CN106924477B - 一种混合菌发酵生产的复合中药发酵制剂及其制备方法 - Google Patents
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Abstract
本发明公开了一种混合菌发酵生产的复合中药发酵制剂及其制备方法,属于中药发酵制剂技术领域。其中药各组分的重量份数组成为:柴胡4~12份、川穹2~10份、香附2~10份、陈皮2~10份、枳壳1~9份、芍药1~9份、甘草1~3份。本发明方法选取的中药组分结构合理、比例适当;本发明选取的凝结芽孢杆菌和植物乳杆菌具有耐酸、耐胆盐、抑制病原菌生长的特性,能够有效提高免疫力,促进生长和改善肠道微环境,具有优异的益生作用。在本发明所选取的凝结芽孢杆菌和植物乳杆菌的作用下,显著地提高了原有药物的药效;本发明的发酵药物可以显著促进动物生长、提高免疫力,而且绿色健康,无毒副作用。
Description
技术领域
本发明属于中药发酵制剂技术领域,具体涉及一种混合菌发酵生产的复合中药发酵制剂及其制备方法。
背景技术
凝结芽孢杆菌是微生态制剂中乳酸菌类的一种益生菌,除了具有与乳酸菌和双岐杆菌一样的保健功能以外,还有耐高温、耐酸、耐胆盐等高抗逆性,具有抑制肠道致病菌生长,调节肠道功能紊乱,维持肠道内菌群平衡和提高机体免疫的能力。但凝结芽孢杆菌不同于其它的益生菌,由于菌体在肠道上皮细胞的粘附性较弱,所以在自然条件下一般很难在肠道中存在。一般来说,凝结芽孢杆菌只能在肠道内做短暂的停留,一次性口服凝结芽孢杆菌后,经过大约4~7天时间肠道内的凝结芽孢杆菌便会通过排便而消失殆尽。
现代微生物发酵多为纯种发酵,即用纯的单一菌落进行的发酵过程。纯培养技术使得研究者摆脱了面对多种微生物共存的复杂局面,能够不受干扰地对单一目的菌株进行研究,从而丰富了人们对微生物形态结构、生理生化和遗传特性的认识。但是,在长期的实验和生产实践中,人们不断地发现很多重要生化过程仅靠单株微生物不能完成或只能微弱地进行,必须依靠两种或多种微生物共同培养来完成,即混菌培养或称为混菌发酵。
发明内容
针对现有技术中存在的问题,本发明的目的是提供一种天然安全的复合中药发酵制剂以及制备该发酵制剂的方法,同时,提供一种含有该复合中药发酵制剂的具有提高家禽生长性能和免疫力的饲料。
为了解决上述技术问题,本发明采用以下技术方案予以实现:
一种复合中药发酵制剂,它由中药成分与菌液混合后经发酵制成;所述菌液为植物乳杆菌与凝结芽孢杆菌的混合菌液。
在上述方案的基础上,所述菌液为植物乳杆菌与凝结芽孢杆菌按1∶1的比例混合的菌液。
在上述方案的基础上,所述凝结芽孢杆菌为凝结芽孢杆菌(Bacillus coagulans)zkng160127,保藏编号为CGMCC NO:12125;所述的植物乳杆菌为植物乳杆菌植物亚种(Lactobacillusplantarumsubsp.Plantarum),编号AS1.557,来源于中国科学院微生物研究所。
在上述方案的基础上,所述菌液中凝结芽孢杆菌和植物乳杆菌的含量均≥108CFU/mL。
在上述方案的基础上,所述中药成分由以下组分按重量份数制备而成:柴胡4~12份、川穹2~10份、香附2~10份、陈皮2~10份、枳壳1~9份、芍药1~9份、甘草1~3份。
在上述方案的基础上,所述中药成分由以下组分按重量份数制备而成:柴胡8份、川穹6份、香附6份、陈皮6份、枳壳4.5份、芍药4.5份、甘草1.5份。
在上述方案的基础上,所述复合中药发酵制剂的制备方法,步骤如下:
(1)按量取中药组分,粉碎,过60目筛网,备用;
(2)将粉碎后的各组分混合均匀,加入中药成分总重量80%的水,搅拌均匀后于100℃蒸汽灭菌30min,冷却至20-30℃;
