CN113180148A - 一种代替抗生素的发酵饲料的制备方法 - Google Patents
一种代替抗生素的发酵饲料的制备方法 Download PDFInfo
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Abstract
本发明涉及由配合料加入复合菌液经固态发酵制成的饲料，其中复合菌液由植物乳杆菌菌液1‑10重量份、米曲霉菌液1‑10重量份和酿酒酵母菌液1‑10重量份混合制成。与普通猪用饲料相比，本发明的饲料配伍合理，疗效显著，能够显著降低猪腹泻率，实验证明对改善猪腹泻有特效；由于饲料中无西药抗生素成分，因此本发明的饲料无毒副作用，无耐药性，无污染。
Description
技术领域
本发明涉及饲料领域，具体涉及一种家畜饲料及其应用。
背景技术
20世纪在饲料中添加抗生素类药物，带来了全球畜牧养殖业的飞速发展。但是，动物长期饲用添加抗生素的饲料，会在体内产生耐药菌株，并造成动物产品的药物残留等问题。随着人民生活水平的逐步提高，消费者对畜禽产品的需求已经从追求数量过渡到追求质量，所以在饲料中添加抗生素的做法越来越受到限制，目前在欧洲、北美、亚洲等地区的许多国家已经禁止在饲料中添加抗生素。寻找绿色环保型抗生素替代物，减轻抗生素对畜禽生产带来的负面影响成为各国畜牧业迫切需要解决的问题。
益生菌制剂具有无毒副作用、无耐药性、无残留以及效果显著、成本低等诸多优点，能有效补充消化道内的有益微生物，杀灭有害微生物尤其是大肠杆菌等病原菌，改善消化道菌群平衡，迅速提高机体抗病、代谢及饲料的吸收能力，从而起到防治消化道疾病和促进生长的双重效果。益生菌制剂以其独特的作用方式，天然的组成成份，明显的作用效果，成为了理想的抗生素替代品。
国内外将益生菌大体上可以分成三类，第一类为一般厌氧即耐氧的乳杆菌属，如嗜酸乳杆菌、干酪乳杆菌等；第二类为严格厌氧的双歧杆菌属，如长双歧杆菌、短双歧杆菌等；第三类为兼性厌氧球菌，如粪肠球菌、乳肠球菌等。此外，还有一些酵母菌与酶亦可归入益生菌的范畴。常用于饲料中的益生菌包括乳酸菌、芽孢杆菌和酵母菌等。
植物乳杆菌属于乳酸菌的一种，广义的乳酸菌是指能从葡萄糖或乳糖的发酵过程中产生乳酸的细菌。狭义的乳酸菌是一类革兰染色呈阳性的杆菌或球菌，不形成芽孢、过氧化氢酶试验呈阴性且对葡萄糖发酵能产生超过50％的乳酸，而且是在缺少氧气的环境中生长良好的兼性厌氧性细菌。植物乳杆菌属于乳杆菌科中的乳杆菌属，革兰阳性兼性厌氧，不产芽孢圆端直杆，单个、成对或短链状，产生D型和L型2种乳酸。可在10℃生长，在45℃不生长，最适生长温度为30～35℃。在pH4.5～9.5生长，最适pH约为6.5。植物乳杆菌最早是从唾液中分离出的，分布很广泛。
植物乳杆菌能利用葡萄糖、麦芽糖、蔗糖、L-山梨糖、乳糖、纤维二糖、木糖、蜜二糖、果糖、核糖和葡萄糖酸钠产酸，但发酵葡萄糖和葡萄糖酸钠均不产气，属于同型发酵类型中的兼性异型发酵群。利用其发酵乳糖产酸处理乳清中乳糖转化时，具有其他菌无法比拟的优越性，它既可以高转化率地利用乳糖转化为乳酸，又可以利用乳清中残留蛋白质。植物乳杆菌在发酵过程中只产生乳酸，是典型的兼性厌氧菌，有很强的发酵糖类的能力，较耐盐，与其他乳酸菌有协同作用。
