CN104367673B - 一种治疗鸡球虫的中药组合物 - Google Patents
一种治疗鸡球虫的中药组合物 Download PDFInfo
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- CN104367673B CN104367673B CN201410655531.4A CN201410655531A CN104367673B CN 104367673 B CN104367673 B CN 104367673B CN 201410655531 A CN201410655531 A CN 201410655531A CN 104367673 B CN104367673 B CN 104367673B
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Abstract
本发明涉及一种治疗鸡球虫的中药组合物,原料药组分如下:青蒿、白头翁、常山、苦参、仙鹤草、黄连、大黄、丹参、地榆炭、白芍、枳壳、黄芪、甘草。本发明制备的治疗鸡球虫的中药组合物抗球虫效果良好,能促进肠道快速修复,从根本上治疗球虫病。且将有可能取代饲料中添加的抗球虫药及其他抗生素,为解决肉鸡产品中的药物残留问题提供了可能。
Description
技术领域
本发明涉及一种治疗鸡球虫的中药组合物,属于兽药技术领域。
背景技术
鸡球虫病是一种原虫病,是由寄生于鸡肠上皮细胞内的艾美耳属多种球虫所引起的。该病发生率高,分布范围广,危害严重,其病死率可达40%~80%。目前,化学合成药物和抗生素类药物仍是养鸡生产中控制鸡球虫病的主要手段。虽然这些药物对鸡球虫病的防治起到了积极的重要作用,但同时也促使鸡群产生耐药性等问题。此外,这些化学药物及抗生素的使用不可避免地会在畜产品和环境中残留,从而对人类健康造成危害。而且还降低了这些产品的质量和安全性,影响了其外贸竞争力。
相对于化学合成类抗球虫药而言,中药的毒副作用小,药物残留低,使用较安全,且球虫对其不易产生耐药性,长期使用不蓄积。不仅有抑杀球虫的作用,而且还能增强鸡体的免疫功能,促进鸡的生长发育,提高增重率和生产性能等。近几年,科研工作者在认真分析和研究古方的基础上,采用提纯分离有效成分,验证成方或自拟处方等,对鸡球虫病进行防治试验,积累了许多宝贵经验。
在单味用药方面,赵爱莲等用蛹虫草治疗人工感染球虫病,抗球虫指数达174,其抗球虫效果比咪唑苯尿、氯苯胍好。刘其容用常山加水煎汁拌料喂鸡,严重者用滴管灌服,用药3d后,血粪基本停止,球虫病得到控制,雏鸡口服常山0.5~0.7g是安全的,可用煎剂或粉剂。在提取中草药有效成分治疗鸡球虫病方面,Youn等提取了柴胡、苦参、白头翁、钩藤等中药的有效成分防治球虫病,发现多种成分有较强抑杀球虫的作用,与对照组相比,用药组的肠道病变、出血、卵囊的排出等显著下降,体重显著增加。El-Abasy.M.等用中药甘蔗提取物(SCE)进行球虫病的防治试验,结果口服SCE组与阳性对照组相比,可以显著改善增重率,降低血便记分和卵囊排出量。还可以缓解因为球虫病感染而引起的盲肠扁桃体内CD4细胞的下降,表明SCE对鸡感染E.tenella有免疫刺激和保护作用。
中国专利文献CN102100788A(申请号200910255666.0)公开了一种预防及治疗鸡球虫病的中药制剂。本发明的药物是青蒿、常山、大黄、苦参、黄芪、黄柏、当归、白头翁、地榆、仙鹤草、金银花等中药为原料,将上述原料按重量份数配好用量,混匀,粉碎过80目筛,制备得到的中药制剂。
发明内容
本发明针对现有技术的不足,提供一种治疗鸡球虫的中药组合物。
一种治疗鸡球虫的中药组合物,原料药组分如下,均为重量份:
青蒿27~33份、白头翁27~33份、常山21~29份、苦参21~29份、仙鹤草16~24份、黄连16~24份、大黄11~19份、丹参11~19份、地榆炭11~19份、白芍5~15份、枳壳5~15份、黄芪5~15份、甘草3~13份。