(3)向冷却后的药物中加入混合菌液,菌液用量占加入后总量的10~15%;置于单向排气塑料袋中,在20℃~30℃条件下发酵30天。
上述的单向排气塑料袋为专利号为:200920100585.9的厌氧发酵饲料、中药及添加剂的单向排气袋。
在上述方案的基础上,所述菌液由以下方法制得:将凝结芽孢杆菌和植物乳杆菌菌种接种于活化培养基进行活化培养,培养温度为37℃,时间24h;将活化好的菌种分别接种于发酵罐中,控制温度37℃~43℃,时间24h~36h。
在上述方案的基础上,所述活化培养基和所述发酵所用培养基为:MRS培养基,成分为(g/L):蛋白胨10.0、肉膏10.0、酵母浸粉5.0、葡萄糖20.0、磷酸氢二钾2.0、柠檬酸氢二铵2.0、乙酸钠5.0、硫酸镁0.58、硫酸锰0.28、半胱氨酸0.5、吐温80 1.0,调pH6.2-6.4。
一种具有提高家禽生长性能和免疫力的饲料,在饲料中添加上述方法制备的复合中药发酵制剂,添加量为2kg/吨。
本发明的提高生长性能和免疫力的中药组合物,各中药的药性药效如下:
柴胡:主要含柴胡皂甙、植物甾醇,另含少量挥发油,茎叶含芦丁,具有疏气、解郁、散火等功效。
川穹:为伞形科草本植物川芎的根茎。辛,温。归肝、胆、心包经,具有活血行气、祛风止痛的功效。
香附:为莎草科多年草本植物莎草的干燥块茎,野生于海、河、溪边沙地上较多。含挥发油,其中主要为香附稀,香附醇,并含脂肪酸等成分,具有理气解郁,调经止痛等功效。
陈皮:为芸香科植物常绿小乔木福柑、朱桔、蜜柑等多种桔柑的成熟干燥果皮,均为栽培。含橙皮甙、中肌醇、挥发油(主要成分为右旋柠檬烯)和维生素B1等主要成分,具有理气健脾,燥湿化痰等功效。
枳壳:原植物主要有酸橙、香橼、代代花及枳四种,幼小的果实称为枳实,成熟的果实成为枳壳。入脾、胃二经,具有破气消积,化痰除痞等功效。
芍药:为毛茛科多年生草本植物芍药的干燥根,多为人工栽培。主要含白芍素,即芍药甙,此外尚含苯甲酸、β-固甾醇、鞣质、挥发油、脂肪油等成分。具有补血敛阴,平肝止痛等功效。
甘草:别名甜甘草,为豆科多年生草本植物甘草和光果甘草等的干燥根和根状茎(芦头),多为野生。含甘草甜素、甘草黄素、葡萄糖、甘露醇、苹果酸、天门冬酰胺等主要成分,具有补脾益气、清热解毒、润肺止咳等功效。
其中所述凝结芽孢杆菌zkng160127为凝结芽孢杆菌(Bacillus coagulans)的一个亚种,保藏编号为CGMCC NO:12125,保藏于中国普通微生物菌种保藏管理中心。
所述的植物乳杆菌为植物乳杆菌植物亚种(Lactobacillusplantarumsubsp.Plantarum),编号AS1.557,来源于中国科学院微生物研究所。
本发明的技术原理:
本方为为理气剂,具有疏肝理气,活血止痛之功效。主治肝气郁滞症。胁肋疼痛,胸闷善太息,情志抑郁易怒,或嗳气,脘腹胀满,脉弦。肝主疏泄,性喜条达,其经脉布胁肋循少腹。若情志不遂,木失条达,则致肝气郁结,经气不利,故见胁肋疼痛,胸闷,脘腹胀满;肝失疏泄,则情志抑郁易怒,善太息;脉弦为肝郁不舒之征。遵《内经》"木郁达之"之旨,治宜疏肝理气之法。方中以柴胡功善疏肝解郁,用以为君。香附理气疏肝而止痛,川芎活血行气以止痛,二药相合,助柴胡以解肝经之郁滞,并增行气活血止痛之效,共为臣药。陈皮、枳壳理气行滞,芍药、甘草养血柔肝,缓急止痛,均为佐药。甘草调和诸药,为使药。诸药相合,共奏疏肝行气、活血止痛之功。
本发明复合中药发酵制剂具有以下特点:(1)本发明选用的凝结芽孢杆菌菌株能耐受消化道内的环境,耐受胃酸和胆盐,抗逆性强(2)本发明选用的凝结芽孢杆菌和植物乳杆菌菌株具有增强动物免疫力和促进生长的功能(3)本发明选用的凝结芽孢杆菌和植物乳杆菌及其代谢产物能有效的补充或增强原有药物的功能;(4)中药经凝结芽孢杆菌和植物乳杆菌转化后产生了新的活性物质,并带来新的保健、预防或治疗功能。
