CN111358850B - 一种用于防治鸡大肠杆菌病的中药组合物及其制备方法 - Google Patents
一种用于防治鸡大肠杆菌病的中药组合物及其制备方法 Download PDFInfo
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Abstract
本发明属于中兽药技术领域,公开一种用于防治鸡大肠杆菌病的中药组合物及其制备方法。所述中药组合物由下列重量份的原料药制成:红景天50~70份、女贞子40~60份、积雪草20~40份、穿心莲20~30份、三颗针20~40份、十大功劳20~40份、地锦草20~40份、侧柏叶20~30份、佛手20~30份、甘草5~15份。重量份以g计、体积份以mL计,各原料药按照比例混合、粉碎,然后提取、除杂、浓缩,定容至1000体积份,制得口服液。本发明系纯中药制剂,安全、无毒副作用,无适口性问题,能提高大肠杆菌病成活率至65%以上且能改善增重。
Description
技术领域
本发明属于中兽药技术领域,具体涉及一种用于防治鸡大肠杆菌病的中药组合物及其制备方法。
背景技术
鸡大肠杆菌病是我国临床最常见、危害最大的细菌病之一,具有发病快、死亡率高、病程长的特点,给养鸡业造成了极大的危害。目前对于大肠杆菌病多采用化学抗菌药物治疗,但由于此类药物的过量使用,已导致大肠杆菌的耐药谱不断扩大,防治效果下降,同时抗菌药物在体内的残留,降低了肉蛋产品品质,影响了人类健康和公共卫生安全。抗大肠杆菌病的疫苗尽管不断研制并投放市场,但致病性大肠杆菌血清型种类繁多,造成实际预防效果低下,极少进入商业化应用。各类微生态、酸化剂、精油以及各种营养性添加剂等在实际生产中的抗大肠杆菌效果不尽人意。
同时,鸡养殖过程中减少抗生素使用,已逐渐成为国家强制性的刚性需求,一系列政策已在终端客户处落实,如鸡蛋和鸡肉中的抗生素残留成为各地食药监局重点监控项目。农业农村部自2018年05月份启动实施兽用抗菌药使用减量化行动以来,在各个省份进行科学合理使用抗菌药活动,全面推进抗菌药减量化活动。饲料禁抗也将于2020年7月正式实施。因此鸡养殖过程中替抗药物的研发日益成为各动保企业的研发热点。
中兽医肇源于经典理论指导,落地于临床实战检核,理实互参,长于知变化、重调节、着眼整体、平衡阴阳,正以其在感冒、腹泻、气血亏虚、脾胃虚弱、肠燥便秘等外感和内伤诸多领域的品类优势日益受到养殖业的认可和青睐,品类优势明显。
大量临床实践表明,中兽药对鸡大肠杆菌病具有较好的防治效果,并具有相对绿色安全、治愈率高、不良反应少、不易复发等特点,符合当今社会倡导的绿色养殖的行业要求,有着广阔的发展前景。然而,目前养殖业中使用的防治鸡大肠杆菌病的中兽药制剂,存在处方组成不科学、用药成本高、给药方式不合理、治疗效果不明显、易复发等弊端。
因此,迫切需要根据传统中兽医药理论,结合当前鸡大肠杆菌病的发病特征,筛选出疗效显著且临床使用性价比高的防治鸡大肠杆菌病的中兽药方剂。
发明内容
为克服现有技术中存在的不足之处,本发明的目的旨在提供一种用于防治鸡大肠杆菌病的中药组合物及其制备方法。
为实现上述目的,本发明采取的技术方案如下:
一种用于防治鸡大肠杆菌病的中药组合物,所述中药组合物由下列重量份的原料药制成:红景天50~70份、女贞子40~60份、积雪草20~40份、穿心莲20~30份、三颗针20~40份、十大功劳20~40份、地锦草20~40份、侧柏叶20~30份、佛手20~30份、甘草5~15份。
