CN106039138B - 一种用于水产养殖的乳酸菌中药发酵抗病毒制剂 - Google Patents
一种用于水产养殖的乳酸菌中药发酵抗病毒制剂 Download PDFInfo
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Abstract
一种用于水产养殖的乳酸菌中药发酵抗病毒制剂，其特征在于：该抗病毒制剂是将乳酸菌种子液与配伍好的中药混合提取物发酵制得的，配伍中药的各组分质量份数如下：板蓝根10～20份，大青叶10～20份，大黄5～10份，黄芪1～5份，鱼腥草3～5份，青蒿1～5份，荆芥2～5份，五味子1～5份，金银花4～7份，连翘1～5份，甘草5～7份，虎杖1～5份，大麦芽1～5份；本发明采用的中药混合物复合依据中药理论，科学配伍，保留了中药方剂辨证论治的组合效应，并运用三次水煎得到提取物，最大程度的保留中药的有效成分，避免了现有技术中中药直接用于水产养殖而吸收不佳的缺点。
Description
技术领域
本发明属于生物发酵中药技术领域，具体涉及一种用于水产养殖的乳酸菌中药发酵抗病毒制剂。
背景技术
乳酸菌是一类无芽孢、革兰氏染色阳性细菌的统称，是动物肠道内的常在菌，具有调节畜禽硝化到菌群平衡，提高动物免疫性能的作用，目前市场上常用的乳酸菌微生态制剂多以普通培养基培养乳酸菌，然后以液体或粉剂形式存在。目前，在防疫和治疗病毒性疾病多采用广谱抗病毒类西药，这类药物，很容易产生抗药性，使病毒变异，水产养殖中的病毒性变异便是最好的证明，所以，这类药物的使用，具有较大的局限性。近年来，随着水产养殖业发展，渔民追求高利益，集约化养殖使得病毒性疾病发病率极高，病毒不断变种，传播快，流行广，严重危害水产养殖业发展。临床应用的抗病毒药物的作用途径很多，如直接抑制或杀灭病毒、干扰病毒吸附、阻止病毒穿入细胞、抑制病毒生物合成、抑制病毒释放或增强宿主抗病毒能力等。但目前国内外治疗病毒有效的西药副作用大，抗病毒中药品种少且有效率低，因此，开发抗病毒的中药新品种应用疾病防治是十分必要的。
发明内容
针对上述问题，本发明的目的提供一种用于水产养殖的乳酸菌中药发酵抗病毒制剂，有效解决现有水产养殖过程中抗病毒制剂易产生抗药性的缺点。
本发明解决上述技术问题所采用的技术方案是：一种用于水产养殖的乳酸菌中药发酵抗病毒制剂，其特征在于：该抗病毒制剂是将乳酸菌种子液与配伍好的中药混合提取物发酵制得的，配伍中药的各组分质量份数如下：
进一步，所述乳酸菌种子液由屎肠球菌与植物乳杆菌按3：7比例混合培养制得，有效活菌数量大于1010个。
进一步，所述中药混合提取物为每25g中药混合物加100g水煎三次合并制得。
进一步，所述抗病毒制剂中还包括MRS液体培养基。
一种制备用于水产养殖的乳酸菌中药发酵抗病毒制剂方法，其特征在于：包括以下步骤：
(1)将如下重量份数的中药放入研磨机中粉碎，并过80～100目筛，得到中药粉
(2)将步骤(1)得到的中药粉每25g中药粉加入100mL水，加热至沸腾，并且持续沸腾23～30分钟，过滤取上清液；滤渣继续加入100mL水，加热至沸腾，并且持续沸腾34～44分钟，取上清液与第一次的上清液混合；然后滤渣继续加入100mL水，加热至沸腾，并且持续沸腾45～50分钟，过滤上清液与前两次的上清液混合，得到中药混合提取物150～200mL；
(3)MRS液体培养基的制备：胰蛋白胨10.0g、牛肉膏10.0g、酵母膏5.0g、柠檬酸铵2.0g、葡萄糖20.0g、KH2PO42.0g、乙酸钠5g、吐温-801.0mL、七水硫酸镁0.58g、硫酸锰0.25g、浓度为0.25～1.0mol/L的KH2PO4缓冲液1000mL，用0.1mol/L的NaOH调节pH至6.5，121℃，灭菌25min，得到所述改良MRS液体培养基；
(4)将步骤(2)得到的中药混合提取物和步骤(3)得到的MRS液体培养基按40～60:60～40的重量比混合，然后加入乳酸菌种子液37～39°厌氧发酵2～3天得到抗病毒制剂。
进一步，所述乳酸菌种子液由屎肠球菌与植物乳杆菌按3：7比例混合培养制得，有效活菌数量大于1010个。
