CN116966245A - 一种防治鸡毒支原体感染的中药复方制剂及其制备方法 - Google Patents
一种防治鸡毒支原体感染的中药复方制剂及其制备方法 Download PDFInfo
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明公开一种防治鸡毒支原体感染的中药复方制剂及其制备方法,属于中兽药技术领域。该中药复方制剂包括葛根15~30份、黄芩15~30份、虎杖15~30份、鱼腥草5~20份、菊花5~20份、板蓝根5~20份、桔梗3~10份、竹叶3~10份、薄荷3~10及甘草3~10份。本发明将上述原料混合,加水煎煮,冷却后过滤,滤液每千克体重的鸡每天饮水或灌服1mL,连喂7天。本发明产品为纯中药制剂,组合配伍合理,既能抑制鸡毒支原体的生长,也具有清热解毒、润肺止咳及抗炎的功效,对鸡毒支原体感染引起的慢性呼吸道病疗效显著。该发明可改善和调节疾病过程中的平衡失调,标本兼治,治愈率高,安全可靠,且不易产生耐药性,确保食品安全,制备方法简单,成本低,应用前景广阔。
Description
技术领域
本发明涉及中医药技术领域,具体涉及一种防治鸡毒支原体感染的中药复方制剂及其制备方法。
背景技术
鸡毒支原体(Mycoplsma gallisepticum,MG)是鸡慢性呼吸道病(ChronicRespiratory Disease,CRD)和火鸡传染性窦炎的病原,引起鸡咳嗽、流泪、呼吸啰音等临床症状。CRD 在世界范围内广泛流行,不同品种、日龄及性别的鸡在各个季节均可发病,且发病率逐年递增,是危害养禽业的重要传染病之一。
MG主要通过膜表面脂质相关蛋白的特殊末端结构吸附于宿主呼吸道粘膜表面,其黏附蛋白与宿主细胞表面的受体结合,使宿主细胞变性,气管纤毛损伤,破坏粘膜天然屏障和免疫系统,不及时防治将导致多系统、多器官病变。MG侵入鸡场,难以被彻底清除,在鸡群中长期存在,引起反复感染和蔓延,导致家禽饲料利用率降低,生长迟缓,死淘率增加,产蛋率下降,畸形蛋率提高等。MG常与大肠杆菌、传染性支气管炎病毒、新城疫病毒等混合或继发感染,对鸡群的危害更加严重,死亡率高达30%以上,给家禽业带来巨大的经济损失。
目前预防鸡群MG感染的最有效方法是接种疫苗。鸡接种疫苗后虽然能产生一定的免疫保护,但不能阻止感染,免疫鸡群在应激情况下仍可大批发病。另外MG不断变异,需要及时研发疫苗来应对变异菌株。生产上,CRD的治疗主要使用抗生素,如强力霉素、泰乐菌素、泰妙菌素、替米考星及阿奇霉素等。抗生素在感染前期可快速杀死鸡毒支原体,减缓鸡体的临床症状,但停药后容易复发,且长期使用抗生素会导致MG的耐药性增加,且毒副作用及药物残留等问题突出。因此,急需探索防治鸡毒支原体感染的理想药物。
中药具有祛邪扶正、清热解毒、止咳平喘等作用,且具有抗菌、抗病毒、增强机体免疫力的功效。中药成分均为生物有机物,其毒副作用小,药物残留低,长期使用安全可靠。因此,发明一种能够有效防治鸡毒支原体感染的中药复方制剂是本领域科研人员急需解决的问题。
发明内容
本发明提供了一种防治鸡毒支原体感染的中药复方制剂及其制备方法。
本发明采取如下技术方案:
本发明的药物,由下述重量份数的原料药组成:葛根15~30份、黄芩15~30份、虎杖15~30 份、鱼腥草5~20份、菊花5~20份、板蓝根5~20份、桔梗3~10份、竹叶3~10份、薄荷3~10 份、甘草3~10份。
本发明进一步优选中药原药重量份组成:葛根17份、黄芩22份、虎杖16份、鱼腥草10份、板蓝根10份、桔梗5份、竹叶5份、薄荷5份和甘草10份。
上述配方中,各原料药的功效如下:
葛根:归肺、脾、胃经,其气轻,善解表发汗,可升发脾胃清阳之气——《本草正义》。
黄芩:归肺、脾经,主泻肺胃上焦之火。主胸中逆气,隔上热痰,消喘止咳——《药品化义》。
虎杖:归肝、胆、肺经,有祛风利湿,止咳化痰之效,可用于肺热咳嗽,痰多喘咳——《中国药典》。