较好地,所述中药组合物由下列重量份的原料药制成:红景天60份、女贞子50份、积雪草30份、穿心莲25份、三颗针30份、十大功劳30份、地锦草30份、侧柏叶25份、佛手25份、甘草10份。
较好地,所述中药组合物的剂型为口服液。
制备方法:重量份以g计、体积份以mL计,红景天、女贞子、积雪草、穿心莲、三颗针、十大功劳、地锦草、侧柏叶、佛手、甘草按照比例混合、粉碎,然后提取、除杂、浓缩,定容至1000体积份,制得口服液。
较好地,重量份以g计、体积份以mL计,步骤如下:
(1)、按所述重量份数取各原料药,粉碎、过筛;
(2)、取原料药总重量6~10倍重量的水加热至80~85 ℃,加入步骤(1)中过筛后的原料药,恒温浸泡10~30 min;
(3)、将步骤(2)所得药液降温至45~50 ℃,调节pH至4.5~5.0,加入复合酶,搅拌均匀,酶解1~3 h,然后升温至75~80 ℃进行酶灭活处理;所述复合酶主要为纤维素酶、果胶酶和淀粉酶的混合物,其中,纤维素酶酶活以20000 U/g计,用量不小于1.0 重量份,果胶酶酶活以50000 U/g计,用量不小于1.0重量份,淀粉酶酶活以30000 U/g计,用量不小于1.0重量份;
(4)、将步骤(3)酶解后药液煎煮两次,合并两次煎煮滤液,离心处理得上清液;
(5)、将步骤(4)所得上清液进行微滤、超滤;
(6)、将步骤(5)超滤后滤液减压浓缩至室温下相对密度为1.04~1.07;
(7)、将步骤(6)减压浓缩后药液加水定容至1000体积份,然后灌装、灭菌即得口服液成品。
较好地,步骤(3)中,所述复合酶还含有半纤维素酶和/或Viscozyme L复合酶;半纤维素酶酶活以30000 U/g计,用量不小于1.0重量份,同时不高于5.0重量份;Viscozyme L复合酶酶活以100 FBG/g计,用量不小于1.0重量份,同时不高于5.0重量份。
较好地,步骤(1)中,过20~40目筛。
较好地,步骤(3)中,选用质量分数为35~45 %的柠檬酸水调节pH值。
较好地,步骤(4)中,所述离心转速为4000~6000 r/min,离心时间为10~20 min。
较好地,步骤(5)中,所述微滤为采用微孔滤膜进行过滤,滤膜孔径为0.45 μm;所述超滤为采用截留分子量为60000道尔顿的聚丙烯中空纤维柱进行过滤。
本发明中药组合物中各原料药皆采购市售品即可,组方原理如下:
红景天:本品为景天科植物大花红景天Rhodiola crenulata(Hook.f.et Thoms.)H.Ohba的干燥根和根茎。甘、苦,平。归肺、心经。功能益气活血,通脉平喘,用于气虚血瘀,倦怠气喘等,有“高原人参”“东方神草”“雪山仙草”等美称,《本草纲目》记载“本经上品,祛邪恶气,补诸不足”,为“已知补益药中所罕见”。以其卓越的抗疲劳、双向调节免疫、抗炎、清除自由基等多种药理作用,在航天医学、军事医学、运动医学等广泛应用,平补心阴(去阴火)而无滋腻之害。
女贞子:为木犀科植物女贞 Ligustrum lucidum Ait.的果实。味苦、甘,性平。入肝、肾经。功能:补肝肾、强腰膝、清热、明目。主治:阴虚内热、头晕、目花、耳鸣、腰膝酸软。现代研究证实,女贞子所含齐墩果酸给大鼠皮下注射,可降低血清谷丙转氨酶,对四氯化碳诱发的肝损伤有明显保护作用,并能促进肝细胞再生,齐墩果酸亦可使肝内甘油三酯蓄积减少、肝细胞变性坏死明显减轻、糖原蓄积增加、血清γ球蛋白下降。
