CN116636577A - 一种利用复合益生菌群协同发酵甘蔗渣制备饲料的方法 - Google Patents
一种利用复合益生菌群协同发酵甘蔗渣制备饲料的方法 Download PDFInfo
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Abstract
本发明提出了一种利用复合益生菌群协同发酵甘蔗渣制备饲料的方法。将地衣芽孢杆菌、枯草芽孢杆菌、酿酒酵母、瑞士乳杆菌、鼠李糖乳杆菌和嗜酸乳杆菌的菌液接种到甘蔗发酵料中进行发酵获得饲料;所述的甘蔗发酵料包括甘蔗渣、海藻干粉、废糖蜜、玉米渣和豆粕。本发明选择6种微生物作为发酵菌种,通过协同发酵的作用相互促进,更好地将蔗渣中的粗纤维和粗蛋白等营养成分转化成蛋白质、氨基酸、抗生素、抗氧化物质、免疫小分子肽等生物活性物质,同时酵母菌和乳酸菌又可增加饲料的发酵酸香味和酒香味,可增加饲料的适口性。
Description
技术领域
本发明属于微生物发酵饲料领域,特别涉及一种利用复合益生菌群协同发酵甘蔗渣制备饲料的方法。
背景技术
随着农业和工业现代化发展,其生产效率大幅度提高的同时也产生了大量的有机废弃物,而目前主要的处理方式仍以填埋、焚烧为主,给环境造成了巨大的负担,因此其生态安全利用问题是我国当前面临的重要机遇和挑战。甘蔗渣是利用甘蔗制糖时压榨后留下的渣滓,其质地较为粗硬,含有丰富的纤维素和半纤维素,分别占比40%和18%,而木质素和粗蛋白含量较少,对于用作纤维原料具有较大的优越性。
我国是养殖业大国,每年的家禽家畜类及养殖水产类总量均占世界前列。随着养殖业的发展,随之带来的饲料资源短缺的问题。因此,我们亟需转向对工业废物等粗纤维资源的合理开发,解决其生态安全利用问题。
目前,甘蔗渣的主要用途以造纸和生产高密度复合材料为主,也有尝试将压榨过后的甘蔗渣直接喂养家禽家畜,但容易造成其消化困难,增加肠道负担,且甘蔗渣中较低含量的蛋白质和能量难以供给家禽家畜的营养需求。基于此,借助微生物手段将粗纤维饲料经发酵制备成营养丰富的适用于蛋鸡喂养的高质量饲料,既能够将甘蔗渣高效、快速、低成本、零污染地增值转化,又给养殖业带来可观的饲料资源。
发明内容:
本发明的目的在于提供一种利用复合益生菌群协同发酵甘蔗渣制备饲料的方法,旨在通过独有的微生物高密度培养及固态发酵核心关键技术,利用分离于乳制品及保健品的益生菌作为发酵菌种,极大缩短甘蔗渣的发酵时间,提高饲料的生物安全性,提高了饲料的适口性、转化效率和抗病能力,有利于缓解养殖业发展与饲料资源短缺的矛盾问题,同时又可促进解决有机废弃物的生态安全利用问题。
为了实现上述目的,本发明提供如下技术方案:
一种利用复合益生菌群协同发酵甘蔗渣制备饲料的方法,包括如下步骤:
将地衣芽孢杆菌、枯草芽孢杆菌、酿酒酵母、瑞士乳杆菌、鼠李糖乳杆菌和嗜酸乳杆菌的菌液接种到甘蔗发酵料中进行发酵获得饲料;
所述的甘蔗发酵料包括甘蔗渣、海藻干粉、废糖蜜、玉米渣和豆粕。
优选,所述的甘蔗发酵料按质量份数计,包括包括甘蔗渣10份、海藻干粉1份、废糖蜜1份、玉米渣1份和豆粕1份。