鱼腥草:归肝、肺经,清热解毒,排脓消痈。治肺痈咳嗽,抗炎消肿——《中华本草》。
菊花:入肺、肝经,散风清热,平肝明目,清三焦郁火,除胸中烦热,安肠胃,利五脉,调四肢——《别录》。
板蓝根:入肝、胃经,清热解毒,凉血利咽,避疫杀虫——《本草便读》。
桔梗:入肺经,开宣肺气,祛痰排脓,引诸药上行,为诸药舟楫——《本草求真》。
竹叶:甘淡微寒,入心、肺、肝、胆经,有凉心缓脾,清痰止咳之效。竹叶生于枝上,可透药,引药上行,治上焦风邪烦热,咳逆喘促——《本草求真》。
薄荷:入肺、肝经,疏风散热,解毒利窍,善走肌表,引诸药入营卫以疏结滞之气——《药品化义》。
甘草:入心、肺、脾、胃经,补脾溢气,润肺,清热解毒,调和诸药——《本草经解》。
葛根性辛甘微寒,与黄岑、虎杖同用可去上焦之火,驱散肺部烦热,达止咳、祛痰平喘之效,发挥主要作用,为君药。鱼腥草、菊花、板蓝根皆清热解毒,辅助君药散风清热,平肝明目,安神利窍,为臣药。桔梗开宣肺气,引药上行;竹叶生于枝,轻浮,可透药而引药上浮也,薄荷引诸药入卫营,此三者皆可载药而行,为佐药也。甘草补脾溢气,润肺止咳,调和诸药,为使药也。诸药配伍,共奏清热解毒,开宣肺气,祛痰止咳,安神明目之效。
本发明中药复方制剂制备方法如下:
按上述配伍比例称取各种原料药,除鱼腥草、薄荷外,其余几种混合均匀,放置煎药机中,并用冷水浸泡30分钟,武火煮沸,文火慢煎30分钟,然后加入鱼腥草和薄荷,继续文火慢煎15分钟。熬制完毕待冷却后过滤,药液装袋,于4℃保存备用,使用前40℃水浴锅预热药液。
本发明的中药药液使用方法为:鸡每千克体重每日灌服或饮水1mL药液(每毫升药液相当于含原生药材250mg),连续使用7天。
本发明的有益效果:(1)本发明提供的中药复方制剂对毒支原体感染疗效显著,能够提高机体免疫力,防止生产性能下降。(2)本复方制剂为纯中药制剂,不添加任何抗生素及其它化学合成的抗菌素,避免了耐药性问题的产生,且本复方制剂对机体无害,畜禽产品中无残留,提高了禽产品的安全性,有益于人类健康。(3)本复方制剂成本低,生产工艺简单,适合实际生产应用。
附图说明
图1为中药复方制剂对鸡毒支原体的最小抑菌浓度。
图2为各组鸡气管切片
图2-1为正常组鸡气管,结构组织完整,纤毛清晰可见;
图2-2为单独用复方中药制剂鸡气管,组织结构完整,粘膜上皮细胞的纤毛连续、清晰可见,与正常组无差别;
图2-3为MG感染模型组鸡气管,纤毛脱落,粘膜增厚,局部粘膜上皮坏死脱落;
图2-4、2-5、2-6分别为复方低剂量预防组、中剂量预防组及高剂量预防鸡气管,组织结构完整,纤毛清晰可见,无脱落,与正常组气管无差异;
图2-7为复方低剂量治疗组鸡气管,纤毛完全脱落,粘膜上皮变性,粘膜层明显增厚,单核淋巴细胞明显增多;
图2-8为复方中剂量治疗组鸡气管,组织结构比较完整,纤毛局部脱落(见黑色箭头),出现淋巴滤泡;
图2-9为复方高剂量治疗组鸡气管,组织结构正常,纤毛完整清晰,与正常组无差异;
图2-10为泰妙菌素治疗组鸡气管,粘膜上皮完整,纤毛可见,但比较稀疏(见黑色箭头)。
具体实施方式
为了便于更好地理解本发明,以下通过具体实施例说明本发明的具体疗效。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,均属于本发明保护的范围。其中所述的中药原料,如无特别说明,均为市售,涉及的仪器若无特别说明均为常规仪器,涉及的实验方法若无特别说明,均为常规方法。
实施例1:称取葛根85g、黄芩110g、虎杖80g、鱼腥草50g、板蓝根50g、桔梗25g、竹叶25g、薄荷25g、甘草50g。将葛根、黄芩、虎杖、板蓝根、桔梗、竹叶、甘草混合均匀,放置于煎药机中,用冷水浸泡30分钟,武火煮沸,文火慢煎30分钟后,然后放入薄荷与鱼腥草煎煮15分钟。待药液冷却后过滤,装袋,4℃保存备用,使用前用40℃水浴锅预热药液。本次所制得的药液浓度为250mg/mL。
实施例2:本发明复方中药制剂的体外抑菌试验