本发明方法选取的中药组分结构合理、比例适当,选用的凝结芽孢杆菌和植物乳杆菌具有活性高、抗逆性强等优势,在本发明所选取的凝结芽孢杆菌和植物乳杆菌混合发酵的作用下,显著提高了原有药物的药效,使用本发明的复合中药发酵制剂可以显著提高家禽生长性能和机体免疫力。
具体实施方式
以下结合较佳实施例,对依据本发明提供的具体实施方式详述如下:
实施例1
菌液的制备:
将本发明的凝结芽孢杆菌和植物乳杆菌菌种分别接种于活化培养基进行培养,培养温度为37℃,时间24h;将活化好的菌种分别接种于发酵罐中,控制温度37℃~43℃,时间24h~36h,制备发酵液。
其中所述凝结芽孢杆菌zkng160127为凝结芽孢杆菌(Bacillus coagulans)的一个亚种,保藏编号为CGMCC NO:12125,保藏于中国普通微生物菌种保藏管理中心。
所述的植物乳杆菌为植物乳杆菌植物亚种(Lactobacillusplantarumsubsp.Plantarum),编号AS1.557,来源于中国科学院微生物研究所。
所述活化培养基为:MRS培养基,成分为(g/L):蛋白胨10.0、肉膏10.0、酵母浸粉5.0、葡萄糖20.0、磷酸氢二钾2.0、柠檬酸氢二铵2.0、乙酸钠5.0、硫酸镁0.58、硫酸锰0.28、半胱氨酸0.5、吐温80 1.0,调pH6.2-6.4。
发酵所用培养基为:MRS培养基,成分同上。
发酵完成后,两种菌液中菌含量均≥108CFU/mL;将两种菌的菌液按1∶1的体积比混合后,待用。
实施例2
一种提高家禽生长性能和免疫力的复合中药发酵制剂,其各组分的重量份数为:柴胡8份、川穹6份、香附6份、陈皮6份、枳壳4.5份、芍药4.5份、甘草1.5份。
具体制备方法如下:
(1)按量取中药组分,粉碎,过60目筛网,备用;
(2)将粉碎后的各组分混合均匀,加入中药成分总重量80%的水,搅拌均匀后于100℃蒸汽灭菌30min,冷却至20-30℃;
(3)向冷却后的药物中加入混合菌液,菌液用量占加入后总量的10~15%;置于单向排气塑料袋中,在20℃~30℃条件下发酵30天。
实施例3
一种提高家禽生长性能和免疫力复合中药发酵制剂,其各组分的重量份数为:柴胡4份、川穹3份、香附3份、陈皮2份、枳壳1份、芍药1份、甘草1份。
制备方法与实施例2相同。
实施例4
一种提高家禽生长性能和免疫力复合中药发酵制剂,其各组分的重量份数为:柴胡12份、川穹9份、香附9份、陈皮10份、枳壳9份、芍药9份、甘草3份。
制备方法与实施例2相同。
一、药效试验:
1.1试验组方
1.1.1传统工艺制备的非发酵中药制剂组(将本发明实施例2所选取的药物进行超微粉碎,粉碎至700目,混合均匀。)
1.1.2实施例2组
1.1.3实施例3组
1.1.4实施例4组
1.1.5空白对照组
1.2动物实验设计
选择同批孵化、体重相近、健康的150只黄羽肉鸡随机分成5组,每组30只,各试验组在基础日粮中分别添加非发酵制剂的超微粉、实施例2的复合中药发酵制剂、实施例3的复合中药发酵制剂和实施例4的复合中药发酵制剂,添加量为每吨饲料2kg;另设一组空白对照组,其他试验条件完全一致。
所述所述基础日粮参照NRC(1994)肉鸡营养需要量标准配制玉米—豆粕型基础日粮,日粮组成及营养水平见表1。
表1基础日粮组成及营养水平
采取笼养方式饲养,自由采食、饮水,常规的日常管理,按正常免疫程序进行免疫接种。每天记录每组鸡的采食量,每周进行1次鸡体重的测量并记录。
1.3指标测定
1.3.1复合中药发酵制剂对21日龄黄羽肉鸡免疫器官指数的影响