米曲霉是一种好气性真菌，属于半知菌亚门、曲霉属，菌丝一般呈黄绿色，后为黄褐色，分生孢子梗生长在厚壁的足细胞上，分生孢子头呈放射形，项囊球形或瓶形，小梗一般为单层，分生孢子球形平滑，少数有刺，培养适温为37℃。米曲霉的菌丝由多细胞组成，是一类产复合酶的菌株，除产蛋白酶外，还可产淀粉酶、糖化酶、纤维素酶、植酸酶等。在淀粉酶的作用下，将原料中的直链、支链淀粉降解为糊精及各种低分子糖类，如麦芽糖、葡萄糖等；在蛋白酶的作用下，将不易消化的大分子蛋白质降解为蛋白胨、多肽及各种氨基酸，而且可以使辅料中粗纤维、植酸等难吸收的物质降解，提高营养价值、保健功效和消化率，广泛应用于食品、饲料、生产曲酸、酿酒等发酵业。
酿酒酵母是单细胞微生物，属于真菌类，耐酸性强，是兼性厌氧菌。活性干酵母、酵母培养物、酵母提取物、酵母细胞壁等都是我国农业部《饲料添加剂品种目录》所允许使用的饲料添加剂。酵母类产品能有效改善畜禽消化道菌群平衡、增强机体免疫力、提供丰富的营养物质，从而达到防治消化道疾病和促进生长等多重作用，具有无毒、无副作用、无污染、不产生抗药性等特点，是一种绿色的饲料添加剂，在饲料工业中有广泛的研究和应用。
目前，将植物乳杆菌、米曲霉和酿酒酵母复配作为猪饲料添加剂的应用尚未见报道。
发明内容
本发明所要解决的技术问题是提供一种能有效改善猪腹泻，且无毒副作用，无污染，无耐药性的猪用饲料。
本发明解决上述问题所采用的技术方案是，提供一种微生物发酵制备的饲料，由配合料加入复合菌液固态发酵制成，其中复合菌液由植物乳杆菌菌液1-10重量份、米曲霉菌液1-10重量份和酿酒酵母菌液1-10重量份混合制成。
优选的，所述复合菌液由植物乳杆菌菌液1-5重量份、米曲霉菌液1-5重量份和酿酒酵母菌液1-5重量份复配制成。
优选的，所述复合菌液由植物乳杆菌菌液1重量份、米曲霉菌液2重量份和酿酒酵母菌液2重量份复配制成。
更优选的，单位重量份的所述三种菌液中活菌数相同。
优选的，所述配合料包括豆粕、玉米蛋白粉、菜籽粕、花生粕、磷酸氢钙和食盐等。
更优选的，所述配合料还包括丝氨酸、缬氨酸和异亮氨酸等。
更优选的，所述配合料还包括维生素C、维生素D、维生素E和维生素B12等。
最优选的，所述配合料由豆粕50重量份、玉米蛋白粉20重量份、菜籽粕10重量份、花生粕5重量份、磷酸氢钙5重量份、食盐5重量份、丝氨酸1重量份、缬氨酸1重量份、异亮氨酸1重量份、维生素C 0.5重量份、维生素D 0.5重量份、维生素E 0.5重量份和维生素B12 0.5重量份混合均匀制成。
本发明还提供一种微生物发酵饲料的制备方法：将植物乳杆菌菌液、米曲霉菌液和酿酒酵母菌液低温浓缩至菌液中活菌数相同，再按照上述复合菌液配比混合后，以一定比例的接种量加入到上述配合料中，在35℃条件下固态发酵7d，即得。
优选的，所述配合料与复合菌液的重量比为(90-95):(10-5)。
更优选的，所述配合料与复合菌液的重量比为95:5。
本发明具有积极有益的效果：
令人惊奇的是，经过反复多次试验，本发明意外发现将植物乳杆菌菌液、米曲霉菌液和酿酒酵母菌液复配后经固态发酵获得的饲料对于防治猪腹泻具有增效的效果，配合其他活性成分使用效果更佳。
