CN110812408A - 一种防控仔猪腹泻的液态发酵中药制剂及其制备方法 - Google Patents
一种防控仔猪腹泻的液态发酵中药制剂及其制备方法 Download PDFInfo
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Abstract
本发明公开了一种防控仔猪腹泻的液态发酵中药制剂，其原料包括以下重量份的组分：白头翁提取物30‑38份、黄连提取物22‑28份、青皮提取物20‑25份、黄柏提取物22‑28份、木香提取物18‑25份、乌梅提取物25‑30份、诃子提取物22‑26份、伏苓提取物18‑22份、木通提取物15‑20份、甘草提取物18‑22份。本发明以中药作为发酵原料，利用优选的丁酸梭菌及乳酸菌对中间物质进行生物转化，制成无毒、无残留、绿色、易吸收、抗菌活性明显增强的中药微生态制剂，使药效得到明显改善，本发明提供的一种中药发酵制剂的制备方法简单合理，对中药进行发酵，使得发酵中药结合了传统中药和益生菌的优点，且生产周期短，成本较低。
Description
技术领域
本发明涉及发酵中药添加剂技术领域，具体是一种防控仔猪腹泻的液态发酵中药制剂及其制备方法。
背景技术
随着我国现代化进程的不断加快，养殖业集约化的程度也越来越高，发生仔猪腹泻病的几率也不断增加，同时，导致仔猪腹泻的病因也越来越复杂。仔猪腹泻通常是由单一或多重因素共同作用引起（大多数为多因素作用）的仔猪消化机能紊乱，进而出现消化吸收不良，仔猪肠道菌群失衡，肠道内聚集大量的水分和电解质，粪便常呈液体状排出或喷射出体外的一种病理现象。据数据统计显示，在规模化大型猪场中仔猪死亡率很高，约占猪死亡总数的65% 左右，而腹泻引起的死亡高达30% 左右。引发该病的病因比较复杂，包括环境因素、饲养管理因素、病原性因素（细菌、病毒、寄生虫等）和其他因素，并且多种因素间相互作用，给临床诊断和治疗带来了很大的困扰。仔猪感染本病后，严重影响仔猪的生长发育，该病的致死率较高，如果不能有效做好预防工作，及时做出诊断和治疗，进而可能会引起并发或 继发其他感染性疫病，给养殖场造成巨大的经济损失，也严重威胁着畜牧业的健康发展。
当仔猪出现因腹泻发生时，通常有以下几种原因与对策：一是细菌感染，常规的做法是选用高效敏感的广谱类抗生素药物，如土霉素、庆大霉素等；二是病毒传染性腹泻，尚没有高效的药物，通常使用一些抗病毒的药物，同时配合使用抗生素药物，防止患病仔猪发生继发感染。三是寄生虫病腹泻，可以使用抗寄生虫类的广谱高效的药物；三是原虫，可以使用磺胺嘧啶等，蠕虫或外表寄生虫可使用阿维菌素或伊维菌素等药物治疗；四是应激因素，做好保温通风工作，强化饲养管理，给予科学合理配比的全 饲料等综合管理措施；五是定期免疫接种，控制蓝耳病、圆环病毒、伪狂犬病发生。但随着养殖环境恶化，疾病趋于更加复杂化。而且现有抗生素滥用结果，造成仔猪腹泻难以控制，二月龄仔猪死淘率居高不下。
发明内容
本发明的目的在于提供一种防控仔猪腹泻的液态发酵中药制剂及其制备方法，以解决上述背景技术中提出的问题。
为实现上述目的，本发明提供如下技术方案：
一种防控仔猪腹泻的液态发酵中药制剂，其原料包括以下重量份的组分：白头翁提取物30-38份、黄连提取物22-28份、青皮提取物20-25份、黄柏提取物22-28份、木香提取物18-25份、乌梅提取物25-30份、诃子提取物22-26份、伏苓提取物18-22份、木通提取物15-20份、甘草提取物18-22份。
作为本发明进一步的方案：其原料包括以下重量份的组分：白头翁提取物30份、黄连提取物25份、青皮提取物20份、黄柏提取物25份、木香提取物20份、乌梅提取物25份、诃子提取物25份、伏苓提取物20份、木通提取物15份、甘草提取物20份。
一种防控仔猪腹泻的液态发酵中药制剂的制备方法，具体步骤如下：