积雪草:本品为伞形科植物积雪草Centella asiatica(L.)Urb.的干燥全草。苦、辛,寒。归肝、脾、肾经。功能清肝热胃火,纾解肝郁,解毒利湿。用于湿热黄疸,中暑腹泻等。现代研究证实,积雪草酸强烈抑制肝星状细胞激活,显著阻断炎性反应所致的肝炎、肝纤维化(《积雪草酸阻断大鼠肝纤维化的实验研究》)。
穿心莲:为爵床科草本植物穿心莲Andrographis paniculata(Burm.f.)Nees的全草或叶。味苦,性寒。入心、肺、大肠、膀胱经。功能:清热解毒、凉血消肿。主治:急性菌痢、胃肠炎等。
三颗针:为小檗科植物拟獠猪刺Berberis soulieanaSchneid.、小黄连刺Berberis wilsonae Hemsl.等的根。苦,寒;有毒。归肝、胃、大肠经。清热燥湿,泻火解毒。用于湿热泻痢,黄疸,湿疹,咽痛目赤,醇耳流脓,痈肿疮毒。
十大功劳:为小檗科植物阔叶十大功劳的根、茎。味苦,性凉。清火解毒,利胆退黄。现代研究证实,阔叶十大功劳中的小檗碱对多种革兰阳性和阴性菌有抑制作用,其中对致病性大肠杆菌、溶血性链球菌、肺炎链球菌、脑膜炎球菌、霍乱弧菌、炭疽杆菌、枯草杆菌、肺炎杆菌、百日咳杆菌、鼠疫杆菌、布氏杆菌、结核杆菌等均有抗菌作用。
地锦草:为大戟科植物地锦草 Euphorbia humifusa Willd.的全草。味辛,性平。功能:清热解毒、活血、止血、利湿、通乳。主治:菌痢、肠炎。地锦草主要含没食子酸、没食子甲脂、槲皮甙、槲皮素、肌醇和鞣酸等,没食子酸为其主要抗菌有效成分。体外实验表明:地锦草鲜汁、煎剂以及水煎浓缩乙醇提取液对于多种致病性球菌及大肠杆菌有明显抑菌作用。
侧柏叶:为柏科植物侧柏 Biota orientalis(L.)Endl.的嫩枝和叶。味苦、涩,性寒。入心、肝、大肠经。功能:凉血、止血、祛风湿、散肿毒。主治:血痢、细菌性痢疾等。
佛手:为芸香科植物佛手Citrus medicaL var. sarcodac-tylis Swingle的干燥果实。辛、苦、酸,温。归肝、脾、胃、肺经。疏肝理气,和胄止痛,燥湿化痰。用于肝胃气滞,胸胁胀痛等。
甘草:为豆科植物甘草 Glycyrrhiza uralensis Fisch. 胀果甘草Glycyrrhizainflata Batal.和光果甘草Glycyrrhiza glabra L. 的干燥根及根茎。味甘,性平。入心、肺、脾、胃经。功能:和中缓急、润肺、解毒、调和诸药。主治:(炙用)脾胃虚弱、食少、腹痛便溏、劳倦发热等。
鸡感染大肠杆菌后常见闭眼缩颈、食少消瘦、拉黄白稀粪,死亡鸡只剖检可见心包炎、肝周炎、腹膜炎等。现代药理研究发现:大肠杆菌感染鸡后,很快会出现急性炎症反应,由于内毒素和大肠杆菌直接作用,肝脏产生急性期蛋白、IL-1,IL-6和肿瘤坏死因子等细胞因子迅速增多,随后血管通透性增加,使组织内渗出液及蛋白质蓄积,浆膜潮湿和水肿,并有液体在体腔内积聚。由于趋化因子作用,异嗜细胞从毛细血管后尾静脉的边缘区游走到周围组织中。感染后6~12 h,组织内可见大量胶冻状渗出物,异嗜细胞脱颗粒或死亡后释放β-防御素(defensins),感染12 h后,炎性细胞逐渐由异嗜细胞转变为巨噬细胞和淋巴细胞,心、肝渗出物持续积聚,最终转变成黄色坚硬、不规则、奶酪样的干酪状坏死团块。显微镜下观察干酪渗出物由外层栅栏状排列的多核巨细胞和巨噬细胞包围的细菌组成。