根据本发明优选的,原料药组分如下,均为重量份:
青蒿30份、白头翁30份、常山25份、苦参25份、仙鹤草20份、黄连20份、大黄15份、丹参15份、地榆炭15份、白芍10份、枳壳10份、黄芪10份、甘草8份。
一种治疗鸡球虫的中药口服液的制备方法,包括如下步骤:
(1)将上述除白头翁以外的原料药与水混合,料水质量比为1:(12~18),加热至沸腾,然后加入白头翁,保持微沸0.8~1.5h,过滤,制得一次药液,药渣与水混合,料水质量比为1:(10~15),加热至沸腾,保持微沸0.6~1h,过滤,制得二次药液,合并一次药液和二次药液,制得药液;
(2)将步骤(1)制得的药液浓缩至相对密度≥1.05,然后加入乙醇溶液,使乙醇的体积浓度达到65%~70%,0~4℃静置20~28小时,经过滤,取上清液,制得一次醇沉液,将过滤的沉淀加入到沉淀的6~10倍体积的乙醇溶液中,乙醇的体积浓度为65%~70%,0~4℃静置10~14小时,过滤,取上清液,制得二次醇沉液,合并一次醇沉液和二次醇沉液,去除乙醇,制得治疗鸡球虫的中药口服液。
根据本发明优选的,所述步骤(2)中的乙醇溶液为体积浓度95%的乙醇溶液。
根据本发明优选的,所述步骤(2)去除乙醇后,还包括添加防腐剂苯甲酸钠的步骤。
药理分析
本发明根据中草药的四气五味及归经、主治等理论,合理配伍,结合中兽医辨证论治的防治法则,根据鸡病的不同情况,选药组方,确定诸药间的最佳比例,制成剂型,调整相关脏腑的功能或由疾病所损伤的其他脏腑,从而达到标本兼治扶正祛邪的目的。
方中青蒿与白头翁性味苦辛、寒,清热解毒,退热杀虫,凉血止痢为君药。
青蒿:主要有效成分有青蒿素(artem isin in),青蒿烯,青蒿醇,黄酮类,香豆精类,挥发油,棕榈酸等。青蒿素是继氯喹、乙氨嘧啶、伯喹和磺胺后最热门的抗疟特效药。
白头翁:白头翁的化学成分主要有三萜及其苷、白头翁素、白头翁灵、白头翁因等强心成分和豆甾醇、β-谷甾醇等。主要成分均可溶于水,所以可以采取煎煮提取有效成分。
常山性味苦辛寒,杀虫,祛痰消积;仙鹤草性味苦涩平,收敛止血,止痢,解毒;苦参性味苦寒,清热燥湿,杀虫去积,利水。以上三药合并为臣药。
常山:主要含有黄酮类、挥发油类、生物碱类、苯丙素类和糖苷类等几大类化学成分。主要成分均可溶于水,所以可以采取煎煮提取有效成分。
苦参:主要有效成分有生物碱类、黄酮类、脂肪酸类、氨基酸类、挥发油类等化合物,主要成分均可溶于水,所以可以采取煎煮提取有效成分。
仙鹤草:为蔷薇科多年生草本植物龙牙草的全草,异名脱力草、马鞭草、龙牙草、石打穿等。始载于滇南草本,主要成分含有仙鹤草素、仙鹤草内酯、鞣质、甾醇、有机酸、酚性成分、皂甙、仙鹤草酚(A、B、C、D)、维生素C、K等。主要成分均可溶于水,所以可以采取煎煮提取有效成分。
黄连性味苦寒,清热燥湿,泻火解毒;大黄与丹参苦寒,泻热通肠,凉血解毒,破积行淤,通经止痛;地榆炭性味苦酸、微寒,凉血解毒,止血敛疮。以上四药并合为佐药。
大黄:大黄为蓼科草本植物,主要含有大黄素、大黄酚苷、大黄酚、没食子酸、香草酸、胡罗卜苷、油酸、三油酸甘油酯、四环三萜苦木内酯类等。主要成分均可溶于水,所以可以采取煎煮提取有效成分。主要成分均可溶于水,所以可以采取煎煮提取有效成分。
丹参:主要含有丹参酮、丹酚酸、丹参单体IH764-3等多种类型的化合物。主要成分均可溶于水,所以可以采取煎煮提取有效成分。
黄连:其主要成分有:黄连素、黄连碱、甲基黄连碱、掌叶防己碱、药根碱、非洲防己碱、黄柏酮、黄柏内酯、木兰花碱、阿魏酸等。主要成分均可溶于水,所以可以采取煎煮提取有效成分。
地榆:主要有效成分三萜及其皂甙类、鞣质和黄酮等。主要成分均可溶于水,所以可以采取煎煮提取有效成分。
白芍:主要含有芍药甙、芍药花甙,牡丹酚、苯甲酸、挥发油、脂肪油、树脂、鞣质、糖、淀粉、粘液质、蛋白质、β-谷甾醇和三萜类等。因主要成分均可溶于水,所以可以采取煎煮提取有效成分。