（1）精选中药提取物，称取相应份数的白头翁、黄连、青皮、黄柏、木香、乌梅、诃子、伏苓、木通、甘草，组成混合粉料；
（2） 将所述混合粉料和发酵培养基混匀后，接种益生菌，并于36℃～38℃发酵24h～48h，得到发酵水剂；
（3）将所述液体发酵混合物分装后入库。
作为本发明进一步的方案：所述步骤（2）中益生菌为嗜酸乳杆菌+丁酸梭菌。
作为本发明进一步的方案：所述步骤（2）中的发酵在发酵罐中进行，发酵的条件为：压强为0.02kPa～0.1kPa，搅拌转速为10r/min～30r/min，pH值为6.0～7.0。
作为本发明进一步的方案：所述步骤（2）中混合粉料和发酵培养基的添加比例为(8～20)g:1L。
作为本发明进一步的方案：所述步骤（2）中接种的益生菌为益生菌菌种的二级种子液,二级种子液的制备步骤如下：
（1）将益生菌菌种活化后，接种于种子培养基中，于36℃～38℃静止培养12h～24h，得到一级种子液；
（2）将所述一级种子液按照2vol％～10vol％的接种量接种于种子培养基中，并于36℃～38℃静止培养12h～24h，得到二级种子液；
（3）按照所述二级种子液与所述发酵培养基的体积比为3vol％～8vol％，接种所述二级种子液。
作为本发明进一步的方案：所述发酵培养基的原料包括以下重量份的组分：精制蛋白胨10份、酵母浸膏5份、葡萄糖20份、醋酸钠5份、柠檬酸铵2份、吐温80合0.1份、硫酸镁0.58份、硫酸锰0.28份、蒸馏水1000份。
其防控仔猪腹泻机理是：
一是中药发酵制剂可以增强仔猪机体免疫力，中药发酵制剂中含有多种免疫活性成分，包括 ：多糖类、黄酮类、糖苷 类和生物碱等，不仅能直接抑制病原菌，还能通过激活免疫系统从而抵抗疾病。发酵中药制剂中含有多种营养物质，不仅包括蛋白质、糖类和脂肪，还含有氨基酸、维生素和矿物质微量元素等机体生长所必需的营养物质，可以补充仔猪生长过程中出现的营养不足。发酵中药制剂还能够增强脾胃功能，促进营养物质代谢，从而促进仔猪机体生长发育。中药发酵制剂含有多种免疫活性物质，缓解断奶仔猪应激。
二是液态发酵中药制剂中含丁酸梭菌、乳酸菌微生态益生菌，其可以定植仔猪肠道并显著增加肠道中乳酸杆菌和双歧杆菌的数量而减少肠道中大肠杆菌的数量，因而可以降低仔猪腹泻的发生率。微生态益生菌还可提高仔猪的免疫机能。微生态制剂是良好的免疫激活剂，可提高干扰素和巨噬细胞的活性，增强吞噬细胞的能力，在抗菌、抗病毒及抵抗霉菌毒素方面表现出显著效果。
白头翁化学成分，近年国内外对白头翁及白头翁属植物化学成分进行 了较系统的研究，主要包括三萜皂苷、三萜酸、木脂素、 胡萝卜苷以及糖蛋白等成分。白头翁保护肠道微生物屏障，纠正肠道菌群紊乱，调节肠道微生态。采用白头翁水提液，小鼠连续灌胃6 d( 100 mg/kg) ，小鼠腹腔巨噬细胞的吞噬率为51.18%，吞噬指数1.10，脾指数35. 63，与对照组比较均有显著性差异，表明白头翁水提货对免疫功能具有增强作用。白头翁水提物能够提高小鼠血清超氧化物歧化酶的活性，给药组小鼠血清 SOD 水平为 517. 86 U/mL，比空白组提高 16. 0%，具有显著性差异，其机制可能与消除氧自由基、中断或终止自由基的氧化反应以及增强机体的总抗氧化能力有关。
黄连又名川连、川黄连、姜制黄连、姜连、萸连等。为 毛茛科植物黄连Coptischinensis Franch.、或云连Coptis teeta Wall. 以及三角叶黄连 Coptis deltoidea C.Y. Chenget Hsiao 的干燥根茎，属于清热类药物，始载于《神农本草经》，且被列为上品。