就治疗策略而言,现有大肠杆菌用药一般按湿热泻痢来治疗,常用白头翁、黄连、黄柏、秦皮、苦参、苍术等药物,多年临床实践发现,此类药物对改善泻痢有一定作用,但对死亡率和采食、体重影响不大,治标不治本。我们通过大量临床筛方试验确认,此病属虚、瘀、热三邪合凑,主要病机为邪盛正虚,不能单纯燥湿止痢而不顾机体气阴两虚的体况,治宜益气养阴、清热理气。
本发明中药组合物中,重用藏药红景天,性平味甘苦,归益气活血、通脉化瘀,有“高原人参”、“东方神草”、“雪山仙草”等美称,《本草纲目》记载“本经上品,祛邪恶气,补诸不足”,为“已知补益药中所罕见”,以其卓越的抗疲劳、双向调节免疫、抗炎、清除自由基等多种药理作用,在航天医学、军事医学、运动医学等广泛应用,平补心阴(去阴火)而无滋腻之害;女贞子,主补肝肾阴虚、内热消渴,辅助君药,扶正祛邪,对阴虚伏邪所致的“内火”釜底抽薪;红景天、女贞子两药共合用益气养阴,共为君药;积雪草,味苦辛寒,清肝热胃火,纾解肝郁,解毒利湿;侧柏叶,味苦、涩性寒,凉血止血;穿心莲、十大功劳凉血解毒;地锦草;利湿解毒;积雪草、侧柏叶、穿心莲、十大功劳、地锦草配伍,泄肝火、清血热、祛脓毒,共为臣药。佛手,疏肝理气为佐药;三颗针,清热燥湿,泻火解毒;甘草,缓急止痛,调和诸药。诸药合用,共奏益气养阴、清热理气之功,则虚得补、热得清、瘀自消。
本发明系纯中药制剂,安全、无毒副作用,无适口性问题,前期通过在蛋鸡、肉鸡主产区筛方验证试验,结果显示:按照0.2~0.4 mL/kg(口服液/鸡只体重)的推荐剂量,连用3~5天,提高大肠杆菌病成活率至65 %以上且能改善增重;且安全、无副作用,具有良好的市场应用价值。
本发明所提供的抗大肠杆菌病口服液制备方法,相较于现有技术,可更为有效地酶解植物细胞壁,从而提高药材有效成分的溶出率;并且通过微滤与超滤技术的联合使用,有效去除了药液中的杂质,溶液澄清度更高,药材有效成分损失也更少,药物临床疗效得到了较好的保证。本发明通过合理搭配组合生物酶,利用生物酶辅助提取药材中有效成分,有效提升了药液中关键药效成分的含量,联动使用超滤技术和减压浓缩技术,使得产品质量得到了较好的提升,因而具有较好的推广应用价值。
具体实施方式
以下结合具体实施例,对本发明做进一步说明。应理解,以下实施例仅用于说明本发明而非用于限制本发明的范围。
实施例中所用部分原料及仪器设备说明如下:所用中药材购自河南禹州中药材市场;所用生物酶均由诺维信(中国)生物技术有限公司生产;所用离心机为GQ145高速管式离心机,分离因数:15900R.C.F,由上海天本离心机械有限公司生产;减压浓缩提取设备为RTN系列热回流提取浓缩机组RTN-200L,由湖南金杯药机有限公司生产。
实施例1
一种用于防治鸡大肠杆菌病的中药口服液的制备方法,以制备1000 mL口服液为例,所采用的原料药为:红景天60 g、女贞子50 g、积雪草30 g、穿心莲25 g、三颗针30 g、十大功劳30 g、地锦草30 g、侧柏叶25 g、佛手25 g、甘草10 g,具体包括如下制备步骤:
(1)、将各原料药粉碎,过20目筛;
(2)、取原料药总重量6倍重量的医用纯化水加热至80 ℃,加入步骤(1)中过筛后的原料药,恒温浸泡10 min;