枳壳:主要含有挥发油和黄酮甙等物质。玳玳花未成熟果实的果皮中含新橙皮甙;果实成熟时,新橙皮甙消失而柚皮甙增多,接近成熟的酸橙果实中含维生素C,从香圆和枸橘中分离得同一种生物碱样物质。因主要成分均可溶于水,所以可以采取煎煮提取有效成分。
黄芪:主要含有黄芪皂苷类、黄芪多糖类、黄酮类、氨基酸类及少量微量元素。因主要成分均可溶于水,所以可以采取煎煮提取有效成分。
甘草:主要含甘草甜素,系甘草酸的钙盐,钾盐。甘草酸水解后产生葡萄糖醛酸,甘草次酸。还含黄酮类化合物甘草甙,甘草甙元,异甘草甙,异甘草甙元及甘草利酮,芒柄黄花素,甘草素,异甘草素,树脂等。因主要成分均可溶于水,所以可以采取煎煮提取有效成分。
有益效果
本发明制备的治疗鸡球虫的中药组合物抗球虫效果良好,能促进肠道快速修复,从根本上治疗球虫病。且将有可能取代饲料中添加的抗球虫药及其他抗生素,为解决肉鸡产品中的药物残留问题提供了可能。
具体实施方式
下面结合实施例对本发明的技术方案做进一步说明,但本发明所保护范围不限于此。
实施例1
一种治疗鸡球虫的中药组合物,原料药组分如下,均为重量份:
青蒿30份、白头翁30份、常山25份、苦参25份、仙鹤草20份、黄连20份、大黄15份、丹参15份、地榆炭15份、白芍10份、枳壳10份、黄芪10份、甘草8份。
将上述治疗鸡球虫的中药组合物制备成中药口服液的方法,包括如下步骤:
(1)将上述除白头翁以外的原料药与水混合,料水质量比为1:15,加热至沸腾,然后加入白头翁,保持微沸1h,过滤,制得一次药液,药渣与水混合,料水质量比为1:12,加热至沸腾,保持微沸0.75h,过滤,制得二次药液,合并一次药液和二次药液,制得药液;
(2)将步骤(1)制得的药液浓缩至相对密度≥1.05,然后加入乙醇溶液,使乙醇的体积浓度达到65%,0℃静置24小时,经过滤,取上清液,制得一次醇沉液,将过滤的沉淀加入到沉淀的8倍体积的乙醇溶液中,乙醇的体积浓度为65%,0℃静置12小时,过滤,取上清液,制得二次醇沉液,合并一次醇沉液和二次醇沉液,去除乙醇,制得治疗鸡球虫的中药口服液。
所述步骤(2)中的乙醇溶液为体积浓度95%的乙醇溶液;
所述步骤(2)去除乙醇后,还包括添加防腐剂苯甲酸钠的步骤。
实施例2
一种治疗鸡球虫的中药组合物,原料药组分如下,均为重量份:
青蒿33份、白头翁27份、常山29份、苦参21份、仙鹤草24份、黄连16份、大黄19份、丹参11份、地榆炭19份、白芍5份、枳壳15份、黄芪5份、甘草13份。
将上述治疗鸡球虫的中药组合物制备成中药口服液的方法,包括如下步骤:
(1)将上述除白头翁以外的原料药与水混合,料水质量比为1:18,加热至沸腾,然后加入白头翁,保持微沸1.5h,过滤,制得一次药液,药渣与水混合,料水质量比为1:15,加热至沸腾,保持微沸1h,过滤,制得二次药液,合并一次药液和二次药液,制得药液;
(2)将步骤(1)制得的药液浓缩至相对密度≥1.05,然后加入乙醇溶液,使乙醇的体积浓度达到70%,4℃静置28小时,经过滤,取上清液,制得一次醇沉液,将过滤的沉淀加入到沉淀的10倍体积的乙醇溶液中,乙醇的体积浓度为70%,4℃静置14小时,过滤,取上清液,制得二次醇沉液,合并一次醇沉液和二次醇沉液,去除乙醇,制得治疗鸡球虫的中药口服液。
所述步骤(2)中的乙醇溶液为体积浓度95%的乙醇溶液;
所述步骤(2)去除乙醇后,还包括添加防腐剂苯甲酸钠的步骤。
实施例3
一种治疗鸡球虫的中药组合物,原料药组分如下,均为重量份:
青蒿27份、白头翁33份、常山21份、苦参29份、仙鹤草16份、黄连24份、大黄11份、丹参19份、地榆炭11份、白芍15份、枳壳5份、黄芪15份、甘草3份。
将上述治疗鸡球虫的中药组合物制备成中药口服液的方法,包括如下步骤:
(1)将上述除白头翁以外的原料药与水混合,料水质量比为1:12,加热至沸腾,然后加入白头翁,保持微沸0.8h,过滤,制得一次药液,药渣与水混合,料水质量比为1:10,加热至沸腾,保持微沸0.6h,过滤,制得二次药液,合并一次药液和二次药液,制得药液;
(2)将步骤(1)制得的药液浓缩至相对密度≥1.05,然后加入乙醇溶液,使乙醇的体积浓度达到68%,0℃静置20小时,经过滤,取上清液,制得一次醇沉液,将过滤的沉淀加入到沉淀的6倍体积的乙醇溶液中,乙醇的体积浓度为68%,0℃静置10小时,过滤,取上清液,制得二次醇沉液,合并一次醇沉液和二次醇沉液,去除乙醇,制得治疗鸡球虫的中药口服液。
所述步骤(2)中的乙醇溶液为体积浓度95%的乙醇溶液;
所述步骤(2)去除乙醇后,还包括添加防腐剂苯甲酸钠的步骤。
对比例1
一种治疗鸡球虫的中药组合物,原料药组分如下,均为重量份:
青蒿12份、常山15份、大黄4份、苦参8份、黄芪6份、黄柏8份、当归8份、白头翁14份、地榆9份、仙鹤草6份、金银花4份。
将上述治疗鸡球虫的中药组合物制备成中药口服液的方法,包括如下步骤:
(1)将上述除白头翁以外的原料药与水混合,料水质量比为1:15,加热至沸腾,然后加入白头翁,保持微沸1h,过滤,制得一次药液,药渣与水混合,料水质量比为1:12,加热至沸腾,保持微沸0.75h,过滤,制得二次药液,合并一次药液和二次药液,制得药液;
(2)将步骤(1)制得的药液浓缩至相对密度≥1.05,然后加入乙醇溶液,使乙醇的体积浓度达到65%,0℃静置24小时,经过滤,取上清液,制得一次醇沉液,将过滤的沉淀加入到沉淀的8倍体积的乙醇溶液中,乙醇的体积浓度为65%,0℃静置12小时,过滤,取上清液,制得二次醇沉液,合并一次醇沉液和二次醇沉液,去除乙醇,制得治疗鸡球虫的中药口服液。
所述步骤(2)中的乙醇溶液为体积浓度95%的乙醇溶液;
所述步骤(2)去除乙醇后,还包括添加防腐剂苯甲酸钠的步骤。
试验例1
稳定性试验
中药治疗疾病的原则为急则治其标,缓则治其本,标本并重则要标本兼治,且中药的治疗效果接近或优于西药,毒副作用小,无药物残留,不产生耐药性,还能调动机体各方面因素,提高机体的免疫力。稳定性试验的目的是考察中药在温度、湿度、光线、微生物的影响下随时间变化的规律。为中药的生产、包装、贮存、运输条件提供科学依据,同时根据试验结果建立药品的有效期。
本试验为考察实施例制备的口服液的化学或物理性质在温度、湿度、光线的影响下随时间的变化规律,同时进行加速试验、光加速试验,预测口服液的稳定性。
1.材料与方法
1.1 材料
1.1.1 实施例1-3制得的口服液供试品
实施例1-3制得的口服液,棕色至棕褐色液体,长久贮存或冷冻后有沉淀析出。青蒿素含量不低于2.50mg/ml,总糖含量(以葡萄糖计)不低于0.160g/ml,pH5.0~7.0,密度不低于1.00g/ml。
1.1.2 主要试剂
标准葡萄糖溶液、5%苯酚溶液、30%硫酸锌试液、15%亚铁氰化钾试液、浓硫酸。
青蒿素标准溶液配制:精确称取青蒿素5.00mg,用50%甲醇溶解,定容于50ml量瓶中,分别吸取0.5、1.0、2.0、4.0、6.0、8.0ml,50%甲醇稀释,定容于10ml量瓶,即得青蒿素标准溶液。
对照品溶液制备:精确称取青蒿素标准品适量,置棕色瓶中,加50%甲醇溶解,制成每ml含40ug的溶液,即得对照品溶液(10℃以下保存)。
供试品溶液制备:精确吸取实施例1-3制得的口服液25ml,加甲醇25ml,加热回流2h,过滤,滤液定容于500ml量瓶,精确吸取1ml,50%甲醇稀释,定容于25ml量瓶,即得供试品溶液。
流动相:乙腈-0.4%磷酸(23:87)。
1.1.3 主要仪器
T6新世纪紫外可见分光光度计,北京普析通用仪器有限责任公司;PHSJ-4A型试验室pH计,上海精密科学仪器有限公司;日本岛津LC-10ATvp高效液相色谱仪、SPD-10Avp可见紫外检测器、GSP-781型隔水式电热恒温培养箱、N-1330照度计、MP500B电子天平、MettlerAE-240分析天平。