在21日龄每组随机抽取体重相近的鸡10只,空腹称个体重后屠宰,摘取脾脏、胸腺和法氏囊,剔除脂肪后称鲜重,计算器官指数(免疫器官鲜重g/宰前空腹活重kg)。如表2所示。
表2添加不同组分对21日龄黄羽肉鸡免疫器官指数的影响
注:同列肩标相同字母表示差异不显著(P>0.05),肩标不同小写字母表示差异显著(P<0.05)。
由表2可知,各试验组的法氏囊指数均高于对照组但无显著性差异(P>0.05)。脾脏指数超微粉组比对照组提高了14.45%(P>0.05);实施例2、3和4组分别比对照组显著提高了42.17%、39.76%和38.55%(P<0.05),比超微粉组分别提高了24.21%、22.10%和21.05%(P>0.05);胸腺指数超微粉组比对照组提高了18.48%(P>0.05);实施例2、3和4组分别比对照组显著提高了63.64%、52.20%和54.54%(P<0.05),比超微粉组显著提高了28.57%、19.58%和21.43%(P<0.05)。
1.3.2增重效果测定
观测并记录1~28日龄的平均日采食量、平均日增重及料肉比。
28日龄空腹称重,并准确称量和记录日供料量、余料量和损失量,计算平均日增重、平均日采食量和料重比。如表3所示。
表3增重指标测定结果
注:同列肩标相同字母表示差异不显著(P>0.05),肩标不同小写字母表示差异显著(P<0.05)。
由表3可知,平均日增重实施例2、3和4组显著高于对照组和超微粉组(P<0.05),分别比对照组显著提高了18.13%、13.42%和11.90%,分别比超微粉组显著提高了9.56%、5.19%和3.78%。平均日采食量各试验组与对照组之间无显著性差异(P>0.05)。料重比实施例2、3和4组显著低于对照组和超微粉组(P<0.05),分别比对照组显著降低了16.09%、13.79%和13.79%;分别比超微粉组降低了14.62%、12.28%和12.28%;超微粉组比对照组降低了1.72%,但无显著性差异(P>0.05)。
1.3.3屠宰性能测定
喂至65日龄,各组抽取体重中等的健康肉鸡10只,进行屠宰性能的测定,内容包括屠宰率、半净膛率、全净膛率、腿肌率、胸肌率,其测定方法参照《家禽生产性能名词术语和度量统计方法》(NY/T823—2004)。
屠宰率、全净膛率、半净膛率、胸肌率、腿肌率的具体计算方法如下:
屠宰率(%)=屠体重/活体重×100
全净膛率(%)=全净膛重/屠体重×100
半净膛率(%)=半净膛重/屠体重×100
胸肌率(%)=胸肌重/屠体重×100
腿肌率(%)=腿肌重/屠体重×100
结果如表4所示。
表4各试验组分对黄羽肉鸡屠宰性能的影响
注:同列肩标相同字母表示差异不显著(P>0.05),肩标不同小写字母表示差异显著(P<0.05)。
由表4可知,与对照组相比,各试验组屠宰率均显著提高(P<0.05),超微粉组、实施例2、3和4组,分别比对照组显著提高了4.85%、5.87%、5.85%和5.36%;各实施例组高于超微粉组但无显著性差异(P>0.05)。
半净膛率各试验组均显著高于对照组(P<0.05),超微粉组、实施例2、3和4组,分别比对照组显著提高了2.03%、3.75%、3.00%和3.08%;实施例2组较超微粉组显著提高了1.69%(P<0.05);实施例3、4组与超微粉组无显著性差异(P>0.05);实施例2组与实施例3、4组无显著性差异(P>0.05)。
全净膛率各试验组均显著高于对照组(P<0.05),超微粉组、实施例2、3和4组,分别比对照组显著提高了4.30%、7.02%、6.00%和5.89%;实施例2、3和4组较超微粉组分别显著提高了2.61%、1.63%和1.53%(P<0.05);实施例2组比实施例3、4组分别显著提高了0.96%和1.06%(P<0.05)。