所述的地衣芽孢杆菌、枯草芽孢杆菌、酿酒酵母、瑞士乳杆菌、鼠李糖乳杆菌和嗜酸乳杆菌的菌液是分别将菌株接种到肉汤培养基进行培养获得,利用MRS肉汤进行瑞士乳杆菌、鼠李糖乳杆菌和嗜酸乳杆菌的增殖,利用YPD肉汤进行酿酒酵母的增殖;利用LB肉汤进行地衣芽孢杆菌、枯草芽孢杆菌的增殖。
优选,所述的菌液的细胞浓度为OD600=15。
优选,所述的地衣芽孢杆菌、枯草芽孢杆菌、酿酒酵母、瑞士乳杆菌、鼠李糖乳杆菌和嗜酸乳杆菌的菌液的接种体积比是1:1:1:1:1:1。
优选,所述的菌液的制备方法是:取菌种于肉汤培养基中37℃,200rpm下培养12小时后,再取菌液相应地转接至肉汤培养基中,于37℃,200rpm下扩大培养,期间通过及时补充一定的碳氮源和调整pH进行菌种的高密度培养,使得菌液的细胞浓度达到OD600=15。
作为优选,进行高密度培养时,通过添加酵母粉和葡萄糖作为菌种的碳氮源,通过滴加5mol/L NaOH来调节菌种的pH。
优选,所述的发酵是在一定的温度(36-40℃)与转速(10-20rpm/min)进行协同发酵1-3天。
发酵菌种均为对宿主有益的活性菌种,分别为地衣芽孢杆菌、枯草芽孢杆菌、酿酒酵母、瑞士乳杆菌、鼠李糖乳杆菌和嗜酸乳杆菌,均分离于益生菌制品的酸奶、饮料和制剂。
本发明与现有的技术相比优点体现在:
本发明的发酵菌种均分离于乳制品及调节人体肠道稳态的制剂的益生菌,使得制备的甘蔗饲料具有绝对的生物安全性;选择枯草芽孢杆菌和地衣芽孢杆菌,在培养过程中产生细菌素和脂肽类活性物质,可抑制饲料制备过程中存在的致病微生物,提高甘蔗饲料的抗病能力;选择瑞士乳杆菌、鼠李糖乳杆菌和嗜酸乳杆菌三种乳酸菌能够促进动物生长、提高动物的免疫调节活性、维持动物肠道内的菌群微生态平衡;通过独有的高密度培养核心关键技术,可大大提高菌种的细胞密度,有利于在发酵饲料过程中有效减少发酵时间,提高发酵效率;选择6种微生物作为发酵菌种,通过协同发酵的作用相互促进,更好地将蔗渣中的粗纤维和粗蛋白等营养成分转化成蛋白质、氨基酸、抗生素、抗氧化物质、免疫小分子肽等生物活性物质,同时酵母菌和乳酸菌又可增加饲料的发酵酸香味和酒香味,可增加饲料的适口性。
附图说明
图1是蛋鸡喂养情况。
具体实施方式:
下面将通过具体实施例进一步清楚地描述本发明。需要指出的是,在本发明的范围内或者在不脱离本发明的内容、精神和范围内了,对本发明进行的替换和改动,对于本领域的技术人员来说是显而易见的,且包含在本发明的范围之内。
实施例1
1.利用复合益生菌群协同发酵甘蔗渣制备饲料的方法,由不同的原料组成:甘蔗渣、海藻干粉、废糖蜜、玉米渣、豆粕、发酵菌种,包括如下步骤:
(1)发酵菌种:地衣芽孢杆菌、枯草芽孢杆菌、酿酒酵母、瑞士乳杆菌、鼠李糖乳杆菌和嗜酸乳杆菌。
(2)发酵菌种种子液的制备:取200uL菌液(地衣芽孢杆菌、枯草芽孢杆菌、酿酒酵母、瑞士乳杆菌、鼠李糖乳杆菌或嗜酸乳杆菌)于肉汤培养基中37℃,200rpm下培养12小时后,分别取5mL的菌液相应地转接至500mL的肉汤培养基中,于37℃,200rpm下扩大培养,培养时间为3天左右,期间通过及时补充一定的酵母粉和葡萄糖作为菌种的碳氮源和通过滴加NaOH(5mol/L)来调节菌种的pH为7进行各菌种的高密度培养,使得菌液的细胞浓度达到OD600=15,由此得到种子液。用MRS肉汤进行瑞士乳杆菌、鼠李糖乳杆菌和嗜酸乳杆菌的增殖,利用YPD肉汤进行酿酒酵母的增殖;利用LB肉汤进行地衣芽孢杆菌、枯草芽孢杆菌的增殖。