1.2 方法
1.2.1 光加速试验
参照兽药稳定性试验技术规范(试行),取实施例1-3制得的口服液供试品三批,装入无色透明容器内,放置在光厨,于照度4500±500Lx的条件下放置十天,在第五、十天定时取样,按口服液稳定性重点考察项目进行检测,分别考虑口服液性状、青蒿素含量、色泽、澄清度、pH值、密度。
1.2.2 加速试验
取实施例1-3制得的口服液,按市售包装,在温度40±2℃条件下放置六个月,每一个月取样一次,按口服液稳定性重点考察项目进行检测,分别考虑口服液性状、口服液总糖含量、色泽、澄清度、pH值、密度。
2.结果
2.1 光加速试验
试验结果表明:实施例1-3制得的口服液在照度4500±500Lx的条件下放置十天,其性状、色泽、澄清度、pH值、密度均在质量标准范围内。
含量在第五天1.72mg/ml,第十天为1.56mg/ml,超出了实施例1-3制得的口服液质量标准范围,表明实施例1-3制得的口服液对光不稳定,应避光贮存,结果见表1-1。
2.2 加速试验
试验结果表明:实施例1-3制得的口服液在40±2℃条件下放置六个月,其性状、色泽、澄清度、pH值、密度均在质量标准范围内,5~6月有轻摇易散的沉淀析出,口服液总糖含量(以葡萄糖计)下降了18.5%~19.2%,未超出青翁球康口服液质量标准范围,表明青翁球康口服液对热不稳定,应在阴凉条件下贮存,结果见表1-2。
表1-1 加速试验结果
表1-2 加速试验结果
3.结果分析
试验结果表明,在光加速第五天后,实施例1-3制得的口服液中青蒿素含量超出了制剂质量标准,其余各项指标变化不明显;在加速试验第5个月时,实施例1-3制得的口服液出现了轻摇易散的沉淀,总糖含量(以葡萄糖计)下降18.5%~19.2%,符合制定的质量标准,其余各项指标无明显变化。实施例1-3制得的口服液对光线、温度敏感,应在避光、阴凉处贮存。
试验例2
l.材料与方法
1.1 虫种的来源及传代试验用的球虫卵囊来源于聊城大学农学院临床教研室,经过滤、沉淀,2.5wt%重铬酸钾28℃保存,经3d孢子化后,经10日龄817肉鸡传代,再经孢子化后备用。
1.2 药物来源及使用浓度
试验用药:实施例1制得的口服液;
实施例1制得的口服液分别以0.15wt%,0.2wt%,0.25wt%混入饮水中施用。
1.3 试验动物 试验用动物为817肉鸡。来源于聊城市畜牧兽医研究所孵化厂。
1.4 试验动物的饲养及分组 1日龄817肉鸡饲养在消毒后的垫料上,饲喂聊城市康达饲料公司的肉雏鸡料。试验前显微镜检查粪便无球虫卵囊。于12日龄称重并随机分成6组,分别是:Ⅰ组为实施例1制得的口服液高剂量组;Ⅱ组为实施例1制得的口服液中剂量组;Ⅲ组为实施例1制得的口服液低剂量组;Ⅳ组为氨丙啉组;Ⅴ组为感染对照组(攻虫);Ⅵ组为健康对照组,Ⅶ组为对比例1制得的口服液高剂量组。每组15只,记录初始重。分组后各组分开饲养在消毒后的不同笼中。各组饮水器、料桶大小一致,便于观察各组饮水量、采食量。Ⅰ组、Ⅱ组和Ⅲ组分别以0.25wt%(实施例1高剂量),0.2wt%(实施例1中剂量),0.15wt%(实施例1低剂量)混入饮水中施用;Ⅳ(氨丙啉组)氨丙啉混入饮水中施用;感染不给药组(感染对照组)和不感染不给药组(健康对照组),饮用未加药的饮水;Ⅶ组以0.25wt%(对比例1高剂量)混入饮水中施用。对每组饲料作称重记录,健康对照组一切用具和其他各组严格分开,以防止健康对照组被试验球虫感染。
1.5 小肠球虫混合卵囊接种14日龄时,4个给药组、阴性对照组的每只鸡分别经口接种小肠球虫混合卵囊1.0×105个。除健康对照组和感染对照组外,其余各组当天按照1.4的用量开始用药。直至8天结束。
1.6 药效判定指标
1.6.1 临床表现