(3)、将步骤(2)所得药液通入冷凝水降温至45 ℃,用质量分数为40 %的柠檬酸水调节pH至4.5,加入纤维素酶3 g(酶活20000 U/g)、果胶酶2 g(酶活50000 U/g)、淀粉酶1 g(酶活30000 U/g)、Viscozyme L复合酶(酶活100 FBG/g)2.5g,搅拌均匀,酶解3 h,然后升温至75 ℃进行酶灭活处理;
(4)、将步骤(3)酶解后药液煎煮两次,第一次煎煮1 h,第二次加入原料药总重量6倍重量的水煎煮0.5 h,合并两次煎煮滤液,5000 r/min离心15 min得上清液;
(5)、将步骤(4)所得上清液先用孔径为0.45 μm的微孔滤膜进行微滤,然后用截留分子量为60000道尔顿的聚丙烯中空纤维柱进行超滤除去大分子杂质;
(6)、将步骤(5)超滤后滤液减压浓缩至室温下相对密度为1.05;
(7)、将步骤(6)减压浓缩后药液加蒸馏水定容至1000 mL,然后灌装、灭菌即得口服液成品。
实施例2
一种用于防治鸡大肠杆菌病的中药口服液的制备方法,以制备1000 mL口服液为例,所采用的原料药为:红景天50 g、女贞子60 g、积雪草40 g、穿心莲20 g、三颗针30 g、十大功劳40 g、地锦草30 g、侧柏叶20 g、佛手30 g、甘草10 g,具体包括如下制备步骤:
(1)、将各原料药粉碎,过20目筛;
(2)、取原料药总重量8倍重量的医用纯化水加热至85 ℃,加入步骤(1)中过筛后的原料药,恒温浸泡30min;
(3)、将步骤(2)所得药液通入冷凝水降温至50 ℃,用质量分数为40 %的柠檬酸水调节pH至5.0,加入纤维素酶3 g(酶活20000 U/g)、果胶酶2 g(酶活50000 U/g)、淀粉酶1 g(酶活30000 U/g),搅拌均匀,酶解1 h,然后升温至80 ℃进行酶灭活处理;
(4)、将步骤(3)酶解后药液煎煮两次,第一次煎煮2 h,第二次加入原料药总重量8倍重量的水煎煮1 h,合并两次煎煮滤液,5000 r/min离心15 min得上清液;
(5)、将步骤(4)所得上清液先用孔径为0.45 μm的微孔滤膜进行微滤,然后用截留分子量为60000道尔顿的聚丙烯中空纤维柱进行超滤除去大分子杂质;
(6)、将步骤(5)超滤后滤液减压浓缩至室温下相对密度为1.06;
(7)、将步骤(6)减压浓缩后药液加蒸馏水定容至1000 mL,然后灌装、灭菌即得口服液成品。
临床疗效试验
1.试验目的
验证本发明实施例1制备的口服液对鸡大肠杆菌病的治疗效果。
2.材料与方法
2.1主要试剂
肉汤培养基、营养琼脂、麦康凯培养基(北京奥博星生物技术有限公司生产)。
2.2实验动物
16日龄健康海兰褐蛋公鸡(购自开封东杨养殖场)。
2.3药物
复方阿莫西林可溶性粉(规格为50 g∶阿莫西林5 g+克拉维酸钾1.25 g,批号为191002,河南牧翔动物药业有限公司生产);实施例1制备的口服液。
2.4菌种
大肠杆菌采自开封东杨蛋鸡养殖场,自病死鸡肝脏分离获得,经生化实验和动物实验鉴定为致病性大肠杆菌,血清型鉴定为O2(大肠杆菌单因子血清购自中国兽医药品监察所)。
将分离的鸡致病性大肠杆菌接种于伊红美兰培养基,37 ℃培养20 h,选择典型菌落接种于灭菌营养肉汤培养基中,37 ℃培养20 h后,采用平板计数法进行菌落计数,用无菌水稀释,获得1×1010 cfu/mL菌液,备用。
2.5体内实验
2.5.1实验菌株半数致死量LD50的确定
