特征在于,所述制备方法按照以下步骤制备:
(1)分别准确按照重量份数称取:大黄、甘草、黄芩、丹参、刺五加、天花粉、青蒿、荆芥、麦冬、板蓝根、白术、滑石;
(2)将步骤(1)中草药原料除滑石外全部投入中药提取机中,随后加入各原料重量份数之和的10-15倍量水,煎煮30-40min后加入滑石,继续煎煮30-40min,过滤收集滤液,在药渣中再次加入各原料重量份数之和的8-10倍水,继续煎煮60-70min,过滤收集滤液,将两次收集的滤液合并,进行喷雾干燥,得到混合物A;
(3)分别准确按照重量份数称取:聚乙烯吡咯烷酮、乳糖、碳酸氢钠、柠檬酸、猪乳香,其中乳糖、碳酸氢钠和柠檬酸分别粉碎过100-200目药筛;将上述各辅料混合得到混合物B;
(4)将混合物A和混合物B投入到混合机组,混合45min,得到混合物C;
(5)将混合物C投入到颗粒剂干法压片机中,经过压片、粉碎、过1号药筛、整粒,即得本发明预防畜禽疫病的中药分散颗粒剂。
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