将本发明实施例所得的复方中药制剂浓缩成500mg/mL,过滤除菌,使用2倍稀释法测定该中药复方制剂的最小抑菌浓度。将鸡毒支原体(MG-HS)培养物浓度稀释到1×104CCU/mL。取13支2mL的灭菌EP管,各管最终药液浓度依次为100、50、25、12.5、6.25、3.13、1.56、 0.78、0.39、0.195、0.098、0.048、0.024mg/mL,然后在各管依次加入鸡毒支原体菌液0.5mL。另取1支2mL无菌EP管,加入1mL培养基作为对照。37℃恒温培养14天,观察颜色变化,发现该复方中药制剂的最小抑菌浓度为25mg/mL。
实施例3:本发明复方中药制剂的药效试验
将180羽青脚麻鸡饲养至11日龄,称重,并随机分为10组,每组18只,每组3个重复,每重复6只。组别分别为A(正常组)、B(复方中药制剂组)、C(MG感染模型组)、D(低剂量预防组)、E(中剂量预防组)、F(高剂量预防组)、G(低剂量治疗组)、H(中剂量治疗组)、I(高剂量治疗组)、J(泰妙菌素治疗组)。其中,A组鸡整个试验期不做任何处理,B 组从第12天开始,鸡每千克体重每日灌服0.5mL复方中药制剂,连续使用21天后停药正常饲养。第12-18天D、E、F组的雏鸡每千克体重鸡每天分别灌服药物0.25mL、0.5mL、1mL, C、G、H、I、J组不做任何处理;第19-23天分别对C、D、E、F、G、H、I、J组人工攻毒,用1×109CCU/mL的MG-HS菌液每只鸡每天滴鼻点眼0.3mL及气囊注射0.7mL,攻毒结束后检查感染情况;第24-30天D、E、F、G、H、I组使用中药治疗,根据鸡体重给药,剂量分别为D、G组0.25mL/kg,E、H组0.5mL/kg,F、I组1mL/kg,J组使用泰妙菌素饮水给药,剂量为350mg/L;第31-44天所有组鸡正常饲养,不做任何处理。整个试验期记录生产性能指标、观察鸡的临床症状,第45天试验结束对所有鸡采血、屠宰,观察内脏器官病变情况。各组鸡取气管的中段用4%甲醛固定制备石蜡切片,用苏木精伊红染色法观察气管损伤程度。结果如下:
(1)鸡的临床表现
当本病呈隐性感染时,其感染症状表现得不够明显亦不够典型。若单纯患本病,气管及支气管等部位出现炎症而发生肿胀,致使呼吸道中的浆性、黏性液体向上逆流而出形成浆黏性鼻液,因气管和咽部等部位发生肿胀,导致咽部发痒,疼痛难忍,进而引起打鸡喷嚏,咳嗽等。当感染程度加重时,会引发鼻窦炎;眼睛有分泌性、黏性浆液流出,难以睁开,严重者脸部肿胀,眼睛失明。病鸡呼吸道中有大量痰块存在,引起呼吸困难,如经常啰音,持续张口呼吸,咳嗽并伴随轻微甚至严重的喘气。患病严重的鸡群精神极度抑郁,羽毛蓬松凌乱,不喜采食饲料,饮水增加,生长出现停滞,甚至出现负增长的情况。
(2)复方中药制剂对鸡生产性能的影响
由表1和表2可知,单独使用复方制剂组和正常组的体增重及料肉比差异均不显著。说明复方中药对鸡的生产性能没有影响。
攻毒以后,MG感染模型组的体增重显著低于其余各组;正常组、复方制剂组和预防高中剂量组的体增重无明显差异,而预防低剂量组的体增重稍高于感染组,但无显著差异。在料肉比方面,感染组显著高于其余各组,预防低剂量组显著低于感染组而高于其余各组。
使用药物治疗7d以后,各组体增重及料肉比见表2治疗期。在体增重方面,MG感染组显著低于其余各组,而与G组(低剂量治疗组)无显著差异;DEF组(预防组)、IH组(治疗高、中剂量组)与J组(泰妙菌素组)之间没有显著差异;D(预防低剂量组)、GHI组(治疗组)和J组(泰妙菌素组)之间没有显著差异;FE组(预防高、中剂量组)、A组(正常组)与B组(复方制剂组)之间没有显著差异。在料肉比方面,MG感染组显著高于其余各组,其余各组之间没有显著差异。
治疗结束,停药观察14d后,各组体增重及料肉比见表2观察期。在体增重方面,MG感染组,显著低于其余各组;DEF组(预防组)、GHI组(治疗组)、J组(泰妙组)之间没有显著差异,而显著低于A组(正常组)和B组(复方制剂组)。在料肉比方面,随着鸡日龄的增长,各组已无显著差异。