黄连的主要组成为小檗碱、黄连碱分子式C19H15NO5，相对分子质量为337.32、掌叶防己碱异名为巴马丁，分子式C21H22NO4，相对分子质量为 352.40、木兰碱、2,3,4-三羟基苯丙（2,3,4-trihudroxybenzenpropanoic acid）、3- 羧基 -4 羟基苯氧葡萄糖（3-carboxy4-hudroxy-phenoxyglucoside）、3,4-二羟基苯乙醇葡萄糖苷（3, 4-dihudroxy-phenylethylalcohol glucoside）等 。黄连的主要功能：一是抗炎、解热 ，盐酸小檗碱对急、慢性炎症均有抑制作用。小檗碱抗炎作用与其抑制炎症过程的某些环节有关，如黄连和小檗碱在体内外均能增强白细胞的吞噬功能。黄连可通过抑制中枢发热介质的生成或释放产生解热作用，如解热黄连注射液对实验性发热就有明显的解热作用，并且能降低脑脊液中 cAMP 含量；二是抗细菌毒素、抗腹泻， 黄连和盐酸小檗碱能够对抗多种病原微生物的毒素，提高机体对细菌内毒素的耐受能力，且能对抗大肠杆菌引起的腹泻。此外，小檗碱还能对抗霍乱毒素引起的腹泻以及非感染性腹泻，并减轻小肠绒毛的水肿、分泌亢进等炎症反应， 降低死亡率；三是抗病原体，黄连及小檗碱具有广谱抗菌的作用，对多种细菌、结核 杆菌及真菌等都有抑制或杀灭作用。已有文献报道，黄连及盐酸小檗碱对溶血性链球菌、金黄色葡萄球菌、绿脓杆菌、 大肠杆菌、痢疾杆菌、阴沟杆菌及伤寒杆菌共 7 种 16 株致病菌均有抑制作用。此外，黄连及三黄注射液（等量黄连、黄芩、黄柏组成）还具有抗真菌作用，黄连煎液及盐酸小檗碱在体外及体内均有一定的抗阿米巴作用，盐酸小檗碱对多种流感病毒均有抑制作用。值得注意的是，黄连低浓度时抑菌， 高浓度杀菌，抗菌作用环节一般是：破坏细菌结构 — 抑制细菌糖代谢（黄连能抑制酵母菌及细菌糖代谢的中间环节丙酮酸的氧化脱羧过程）— 抑制核酸、蛋白质合成。
青皮 ,亦称青桔皮或青柑皮 ,为芸香科柑橘属植物橘 Citrus reticulataBlanco及其变种自行落地的未成熟果实。 青皮药材包括个青皮、四花青皮。青皮为常用中药 ,性味苦辛 ,微温 ,有破气行痰 ,消痞除满之功效。化成成分：右旋柠檬烯 (dlimonen,53. 3% ) 、黄酮类化合物（青皮内总黄酮为 24. 60%）、氨基酸类（谷氨酸、脯氨酸、甘氨酸、丙氨酸、胱氨酸、缬氨酸、异亮氨酸、亮氨酸、苯丙氨酸、组氨酸、精氨酸、酪氨酸等）。
所含挥发油有祛痰作用，其有效成分为柠檬烯。麻醉猫静脉注射自青皮甲醇浸膏中分得的对羟福林草酸盐lmg／kg，可完全对抗组胺引起的支气管收缩，作用持续约l小时。
青皮注射液能降低离体豚鼠胃、肠、胆囊及小鼠子宫的紧张性收缩，并使膀胱平滑肌兴奋。对乙酰胆碱引起的豚鼠离体胃、肠及家兔在体胃平滑肌以及氨甲酰胆碱引起的胆囊收缩，有显著的解痉作用。
丁酸梭菌作为饲用益生菌种， 在动物肠道内呈现出五大生物特性： 1）丁酸梭菌作为厌氧芽孢杆菌，不受胃酸、胆汁酸等影响 ； 2）丁酸梭菌产生的丁酸、乳酸，能够促进动物肠道中乳酸菌、双歧杆菌等有益 菌群的增殖与发育，抑制肠道内有 害菌和腐败菌的生长、繁殖，纠正肠道菌群紊乱，调节肠道微生态平衡，减少肠毒素的产生 ； 3）丁酸梭菌在动物肠道内产生的淀粉酶、纤维素酶、蛋白酶等消化酶以及B 族 维生素、维生素K 等维生素，这些物质连同丁酸等对动物具有营养保健作用；4）丁酸、乙酸、氢气等丁酸梭菌的代谢产物，能够减轻 肠道炎症，加速肠道损伤修复，尤其是主要代谢产物丁酸，它是肠道上皮组织细胞再生和修复的主要营养物质 ； 5）对多种饲用抗生素 具有较强的耐受性，可以与之配伍使用。由此可见，丁酸梭菌作为饲用益生菌，集芽孢杆菌与乳酸菌的优点于一身，既具有优良的生产性 能，又具有耐热、易贮藏、适应动物肠道微生态环境（厌氧、酸、胆盐） 的特性，对于减少当前动物生产中饲用抗生素的滥用、减少药物在肉、蛋、乳等畜禽产品中的残留、降低 动物细菌的耐药性与保障动物健康 方面有着重要的意义，是一个极具 潜力的饲用益生菌种，有着广阔的 市场前景。