胸肌率各试验组均显著高于对照组(P<0.05),超微粉组、实施例2、3和4组,分别比对照组显著提高了6.90%、15.32%、14.91%和14.01%;实施例2、3和4组较超微粉组分别显著提高了7.88%、7.49%和6.65%(P<0.05)。
腿肌率各试验组均显著高于对照组(P<0.05),超微粉组、实施例2、3和4组,分别比对照组显著提高了6.28%、18.72%、14.14%和14.40%;实施例2、3和4组较超微粉组分别显著提高了11.71%、7.39%和7.64%(P<0.05);实施例2组比实施例3、4组分别显著提高了4.02%和3.78%(P<0.05)。
1.4血清抗氧化指标的测定
1.4.1血清样品采集
饲养试验结束时,每个试验组选取接近平均体重的10只鸡进行翅静脉采血,静置,3000rpm离心10min,离心后血清分装于离心管中,-30℃保存待测。
1.4.2血清抗氧化指标及其测定方法
解冻血清用于测定谷胱甘肽含量(GSH)、总抗氧化能力(T-AOC)和超氧化物歧化酶(T-SOD),所有指标均采用南京建成生物工程研究所试剂盒测定。
表5各试验组分对黄羽肉鸡血清抗氧化指标性能的影响
注:同列肩标相同字母表示差异不显著(P>0.05),肩标不同小写字母表示差异显著(P<0.05)。
由表5可知,血清中GSH和T-SOD水平各试验组之间无显著性差异(P>0.05)。血清中T-AOC水平实施例2、3和4组分别比对照组显著提高了61.58%、60.14%和57.28%(P<0.05);实施例2、3和4组高于超微粉组但无显著性差异(P>0.05),分别比超微粉组提高了23.32%、22.22%和20.04%;超微粉组比对照组提高了31.03%(P>0.05)。
1.5不同发酵制剂对黄羽肉鸡肠道中大肠杆菌和乳酸杆菌含量的影响
菌群的测定:称黄羽肉鸡宰前活体重量后,颈静脉放血致死,立即剖腹,取出盲肠内容物,加无菌生理盐水匀浆,然后逐级10倍稀释。
大肠杆菌用麦康凯培养基37℃有氧培养24h后进行菌落计数。
乳酸杆菌用MRS培养基平皿上37℃厌氧培养48h后进行菌落计数。
细菌数量采用平板菌落计数法进行统计。结果如表6所示。
麦康凯培养基(g/L):蛋白胨20.0、乳糖10.0、牛胆盐5.0、NaCl 5.0、1%结晶紫0.1mL、琼脂12.0,调pH至7.2,121℃高压灭菌15min,备用;临用前加入1%中性红溶液(4mL/L)。
表6各试验组分对黄羽肉鸡盲肠微生物的影响
注:同列肩标相同字母表示差异不显著(P>0.05),肩标不同小写字母表示差异显著(P<0.05)。
由表6可知,实施例2、3和4组显著降低了盲肠中大肠杆菌的数量且较对照组显著增加了盲肠中乳酸杆菌数量(P<0.05)。
二、药理学毒性试验
2.1急性毒性试验资料
检测试剂:血清AST、ALT检测试剂盒购自南京建成生物工程研究所。
检测设备
Micro-200半自动生化检测仪,分析天平,解剖器械,注射器,试管等。
2.2试验方法
选取体重为18~22g的ICR鼠100只,经3天适应性饲养后,随机均分为5组(雌雄各半),超微粉组、实施例2组、实施例3组、实施例4组和空白对照组,每组各20只;空白对照组灌胃生理盐水,超微粉组所用超微粉选用本发明实施例2中的中药组合物进行超微粉碎得到的超微粉;超微粉组将超微粉溶于水中灌服,实施例2组、实施例3组、实施例4组分别将各实施例制备的发酵制剂溶于水中灌服,超微粉组、实施例2组、实施例3组、实施例4组的药物添加量为0.1g/mL,0.4mL/次,一天3次,连续7天。
2.3测定指标及方法
(1)临床症状观察:每日观察试验鼠的精神状态、饮食情况,每日记录发病死亡情况。