本发明的优点在于：1、本发明采用的中药混合物复合依据中药理论，科学配伍，保留了中药方剂辨证论治的组合效应，并运用三次水煎得到提取物，最大程度的保留中药的有效成分，避免了现有技术中中药直接用于水产养殖而吸收不佳的缺点；2、本发明在中药混合提取物和MSR液体培养基混合用乳酸菌发酵直接得到抗病毒制剂，所得到的抗病毒制剂为液态的，可以直接用于水产养殖；3、本发明含有的中药成分经过益生菌转化，有利于机体吸收；能提速疗效及提高药效；此外发酵过程分解毒性，与使用化学药剂相比，也避免了化学药物的残留，具有清热解毒、祛风燥湿的作用。
具体实施方式
下面通过具体实施例对本发明作进一步详述，以下实施例只是描述性的，不是限定性的，不能以此限定本发明的保护范围。
实施例一：
将上述重量的中药放入研磨机中粉碎，并过80～100目筛，得到中药粉，然后加入300mL水，加热至沸腾，并且持续沸腾30分钟，过滤取上清液；滤渣继续加入300mL水，加热至沸腾，并且持续沸腾44分钟，取上清液与第一次的上清液混合；然后滤渣继续加入300mL水，加热至沸腾，并且持续沸腾50分钟，过滤上清液与前两次的上清液混合，得到中药混合提取物450mL；
MRS液体培养基的制备：胰蛋白胨10.0g、牛肉膏10.0g、酵母膏5.0g、柠檬酸铵2.0g、葡萄糖20.0g、KH2PO42.0g、乙酸钠5g、吐温-80 1.0mL、七水硫酸镁0.58g、硫酸锰0.25g、浓度为0.25～1.0mol/L的KH2PO4缓冲液1000mL，用0.1mol/L的NaOH调节pH至6.5，121℃，灭菌25min，得到所述改良MRS液体培养基；
将中药混合提取物和MRS液体培养基按40:60的重量比混合，然后加入乳酸菌种子液37～39°厌氧发酵2～3天得到抗病毒制剂。
实施例二：
参照实施例一制备抗病毒制剂，区别在于中药组分如下：
将实施例一和实施例二得到的抗病毒制剂投放至鲤鱼养殖池中，对于患鲤鱼疱疹病毒病的鲤鱼，按1.0mL/每公斤鱼体重计算药量，拌饲料连续投喂5天治愈率达88％和90％。
实施例一和实施例二中的有效活菌数量分别为6×1010,7×1010，说明中药混合提取物可以增效乳酸菌的发酵。
最后应说明的是：以上各实施例仅用以说明本发明的技术方案，而非对其限制；尽管参照前述各实施例对本发明进行了详细的说明，本领域的普通技术人员应当理解：其依然可以对前述各实施例所记载的技术方案进行修改，或者对其中部分或者全部技术特征进行等同替换；而这些修改或者替换，并不使相应技术方案的本质脱离本发明各实施例技术方案的范围。
Claims (1)
1.一种用于水产养殖的乳酸菌中药发酵抗病毒制剂，其特征在于：该抗病毒制剂是将乳酸菌种子液与配伍好的中药混合提取物发酵制得的，配伍中药的各组分质量份数如下：
所述乳酸菌种子液由屎肠球菌与植物乳杆菌按3：7比例混合培养制得，有效活菌数量大于1010个；
所述的用于水产养殖的乳酸菌中药发酵抗病毒制剂的制备方法具体包括以下步骤：
(1)将上述重量份数的中药放入研磨机中粉碎，并过80～100目筛，得到中药粉；
(2)将步骤(1)得到的中药粉每25g中药粉加入100mL水，加热至沸腾，并且持续沸腾23～30分钟，过滤取上清液；滤渣继续加入100mL水，加热至沸腾，并且持续沸腾34～44分钟，取上清液与第一次的上清液混合；然后滤渣继续加入100mL水，加热至沸腾，并且持续沸腾45～50分钟，过滤上清液与前两次的上清液混合，得到中药混合提取物150～200mL；
(3)MRS液体培养基的制备：胰蛋白胨10.0g、牛肉膏10.0g、酵母膏5.0g、柠檬酸铵2.0g、葡萄糖20.0g、KH2PO42.0g、乙酸钠5g、吐温-801.0mL、七水硫酸镁0.58g、硫酸锰0.25g、浓度为0.25～1.0mol/L的KH2PO4缓冲液1000mL，用0.1mol/L的NaOH调节pH至6.5，121℃，灭菌25min，得到MRS液体培养基；
(4)将步骤(2)得到的中药混合提取物和步骤(3)得到的MRS液体培养基按40～60:60～40的重量比混合，然后加入乳酸菌种子液37～39℃厌氧发酵2～3天得到抗病毒制剂。
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