把步骤2.4所得菌液浓度调节至2×109 cfu/mL,依次稀释至2×108 cfu/mL、2×107 cfu/mL、2×106 cfu/mL,备用。把实验鸡按完全随机法平均分成5组,每组18只,第1~4组按以上浓度梯度攻毒,各组每只鸡胸肌注射0.2 mL,第5组为对照组,用相同方法胸肌注射灭菌肉汤培养基0.2 mL。给予充足饮水和饲料。攻毒后2 h即出现精神沉郁,12 h即开始出现死亡,观察至攻毒后第120 h,记录死亡数,计算出死亡率。按Karber法计算半数致死量,即lgLD50=L+d×(S-0.5),其中L为最低稀释度的对数;d为组距(稀释系数即稀释倍数的对数);S为死亡率之和,由此可得出半数致死量为2×108 cfu/mL。半数致死量确定的试验结果如下表1。
2.5.2实验分组
预防实验:将16日龄蛋公鸡,剔除残弱异常鸡只,将体重接近的群体144只用完全随机法平均分成6组,每组24只,每组再按随机分组法平均分成3个重复小组,每重复小组8只。
治疗实验:分组方法同预防实验。
具体分组及用药方法和剂量见下表2。
2.5.3攻毒和给药方式
预防实验:(1)、攻毒方法:空白对照组胸肌注射灭菌肉汤培养基0.2 mL/只,其他各组均胸肌注射浓度为2×108 cfu/mL的菌液0.2 mL/只;(2)、给药方式:按口服灌胃2 d后攻毒,攻毒之后再给药3 d(每天给药1次),停药后观察3 d然后剖检。
治疗实验:(1)、攻毒方法同预防实验;(2)、给药方式:攻毒后30 min灌胃给药,每天给药1次,连续给药5 d,停药后观察3 d然后剖检。
2.5.4疗效评价
大肠杆菌感染发病的标准:精神沉郁,排黄、白绿稀便,采食停止或减慢,剖检死鸡呈现心包炎或肝周炎、腹膜炎、气囊炎,病变部位可分离到大肠杆菌。
实验前各组鸡只逐只称重,记录,实验过程死亡鸡只记录体重,计入终末体重中,实验结束存活鸡逐只称重记录。增重率(%)=(结束时平均体重-初始平均体重)/初始平均体重×100%。
实验结束时记录各组存活鸡只数量。存活率%=结束时存活数/初始样本数量×100%。
实验过程中死亡鸡只剖检,取肝脏涂片染色,镜检,如不能确定可用分离培养鉴定,确定可检出大肠杆菌。实验结束时,宰杀存活鸡只,剖检取肝脏,分离大肠杆菌,确定是否可检出大肠杆菌。大肠杆菌检出率=剖检后取肝脏经分离培养鉴定出大肠杆菌的鸡只个数/样本总数×100%。
通过存活率和增重率综合评价待测药物对大肠杆菌病的效果。
2.5.5数据分析与处理
采用spss17.0统计处理软件进行数据分析,比较各组增重、成活率的差异性。
3.结果
3.1预防实验结果
3.1.1口服药物后对鸡增重的影响
实施例1口服液中、高剂量组经攻毒后体重下降幅度略低于感染对照组,略高于复方阿莫西林对照组,但差异均不显著(p>0.05)。
3.1.2口服药物后对发病率和存活率的影响
各攻毒实验组均100%发病。实施例1口服液各剂量组存活率均高于感染对照组,其中中、高剂量组和复方阿莫西林组与感染对照组相比,差异显著(p<0.05),低剂量组与感染对照组相比差异不显著(p>0.05);实施例1口服液各剂量组与复方阿莫西林组相比均无显著性差异(p>0.05)。至实验结束时,各组存活鸡只精神状态已经恢复,开始进食和饮水。
3.2治疗实验结果
3.2.1口服药物后对鸡增重的影响