与普通猪用饲料相比，本发明的饲料配伍合理，疗效显著，能够显著降低猪腹泻率，实验证明对改善猪腹泻有特效；由于饲料中无西药抗生素成分，因此本发明的饲料无毒副作用，无耐药性，无污染。
具体实施方式
下面结合实施例对本发明作更进一步的说明，但本发明的实施方式不限于此。下述实施例中所使用的实验方法如无特殊说明，均为常规方法。
实施例1、植物乳杆菌菌液的制备
将植物乳杆菌(菌株号ACCC11118)接种至装有MRS培养基的厌氧管中，37℃恒温厌氧培养48h，得到副干酪乳杆菌菌液。
所述MRS培养基为：蛋白胨10.0g/L、牛肉膏8.0g/L、酵母膏4.0g/L、葡萄糖20.0g/L、吐温801ml/L、磷酸氢二钾2.0g/L、三水乙酸钠5.0g/L、柠檬酸三铵2.0g/L、七水硫酸镁0.2g/L、硫酸锰0.05g/L，pH为6.2-6.6。
实施例2、米曲霉菌液的制备
将米曲霉菌(菌株号CICC40188)接种至Czapek’s琼脂培养基上，28℃培养72h，再接种到Czapek’s液体培养基中，28℃恒温培养72h，得到米曲霉菌液。
所述Czapek’s琼脂培养基为：蔗糖3％、硝酸钠0.3％、七水合硫酸镁0.05％、氯化钾0.05％、四水合硫酸亚铁0.001％、磷酸氢二钾0.1％、琼脂1.5％，pH值6.0-6.5。不加琼脂即为Czapek’s液体培养基。
实施例3、酿酒酵母菌液的制备
将酿酒酵母(菌株号CGMCC No.2388)接种至装有100ml YPD培养基的250ml锥形瓶中，30℃恒温摇床中培养16h，摇床转速为250rpm。将制备好的酿酒酵母菌种子液按照1％的接种量接种至装有100mlYPD培养基的250ml锥形瓶中，30℃恒温摇床中培养16h，摇床转速为250rpm。
所述YPD液体培养基为：酵母粉5.0g/L，蛋白胨10.0g/L，葡萄糖20.0g/L，丙酸钠2.5g/L，pH 4.0±0.1。
实施例4、配合料的制备
取豆粕50重量份、玉米蛋白粉20重量份、菜籽粕10重量份、花生粕5重量份、磷酸氢钙5重量份、食盐5重量份、丝氨酸1重量份、缬氨酸1重量份、异亮氨酸1重量份、维生素C 0.5重量份、维生素D 0.5重量份、维生素E 0.5重量份和维生素B120.5重量份混合均匀即得配合料。
实施例5、饲料的制备(A1-A6)
将实施例1制备得到的植物乳杆菌菌液低温浓缩至菌液中活菌数约为108个/g，按照5％的接种量加入到实施例4制备得到的配合料中，在35℃条件下固态发酵7d，即得对照饲料A1。
将实施例2制备得到的米曲霉菌液低温浓缩至菌液中活菌数约为108个/g，按照5％的接种量加入到实施例4制备得到的配合料中，在35℃条件下固态发酵7d，即得对照饲料A2。
将实施例3制备得到的酿酒酵母菌液低温浓缩至菌液中活菌数约为108个/g，按照5％的接种量加入到实施例4制备得到的配合料中，在35℃条件下固态发酵7d，即得对照饲料A3。
将实施例1-3制备得到的植物乳杆菌菌液、米曲霉菌液和酿酒酵母菌液低温浓缩至三种菌液中活菌数均约为108个/g，再以1:2:2的重量比混合制成复合菌液，按照5％的接种量加入到实施例4制备得到的配合料中，在35℃条件下固态发酵7d，即得本发明饲料A4。