试验期间观察和记录试验鸡的精神、食欲及粪便排出情况。感染后第5d开始每天检查粪便,记录血粪情况,直至扑杀。从感染后第5d开始每天进行克粪便卵囊计数,连续三天,第8d剖检,重点检查肠道病变,根据Johnson的方法评分。
1.6.2 相对增重率
14日龄早晨空腹称重,记为初重。试验结束时称末重,计算相对增重率。相对增重率(%)=(感染用药组的平均增重/健康对照组的平均增重)×100%。
1.6.3 存活率
存活率=(存活只数/试验只数)×100%。
1.6.4 血便记分
接种卵囊后第5、6、7、8d,参考Morehouse的方法,按以下标准记分:正常粪便0分;少量血便(1%~25%)1分;中等量血便(25%~50%)2分;许多血便(50%~75%)3分;无正常粪便4分。0分不作记录次数计。平均粪便记分=每组鸡粪便记分之和/记录次数。
1.6.5 病变计分
按Johnson等的方法,做鸡的肠道病变记分。病变值=每组平均病变记分×10。摘取肠道,剪开洗净。根据剖检后肠道病变的严重程度记分。无病变记0分;轻微病变者记1分;肠壁增厚,有小到中等出血记2分;内有很多出血块记3分;因球虫病而死亡记4分。
1.6.6 卵囊值
感染后第5~7d,收集粪便,计算每克粪便中的卵囊数(O.P.G)。以第六天的卵囊数为标准,计算卵囊值,方法见角田清的标准换算。即感染后的O.P.G值与感染对照组的O.P.G值的比值小于1%,卵囊值为0;在1%~25%时,卵囊值为5;在25%~50%时,卵囊值为10;在50%~75%时,卵囊值为20;在75%~100%时,卵囊值为40。
1.6.7 抗球虫指数(ACI)
按照美国默克(Merck)公司的计计算公式,ACI=(存活率+相对增重率)一(病变值+卵囊值+血便计分)。ACI≤120为无效,ACI=120~160为中等有效,ACI=160~180为良好,ACI>180为优秀。
1.6.8 中药对球虫感染雏鸡血清乳酸脱氢酶和碱性磷酸酶活性的影响分别于感染后0,3,6,8d,每组随机抽取5只鸡,心脏采血,分离血清-20°保存,用于血清LDH和ALP活性的测定。测定方法采用酶联免疫吸附试验(ELISA),试剂盒购自联星泰安生物技术有限公司。
2.结果
2.1 临床症状
感染孢子化卵囊第4d,感染对照组有少部分雏鸡精神出现沉郁,两三成群聚堆,闭眼静立,地面上有零星白色或褐色的稀粪出现。用药组和正常对照组精神要好于感染对照组,偶见个别闭眼静立的鸡。第5天,除正常对照组外,所有组均能见到精神沉郁,闭眼呆立的鸡,鸡活动范围和食欲下降明显,特别是感染对照组和低剂量组扎堆的鸡很多,和正常对照组相比,食料与饮水剩余均明显增多,部分鸡表现为羽毛蓬乱,闭眼静立,地上见到咖啡色粘稠样稀粪。高剂量组无论在精神状态,还是采食量上,要比低剂量组强。第六天,鸡的精神状态和采食量没有好转的变化,特别是感染对照组的部分鸡精神沉郁,不愿动,极个别的鸡喘气,卧在一腢,见人后也不动。而高剂量和中剂量的鸡精神状态明显好于其他组。各组差别最大的是地上血便的数量,感染对照组最多,高剂量和中剂量组相对少一些。第7天,感染对照组的鸡死亡2只,精神状态变化不大。低剂量组的精神状态好于感染对照组,其他组的精神状态和采食量要有一定好转,地上的血便数量不如第6天多。
健康对照组雏鸡精神状态、饮食欲、粪便状态一直正常。
2.2试验前后各试验组体重的变化
试验前后各试验组鸡体重的变化见表2,和健康对照组相比,球虫感染后引起了试验鸡增重及增重率下降。实施例1制得的口服液高、中剂量组增重率及相对增重率明显高于感染对照组,特别是实施例1制得的口服液低剂量组与感染对照组相比,增重率及相对增重率也大于感染对照组。说明实施例1制得的口服液有防止感染球虫鸡体重下降的作用,且低剂量组的效果与氨丙啉组相当。
对比例1制得的口服液增重率及相对增重率高于感染对照组,但均低于实施例1组和氨丙啉组。说明实施例1的中药配伍是科学的。
表2 各试验组增重情况及增重率
2.3 血便计分