实施例1口服液中、高剂量组的平均增重显著高于感染对照组(p<0.05),且略优于复方阿莫西林组,但差异不显著(p>0.05)。
3.2.2口服药物后对鸡发病率、发病率的影响
各攻毒实验组均100%发病。实施例1口服液高剂量组存活率为70.83%,远高于感染对照组的50.00%,差异显著(p<0.05),同时也高于复方阿莫西林组66.67%,但差异不显著(p>0.05)。实施例1口服液中、低剂量组及复方阿莫西林组的存活率也高于感染对照组,但差异不显著(p>0.05)。
4.试验结论及分析
试验结果显示:实施例1口服液按0.2 mL/kg或0.4 mL/kg体重口服用药用于预防或治疗人工感染蛋雏鸡大肠杆菌病,其存活率和增重率均优于感染对照组,且差异显著(p<0.05);与复方阿莫西林相比都无显著性差异(p>0.05),高剂量组甚至略优于复方阿莫西林。但所有用药组的存活率和增重率均低于空白对照组,且差异显著(p<0.05)。
用中医理论分析,伴随着家禽育种技术、动物营养科学和饲养管理技术的不断发展和进步,家禽的生长速度和生产能力得到大幅提高。家禽个体则表现为生发之力旺盛,是谓木旺。五行木旺则土不及,表现为容易出现消化不良、过料拉稀的现象,同时家禽新鲜粪便中蛋白质量分数为11.2~15.0%,数倍于牛粪和猪粪,说明鸡的体质存在木旺土弱的平衡特点。木旺生火,火克肺金,故多有热病伤肺,心力衰竭而死者,现代生理学发现家禽心率为280~350次/min,也远高于猪和牛等。肾水为肝木生发之源,生发过旺而致肝肾阴虚,临床所见各种病鸡,几无肝肾完好者。所以综上分析,家禽属风木体质,脾胃相对不足,易阴虚,易肺热。尤其本次实验中建立的攻毒模型,鸡群表现的症状发热、肌肤失水,爪干毛燥,剖检出现典型心包炎、肝周炎、肠内有硬结的盲肠芯,十分符合阴虚内热特点。而本发明组方中,红景天、女贞子益气养阴为君药,积雪草、侧柏叶、穿心莲、十大功劳、地锦草清热解毒、凉血止血、利湿解毒,共为臣药,君臣相辅,益气而不助热,清热而不伤阴。
本试验在动物模型建立过程中发现攻毒后鸡群几乎停止采食饮水,直到用药第3天或第4天,多数存活鸡才开始恢复采食饮水,因此至实验结束时,各实验组鸡的体重均呈负增长。与文献报道的人工感染大肠杆菌动物实验中体重都呈正增长有不同之处,分析可能是实验鸡品种及实验条件差异所致。
根据试验中药物的实际临床疗效、作用特点等,建议本品在临床中连续使用4天即可。
Claims (6)
1.一种用于防治鸡大肠杆菌病的中药组合物,其特征在于,所述中药组合物由下列重量份的原料药制成:红景天60份、女贞子50份、积雪草30份、穿心莲25份、三颗针30份、十大功劳30份、地锦草30份、侧柏叶25份、佛手25份、甘草10份;重量份以g计、体积份以mL计,制备步骤如下:
(1)、按所述重量份数取各原料药,粉碎、过筛;
(2)、取原料药总重量6倍重量的水加热至80 ℃,加入步骤(1)中过筛后的原料药,恒温浸泡10 min;
(3)、将步骤(2)所得药液降温至45 ℃,调节pH至4.5,加入复合酶,搅拌均匀,酶解3 h,然后升温至75 ℃进行酶灭活处理;所述复合酶为纤维素酶、果胶酶、淀粉酶和Viscozyme L复合酶的混合物,其中,纤维素酶酶活以20000 U/g计、用量3 重量份,果胶酶酶活以50000U/g计、用量2重量份,淀粉酶酶活以30000 U/g计、用量1.0重量份;Viscozyme L复合酶酶活以100 FBG/g计、用量2.5重量份;