(2)病理学观察:对于死亡鼠进行剖检观察内脏变化。
(3)生长指标观察:对试验鼠进行定期称重;试验结束,扑杀鼠称量体重、脏器重脏器指数测定:剖检大鼠,取心脏、脾脏、肝脏、肾脏、睾丸、卵巢,称重,按照下式计算脏器指数:脏器指数=脏器重/体重×100。
(4)肝脏功能变化观察:试验结束后,扑杀鼠,采取血液分离血清,测定AST、ALT。
2.4数据处理与分析
所有采集数据采用Excel软件进行前期处理,然后用SPSS19.0进行统计分析,LSD进行多重比较,结果采用平均值±标准差表示。
2.5临床症状观察
对照组和试验组鼠,精神状态和饮食欲、对外界反应能力未见明显的异常。各组鼠发病和死亡情况如下表7。
表7鼠发病和死亡情况
分组 | 总数 | 死亡(只) | 死亡率(%) |
超微粉 | 20 | 0 | 0 |
实施例2组 | 20 | 0 | 0 |
实施例3组 | 20 | 0 | 0 |
实施例4组 | 20 | 0 | 0 |
对照组 | 20 | 0 | 0 |
病理学观察:对照组和试验组,鼠扑杀后进行剖检眼观未见观察内脏器官明显的病理变化。
生长指标观察:试验结束,扑杀鼠称量体重、脏器重分析见表8。
表8鼠脏器重分析
分组 | 肝脏指数 | 心脏指数 | 肾脏指数 | 肺脏指数 | 脾脏指数 |
超微粉 | 53.96±1.22<sup>a</sup> | 4.33±0.18<sup>a</sup> | 13.42±0.28<sup>a</sup> | 6.71±0.52<sup>a</sup> | 4.88±0.43<sup>ab</sup> |
实施例2组 | 54.19±1.05<sup>a</sup> | 4.68±0.24<sup>a</sup> | 13.57±0.25<sup>a</sup> | 6.83±0.39<sup>a</sup> | 5.59±0.32<sup>b</sup> |
实施例3组 | 54.13±1.24<sup>a</sup> | 4.53±0.22<sup>a</sup> | 13.53±0.28<sup>a</sup> | 6.77±0.42<sup>a</sup> | 5.52±0.42<sup>b</sup> |
实施例4组 | 54.11±1.39<sup>a</sup> | 4.52±0.25<sup>a</sup> | 13.49±0.26<sup>a</sup> | 6.80±0.44<sup>a</sup> | 5.51±0.35<sup>b</sup> |
对照组 | 53.88±1.58<sup>a</sup> | 4.27±0.15<sup>a</sup> | 13.30±0.24<sup>a</sup> | 6.64±0.45<sup>a</sup> | 4.14±0.70<sup>a</sup> |
注:同列肩标字母相同或无字母表示差异不显著(P>0.05),肩标字母不同小写字母表示差异显著(P<0.05)。
由表8可知,实施例2、3和4组显著增加了ICR鼠的脾脏指数(P<0.05),超微粉组与实施例2、3和4组差异不显著;其它指数对照组与试验组无显著性差异(P>0.05)。
肝脏功能变化观察
试验结束后,扑杀鼠,采取血液分离血清,测定AST(谷草转氨酶)、ALT(谷丙转氨酶)见表9。结果显示各组数据与空白对照组相比差异不显著,都处于正常范围内。
表9 AST、ALT的测定结果
分组 | 总数 | ALT | AST |
超微粉 | 20 | 39.11±2.23<sup>a</sup> | 105.43±4.18<sup>a</sup> |
2组 | 20 | 39.29±2.44<sup>a</sup> | 105.90±3.75<sup>a</sup> |