综上所述,使用高、中剂量复方中药对MG感染进行预防,可有效防止鸡的生产性能下降;MG感染以后,如需短时间内有效的提高鸡的生产性能,可使用高、中剂量的复方中药进行治疗;而低剂量复方中药在较长时间内亦可提高感染鸡的生产性能。
表1复方制剂预防MG感染对鸡生产性能的影响
表2复方制剂防治MG感染对生产性能的影响
(3)复方中药对气囊和气管的保护效果
实验结束后,所有鸡全部屠宰,进行气囊损伤评分。气囊薄而透明,带有光泽记0分;气囊出现黄色斑点记1分;气囊表面的黄色干酪物较少,有泡沫,记2分;气囊表面出现较多的黄色干酪物,明显增厚,变成浑浊记为3分;气囊表面附着大量的黄色干酪状渗出物记4分。气囊损伤减少率=(感染组气囊评分-实验组气囊评分)/感染组气囊评分。评分结果见表3。
MG感染组的气囊评分显著高于其它组;预防高中剂量组、治疗高剂量组与正常组和复方组无差异,显著低于其余各组;预防低剂量组、治疗中低剂量组和泰妙组之间无显著差异。可以看出在停药继续观察14d后,预防高中剂量组和治疗高剂量组的鸡的气囊损伤减少率最高,达100%,其次是预防低剂量组,效果均优于泰妙组;治疗中低剂量组的气囊损伤减少率也达70%以上。说明使用复方中药进行防治,可以有效保护鸡的气囊。
表3各组鸡气囊损伤评分及气囊损伤减少率
根据切片结果表明(附图图2),正常组的粘膜上皮细胞的纤毛完整、清晰可见,而MG 感染组的纤毛几乎完全脱落,粘膜层明显增厚,局部粘膜上皮出现坏死并脱落;复方制剂组、预防高、中、低剂量组和治疗高剂量组的组织结构均完整,纤毛浓密连续、清晰可见,与正常组无差别;治疗中剂量组的组织结构比较完整,纤毛局部脱落,出现淋巴滤泡;治疗低剂量组的纤毛严重脱落,粘膜上皮变性,粘膜层明显增厚,单核淋巴细胞明显增多;泰妙组粘膜上皮完整,纤毛可见,但部分较稀疏。以上结果表明,MG感染会破坏气管的纤毛结构,并导致气管粘膜增厚,造成粘膜上皮细胞坏死脱落,而使用复方中药进行防治能有效修复MG 对气管造成的炎症损伤。
(4)复方中药制剂对鸡毒支原体感染的防治效果
复方中药对人工感染鸡慢性呼吸道病的防治效果见表4。结果表明,攻毒后,感染组、预防组、治疗组、泰妙组均普遍感染,但预防组的致病率明显较低,说明预防有一定效果;其中MG感染模型组的致死率最高,其余各组均无MG致死情况;高、中剂量预防组的治愈率最高,达90%以上,效果与泰妙菌素相当;其次是高、中剂量治疗组,治愈率达87.5%;再然后是预防、治疗低剂量组,但其治愈率也达80%以上。
表4复方中药对人工感染鸡慢性呼吸道病的防治效果
综上所述,本发明提供的复方中药制剂具有很高的抑菌效果,对人工造模的慢性呼吸道病鸡有明显的防治作用。且制备工艺简单、用量少、成本低,不易产生耐药性,减少了兽药残留,提高了畜禽产品的安全性,可以用于鸡毒支原体引起的慢性呼吸道病的防治。
Claims (3)
1.一种防治鸡毒支原体感染的中药复方制剂,其特征在于由以下重量份的原料药制成:葛根15~30份、黄芩15~30份、虎杖15~30份、鱼腥草5~20份、菊花5~20份、板蓝根5~20份、桔梗3~10份、竹叶3~10份、薄荷3~10份、甘草3~10份。
2.根据权利要求1所述一种防治鸡毒支原体感染的中药复方制剂,各原料药的重量份数是:葛根17份、黄芩22份、虎杖16份、鱼腥草10份、板蓝根10份、桔梗5份、竹叶5份、薄荷5份及甘草10份。
3.一种根据权利要求1或2所述的一种防治鸡毒支原体感染的中药复方制剂的制备方法如下:
(1)按照权利要求1或2所述重量份数称取各原料药;
(2)将葛根、黄芩、虎杖、菊花、板蓝根、桔梗、竹叶、甘草等混匀,放入煎药机中,用冷水浸泡30分钟,以水没过药3~4厘米为宜;
(3)武火煮沸,文火慢煎45分钟;
(4)煮沸30分钟后,放鱼腥草和薄荷继续熬煮15分钟;
冷却后过滤,袋装,室温或冷藏保存备用;室温可保存7天,4℃冷藏可保存15天。
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