将实施例1-3制备得到的植物乳杆菌菌液、米曲霉菌液和酿酒酵母菌液低温浓缩至三种菌液中活菌数均约为108个/g，再以2:2:1的重量比混合制成复合菌液，按照5％的接种量加入到实施例4制备得到的配合料中，在35℃条件下固态发酵7d，即得本发明饲料A5。
将实施例1-3制备得到的植物乳杆菌菌液、米曲霉菌液和酿酒酵母菌液低温浓缩至三种菌液中活菌数均约为108个/g，再以2:1:2的重量比混合制成复合菌液，按照5％的接种量加入到实施例4制备得到的配合料中，在35℃条件下固态发酵7d，即得本发明饲料A6。
试验例1、本发明饲料在防治猪腹泻中的应用
为验证本发明的应用效果，本发明的饲料添加剂在健康、体重相近的猪仔中进行试验。将猪分为实验组(A1-A6)和对照组，每组各为50只。其中，实验组的日粮为实施例5制备得到的饲料A1-A6，对照组的日粮为实施例4制备的相同重量的配合料，各组猪每日投喂3次，试验期内自由采食，同时提供充足的饮水，其他各项日常管理措施按照养殖场的常规管理进行，定期进行免疫和驱虫，30天后检测粪便中的大肠杆菌数量，同时统计试验期内实验组和对照组中猪的发病情况，计算各组的猪腹泻率(见表1)。
表1本发明饲料对猪腹泻的防治效果
由试验例1的对比实验可以看出，本发明的饲料能显著降低仔猪体内大肠杆菌数量，减少仔猪感染疾病的机会，降低仔猪发病率和腹泻率。此外，在菌液总量保持不变的情况下，由植物乳杆菌菌液、米曲霉菌液和酿酒酵母菌液复配制成的复合菌液制得的饲料的抗猪腹泻效果明显优于单一菌液，其中植物乳杆菌菌液、米曲霉菌液和酿酒酵母菌液以1:2:2的重量比混合后制成的饲料防治仔猪腹泻的效果最为突出，将仔猪腹泻率降低至4％，产生了难以预期的优异效果。
Claims (8)
1.一种微生物发酵制备的饲料，由配合料加入复合菌液经固态发酵制成，其特征在于，其中复合菌液由植物乳杆菌菌液1-10重量份、米曲霉菌液1-10重量份和酿酒酵母菌液1-10重量份混合制成。
2.根据权利要求1所述的饲料，其特征在于，所述复合菌液由植物乳杆菌菌液1-5重量份、米曲霉菌液1-5重量份和酿酒酵母菌液1-5重量份复配制成。
3.根据权利要求1所述的饲料，其特征在于，所述配合料包括豆粕、玉米蛋白粉、菜籽粕、花生粕、磷酸氢钙和食盐。
4.根据权利要求3所述的饲料，其特征在于，所述配合料还包括丝氨酸、缬氨酸和异亮氨酸。
5.根据权利要求3所述的饲料，其特征在于，所述配合料还包括维生素C、维生素D、维生素E和维生素B12。
6.根据权利要求1所述的饲料，其特征在于，所述配合料由豆粕50重量份、玉米蛋白粉20重量份、菜籽粕10重量份、花生粕5重量份、磷酸氢钙5重量份、食盐5重量份、丝氨酸1重量份、缬氨酸1重量份、异亮氨酸1重量份、维生素C 0.5重量份、维生素D 0.5重量份、维生素E0.5重量份和维生素B12 0.5重量份混合均匀制成。
7.根据权利要求1所述的饲料，其特征在于，所述配合料与复合菌液的重量比为(90-95):(10-5)。
8.根据权利要求7所述的饲料，其特征在于，所述配合料与复合菌液的重量比为95:5。
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