乳酸菌是一类能利用可发酵碳水化合物产生大量乳酸细菌的通称，革兰阳性，在自然界分布极为广泛且物种丰富多样。目前乳酸菌主要包括 23 个属， 200多个种。在传统观念上，典型的乳酸菌由4 个属组成: 乳杆菌属 、明串珠菌属、片球菌属和链球菌属 。乳杆菌属作为乳酸菌中重要的一个属，也是最大的一 个属，迄今为止已报道有 155 个种和亚种。目前常用于食品发酵、工业乳酸菌发酵以及医疗领域的乳杆菌有德氏乳杆菌保加利亚亚种、干酪乳杆菌、瑞氏乳杆菌、植物乳杆菌、短乳杆菌、嗜酸乳杆菌和副干酪乳杆菌等。乳酸菌作为益生菌的主要类型之一，常占据并定殖于宿主肠道，调控并保持着宿主和体内微生物群之间的动态平衡并实现着对宿主的益生功能。
复合组方中药提取物经过复合微生态制剂丁酸梭菌与乳酸菌发酵后，大分子有效成分变成小分子有效成分，更加容易被消化吸收，提高了药性。同时，复合微生态制剂本身也是维护肠道健康最忠诚卫生，无论多复杂的仔猪腹泻问题都可以得以迅速解决。为了找到最佳中药配方组合与微生态组合，开展大小试验100余次，最终获得上述的制备数据。
与现有技术相比，本发明的有益效果是：
本发明具有补元阳、暖脾胃，清热解毒、理气止痛，杀虫杀菌，涩肠止泻之功效，乳仔猪服用后可以快速止泻，一般在1-2小时见效，1-2天既可痊愈，不会反复发作。可以有效解决仔猪腹泻防控问题，且不留下任何药物残留。
本发明以中药作为发酵原料，利用优选的丁酸梭菌及乳酸菌对中间物质进行生物转化，制成无毒、无残留、绿色、易吸收、抗菌活性明显增强的中药微生态制剂，使药效得到明显改善，本发明提供的一种中药发酵制剂的制备方法简单合理，对中药进行发酵，使得发酵中药结合了传统中药和益生菌的优点，且生产周期短，成本较低。
具体实施方式
下面将结合本发明实施例，对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。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，都属于本发明保护的范围。
本发明实施例中，本发明提供的一种液态发酵中药微生态制剂，其中药复方的基料组成数据如表1、
一种防控仔猪腹泻液态发酵中药制剂
S1: 按照表1要求分别称取中药提取物，并按下述步骤，制备一种防控仔猪腹泻液态发酵中药制剂；
S2: 将所述混合粉料和发酵培养基混匀后，接种益生菌，并于36℃～38℃发酵24h～48h，得到液态中药发酵制剂；
S3:分别将所述液态中药发酵制剂分装，得到所述液态中药发酵制剂，分开装包并标识待用。
实施案例：安徽省霍邱县新店镇砟巴集村张** 1390564**
张老板的养猪场，2019年5月，存栏仔猪（1-25日龄）120头。分成3组，试验1组分3次预防性口服料1（1-3日龄，每头每天口服3ml；7-10天龄，每头每天口服5ml；15-20天龄，每头每天口服8ml）；试验2组饮水中添加料2（1-3日龄，每头每天口服3ml；7-10天龄，每头每天口服5ml；15-20天龄，每头每天口服8ml）,试验3组为对照组（按常规保健，饮水定期加抗生素）。
从表1看出：试验开始时，各组数量一致，体重大小一致（差异不显著）。
从表2看出：25天龄时，试验组体重超过对照组（差异显著）；试验组成活率高于对照组（差异显著）。
从表3看出：25天龄时，试验组仔猪腹泻发生率为0,而对照组仔猪腹泻发生率为32.9%。试验组仔猪腹泻发生率低于对照组（差异显著）。
综上所述：本发明提供的一种中药微生态制剂成分配比合理，以中药作为发酵原料，利用优选的丁酸梭菌及乳酸菌对中间物质进行生物转化，制成无毒、无残留、绿色、易吸收、抗菌活性明显增强的中药微生态制剂，使药效得到明显改善，本发明提供的一种中药发酵制剂的制备方法简单合理，对中药进行发酵，使得发酵中药结合了传统中药和益生菌的优点，且生产周期短，成本较低。