感染对照组从感染后第4d开始见血便,一直持续到试验结束。其他组在感染后第五天开始零星出现血便。感染后6~7天,各组血便数量均很多,但感染对照组血便程度最严重。实施例1制得的口服液高、中剂量组血便数量要少于与实施例1制得的口服液低剂量(见表3)。对比例1制得的口服液组在感染后第五天出现了血便,感染后6~7天,血便数量增多。
表3 试验鸡只的血便记分
2.4 卵囊数
感染后5~7d各组卵囊计数结果见表4。从表4可知,感染后第6天,卵囊排出数量最多,第7天开始下降。高剂量组卵囊排出数量一直低于其他试验组,感染对照组卵囊排出数量一直高于其他试验组。第6天,氨丙啉组排出的卵囊数量接近于青翁球康口服液的中剂量组,低于实施例1制得的口服液的低剂量组;但是到第7天氨丙啉组排出的卵囊数量接近于实施例1制得的口服液的低剂量组,而高于实施例1制得的口服液的中剂量组。对比例1制得的口服液卵囊排出数量接近于青翁球康口服液的中剂量组,低于实施例1制得的口服液的低剂量组和氨丙啉组。
表4 感染后不同天数各试验组卵囊计数结果
组别 | 感染后第5d | 感染后第6d | 感染后第7d |
Ⅰ组 | 2.29×104 | 8.67×104 | 2.01×104 |
Ⅱ组 | 4.02×104 | 3.52×105 | 5.15×104 |
Ⅲ组 | 1.38×105 | 4.58×105 | 6.34×104 |
Ⅳ组 | 1.32×105 | 3.86×105 | 8.90×104 |
Ⅴ组 | 6.08×105 | 8.60×105 | 3.48×105 |
Ⅶ组 | 6.12×104 | 8.89×105 | 9.87×104 |
2.5 病理变化
试验结束时(22日龄)处死各组试验鸡进行剖检。感染鸡球虫的主要病变分布在小肠和盲肠。从小肠外观看,在小肠前端和中段的浆膜面都有弥漫性出血点、出血斑和白色斑,部分肠段外观肿胀,颜色变红。从盲肠外边看,盲肠两侧或一侧肿胀或鼓气,部分盲肠空虚。剖解开小肠后,可以见到肠粘膜肿胀,内容物呈糜烂样的食糜状,用剪刀刮去,可看到肠壁变薄,有出血点。剖解开盲肠后,可以看到盲肠内的红色粪便。用药组肠道病变的程度从轻到重的顺序依次是实施例1制得的口服液高剂量组、中剂量组和低剂量组,其中实施例1制得的口服液高、中剂量组的病变较轻,氨丙啉组的病变较重。对比例1制得的口服液组病变要比实施例1制得的口服液高、中剂量组的病变重,但是比氨丙啉组的病变要稍轻,说明中药可以提高机体的免疫力。
2.6 抗球虫药效综合判定结果
4种抗球虫药试验组的存活率、相对增重率、卵囊值、病变值和抗球虫指数结果见表5。可以看出,实施例1制得的口服液高剂量组抗球虫指数大于160,达到良好。实施例1制得的口服液的中、低剂量组和氨丙啉组的抗球虫指数均小于160,但大于120,属于有效范围,但从数值上看,抗球虫指数从大到小分别是实施例1制得的口服液高剂量组、中剂量组、氨丙啉组和实施例1制得的口服液低剂量组。对比例1制得的口服液组的抗球虫指数小于160,大于120,属于有效范围。
表5 4种抗球虫药的抗球虫效果
2.7 试验药物对血清乳酸脱氢酶(LDH)活性的影响
LDH广泛存在于机体的组织器官中,各组织器官病变都可释放LDH至血液中,使其活性增强。在感染后3,6d,感染对照组鸡的血清LDH活性比正常组鸡的LDH分别提高了50.30%和84.01%,差异均达到极显著(p<0.01);Ⅰ组鸡的血清LDH活性与Ⅵ组鸡相比,Ⅰ组鸡的血清LDH活性在感染后3,6d分别升高了7.06%和19.59%,差异不显著(p>0.05);Ⅱ组鸡的血清LDH活性与Ⅵ组鸡相比,Ⅱ组鸡的血清LDH活性在感染后3,6d分别提高了8.51%和28.82%,差异不显著(p>0.05);Ⅲ组鸡的血清LDH活性与Ⅵ组鸡相比,Ⅲ组鸡的血清LDH活性在感染后3,6d分别提高了8.42%和40.76%,第六天差异显著(p<0.05);Ⅳ组鸡的血清LDH活性与Ⅵ组鸡相比,Ⅳ组鸡的血清LDH活性在感染后3,6d分别提高了21.69%和60.64%,第六天差异显著(p<0.05)。到第8天时,各组的LDH活性均开始下降(见表6)。对比例1制得的口服液组鸡的血清LDH活性与Ⅵ组鸡相比,对比例1组鸡的血清LDH活性在感染后3,6d分别提高了8.05%和38.36%,第六天差异显著(p<0.05)。