3组 | 20 | 39.19±2.81<sup>a</sup> | 105.77±4.09<sup>a</sup> |
4组 | 20 | 39.25±2.35<sup>a</sup> | 105.71±3.27<sup>a</sup> |
对照组 | 20 | 39.04±2.39<sup>a</sup> | 105.40±5.22<sup>a</sup> |
注:同列肩标字母相同或无字母表示差异不显著(P>0.05),肩标字母不同小写字母表示差异显著(P<0.05)。
由表9可知,各组数据与空白对照组相比差异不显著,都处于正常范围内。
2.6长期毒性试验资料
根据实施例2制备的本发明发酵制剂,20g/kg、100g/kg给1月龄ICR鼠连续灌胃给药1个月,动物的皮毛、肤色、摄食、活动、尿、便等均无异常动物的血常规、肝、肾功能等血液生化指标和动物的主要脏器指数均在正常范围,且正常动力无差异病理组织学检查表明,心、肝、脾、肺、肾、肾上腺、气管、睾丸、卵巢等重要脏器无病理改变。停药两周后检查以上各项指标均无明显毒性反应,表明无蓄积毒性。
以上所述,仅是本发明的较佳实施例而已,并非是对本发明作其它形式的限制,任何熟悉本专业的技术人员可能利用上述揭示的技术内容加以变更或改型为等同变化的等效实施例。但是凡是未脱离本发明技术方案内容,依据本发明的技术实质对以上实施例所作的任何简单修改、等同变化与改型,仍属于本发明技术方案的保护范围。
Claims (5)
1.一种复合中药发酵制剂,其特征在于:它由中药成分与菌液混合后经发酵制成;
所述菌液为植物乳杆菌与凝结芽孢杆菌的混合菌液;
所述菌液为植物乳杆菌与凝结芽孢杆菌按1∶1的比例混合的菌液;
所述凝结芽孢杆菌为凝结芽孢杆菌(Bacillus coagulans)zkng160127,保藏编号为CGMCC NO:12125;所述的植物乳杆菌为植物乳杆菌植物亚种(Lactobacillusplantarumsubsp.Plantarum),编号AS1.557,来源于中国科学院微生物研究所;
所述菌液中凝结芽孢杆菌和植物乳杆菌的含量均≥108CFU/mL;
所述中药成分由以下组分按重量份数制备而成:柴胡8份、川芎 6份、香附6份、陈皮6份、枳壳4.5份、芍药4.5份、甘草1.5份。
2.根据权利要求1所述复合中药发酵制剂的制备方法,其特征在于:步骤如下:
(1)按量取中药组分,粉碎,过60目筛网,备用;
(2)将粉碎后的各组分混合均匀,加入中药成分总重量80%的水,搅拌均匀后于100℃蒸汽灭菌30min,冷却至20-30℃;
(3)向冷却后的药物中加入混合菌液,菌液用量占加入后总量10~15%;置于单向排气塑料袋中,在20℃~30℃条件下发酵30天。
3.根据权利要求2所述复合中药发酵制剂的制备方法,其特征在于:所述菌液由以下方法制得:将凝结芽孢杆菌和植物乳杆菌菌种接种于活化培养基进行活化培养,培养温度为37℃,时间24h;将活化好的菌种接种于发酵罐中,控制温度37℃~43℃,时间24h~36h。
4.根据权利要求3所述复合中药发酵制剂的制备方法,其特征在于:所述活化培养基和所述发酵所用培养基为:MRS培养基,成分为(g/L):蛋白胨10.0、肉膏10.0、酵母浸粉5.0、葡萄糖20.0、磷酸氢二钾2.0、柠檬酸氢二铵2.0、乙酸钠5.0、硫酸镁0.58、硫酸锰0.28、半胱氨酸0.5、吐温80 1.0,调pH 6.2-6.4。
5.一种具有提高家禽生长性能和免疫力的饲料,其特征在于:在饲料中添加如权利要求2方法制备的复合中药发酵制剂,添加量为2kg/吨。
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