对于本领域技术人员而言，显然本发明不限于上述示范性实施例的细节，而且在不背离本发明的精神或基本特征的情况下，能够以其他的具体形式实现本发明。因此，无论从哪一点来看，均应将实施例看作是示范性的，而且是非限制性的，本发明的范围由所附权利要求而不是上述说明限定，因此旨在将落在权利要求的等同要件的含义和范围内的所有变化囊括在本发明内。
此外，应当理解，虽然本说明书按照实施方式加以描述，但并非每个实施方式仅包含一个独立的技术方案，说明书的这种叙述方式仅仅是为清楚起见，本领域技术人员应当将说明书作为一个整体，各实施例中的技术方案也可以经适当组合，形成本领域技术人员可以理解的其他实施方式。
Claims (9)
1.一种防控仔猪腹泻的液态发酵中药制剂，其特征在于，其原料包括以下重量份的组分：白头翁提取物30-38份、黄连提取物22-28份、青皮提取物20-25份、黄柏提取物22-28份、木香提取物18-25份、乌梅提取物25-30份、诃子提取物22-26份、伏苓提取物18-22份、木通提取物15-20份、甘草提取物18-22份。
2.根据权利要求1所述的防控仔猪腹泻的液态发酵中药制剂，其特征在于，其原料包括以下重量份的组分：白头翁提取物30份、黄连提取物25份、青皮提取物20份、黄柏提取物25份、木香提取物20份、乌梅提取物25份、诃子提取物25份、伏苓提取物20份、木通提取物15份、甘草提取物20份。
3.一种如权利要求1-2任一所述的防控仔猪腹泻的液态发酵中药制剂的制备方法，其特征在于，包括如下步骤：
（1）精选中药提取物，称取相应份数的白头翁、黄连、青皮、黄柏、木香、乌梅、诃子、伏苓、木通、甘草，组成混合粉料；
（2） 将所述混合粉料和发酵培养基混匀后，接种益生菌，并于36℃～38℃发酵24h～48h，得到液态发酵水剂；
（3）将所述液体发酵混合物分装后入库。
4.根据权利要求3所述的防控仔猪腹泻的液态发酵中药制剂的制备方法，其特征在于，所述步骤（2）中益生菌为嗜酸乳杆菌+丁酸梭菌。
5.根据权利要求3所述的防控仔猪腹泻的液态发酵中药制剂的制备方法，其特征在于，所述步骤（2）中发酵培养基的原料包括以下重量份的组分：精制蛋白胨10份、酵母浸膏5份、葡萄糖20份、醋酸钠5份、柠檬酸铵2份、吐温80合0.1份、硫酸镁0.58份、硫酸锰0.28份、蒸馏水1000份。
6.根据权利要求3所述的防控仔猪腹泻的液态发酵中药制剂的制备方法，其特征在于，所述步骤（2）中的发酵在发酵罐中进行，发酵的条件为：压强为0.02kPa～0.1kPa，搅拌转速为10r/min～30r/min，pH值为6.0～7.0。
7.根据权利要求3所述的防控仔猪腹泻的液态发酵中药制剂的制备方法，其特征在于，所述步骤（2）中混合粉料和发酵培养基的添加比例为(8～20)g:1L。
8.根据权利要求3所述的防控仔猪腹泻的液态发酵中药制剂的制备方法，其特征在于，所述步骤（2）中接种的益生菌为益生菌菌种的二级种子液,二级种子液的制备步骤如下：
（1）将益生菌菌种活化后，接种于种子培养基中，于36℃～38℃静止培养12h～24h，得到一级种子液；
（2）将所述一级种子液按照2vol％～10vol％的接种量接种于种子培养基中，并于36℃～38℃静止培养12h～24h，得到二级种子液；
（3）按照所述二级种子液与所述发酵培养基的体积比为3vol％～8vol％，接种所述二级种子液。
9.根据权利要求8所述的防控仔猪腹泻的液态发酵中药制剂的制备方法，其特征在于，一级种子液与二级种子液制备所用的种子培养基配方相同，其原料包括以下重量份的组分：精制蛋白胨10份、酵母浸膏5份、葡萄糖20份、醋酸钠5份、柠檬酸铵2份、吐温80合0.1份、硫酸镁0.58份、硫酸锰0.28份、蒸馏水1000份。
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