(4)、将步骤(3)酶解后药液煎煮两次,合并两次煎煮滤液,离心处理得上清液;
(5)、将步骤(4)所得上清液进行微滤、超滤;
(6)、将步骤(5)超滤后滤液减压浓缩至室温下相对密度为1.05;
(7)、将步骤(6)减压浓缩后药液加水定容至1000体积份,然后灌装、灭菌即得口服液成品。
2.一种如权利要求1所述用于防治鸡大肠杆菌病的中药组合物的制备方法,其特征在于:
重量份以g计、体积份以mL计,步骤如下:
(1)、按所述重量份数取各原料药,粉碎、过筛;
(2)、取原料药总重量6倍重量的水加热至80 ℃,加入步骤(1)中过筛后的原料药,恒温浸泡10 min;
(3)、将步骤(2)所得药液降温至45 ℃,调节pH至4.5,加入复合酶,搅拌均匀,酶解3 h,然后升温至75 ℃进行酶灭活处理;所述复合酶为纤维素酶、果胶酶、淀粉酶和Viscozyme L复合酶的混合物,其中,纤维素酶酶活以20000 U/g计、用量3 重量份,果胶酶酶活以50000U/g计、用量2重量份,淀粉酶酶活以30000 U/g计、用量1.0重量份;Viscozyme L复合酶酶活以100 FBG/g计、用量2.5重量份;
(4)、将步骤(3)酶解后药液煎煮两次,合并两次煎煮滤液,离心处理得上清液;
(5)、将步骤(4)所得上清液进行微滤、超滤;
(6)、将步骤(5)超滤后滤液减压浓缩至室温下相对密度为1.05;
(7)、将步骤(6)减压浓缩后药液加水定容至1000体积份,然后灌装、灭菌即得口服液成品。
3.如权利要求2所述用于防治鸡大肠杆菌病的中药组合物的制备方法,其特征在于:步骤(1)中,过20~40目筛。
4.如权利要求2所述用于防治鸡大肠杆菌病的中药组合物的制备方法,其特征在于:步骤(3)中,选用质量分数为35~45 %的柠檬酸水调节pH值。
5.如权利要求2所述用于防治鸡大肠杆菌病的中药组合物的制备方法,其特征在于:步骤(4)中,所述离心转速为4000~6000 r/min,离心时间为10~20 min。
6.如权利要求2所述用于防治鸡大肠杆菌病的中药组合物的制备方法,其特征在于:步骤(5)中,所述微滤为采用微孔滤膜进行过滤,滤膜孔径为0.45 μm;所述超滤为采用截留分子量为60000道尔顿的聚丙烯中空纤维柱进行过滤。
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Address after: No. 9, Yugang Road, Airport Economic Comprehensive Experimental Zone, Zhengzhou City, Henan Province, 451162 Patentee after: Henan Muxiang Biotechnology Co.,Ltd. Country or region after: China Address before: 451162 airport road five, Zhengzhou, Henan Patentee before: HENAN SOAR VETERINARY PHARMACEUTICAL Co.,Ltd. Country or region before: China Patentee before: HENAN MUXIANG BIOTECHNOLOGY Co.,Ltd. |