表6 试验药物对LDH活性的影响
2.8 试验药物对血清碱性磷酸酶(ALP)活性的影响
在试验期间,除Ⅵ组外,其他各组的血清ALP活性都明显提高,特别是在感染后第6天,ALP活性提高到最高值,Ⅴ组升高的幅度最大,达到初始量的164.92%。在第6天,Ⅰ组、Ⅱ组、Ⅲ组、Ⅳ和Ⅴ组分别比Ⅵ组提高了8.18%(p>0.05)、9.32%(p>0.05)、15.54%(p>0.05)、41.04%(p<0.05)。第8天,血清碱性磷酸酶的活性开始下降,说明肠道开始修复。对比例1制得的口服液组血清ALP活性比Ⅵ组提高了16.20%(p>0.05)。
表7 试验药物对ALP活性的影响
3、结果与讨论
实施例1制得的口服液与对比例1制得的口服液中主要药物皆具有驱虫、增强机体胃肠道功能等作用。但是通过上述研究数据可以看出,实施例1制得的口服液组在抗球虫、提高生产性能方面要显著好于对比例1制得的口服液组。根据中医辩证施治的原则,实施例1组方更加科学。两个组方中均含有青蒿、常山、大黄、苦参、黄芪、白头翁、地榆、仙鹤草8味药,与对比例1制得的口服液组组方相比,实施例1制得的口服液组方中白头翁加大了用量与青蒿二药为君药,去掉了黄柏、当归和金银花配伍了黄连、丹参、白芍和甘草。黄连性味苦寒,清热燥湿,泻火解毒;丹参苦寒,凉血解毒,破积行淤,通经止痛;白芍性凉,味苦酸,微寒,具有补血养血、平抑肝阳、柔肝止痛的功效。根据球虫的发病机理及病症表现,选取黄连、丹参着重用于血便的治疗,选取白芍主要针对肠粘膜进行修复,配伍甘草的目的是与黄芪共同发挥提高机体免疫力的作用。
Claims (5)
1.一种治疗鸡球虫的中药组合物,其特征在于,原料药组分如下,均为重量份:
青蒿27~33份、白头翁27~33份、常山21~29份、苦参21~29份、仙鹤草16~24份、黄连16~24份、大黄11~19份、丹参11~19份、地榆炭11~19份、白芍5~15份、枳壳5~15份、黄芪5~15份、甘草3~13份。
2.如权利要求1所述的治疗鸡球虫的中药组合物,其特征在于,原料药组分如下,均为重量份:
青蒿30份、白头翁30份、常山25份、苦参25份、仙鹤草20份、黄连20份、大黄15份、丹参15份、地榆炭15份、白芍10份、枳壳10份、黄芪10份、甘草8份。
3.一种权利要求1所述的治疗鸡球虫的中药组合物的制备方法,其特征在于,包括如下步骤:
(1)将除白头翁以外的原料药与水混合,料水质量比为1:(12~18),加热至沸腾,然后加入白头翁,保持微沸0.8~1.5h,过滤,制得一次药液,药渣与水混合,料水质量比为1:(10~15),加热至沸腾,保持微沸0.6~1h,过滤,制得二次药液,合并一次药液和二次药液,制得药液;
(2)将步骤(1)制得的药液浓缩至相对密度≥1.05,然后加入乙醇溶液,使乙醇的体积浓度达到65%~70%,0~4℃静置20~28小时,经过滤,取上清液,制得一次醇沉液,将过滤的沉淀加入到沉淀的6~10倍体积的乙醇溶液中,乙醇的体积浓度为65%~70%,0~4℃静置10~14小时,过滤,取上清液,制得二次醇沉液,合并一次醇沉液和二次醇沉液,去除乙醇,制得治疗鸡球虫的中药口服液。
4.如权利要求3所述的制备方法,其特征在于,所述步骤(2)中的乙醇溶液为体积浓度95%的乙醇溶液。
5.如权利要求3所述的制备方法,其特征在于,所述步骤(2)去除乙醇后,还包括添加防腐剂苯甲酸钠的步骤。
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