(3)协同发酵甘蔗渣制备饲料:将制备的地衣芽孢杆菌、枯草芽孢杆菌、酿酒酵母、瑞士乳杆菌、鼠李糖乳杆菌和嗜酸乳杆菌种子液,按照体积比为1:1:1:1:1:1于容器内混匀,利用5mol/L NaOH调节pH为7,得到混合种子液。接着将甘蔗渣与混合种子液按质量比1:1在处理器中进行充分的搅拌混匀,随后加入适量的海藻干粉、废糖蜜、玉米渣、豆粕(海藻干粉、废糖蜜、玉米渣、豆粕与甘蔗渣的质量比都为0.1:1),设置好一定的温度(37℃)与转速(20rpm/min)进行协同发酵3天。
由此得到利用复合益生菌群协同发酵甘蔗渣制备的饲料。
参照2020年国家新发布的蛋鸡饲料标准,依托广州汇标检测中心检测,结果如表1所示,我们的利用复合益生菌群协同发酵甘蔗渣制备的饲料成分完全符合国家的标准,尤其是20.3%的粗蛋白的含量显著高于国标所规定的16-17%,表明我们制备的饲料营养成分丰富且蛋白质含量充足。
表1.实施例1的饲料成分检测表
图1A为实施例1通过造粒机制备的长条颗粒状蛋鸡饲料,并于专业的养鸡厂现场评测蛋鸡的食用情况(图1B),由此可见,蛋鸡的喂养情况十分良好,表明制备的饲料具备非常高的适口性。
Claims (8)
1.一种利用复合益生菌群协同发酵甘蔗渣制备饲料的方法,其特征在于,包括如下步骤:
将地衣芽孢杆菌、枯草芽孢杆菌、酿酒酵母、瑞士乳杆菌、鼠李糖乳杆菌和嗜酸乳杆菌的菌液接种到甘蔗发酵料中进行发酵获得饲料;
所述的甘蔗发酵料包括甘蔗渣、海藻干粉、废糖蜜、玉米渣和豆粕。
2.根据权利要求1所述的方法,其特征在于,所述的甘蔗发酵料按质量份数计,包括甘蔗渣10份、海藻干粉1份、废糖蜜1份、玉米渣1份和豆粕1份。
3.根据权利要求1或2所述的方法,其特征在于,所述的地衣芽孢杆菌、枯草芽孢杆菌、酿酒酵母、瑞士乳杆菌、鼠李糖乳杆菌和嗜酸乳杆菌的菌液是分别将菌株接种到肉汤培养基进行培养获得,利用MRS肉汤进行瑞士乳杆菌、鼠李糖乳杆菌和嗜酸乳杆菌的增殖,利用YPD肉汤进行酿酒酵母的增殖;利用LB肉汤进行地衣芽孢杆菌、枯草芽孢杆菌的增殖。
4.根据权利要求3所述的方法,其特征在于,所述的菌液的细胞浓度为OD600=15。
5.根据权利要求1或4所述的方法,其特征在于,所述的地衣芽孢杆菌、枯草芽孢杆菌、酿酒酵母、瑞士乳杆菌、鼠李糖乳杆菌和嗜酸乳杆菌的菌液的接种体积比是1:1:1:1:1:1。
6.根据权利要求3所述的方法,其特征在于,所述的菌液的制备方法是:取菌种于肉汤培养基中37℃,200rpm下培养12小时后,再取菌液相应地转接至肉汤培养基中,于37℃,200rpm下扩大培养,期间通过及时补充一定的碳氮源和调整pH进行菌种的高密度培养,使得菌液的细胞浓度达到OD600=15。
7.根据权利要求6所述的方法,其特征在于,进行高密度培养时,通过添加酵母粉和葡萄糖作为菌种的碳氮源,通过滴加5mol/L NaOH来调节菌种的pH。
8.根据权利要求1所述的方法,其特征在于,所述的发酵获得饲料是在温度36-40℃与转速10-20rpm/min进行协同发酵1-3天。
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